Co So Di Truyen Cua Tinh Chong Chiu Voi Moi Truong Cua Cay Lua

Tai lieu chia se tai: wWw.SinhHoc.edu.
vn
BI CH BU - NGUYN TH LANG
C S DI TRUYN TNH CHNG CHU I VI THIT HI DO MI TRNG CA CY LA
NH XUT BN NNG NGHIP Thnh ph H ch Minh 2003
Tai lieu chia se tai: wWw.SinhHoc.edu.vn
Bi ch Bu Ph Gio S, Tin S Vin Trng Nguyn th Lang Tin s Trng B mn Di truyn Qy Gen VIN LA NG BNG SNG CU LONG

LI TA
Nhng thit hi do mi trng bao gm cc yu t t ai, nc, nhit (nng hoc lnh) nh hng rt ln n nng sut cy trng ni chung v cy la ni ring. Trn th gii, cc nh khoa hc s dng thut ng abiotic stress khi qut tt c nhng stress do yu t khng phi sinh hc gy ra. Phn ln c ch chng chu v sinh l hc ca nhng thit hi nh vy i vi cy trng ang c tp trung nghin cu vi s phi hp gia cc nh sinh l, sinh ha, di truyn, chn ging. Nhng y l lnh vc rt kh, mt s c ch chng chu vn cha c bit r, mc d ngi ta phn tch s kin mc sinh hc phn t. Vic ci tin ging c nng sut cao kt hp vi kh nng chng chu stress nh vy vn ang din bin rt chm, v nhng kin thc c bn v di truyn v sinh l thc vt cn hn ch, k thut thanh lc rt phc tp v tn km. Quyn sch ny ch yu tp trung cc vn chnh trn cy la, bao gm: tnh chng chu kh hn, tnh chng chu mn, tnh chng chu ngp hon ton, tnh chng chu c nhm, tnh chng chu thiu ln, tnh chng chu c st v tnh chng chu lnh. Cc phng php nghin cu di truyn c gii thiu trong chng u tin bao gm nhng thnh tu v genomics, chc nng genome hc, ng dng microarray, bn ng dng trn c s hin tng synteny ca Tin s Gale (Rockefeller Fondation), v c bit l phn tch QTL i vi nhng tnh trng di truyn s lng, vi cc phn mm rt c ch cho nhiu mc tiu gii thch khc nhau. S khan him v ngun nc ti cho nng nghip hin nay v trong tng lai l vn ngy cng tr nn nghim trng c tnh cht ton cu. Do , cc d n nghin cu v cy trng chng chu kh hn ang l hng u tiu u t ca cc d n quc t v quc gia. Ging la chng chu kh hn phi c nghin cu trn c s hiu bit r rng v c ch chng chu v kh nng di truyn ca ging, trc nguy c khng hong thiu nc trong tng lai gn. C ch trnh n, c ch thot v c ch chng chu c cp mt cch h thng trn c s di truyn s lng vi nhng QTL c tnh cht gi nh v gen iu khin hin tng chng chu rt phc tp ny. Bn cnh , nh hng ca mi trng tc ng vo tnh trng di truyn s lng nh vy cng cn c gii thch. C ch iu tit p sut thm thu v qa trnh nhn tn hiu ca stress do kh hn, mn, v lnh tng i ging nhau v nguyn tc chung. Ngi ta t ra mt cu hi: tn hiu y c s cm nhn ca di truyn nh th no iu chnh gen mc tiu p ng vi kh nng chng chu ca cy trong tng hon cnh khc nhau nh vy? l mt hin tng kh th v trong thin nhin, khi chng ta nghin cu s iu tit rt tinh vi ca gen (gene regulation). Di truyn tnh chng chu mn c Vin La ng bng sng Cu Long (BSCL) tin hnh nghin cu t di truyn c bn n ng dng chn to ra mt vi ging ang pht trin trong sn xut. Nhng tng tc gia kiu gen x mi trng vn cn l mt th thch rt ln, v ngng chng chu phi c xc nh r (EC khong 4-6 dS/m). Tp th tc gi rt cm n s hng dn ca TS Ning Huang (IRRI) v phn tch genome, s cng tc ca TS Yanagihara (JIRCAS), TS Zhikang Li (IRRI) cung cp vt liu v phng tin, TS Kenneth McNally (IRRI) trong phn tch m hnh ton gii thch kt qa. Di truyn tnh chng chu ngp hon ton c thc hin vi s nm tham gia nghin cu lu nht ca tp th tc gi. Chng ti cm n anh bn ngi Thi Lan Sripongpankul cho php s dng s liu trong lun n Tin s minh ha thm trong t liu ny, c bit TS Senadhira, TS Derk HilleRisLamber (IRRI), TS V tng Xun, TS Nguyn vn Lut hng dn chng ti trong nhng nm u tin tham gia nghin cu, TS Puckridge i din ca IRRI ti Thi Lan to iu kin cho chng ti c kho st rt nhiu vng la nc
su Chu v tip cn cc nh khoa hc chuyn ngnh v lnh vc ny. Chng ti xin cm n TS Mackill (IRRI) vi nhng tho lun rt l th v kh nng chn to ging chng chu. Di truyn tnh chng chu c nhm c thc hin vi s cng tc ca tp th cc nh khoa hc TS Brar (IRRI), TS Henry Nguyn (H Texas Tech), cng cc ng nghip chng ti ti Vin La BSCL, nhm khai thc ngun gen mc tiu t ging la hoang Oryza rufipogon ng Thp Mi. Anh Nguyn duy By n lc hon thnh lun n Tin s vi cng trnh ny, lm c s khoa hc cho cc ng nghip Mn pht trin thnh cng ging la AS996 (ging quc gia) chng chu phn v nng sut cao BSCL. Chng ti cm n TS HilleRisLamber, TS Akita, TS Chang (IRRI) to iu kin cho chng ti thc hin thanh lc nhm tp on ging la bn a ca Vit Nam trong nm 1985, TS Vaughan Duncan (JIRCAS) trong qa trnh thu thp la hoang vi chng ti BSCL xc nh vt liu cho gen mc tiu. Chng ti xin by t lng bit n i vi TS John OToole gip v kinh ph v chng trnh o to thng qua d n cng ngh sinh hc ca Rockefeller Foundation. Trong t phn, ngoi c nhm v st, nh hng thiu ln cng l vn rt ln cho cy la. Chng ti thc hin cng trnh nghin cu ny trn c s mt th nghim di hn ca B Mn Canh tc thuc Vin La BSCL. Kh nng nhnh c th c xem l mt tiu chun chn lc rt c gi tr, do a gen iu khin trong iu kin thiu ln, trong hot ng ca gen khng cng tnh v gen cng tnh u c nh hng quan trng nh nhau. Chng ti rt cm n TS JJ Ni (Trung Quc) cho php chng ti s dng t liu ca anh trong thuyt minh c s di truyn ca tnh chng chu v nhng tho lun b ch trong thi gian cng nhau lm vic ti IRRI. Tnh chng chu c st rt t c cng b v vic nh gi kiu hnh c th c xem nh kh khn nht trong cc tnh trng lit k, v Fe++ chuyn i sang Fe+++ rt d dng, lm sai lch kt qa quan st. Nhim sc th s 1 c xem nh nhim sc th cha nhiu gen ng c vin cho tnh trng lin quan n hin tng chng chu. Tnh chng chu lnh c rt nhiu kt qa nc ngoi c cng b c tng hp trn c s di truyn v chn ging, cha c cng trnh c th t Vin La BSCL. Chng ti hi vng quyn sch ny s cung cp nhng t liu cn thit cho sinh vin, nghin cu sinh, cc nh nghin cu quan tm n nhng lnh vc chng chu bt li do mi trng, nhng cn b nng nghip ang phc v trong lnh vc chn to ging cy trng. Knh mong bn c thng cm nhng thiu st trong qa trnh bin son, in n v ng gp kin cho tc gi. Ch bin BI CH BU Vin trng Vin La ng Bng Sng Cu Long
NI DUNG
Trang
Li ta Chng 1: Gii thiu tng qut ng dng cng ngh sinh hc trong ci tin ging cy trng chng chu iu kin bt li ca mi trng 1-1. Kh hn 1-2. t mn 1-3. t acid v t bc mu 1-4. Nhit lnh v nhit nng 1-5. Bn gen v s pht trin marker phc v phn tch di truyn 1-6. C ch sinh l hc v di truyn i vi hin tng chng chu stress 1-7. Tin ci tin ging chng chu 1-8. K thut thanh lc c tnh kh thi vi qui m ln 1-9. Genome hc, ngnh hc di truyn mi 1-10. Hin tng synteny v genomics c tnh cht so snh 1-11. ng dng genomics trong ci tin ging cy trng 1-11-1.Th vin DNA 1-11-2. K thut cloning cc gen 1-11-3. Microarray 1-11-4. Qun th knockout 1-11-5. K thut chuyn np gen 1-12. Phn tch QTL 1-12-1. Nhng m hnh v di truyn s lng 1-12-1-1. M hnh QTL n (single-QTL) 1-12-1-2. M hnh multiple-locus 1-12-2. Phng php phn tch marker n (SMA) 1-12-2-1. SMA trn qun th hi giao 1-12-2-2. Kt hp hin tng phn ly ca QTL v marker 1-12-2-3. Php th t n gin trong qun th hi giao 1-12-2-4. Phn tch phng sai trong qun th hi giao 1-12-2-5. M phng trong qun th hi giao 1-12-3. Phng php SMA trong qun th F2 1-12-4. Phn tch QTL trn c s bn cch qung (interval mapping) 1-12-4-1. Bn cch qung trong qun th hi giao (BC) 1-12-4-2. Bn cch qung trong qun th F2 1-12-5. Kh nng gii thch v thng k sinh hc ca bn QTL Chng 2: C s di truyn tnh chng chu mn 2-1. t mn 2-2. C ch chng chu mn 2-3. Di truyn tnh chng chu mn 2-3-1. Nghin cu di truyn s lng tnh chng chu mn 2-3-2. Nghin cu di truyn phn t tnh chng chu mn 2-4. S th hin gen chng chu mn 2-4-1. Ph th hin transcript 2-4-2. Phn tch microarray 2-4-3. c im th transcript ca ging la chng chu mn trong iu kin b stress 2-4-4. Vai tr ca abscisic acid, jasmonate, proline

2-5. Nghin cu chuyn Ci tin ging la chng chu mn BSCL 2-5-1. Vt liu & phng php 2-5-2. Kt qa & tho lun 2-5-2-1. Xc nh vt liu lai to 2-5-2-2. Nghin cu c ch chng chu mn 2-5-2-3. Phn tch bn di truyn QTL tnh chng chu mn 2-5-2-4. Chn ging chng chu mn nh marker phn t (MAS) 2-5-2-5. Pht trin ging la trin vng cho vng b nhim mn Chng 3: C s di truyn tnh chng chu kh hn 3-1. Xc nh tnh trng thnh phn trong chng chu kh hn 3-2. Nhng marker phn t DNA v bn QTL 3-3. Bn QTL i vi tnh trng r la 3-4. Bn QTL i vi tnh trng iu tit p sut thm thu (OA) 3-5. Bn QTL i vi tnh trng biu hin mu xanh cao lng (STG) 3-6. Bn QTL cc tnh trng hnh thi quan trng 3-7. Chuyn np gen mc tiu 3-8. C ch truyn tn hiu 3-9. Gen & s khm ph l trnh thng qua genome hc chc nng Chng 4: C s di truyn tnh chng chu ngp ng 4-1. Tng quan nghin cu trc y 4-2. Hnh thi hc ca cy la thch nghi vi vng b l lt 4-3. Kh nng vn lng 4-3-1. Di truyn v kh nng vn lng 4-3-2. Nghin cu la ni BSCL 4-4. Di truyn tnh chng chu ngp hon ton 4-5. C ch chng chu ngp v sinh l hc 4-5-1. Dip lc t 4-5-2. Carbohydrate 4-5-3. Hm lng nitrogen 4-5-4. Hot ng ca peroxidase 4-6. Nghin cu bn di truyn tnh trng vn lng 4-7. Clone ha gen OsGAPDH iu khin tnh chng chu ngp 4-7-1. Phn tch Southern 4-7-2. Phn tch Northern 4-7-3. S th hin dung hp GST-OsGAPDH Chng 5: C s di truyn tnh chng chu c nhm 5-1. Ging la nc su chng chu c nhm BSCL 5-2. Xc nh QTL iu khin tnh chng chu c nhm 5-2-1. iu tra a hnh trong b m 5-2-2. Phn ly v thit lp bn 5-2-3. Bn QTL 5-2-4. Phn tch epistasis 5-2-5. ng dng marker trong chn ging chng chu v gen ng c vin 5-2-6. To ra clone ca nhng QTL chng chu nhm t la hoang

5-3. Chn to ging la chng chu nhm Chng 6: C s di truyn tnh chng chu thiu ln 6-1. Gii thiu chung 6-1-1. t thiu ln 6-1-2. Hin tng thiu ln trn cy la 6-1-3. Biu hin ca ging la chng chu thiu ln 6-1-4. K thut thanh lc 6-2. C ch chng chu thiu ln 6-3. Di truyn tnh chng chu thiu ln 6-3-1. Lp bn QTL bng AFLP 6-3-2. Lp bn QTL bng RFLP 6-3-3. Gen Pup-1: QTL ch lc lm gia tng kh nng hp thu ln 6-4. Phn tch di truyn s lng Chng 7: C s di truyn tnh chng chu c st 7-1. Gii thiu chung 7-2. K thut thanh lc 7-3. Nghin cu di truyn phn t tnh chng chu c st 7-3-1. Tnh chng chu c st trong qun th n bi kp IR64/Azucena 7-3-2. Tnh chng chu c st trong qun th cn giao Nipponbare/Kasalath // Nipponbare Chng 8: C s di truyn tnh chng chu nhit lnh 8-1. Gii thiu chung 8-2. Di truyn tnh chng chu lnh 8-2-1. Bn QTL trn nhim sc th s 4 8-2-2. V tr trn bn ca Ctb-1 v Ctb-2. 8-2-3. Tnh chng chu lnh v chiu di ti phn 8-2-4. Bn QTL tnh trng chng chu lnh giai on lm ng 8-3. S truyn tn hiu v phn ng iu tit p sut thm thu 8-3-1. iu tit phn ng pht quang sinh hc ca Arabidopsis 8-3-2. Phn lp dng t bin c mc pht quang sinh hc thay i 8-4. S bin i protein theo nhit , mn & kh hn Bn ch dn (index)

Chng 1
GII THIU TNG QUT NG DNG CNG NGH SINH HC TRONG CI TIN GING CY TRNG CHNG CHU
IU KIN BT LI CA MI TRNG
Din tch t trng trt trn th gii chim 10% trong tng s 13 t ha. Trong , 11,5 triu ha t ang c canh tc c xem nh khng thun li cho nng nghip (FAO 2002). Hu ht t trng trt c xp vo nhm di mc ti ho cho cy trng. Bn cnh nhng thit hi do su bnh gy ra, ngi ta c on c 70% tim nng v nng sut b mt i do iu kin bt li ca mi trng, ngay c trong nhng quc gia c nn nng nghip pht trin (FAO 2002) Nhng thit hi do su bnh gy ra c gi vi thut ng "thit hi c tnh cht sinh hc" (biotic stresses). Nhng thit hi do iu kin bt li ca mi trng, th d nh kh hn, ngp ng, mn, phn, nng, lnh, v.v.., c gi vi thut ng "thit hi c tnh cht khng phi sinh hc (abiotic stresses) S khan him v nc ti phc v cho nng nghip c bo ng trong nhiu hi ngh khoa hc ca th gii gn y. Do s thay i kh quyn vi hiu ng nh knh, nhit ca kh quyn m dn ln, bng tan hai cc s to s ngp lt cc vng t thp (nh ng bng sng Cu Long). Nh vy, l lt v s xm nhp mn s tr thnh vn ln trong nhiu nm sau. Vi tm quan trng nh vy, ngi ta hoch nh mt th t u tin trong u t nghin cu tnh chng chu kh hn v chng chu mn trn ton th gii, trong lnh vc ci tin ging cy trng, sau l tnh chng chu lnh, chng chu ngp ng, chng chu t c vn (acid, thiu ln, c st, c nhm, thiu km, Mg, Mn v mt s cht vi lng khc nh Cu,) 1-1. KH HN Kh hn s l yu t quan trng bc nht nh hng n an ton lng thc ca th gii, v iu ny tng xy ra trong qa kh. Ti nguyn nc phc v cho nng nghip khng phi v tn, bn cnh l p lc dn s km theo s pht trin th, s kin y s lm gia tng nhu cu nc phc v dn sinh v cho pht trin cng nghip. Do , s khan him nc phc v nng nghip l vn ang c d bo rt cp thit trn qui m ton cu. Hin nay, nc phc v nng nghip chim 70% ngun nc phc v dn sinh ca ton th gii. Mun sn xut c 1 kg thc, ngi ta phi cn 5000 lt nc. Nhiu quc gia nh Ai Cp, Nht Bn, c c gng ci tin nhm nng cao hiu qa s dng nc, gim xung 1,3 m3 / kg thc. Trung Quc, cc nh chn ging ang th nghim m hnh cy la canh tc trn t thong kh, vi thut ng "aerobic rice", khng phi nh cy la ngp nc truyn thng. B r la s pht trin nh cy trng cn, vi ch ti ci tin, nhm tit kim nc ti a. Hn hn c xem nh mt trong nhng hu qa nghim trng do s suy gim ca ngun nc. Do vy, ngi ta qui nh ngy 22 thng 3 hng nm l Ngy NcTh Gii. Hin nay, mc m bo nc trung bnh cho mt ngi trong mt nm gim t 12.800 m3 / ngi vo nm 1990 xung cn 10.900 m3 / ngi vo nm 2000 v c kh nng ch cn khong 8.500 m3 / ngi vo nm 2020. Theo Hi Nc QucT (IWRA), tiu chun cng nhn quc gia c mc bo m nc cho mt ngi < 4000 m3 / nm, c xem nh thiu nc, v < 2000 m3 / nm, thuc loi him nc. Kt qa nh gi ca chng trnh KC12
Vit Nam cho thy: tng lng nc cn dng c nm ca nc ta chim 8,8% tng lng dng chy, nm 1999, tng ln 12,5% trong nm 2000, v c d bo s tng 16,5% vo nm 2010. Tng lng nc phc v ti trong nng nghip ca Vit Nam: 41 km3 nm 1985, tng ln 46,9 km3 nm 1999, v 60 km3 nm 2000. Lng nc cn dng cho ma kh s tng ln 90 km3 vo nm 2010, chim 54% tng lng nc c th cung cp. Xt trn qui m ton cu, nhit tri t nng ln s c kh nng lm mt 1/3 ngun nc ang s dng ca th gii trong 20 nm ti, nh d bo ca Lin Hp Quc. Khng hong thiu nc trn th gii hin nay c nhn nh khng ch do nc qa t so vi nhu cu m cn do qun l ngun nc qa km. Hng nm c 2,2 triu ngi cht do cc cn bnh lin quan n ngun nc nhim v iu kin v sinh km, vi 12.000 km3 nc sch hin b nhim nghim trng. Do , t nm 2000 tr i, cc d n quc t v nng nghip thuc h thng CGIAR nhn mnh n ging cy trng chng chu kh hn, nc sch cho nng thn, th, xem nhng ni dung ny l mt u tin c bit. Cc trung tm nghin cu trn th gii c FAO phn cng ph trch nhng cy trng lin quan n chng chu kh hn l: IITA ph trch nghin cu u cowpea Sahel, u tng v ng Dry Savana ICRISAT ph trch nghin cu cao lng, k, u chickpea, u phng v u pigeon pea n v Sahel CIAT ph trch nghin cu cc loi u Mexico, Trung M v ng bc Brazil IRRI ph trch nghin cu la Bangladesh, ng n , Thi Lan v Indonesia CIP ph trch nghin cu khoai ty Trung Quc. n , Nam Phi, Kazakhstan, Afghanistan CIMMYT ph trch nghin cu la m Trung , Ty , Bc Phi, cy ng vng cn Sahara
1-2. T MN t trng trt b nh hng mn c khong 380 triu ha, chim 1/3 t trng trn ton th gii. N thng i km theo hin tng t kim v ngp nc (Gale 2002). Trong , 60 triu ha l t mn do thy cp a mn ln t mt, ngi ta thng dng thut ng "inland salinity". Hin tng ny do thiu nc ti, lm t tr nn mn ha Chu , Chu Phi v Nam M. Cc trung tm nghin cu trn th gii c FAO phn cng ph trch nhng cy trng lin quan n chng chu mn l: IRRI ph trch nghin cu la vng ven bin Bangladesh, Orissa, Vit Nam, Philippines, vng mn trong t lin chu th sng Hng (n ) v vng ng bc Thi Lan ICARDA ph trch nghin cu la Trung
1-3. T ACID & T BC MU t acid chim 40% din tch t trng trt trn ton th gii, vi pH < 5, trong yu t hn ch ch yu cho cy trng l hm lng cao ca nhm v manganese. Vn ny c bit quan trng ti Nam M (380 triu ha), bao gm khu vc chu th sng Amazone. Yu t hn ch do hm lng st cao, gy c cho cy c ghi nhn Ty Phi. t phn Vit Nam v Thi Lan thuc nhm khc, rt c bit, bi v c cht thuc hp cht sulfate st, nhm, cng thm hin tng thiu ln, pH thp, v rt giu hu c. Cc trung tm nghin cu trn th gii c FAO phn cng ph trch nhng cy trng lin quan n chng chu t acid v bc mu l: IITA ph trch nghin cu u cowpea, u tng vng rng m t CIAT ph trch nghin cu cc loi u Chu Phi, Chu Phi La Tinh

1-4. NHIT LNH & NHIT NNG
IRRI ph trch nghin cu la Bangladesh, Indonesia, Philippines CIMMYT ph trch nghin cu ng Chu M La Tinh, ng Nam v Chu Phi, ph trch nghin cu la m CWANA
Nhit qa nng hoc qa lnh s lm hn ch tim nng pht trin ca cy trng. Hin nay, 70% vng t trng khoai ty ca th gii c kh nng b ri ro do nhit lnh. Cy la trng Hn Quc, Nepal thng b thit hi do lnh Cc trung tm nghin cu trn th gii c FAO phn cng ph trch nhng cy trng lin quan n chng chu nhit bt thun l: CIP ph trch nghin cu khoai ty chu lnh Andes, chu nng Nam ICARDA ph trch nghin cu la mch, u chickpea, cy gai chu lnh IITA ph trch nghin cu u cowpea chu nng Sahel
1-5. BN GEN & S PHT TRIN MARKER PHC V PHN TCH DI TRUYN Bn gen l yu cu trc ht cho phn tch di truyn tnh trng chng chu cc thit hi khng phi sinh hc, ng thi n cng l tiu chun trong chn ging cy trng hin i. Nhm T Vn v Nghin Cu Nng Nghip Quc T (CGIAR) ca FAO ch o cc Vin, Trung Tm trc thuc, hon thnh cc bn mc phn t i vi nhng loi cy trng chnh. Trong , c nhng cng trnh mang tnh cht hp tc quc t rt rng nh: bn gen cy la, la m, khoai ty c th c s dng ph bin. Nhng bn cn bn i vi cy trng c mc ph bin thp cng c xem xt, v c s phn cng ca CGIAR, tuy rng chng c tnh cht quc t ha rt thp. Ch cn mt vi loi cha c xy dng bn . Bn di truyn (genetic map) cn c hiu nh bn lin kt (linkage map) gia marker v gen mc tiu. Bn cnh , ngi ta thc hin nhng hp phn quan trng xy dng bn vt l (physical map) ca nhng gen ny. K thut xy dng bn i vi tnh trng s lng (QTL) thng c rt t thng tin v s kim sot ca gen, bi v n da trn nhng gi nh c tnh cht ton hc. Nhng n v cng quan trng, v hu nh cc tnh trng chng chu vi "stress" u yu cu "QTL mapping". Ngi ta cn phi qut t u n cui b genome vi nhng marker bao ph ton b cc nhim sc th, vi mt trung bnh 10cM gia 2 marker. Thng qua , ngi ta xc nh nhng khu vc gi nh c cha cc gen iu khin tnh trng s lng m ta ang nghin cu. Ngi ta phi da trn c s bin ng ca tnh trng kt hp vi s thay i ca marker tng ng. Mt marker cng dy c, cng tt cho s gi nh, vi mc chnh xc cao, trn mt qun th con lai no ang c s dng phn tch di truyn. Nhng v tr c xc nh nh vy v cng cn thit cho chng trnh chn ging nh marker (MAS = marker-aided selection) i vi tnh trng chng chu, v rt cn thit cho k thut cloning trn c s bn di truyn (map-based cloning) ca nhng gen thuc v QTL Nhng marker c ng dng trong chn ging cy trng phi lin kt cht vi gen mc tiu, trn c s bn di truyn phn t. Hin nay, marker c hiu qa ng tin cy l "microsatellite" vit tt SSR. Ngi ta ang chun b a vo s dng rng ri marker SNP trong vi nm ti (ch SNP c vit tt t thut ng "single nucleotide polymorphisms" = cc a hnh nucleotide n). i vi cy la, thun li ln nht trong ng dng marker l bn genome ca n c bn c gii m, marker khng cn l vn . Theo cng trnh ca Goff v 30 tc gi khc trong nm 2002, chui k t ca SSR v SNP hin c thit k c khong 40.000 marker, k c nhng phn t mt on, hay xen on. y l nhng chui m ng nht mc 1%, mt 24 / mi gen (Gale 2002) 1-6. C CH SINH L HC V DI TRUYN HC I VI HIN TNG CHNG CHU STRESS
C ch sinh l gii thch hin tng p ng ca cy trng i vi stress, v nhm mc ch ci tin cu trc, hot ng sinh l, sinh ha ca cy, gip cy thot khi, hoc n trnh, hoc chng chu s thit hi do stress gy ra. C hai phng php lin quan n ci tin ging cy trng chng chu stress: (1) Phng php da trn kinh nghim (empirical approach) c bt u t vic khai thc bin d di truyn, kt hp vi ngun vt liu cung cp gen chng chu tt nht ca ging cy trng, hoc ca loi hoang di. Ngun vt liu l tng nht i vi tnh trng chng chu c phn lp v xem xt gen iu khin tnh chng chu thng qua phn tch QTL i vi qun th dng con lai ang phn ly, trn hai kiu hnh chng chu cao v chng chu thp. Mc d c ch sinh l vn c xem l c ch chnh c u tin nghin cu, nhng s di truyn tnh trng chng chu ny n ging cy trng mi l mt tin trnh v cng cn thit, vi yu cu chn lc kht khe v tch ly nhng alen c li (2) Phng php lai to ging c kiu hnh l tng (ideotype breeding approach), trong c im v hnh thi hc v sinh l hc u phi c kh nng tham gia vo mc tiu ci tin ging trong iu kin b stress. Gen mc tiu t ging cy trng v t loi hoang di u c quan tm khai thc, chuyn n vo ging cy trng mi. Khi thc hin hai phng php ny, ngi ta c phi s dng nhng k thut lai, k thut chng gen khng nh marker phn t, nhm xc nh nhng alen c li. Trng hp stress l kh hn, mn v lnh, nhng hin tng ny c chc nng v sinh l thc vt lin quan vi nhau, trn c s iu tit p sut thm thu ca t bo. Nhiu chin lc nghin cu nhm ci tin tnh chng chu ny mang tnh cht a ng dng. Chng bao gm hin tng iu tit p sut thm thu r, l, nhm duy tr nc, loi b cc yu t ro cn c tnh cht k nc (hydrophobic barriers) trong r v l, ci tin s thng thng mch dn, nhm thc y s di chuyn nc trong cy. Nhng c ch trnh n (avoidance), thot (escape), v chng chu (tolerance) c nt rt ging nhau i vi kh hn, mn v lnh. Th d nh ci tin tnh trng rt ngn thi gian t gieo n tr, tnh trng r pht trin mc su hn trong t, tnh trng nhy cm trong phn ng co nguyn sinh ca t bo khi gp stress. Di truyn tnh chng chu i vi stress khng phi sinh hc thng c kim sot bi a gen v rt phc tp, cc gen kim sot tnh chng chu c th trng lp nhau i vi nhng stress khc nhau. Trong genome ca la m v la mch, ngi ta nhn thy cc nh hng di truyn kim sot s p ng ca cy i vi kh hn, mn v lnh nm trn cng bn di truyn nhim sc th tng ng. C t nht 10 QTL c tm thy i vi tng tnh trng chng chu ny v chng nm chng ln nhau ti mt s vng ca nhim sc th. Trng hp s dng "phng php da trn kinh nghim" i vi tnh trng chng chu c nhm, ngi ta ghi nhn k thut thanh lc nhm thng ch ci tin s pht trin ca r v chi trong mi trng dinh dng Yoshida c cha Al, iu kin pH thp, v l mt hin tng di truyn rt phc tp. Nhng gen kim sot tnh trng chng chu ny thng tp trung ti mt locus c gi tr ng gp vo tn sut bin d di truyn cao nht. Nhng kt qa nh vy xc nh hng u tin ca MAS (chn ging nh marker phn t), vi s c lp gen c tnh cht quyt nh (key gene) thng qua c hai bin php h tr, l "k thut cloning" v "sn xut cc dng ng gen" (Gale 2002). S c lp gen (gene isolation) v kin thc v chui m ca gen c th c xem l nhng bc pht trin c tnh cht quyt nh lm r thm c ch di truyn tnh chng chu ny. Khng phi tt c cc tnh trng chng chu stress do a gen iu khin. Mt vi tnh trng chng chu vi stress c th hin do s iu khin ca gen ch lc. Th d nh tnh trng chng chu ngp ca cy la vng ng Nam , ni c din tch canh tc b nh hng ca l lt l 25 triu ha, do l n bt ng lm cy la b ngp hon ton (7 n 10
ngy), gen iu khin Sub1 nh v ti mt locus gip cho cy la chng chu c ngp v phc hi bnh thng Kh nng ca cc loi hoang di c quan h gn vi cy trng c nh gi rt cao, trong chin lc du nhp gen mc tiu t loi hoang di vo loi cy trng, nhm gia tng tnh chng chu mt cch c hiu qa. Gen chng chu mn c ghi nhn t mt loi hoang di Porteresia coarctata ca n , Sri Lanka c th chuyn vo la trng. Gen chng chu mn ca Thinopyrum bessabaricum gip n pht trin trong iu kin mn 250 mM NaCl, c th c s dng trong chng trnh ci tin cy la m. i vi tnh trng chng chu nhm, loi c lng ty, Brachiaria decumbens, thuc vng nhit i, c xem l ngun vt liu trong chng trnh ci tin ging la m n i, ging ng n i (Gale 2002). Chin lc ci tin ging nh vy c tn l "crop replacement". 1-7. TIN CI TIN GING CHNG CHU (chm nhng kh thi) Hin nay, tin ci tin ging chng chu i vi stress rt chm bi chng ta cha c s hp tc nghin cu mt cch hiu qa gia nh sinh l thc vt, sinh ha v nh di truyn. Mt vi tin b c ghi nhn trong ci tin ging chng chu (tolerance) hoc trnh n (avoidance) kh hn, mn, c nhm. Nhng hiu bit v c ch chng chu cho tng i tng cn qa hn ch. Nhng ging cy trng c cc c quan thuc h thng CGIAR quan tm u t nghin cu v pht trin: Ging ng nhit i chng chu t acid: Corpoica H-108, H-111 pht trin ti Colombia, ging Pool 25 ti Brazil, ging ZM421, 521 v 621 va c cng nhn cho pht trin ti Nam Phi, n cn chng chu hin tng t ngho N v kh hn gia v trng. Cc ging ny do CYMMIT lai to v hp tc vi cc a phng ni trn Ging la chng chu mn PSBRc 84, 86 v 88 pht trin ti Bangladesh, trong PSBRc 88 c phm cht go tt. Ging la ny do IRRI lai to v hp tc vi cc quc gia trong mng li Ging chui chng chu hn FHIA 01 do INIBAP lai to v chn lc, hin pht trin ti Honduras, Tanzania v 50 quc gia khc Ging khoai ty chng chu nng Unica pht trin ti Peru do CIP lai to v chn lc
Tin ci tin ging cy trng phc v mc tiu chng chu ny hin gp tr ngi do tnh trng nng sut cao v tnh trng chng chu c kh nng tng hp thp. Nng sut cao trong iu kin b stress cng khng tng hp vi nng sut cao trong iu kin bnh thng. iu ny cho thy cn c mt chng trnh ci tin ging c lp nhm to ra nhng ging c yu cu c bit, i vi tng loi stress ring bit. Chng ta vn cha hiu r: liu c s i khng gia nng sut v tnh chng chu hay khng? Gen iu khin nng sut cao trong iu kin b stress v trong iu kin bnh thng cng mt nhm nh nhau. Nhng s tng tc gia kiu gen v mi trng (GxE) vn cn l iu cha c hiu mt cch y . Vi s pht trin ca cng ngh sinh hc, c bit l k thut di truyn, ngi ta ang u tin nghin cu ging chng chu kh hn v chng chu mn, vi qui m hp tc quc t rt tch cc. 1-8. KTHUT THANH LC C TNH KH THI VI QUI M LN Vic nh gi kiu hnh (phenotyping) cc dng con lai i vi cc stress i hi chng ta khng ngng hon thin cc k thut thanh lc r tin, chnh xc, d ng dng khi thc hin vi s lng ln ca con lai. Thanh lc ngoi ng rung thng gp nhng vn pht sinh do cc yu t mi trng m chng ta cha kim sot c, chng c th nh hng lm sai lch kt qa thanh lc. Mun khc phc hn ch ny, ngi ta c gng ci tin phng php b tr th nghim i vi tng vn stress khc nhau
Thanh lc trong phng th nghim hoc trong nh li vi cc yu t tham gia thng nht, c kim sot, thng b hn ch do qui m b, v rt tn km. Hn na c ch chng chu in vitro v c ch chng chu ngoi ng thng khng thng nht, th d nh tnh chng chu mn, tnh chng chu kh hn. Trng hp thanh lc vi c do st trong t phn, chng ta rt kh kim sot s chuyn i t Fe++ sang Fe+++ trong mi trng dinh dng. Trng hp thanh lc tnh trng chng chu ngp hon ton, chng ta phi kim sot c ca nc sao cho gn ging nh iu kin ngoi ng. Trng hp th nghim tnh trng chng chu s thiu ln, phng php b tr th nghim phi gii quyt c hai vn : (1) c ch pht trin trong iu kin t c hm lng ln ngng thp, c th nhng ging nghin cu pht trin tt trong iu kin ny, nhng khng chng chu s thiu ln, (2) c ch pht trin trong iu kin thiu ln thc s. ng dng cng ngh sinh hc, ngi ta xut phng php nh gi kiu gen (genotyping), vi s tr gip ca marker phn t thanh lc con lai c gen chng chu vi stress mc tiu. Marker SSR hin c khuyn co v hiu qa cao, ng tin cy, s lng marker ln. Vn tr ngi ln nht ca tnh chng chu vi stress l a gen iu khin vi nhng gi nh QTL. V vy, cng vic "fine mapping" phi c thc lin lin tc nhm xc nh marker ng tin cy nht cho chin lc MAS (marker-assisted selection) trn c s phn tch vi qun th hi giao ci tin ca vt liu b m c chn lc cn thn. 1-9. GENOME HC - NGNH HC DI TRUYN MI T nm 1998, ngi ta bt u lm quen vi thut ng "Genomics" (genome hc) thng qua chng trnh nghin cu v genome ca ngi. Trong thc vt, genome ca cy Arabidopsis thaliana c gii m c bn vo nm 2000. Genome cy la c cng b gii m trong nm 2001 v hon thin trong nm 2002 ca nhm nghin cu Bc Kinh (Trung Quc) v Tsukuba (Nht Bn) cng vi nhng thng bo ng ch ca Syngenta v Monsanto. Kh nng genome ca cy ng s l mc tiu tip theo c gii m, t nht l ti cc vng trn nhim th tp hp cao cc gen (gene-rich regions) ca b genome. Cng ngh bao trm ton b hot ng ca ngnh genome hc l: (1) c chui m DNA t ng, vi kh nng c ca my l 2 triu cp base mt ngy, vi s knh mao dn t 16, n hn 98 knh, v n ang c tip tc ci tin vi tc pht trin cc nhanh (2) microarray v chip sinh hc DNA, trong 10.000 gen c th c "scan" trong cng mt ln xem xt, cc my nh gi kiu gen t ng c kh nng xt nghim 10.000 im chn on DNA trong mt ngy. Thc vy, kh nng kim tra ton b genome i vi nhng marker di truyn, hoc s th hin gen trn tng chip n l l hin thc trong mt tng lai rt gn. K thut chuyn np gen s n gin v d dng hn nh cc tin b mi trong cng ngh sinh hc, qua , vic ci bin di truyn tr nn hiu qa hn i vi mc tiu ci tin ging cy trng, gc cng ngh c lin quan n "genomics". 1-10. HIN TNG SYNTENY & GENOMICS C TNH CHT SO SNH Trong cui thp k 1990, mt s kin khoa hc quan trng rt ng ghi nh l khm ph thnh phn ca gen, v tr th t ca gen c ghi nhn c quan h rt cht gia cc loi thc vt khc nhau. l hin tng "synteny". Thut ng "synteny" c ngun gc t ch Hi Lp, trong "syn" c ngha l quan h vi nhau, "taenia" c ngha l di bng (ribbon) Thut ng ny c dng trong di truyn m ch s hin din ca hai hoc nhiu hn hai loci trn cng mt nhim sc th. Ngi ta cn s dng thut ng lp bn so snh (comparative mapping) v xc nh gen trn c s "gen pht trin ng dng" (homeologybased gene isolation) minh ha hin tng synteny c trong t nhin (McCouch 2001). Khi nim "synteny" c m rng trn khi nim ng dng ca nhim sc th tng ng. Nghin cu trn h Solanacea, Bonierbale v ctv. (1998) chng minh rng cDNA markers trn 12 nhim sc th ca c chua v khoai ty c tnh cht ng tuyn (collinear), chng ch khc nhau 5 o on khng v tr trung tm, trong khi , cy t c mt s sp xp tng
ng nhng to ln hn hai genome ny. Trong h Graminae, ngi ta nhn thy chng c mc pht trin c th c nh gi cao nht v hin tng synteny, bi v s th hin rt r rng ca cc loi trong h. Hu ht cc gen trong h c chc nng c bit hay cha c bit u c nhng mt m protein tng ng vi cc gen ca chui m cy la. Bn genome ca nhng loi thuc h Graminae bao gm nhng cy m cc c th c ni vi nhau thnh trc ng thng cc gen mc tiu nh v trn genome (vi ln khc nhau, theo vng trn ng tm) (hnh I-1). Nu mt gen ca mt loi c bit v chc nng, ngi ta c th d on trn tt c nhng loi cn li. Theo hnh I-1, cy la c genome nh nht, th hin vng trn trung tm.
Hnh I-1: nh v cc gen c quan h gia cc loi khc nhau bao gm la (rice), k ui chn (foxtail millet), cao lng (sorghum), k ht ngc (pearl millet), ng (maize), nhm la m (Triticeae) theo nghin cu ca Gale v Devos (2002). Gen waxy :amylose thp, lm tinh bt do, dnh, photoperiod sensitivity: cm quang, liguleless : khng c tai l, shattering: d rng ht, d4-dwarfing: gen ln d4, Ga-ins dwarfing: gen ln khng mn cm vi gibberellin. (CT)8 La (GA)20 Gen 1 2 3 4a 4b (TC)2
(TG)8 La mch
10kb
(GGC
(GT)1
Hnh I-2.: Thnh phn gen v v tr th t gen c bo tn gia hai cy la v cy la mch. Mt gen c nhn thnh i trong mt ca hai genome. Gen ca la mch vi kch thc genome ln hn la nc, nh v ri rc vi nhng on phn t lp li kh ln (Dubcovsky v ctv. 2001)
Chc nng ca gen trong iu khin tnh trng no c th c d on trong tt c cc cy thuc nhm m cc. S tng ng gia cc genome nh vy qui kt nhng gen li vi nhau trong mt gii hn c th bit c. Hnh I-2 cho thy cc vng c tnh cht lin gen (intergenic) rt khc nhau gia hai loi c quan h rt gn nhau, v rt khc nhau v kch thc genome Chnh nh hin tng synteny, vic ng dng nhng cng c chnh ca genomics tr nn thun tin hn, t kt qa phn tch genome ca cy la, ngi ta c th ng dng trc tip trong phn tch di truyn genome ca la m, la mch, k, cao lng, v ng. Bn so snh th h mt c cng b i vi cy la v tt c nhng genome ging nh vy (Gale 2002). Mt b su tp cc phn t "probe" ng vai tr neo c s bn sao cDNA thp (anchor probes) c s dng lm ra bn c tnh cht so snh trong cc loi khc nhau (# 7 loi) ca h Ho Bn. Cc vng c gen xp theo th t c bo tn c tnh cht v tr tng ng vi kiu hnh th hin ra bn ngoi c t liu ha mt cch h thng, thng qua s ng gp ca cc phn t "mutant" v nhng QTL. Tuy nhin, c rt nhiu trng hp ngoi l th d nh marker lin kt khng c thy trn bn v tr d on gia hai ng thng xc nh vng mc tiu, v t ra cho chng ta nhiu cu hi cha gii thch c. Kilian v ctv. (1997) ln u tin c gng "clone" mt gen trong mt loi cy trng trn c s thng tin v chui m v thng tin c tnh cht v tr (positional) (thut ng chuyn mn gi l "microsynteny"), hin nay ngi ta s dng thut ng "vng ng dng" (homeologue region) ca mt genus khc vi genus ang nghin cu. Cho d cc on tng ng ca nhim th cy la mch (Hordeum vulgare) v cy la (Oryza sativa) th hin kh r nt v tnh cht ng dng ti vng m gen khng bnh r st ca la mch Rpg1 nh v, nhng gen mc tiu ny khng h c tm thy vng d on trn genome cy la (McCouch 1997). Chong 5 minh ha hin tng synteny trong nghin cu ging la chng chu c nhm 1-11. NG DNG GENOMICS TRONG CI TIN GING CY TRNG CHNG CHU VI STRESS 1-11-1. Th vin DNA (DNA library) Nhm p ng mc tiu ng dng genomics trong ci tin ging cy trng chng chu vi stress khng phi sinh hc, ngi ta rt quan tm n kh nng khai thc hin tng synteny v genomics c tnh cht so snh, hn l ni dung ra cc gii php trn c s thit k chng trnh lai to vi database kh phong ph (modelling) ging nh chng trnh la dng hnh mi c nng sut vt trn, bi v c ch chng chu vn cha c hiu r rng. Nh vy, ngi ta rt cn c mt c s vt cht v genome mt cch cn bn v y . l th vin cc DNA clone. Yu cu ti thiu phi c l (1) bn lin kt gen mc phn t trn tng nhim sc th, (2) mt th vin DNA ln, (3) mt h thng chuyn np c kh nng mang mt s lng ln gen mc tiu chuyn vo cc cy c ci bin v di truyn. Bn phi p ng iu kin ph kn trn nhim th vi 2-3 cM gia hai marker k cn, v mt s lng ln cc loci mang tnh cht neo (anchor). Bn RFLP, EST, SSR, SNP s cung cp cho chng ta nhng so snh v kt qa p dng chng ta la chn. Th vin DNA s c th cho hiu qa tt hn gp i nu l th vin BAC vi kch thc DNA gn vo vect ln hn 100 kb (Gale 2002). Ngi ta cn d tnh s khai thc b su tp EST (nhng chui k t ca gen c chuyn m), mt bn so snh, v mt qun th c tn gi chuyn mn l knockout populations. Nhng EST thng l nhng su tp t cc m thc vt b stress, hoc cha b stress. i vi bn c tnh cht so snh, ngi ta s sp xp nhng nhim sc th c tnh cht ng dng theo mt model chuyn bit no . i vi qun th knockout, ngi ta s to ra thng qua th vin DNA t bin, hay th vin DNA b mt on, trong cc gen nh vy tr nn khng bnh thng mt cch ngu nhin. Hin nay, ngi ta nghin cu T-
DNA c nh du, hay transposon c nh du (Bu 2002), kt qa c minh ha trong trng hp cy la v cy Arabidopsis thaliana. Lnh vc genomics ca cy la, la m v cy ng hin rt phong ph. Bn cnh , CGIAR cng khuyn khch pht trin nghin cu genome ca cy k, cao lng, u , khoai m 1-11-2. K thut cloning cc gen C nhiu cch clone nhng gen mc tiu, tuy nhin, chng ta phi nh rng nu mi hot ng xc nh mt v tr no trn bn ca mt gen mc tiu iu khin tnh chng chu stress, nhng chng ta cha ghi nhn mt chc nng c th ca n, th tin trnh phn lp gen ng c vin ny s gp tr ngi, s th hin gen trong mt iu kin nht nh s khng c hiu r (nht l tng tc ca tnh trng chng chu vi mi trng). Chin lc nghin cu map-based cloning c xut. Ngi ta xp hng cy trng theo quan h huyt thng v xc nh nhng bn c tnh cht kiu mu (model maps) cha ng cc gen ng c vin nh v trong khu vc d on, trn c s phn tch QTL. Vi tn sut khong 30 gen trn mt n v bn ca cy la, ngi ta s c th nh v mt cch chi tit hn nhng QTL, m nhng sng lc nh vy lm cho QTL c xem xt nh nhng gen ch lc trong di truyn Mendel. Do , c mt thut ng mi dng din t s kin ny c gi l Mendelisation, trong mt qun th phn ly rt ln. Nhng BAC contig chng lp nhau to ra series ca nhng clone. Thng qua bn vt l v thng qua chui k t c tnh cht model, cc series ca clone mong mun c th c to ra. Mt k thut mi v map-based cloning c pht trin gn y, l k thut deletion tilling. K thut ny bao gm: (1) to ra s lng phn t b mt on, trong c gen mc tiu, (2) s dng on phn t chng lp ti thiu xc nh gen ng c vin trong khi m phng (trng hp genome la m). Phng php ny c thun li l khng cn bin d ca gen mc tiu hoc vng k cn ca n. Hin nay CGIAR rt ch n vic ng dng ca k thut tilling 1-11-3. Microarray Nhng gen ng c vin cng c th c th hin trong k thut phn tch microarray. Nhng gen nhy cm vi stress, s th hin gen xy ra khi b stress, tr nn rt l tng cho nghin cu microarray. Th nghim c tnh cht in hnh v array ca EST vi RNA c ly trch trong m cy b tn thng do stress, v m cy khng b stress, c thit k tm hiu v hiu qa ca phn tch microarray. Ngi ta d on c khong 25.000 gen trong genome cy Arabidopsis v 50.000 gen trong genome cy la s c pht hin y trong mt tng lai gn (Gale 2002). Microarray c hnh thnh t b su tp EST t th vin cDNA, hoc cDNA c su tp trn m b stress (do kh hn, mn, thiu ln, c nhm, v.v...). Th d trong cy la, 10% gen s iu tit up hoc down trong vng 1 gi sau khi chng b x l trong mi trng mn (Kawasaki v ctv. 2001). Ngi ta s pht trin cng ngh to cc "microarray" hay "gene chips" trong nghin cu genome v chc nng. Nhng chips sinh hc ny rt hu dng trong tng lai gn tm ra nhng gen mc tiu c tnh cht ng c vin (candidate genes) i vi tng tnh trng mong mun. y l bc t ph c tnh cht lch s trong qa trnh pht trin ngnh di truyn phn t ca loi ngi. Nhiu dng t bin mt on, dng du nhp gen cho nng sut cao, c th trng ni thiu nc. Nhiu dng th hin tnh chng chu hn v mn rt tt. Nhng nghin cu v chc nng nh vy cho php chng ta hiu r hn: lm th no cy la c th thch ng vi cc stress, tm ra cc gen hu ch cho cng tc lai to ging la. Theo Tin s Leung, c hn 100 gen gip cy la kim sot tnh khng bnh hi c tm thy to ra ging la khng bnh tt hn. K thut mi v "microarray" bao gm mt s tp trung khong 20.000 gen trn mt "slide". Ngi ta cn gi l "chip" ng vai tr nh mt "sensor" tm ra nhng thng tin di truyn cn thit. Phng php ny cho php chng ta c
th lai cng mt lc vi rt nhiu "probe". "Probe" l nhng chui k t ca cDNA, c ngun gc t cc gen chng chu vi mc stress khc nhau. Phn t mRNA i vi tnh trng chng chu stress no c chuyn m ngc thnh cDNA. Phn t cDNA ny c dng lai vi "microarray", sau ngi ta xc nh nhng "clone" dng tnh. Thng qua nhiu giai on pht trin, sau qa trnh bnh thng ha, ngi ta phi m bo rng nhng chui k t ny ng nht, sn sng c xc nh trn slide hoc trn mng. Chip sinh hc ny c th c ng hoc m khi cy iu kin bnh thng hoc b stress. Phng php ny khng ch xc nh gen ng c vin m cn tm hiu c qa trnh th hin gen trong iu kin b stress 1-11-4. Qun th knockout Ngi ta thc hin nhiu qun th khc nhau c nh du bi transposon hoc T-DNA phc v cho nghin cu genome cy la v cy Arabidopsis. Nhng qun th nh vy cn c gi vi thut ng l gene machines. Nhng my gen ny vi hnh thc di truyn b o ngc s gip chng ta xc nh bt c mt gen no rt cn cho mc tiu nghin cu, trn c s gen b t bin xen on, hoc gen nhy. Nhng dng trong qun th knockout co th c nghin cu theo mt kiu hnh c quan h vi chc nng ca gen mc tiu. Gn y, ngi ta pht trin mt phng php mi, l TILLING, thut ng ny c vit tt t ch targeted induced local lesions in genomes. Phng php TILLING gip chng ta to ra nhng knockout mc tiu v t sng to ra cc series c tnh cht alen vi nhau ca bt c gen no cn nghin cu. Nhng qun th TILLING hin c thnh lp phc v cho nghin cu genome cy la v cy Arabidopsis. Qun th t bin do ha cht hay phng x ca cy trng tr thnh mt yu cu cn thit trong nghin cu di truyn trong tng lai. Nhng dng chng chu vi stress nh kh hn, mn, nhit lnh, ngp ng u c th c xc nh. Th thch trc mt i vi cc nh khoa hc l lin kt kiu hnh vi gen mt on, chui m di truyn ca gen ng c vin c xem nh mc khi ng rt tt 1-11-5. Chuyn np gen K thut chuyn np gen hin tr nn thng dng cho hu ht cc loi cy trng, nhng hiu qa ca n vn cn l vn cn c tip tc nghin cu ci tin. c bit i vi cy mt l mm, hiu qa chuyn np gen t c kh hn so vi cy hai l mm. Nhng c gng u tin s l cng vic kin trc alen ca gen mc tiu gn vi promoter c chc nng kin to, th d CaMV35S trong qa trnh chuyn np gen. Nhng xt nghim v transgenic u tin s l ni dung bao gm kin trc dy antisense, sao cho thng tin c th hin nh mong mun. Nhng xt nghim sau cng s l xem xt kh nng ca nhng alen c bit ca gen trong kin trc tng thch vi nhng promoter th hin chc nng hot ng m, th d m r, m ht trong mt giai on pht trin cc trng no , nhm t c yu cu chng chu stress theo mc tiu ra t ban u. Thit k theo m hnh cy mu Genome cy Arabidopsis thaliana c th c xem nh m hnh cy lm mu (model species) trong nghin cu sinh hc phn t v tnh chng chu i vi stress khng phi sinh hc Tnh trng chng chu lnh c iu khin bi CBF1, mt regulator ca genome cy Arabidopsis. S th hin gen CBF1 lm kch hot mc hot ng ca hng lot cc gen iu khin tnh chng chu lnh, bo v cy chng li s thit hi do gi lnh Mt yu t c tnh cht gii m khc l DREB1A iu tit s th hin hng lot cc gen chng chu vi stress c trong genome cy Arabidopsis. S th hin DREB1A lm kch hot cc gen chng chu kh hn, gen chng chu mn v chng chu lnh. Khi DREB1A c khi ng bi CaMV35S, s pht trin bnh thng ca cy trong iu kin khng c stress b
nh tr mt cch nghim trng (Gale 2002), nhng khi n c t trong mi trng c stress, tnh trng chng chu s c ci tin rt tt. Thu thp qy gen - k thut allele mining v di truyn phi hp Vic thu thp ngn hng gen, a dng ngun vt liu c gen iu khin tnh chng chu vi stress l mt chin lc lu di ci tin cy trng. Ngi ta rt ch mt phng php c tn gi l khai thc m alen (allele mining). Phng php ny bao gm qa trnh thc hin PCR v c chui k t DNA (sequencing) ca hng lot cc gen c tm thy trong ging cy trng bn a, trong cc loi hoang di c quan h huyt thng gn gi. Bin d trong chui m c th cho chng ta mt kt qa tng ng vi tnh chng chu stress ca mu ging, qua , chng ta c th xc nhn alen no l tt nht cho nhng th nghim v chuyn np gen sau ny. Ngi ta cng ang c gng tip cn mt phng php c gi l di truyn phi hp (association genetics). l mt thut ng pht trin t ni dung thu thp qy gen m CGIAR ngh cc nh khoa hc nn khai thc. Lnh vc mi ca khoa hc ny, trn c s thnh tu di truyn hc ca con ngi vi mc phn tch cao trong nhng qun th phn ly v cng to ln, khng cho php chng ta thc hin ni dung nh vy. Ngi ta bn ngh n cc gen c phi hp vi mt tnh trng no , nhng gen ny c xc nh thng qua tng quan gia kiu hnh vi nhng alen chuyn bit mc marker phn t lin kt cht ch vi gen. Trong thc vt, l s th hin tng qut ca nhng mu ging c su tp trong ngn hng gen i vi hin tng bin d ti cc loci ca marker, phn b ri rc trong genome (nh gi kiu gen). Ngi ta pht trin ni dung ny bng cch tm mi tng quan gia nhng kiu gen trong iu kin stress vi hin tng khng cn bng ca alen trong genome (allele dis-equilibrium). y l mt ngnh khoa hc tr trong lnh vc sinh hc thc vt, c tim nng rt ln khm ph ra cc gen mi Ngnh hc mi v sinh hc phn t ca thc vt ang bt u pht huy tc dng trong lnh vc chn to ging cy trng chng chu vi nhng thit hi khng phi sinh hc. Genome hc c tnh cht so snh l mt v d, n rt c trin vng pht trin nhanh hn na (Gale 2002). Mc tiu ca Nhm T Vn v Nghin Cu Nng Nghip Quc T (CGIAR) phi t l to ra cc ging cy trng pht trin rng khp trn t c kh nng trng trt, nng sut cao ngay c trn t c vn , vng kh hu bt thun, m bo yu cu an ton lng thc, c bit cho ngi ngho trn ton th gii Trong chng trnh ci tin ging cy trng, ngi ta phn u p dng nhng thnh tu mi nht ca khoa hc, vi nhng cng c tt nht gii quyt cc vn khc nhau, l: Gen mi v gen ci tin trong trng hp tnh chng chu vi stress khng phi sinh hc Cng c ci tin ging c hiu qa cao trong khi a cc gen ny vo ging cy trng mi, th d phng tin thanh lc ging tt hn, marker phn t lin kt vi tnh trng mc tiu cht ch hn, qui trnh chuyn np gen hiu qa hn Chin lc ci tin ging cy trng i vi tnh chng chu no phi c th hin trong chng trnh trng im quc gia Kin thc c bn v sinh l thc vt, sinh ha i vi c ch chng chu stress phi c ci tin khng ngng
1-12. PHN TCH QTL Phn ln nhng tnh trng chng chu vi iu kin bt li do mi trng l tnh trng di truyn s lng. Do , phn tch nhng loci ca tnh trng s lng QTL (quantitative
trait locus-s t, quantitative trait loci-s nhiu) c pht trin vi nhiu m hnh nhm p ng yu cu nghin cu (Liu 1998) Tnh trng s lng c nh ngha mt cch kinh in l tnh trng c phn b lin tc (continuous distribution), tnh trng ny c iu khin bi nhiu gen, mi gen c mt nh hng nh i vi tnh trng mc tiu. Bn QTL bao gm kin trc ca nhng bn genome v tm kim mi quan h gia tnh trng vi nhng marker a hnh, minh chng mt QTL nh v k cn nhng marker. Di truyn tnh trng s lng rt phc tp do vi tnh trng n gen trong di truyn Mendel, bi v n cn chu s tc ng rt mnh ca mi trng. Danh sch nhng ti liu tham kh v bn QTL hin nay kh phong ph, c bit l cng trnh ca Lander v Schork (1994) Lander v Zheng (1994) Stuber v ctv. (1992) Tanksley (1993) Weller (1998) Nhng tc gi s dng php th T (t-test), phng trnh tuyn tnh, phng trnh a tuyn, phng trnh phi tuyn tnh v php th cch qung xy dng m hnh. Thnh tu ni bc l phn mm MAPMARKER/QTL ca Lander v ctv (1987) QTLSTAT ca Liu v Knapp (1992), QGENE ca Tanksley v Nelson (1996) gip cho vic nghin cu phn tch QTL c din gii mt cch r rng hn Bng 1: Danh mc nhng computer software lin quan n phn tch v lp bn QTL Phn mm MAPMARKER/QTL QTLSTAT LINKAGE PGRI QTL cartographer MAPQTL Map Manager QT QGENE Ngun Lander v ctv. 1987, Lander & Bostein 1989 Knapp v ctv. 1992 Terwilliger & Ott 1994 Liu 1998 Basten, Weir v Zeng 1996 van Ooijen & Maliepard 1996 Manly & Cudmore 1996 Tanksley & Nelson 1996, Nelson 1997
1-12-1. Nhng m hnh v di truyn s lng 1-12-1-1. M hnh QTL n (single-QTL) Mt trong nhng mc tiu ca bn QTL l tm kim trong ton b genome ni nh v ca gen mc tiu thng qua mt trc nghim gi nh i vi mt marker n hoc mt v tr nht nh, sau , ngi ta s xy dng n mt model c tnh cht a QTL (multiple QTL) Nh vy, nh hng tnh cng (a) v nh hng tnh tri (d) c biu th trong bng 2 trong m hnh qun th phn ly F2, v gi tr di truyn trong qun th hi giao, vi cc nh hng nh sau: nh hng chnh, nh hng tng tc 2 chiu, 3 chiu, 4 chiu v tng tc i-way. Cui cng l nh hng tng qut.
Bng 2: M hnh QTL n trong qun th hi giao (backcross) v F2, nQQ, nQq, v nqq biu th kiu gen ca QQ, Qq, v qq M hnh Hi giao (Qq x QQ) Kiu gen QQ Qq nh hng di truyn QQ Qq qq nh hng tnh cng nh hng tnh tri Gi tr 1 2 g = 0,5 (1 - 2) 1 2 3 a = 0,5 (1 - 3) d = 0,5 (22 -1 - 3) Phng sai 2 2 2 (1 / nQQ + 1 /nQq)/4 2 2 2 2 (1 / nQQ + 1 /nqq)/ 4 2(1 / nQQ + 4nQq + 1/nqq)/ 4
F2 (Qq x Qq)
1-12-1-2. M hnh multiple-locus M hnh di truyn tnh trng s lng c nh ngha theo s gen, cc nh hng ca gen, tn sut gen, tng tc gia cc gen, tng tc gia gen x mi trng. Gi nh c n gen iu khin mt tnh trng s lng no . Hy xem xt qun th F2, nhng nh hng di truyn c th xy ra: nh hng chnh, nh hng tng tc 2 chiu, 3 chiu, 4 chiu v tng tc i-way. Cui cng l nh hng tng qut. Bng 3: Cc h s thuc bin s dummy v nh hng ca gen Kiu gen QTL AA Aa Aa Tn sut f1 f2 f3 C1 1 0 -1 C2 1/2 -1/2 1/2 nh hng a + d/2 - d/2 -a + d/2
1-12-2. Phng php phn tch marker n (SMA = single marker analysis) ng dng nguyn tc trong phn tch lin kt gen, chng ta c th xc nh cch thc ca tnh cht di truyn marker v v tr m n hin din trong genome Gen iu khin tnh trng s lng c th c lp bn ging nh bn di truyn ca marker Khi nhng marker ph trn mt on kh ln ca genome, n s to ra kh nng rt tt chng ta tm ra nhng gen iu khin tnh trng s lng Nu cc gen v cc marker ng phn ly trong mt qun th di truyn, tng quan lin kt gia chng c th c pht hin
Phng php phn tch marker n l mt bc khi u, khng nhng ch ra bn QTL, m cn phn tch c c th cc s liu Nhng khm ph u tin ca m hnh c cn c trn hm tuyn tnh Yj = + f(markerj) + j Trong Yj l gi tr ca tnh trng i vi c th th jth, v l gi tr trung bnh ca qun th, f(markerj) l hm s ca kiu gen marker, j l sai s
Th d nh mt gen Q nh v gn marker A v tnh trng mc tiu c iu khin bi gen Q c th c m phng thng qua marker A Yj = + f (A) + j Bi ton l gii p xc sut tin cy c ca f (A) c cha gi tr di truyn ca kiu gen Q v gi tr lin kt gia A v Q Phng php SMA c th c thc hin di dng php th t n gin phn tch phng sai phng trnh tuyn tnh php th t l m phng v c on m phng ti a
Phng php SMA rt n gin trong phn tch s liu v thc hin cc bc tnh ton, ngi ta c th s dng phn mm SAS thit lp m hnh. 1-12-2-1. SMA trn qun th hi giao B m Hi giao (BC) AAQQ x aaqq aaqq x AaQq x AAQQ Tn sut hi vng Con lai BC AaQq Aaqq aaQq aaqq AAQQ AAQq AaQQ AaQq 0,5 (1 r) 0,5 r 0,5 r 0,5 (1 r)
Gi tr r l tn sut ti t hp gia A v Q Hnh 1-3: Con lai hi giao trong SMA 1-12-2-2. Kt hp hin tng phn ly ca QTL v marker Bng 4: Tn sut kiu gen QTL hi vng nh v k cn marker trong qun th hi giao, khng c hin tng qun cho (Liu 1998) Kiu gen marker AA Aa AA Aa Gi tr quan st n1 n2 n1 n2 Tn sut bin 0,5 0,5 0,5 0,5 Kiu gen QTL QQ Qq Tn sut kt hp 0,5 (1-r) 0,5 r 0,5 r 0,5 (1-r) iu kin kt hp 1-r r r 1-r Gi tr l thuyt
(1-r)1 + 2 r1 + (1-r)2
1-12-2-3. Php th t n gin trong qun th hi giao
Bng 5: Phn tch phng sai SMA trong qun th hi giao vi N l qui m qun th con lai, b l s ln lp li v c l h s d on c = N [n12 + n22] / N Ngun Di truyn tng qut QTL G(QTL) S cn df N-1 1 N-2 N(b-1) MS MSG MSQ MSG(Q) MSE MS l thuyt e2 + b2G(QTL) + bcg2 e2 + b2G(QTL) e2 F-test MSQ/MSG(Q)
1 - 2 tQ = ------------------------------------[s2 [ 1/n1 + 1/n2) ]1/2 1-12-2-4. Phn tch phng sai trong qun th hi giao Bng 6: Phn tch phng sai SMA in hnh Ngun Di truyn Marker G(marker) S cn df N-1 1 N-2 N(b-1) MS MSG MSM MSG(M) MSE E(MS) e2 + b2G e2 + b[2G(QTL)+4r(1-r)a2]+bc(1-2r)2.a2 e2 + b[2G(QTL)+4r(1-r)a2] e2
Php th F = MSM / MSG(M) 1-12-2-5. M phng trong qun th hi giao
Hnh 1-4: ng phn b l thuyt ca gi tr tnh trng i vi kiu gen QTL trong tng kiu gen marker AA v Aa trn c s qun th hi giao. Marker A lin kt vi QTL vi r=0,2 (Liu 1998) Phng php tip cn theo kiu m phng (likelihood) c p dng trong SMA (Weller 1986). Mt marker A no c gi nh lin kt vi mt QTL vi tn sut ti t hp r = 0,2. QTL ny c mt nh hng di truyn g = 0,5 (1 - 2) = 0,5 . Phng sai tnh trng ca 2 kiu gen QTL (QQ v Qq) l 2. i vi hai nhm marker AA v Aa, phn b ng biu bin ca gi tr tnh trng l mt kt hp gia hai ng chun nh vy (Hnh 14)
1-12-3. Phng php SMA trong qun th F2 B m F1 T th AAQQ, Aaqq, AAQq AaQq AAQQ x AaQq

aaqq
Con lai F2
aaqq aaQQ AaQQ, Aaqq, aaQq
Tn sut l thuyt 0,25 (1-r)2 0,25 r2 0,5 (1-r) 0,5 [r2 + (1-r)2]
r l gi tr ti t hp gia A v Q Hnh 1-5: Qun th con lai F2 trong SMA Bng 7: Phn tch phng sai SMA in hnh trong qun th F2. Ngun Di truyn Marker A D G(marker) S cn df N-1 2 1 1 N-3 N(b-1) MS MSG MSM MSA MSD MSE F MSM/MSG(M) MSA/MSG(M) MSD(MSG(M)
Bng 8: Tng phn tuyn tnh i vi nhng kiu gen ca marker trong qun th F2. Tng phn Tnh cng Tnh tri Tn sut l thuyt AA 1 1 0,25 Kiu gen marker Aa 0 -1 0,5 Aa -1 1 0,25
Codominant marker: FM = MSM / MSG(M) E (tnh cng) = [ + (1-2r)a + 2r(1-r)d] [ - (1-2r)a + 2r(1-r)d] = 2(1-2r)a E (tnh tri) = [ + (1-2r)a + 2r(1-r)d] 2{[ + [1-r)2 +r2]d} + [ + (1-2r)a + 2r(1-r)d] = 2(1-2r)2 d Dominant marker: ngi ta phi dng l thuyt gi nh H0 = A- - aa E (A- - aa) = [ + 1/3 (1-2r)a + 2/3(1-r+r2)d] - [ - (1-2r)a + 2r(1-r)d] = 1/3[4(1-2r)a + 2(1-2r)2d]
1-12-4. Phn tch QTL trn c s bn cch qung (interval mapping) r A (marker) r1 Q r2 B (marker)
(QTL gi nh) Hnh 1-6: Mi quan h lin kt gia mt QTL v hai marker k cn QTL lin kt vi marker A c gi tr ti t hp (recombination fraction) l r1 QTL lin kt vi marker B c gi tr ti t hp l r2 Mi quan h lin kt c gi tr ti t hp r = r1 + r2 2r1r2 Khi r c gi tr rt nh, r = r1 + r2 V tr ca QTL c th hin bi mt v tr tng ng vi qung gia A v B = r1 / r 1 - = r2 / r 1-12-4-1. Bn cch qung trong qun th hi giao (BC) AAQQBB (1) AaQqBb x aaqqbb (2) x AAQQBB (1) Tn sut l thuyt Con lai BC AAQQBB, AAQqBb, AAQQBb, AAQqBB, AaQqBb AaQQBB AaQqBB AaQQBb 0,5 (1-r) 0,5 r1 0,5 r2 0
Hnh 1-7: Qun th con lai BC trong phn tch bn cch qung vi 2 marker A v B Bng 9: Tn sut hi vng ca QTL v marker trong qun th BC, khng c qun cho Kiu gen marker Gi tr c on p (Qi/Mi) (gi) QQ Qq AABB 0,5 (1-r) 1 0 1 AABb 0,5 r r1 / r = (1- )1 + 2 r2 / r = 1- AaBB 0,5 r r2 / r = 1- 1 + (1-)2 r1 / r = AaBb 0,5 (1-r) 0 1 2 Trung bnh 0,25 2 0,5 (1 + 2) 1 Phng php m phng bn QTL trn qun th BC c thc hin theo hm s 1 (yi -)2 L = ------------------- p (Qi/Mi) exp [-----------] 22 ([2]1/2)N Tn sut pi

Sau , chng ta tnh gi tr Log (L) (logarith likelihood)
Hnh 1-8: Phn b l thuyt ca gi tr tnh trng i vi kiu gen QTL (QQ vq Qq) trong kiu gen marker (AABB, AABb, AaBB v AaBb), trn c s qun th BC. Marker A v B lin kt vi gi tr r = 0,3 v QTL nghin cu nh v gia hai marker ny (Liu 1998) 1-12-4-2. Bn cch qung trong qun th F2 Th h F1 biu th u th lai gia hai dng cn giao AAQQBB x aaqqbb. Bng 10 cho thy tn sut hi vng c c ca kiu gen QTL v marker trong F2 vi gi nh l khng c trao i cho. Tn sut ti t hp gia marker A v B l r, gia QTL v marker A l r1. Phng php m phng bn QTL trn qun th BC c thc hin theo hm s 1 (yi -)2 L = ------------------- p (Qi/Mi) exp [-----------] trong trng hp codominant marker 22 ([2]1/2)N Bng 10: Tn sut hi vng c c ca kiu gen QTL v marker trong F2 Kiu gen marker AABB AABb Aabb AaBB AaBb Aabb AaBB AaBb aabb Tn sut pi 0,25 (1-r) 0,5 (1-r) 0,25 r2 0,5 r (1-r) 0,5 (1-r)2 + r2 0,5 r (1-r) 0,25 r2 0,5 r (1-r) 0,25 (1-r)2
2
QQ 0,25 (1-r)2 0,5 r2 (1-r) 0,25 r22 0,5 r1 (1-r) 0,5 r1r2 0 0,25 r12 0 0
pij Qq 0 0,5 r1 (1-r) 0,5 r1r2 0,5 r2 (1-r) 0,5[(1-r)2+r12+r22] 0,5 r2 (1-r) 0,5 r1r2 0,5 r1 (1-r) 0
qq 0 0 0,25 r12 0 0,5 r1r2 0,5 r1 (1-r) 0,25 r22 0,5 r2 (1-r) 0,25 (1-r)2
LOD score l gi tr chng minh mc c ngha ca phn tch, c tnh ton nh sau:

LOD1 = log10 L (1, 2, 3, 2, r1) log10 L (, 2) LOD2 = log10 L (1, 2, 3, 2, r1) log10 L (1, 3, 2, r1) LOD3 = log10 L (1, 2, 3, 2, r1) log10 L (1, 2, 2, r1) Thng thng bn QTL c ngha khi gi tr LOD 3. 1-12-5. Kh nng gii thch v thng k sinh hc ca bn QTL Nhng yu t sau y c th nh hng n bn QTL: S gen iu khin tnh trng v v tr ca n trong genome S phn b ca cc nh hng di truyn v cch thc tng tc gia cc gen H s di truyn ca tnh trng mc tiu S gen phn ly trong qun th lp bn Loi qun th con lai v ln ca qun th Mt ca bn lin kt gen, s marker ph trn genome Phng php thng k c ng dng, mc c ngha trong phn tch
Trong phng php SMA, kh nng thng k c da trn php th t (t-test), phn tch phng sai v trc nghim m phng theo dng tuyn tnh Trong phng php bn cch qung (IMA: interval mapping analysis), kh nng thng k da trn mt h phng trnh tuyn tnh vi con lai hi giao, i thuyt H0 = 1- 2 = 0 (khng c nh hng QTL trong qung), trc nghim m phng thng qua gi tr log likelihood Trong phng php multiple QTL, kh nng thng k da trn l thuyt mi gen l tp hp ca {p1, p2, p3,.. pq} mc tin cy , nhm pht hin c t nht qe QTL, trong , qe qi (Liu 1998)
Hnh 1-9: Tng quan gia kiu hnh, kiu gen v mi trng (Xu 1997)
Tn sut
Hnh 1-10: Pht trin qun th cn giao ti t hp (RILs) thng qua k thut SSD (single seed descent: thu thp tng ht trn mi c th) (McCouch v Doerge 1995)
Hnh 1-11: Th d bn QTL cy la trn nhim sc th s 8. (a) V tr v kch thc ca mt on ng dng (homeologous segment) ca nhim th 7A la m trn c s cDNA marker chung gia la v la m. (b) Tnh trng cun l (leaf rolling) trong c ch thot kh hn. (c) Tnh trng chiu dy ca r. (d) Tnh trng iu tit p sut thm thu OA. (e) S vt bnh trong tnh khng khng hon ton (partial resistance) bnh o n. (f) Gen Pi-zb (khng o n hon ton. (g) Ngy gieo n tr (x l 10 gi sng). (h) Ngy gieo n tr (x l 14 gi sng). (i) Ngy gieo n tr theo Li v ctv. (j) Ngy gieo n tr theo Xiao v ctv (McCouch v Doerge 1995) TI LIU THAM KHO Basten CJ, BS Weir, ZB Zeng. 1994. ZMAP- A QTL cartographer. Proceedings of the 5th World Congress on Genetics Applied to Livestock Production 22:65-66 Bonierbale MW, RL Plaister, SD Tanksley. 1998. Genetics 120: 1095-1103 Bi ch Bu, Nguyn th Lang. 2002. Chuyn v trong nghin cu chc nng genome thcvt. Bi ch Bu (ch bin). C s di truyn tnh khng su bnh hi cy trng. Nh xut bn Nng nghip, TP H ch Minh. Tr. 53-88 FAO. 2002. FAO statistics, http://apps.fao.org/(2002) Gale M. 2002. Applications of molecular biology and genomics to genetic enhancement of crop tolerance to abiotic stress a discussion document. FAO Consultative Group on International Agricultural Research Interim Science Council. Rome, Italy 26-30 August 2002. 26 p. Kawasaki S, C Borchert, M Deyholos, H Wang, S Brazille, K Kawai, D Galbraith, HJ Bohnert. 2001. Gene expression profiles during the initial phase of salt stress in rice. The Plant Cell 13:889-905 Kilian A, J Chen, F Han, B Steffenson, A Kleinhofs. 1997. Plant Mol Biol 35: 187-195 Knapp SJ, WC Bridges, BH Liu. 1992. Mapping quantitative trait loci using nonsimultaneous and simultaneous estimators and hypothesis tests. In: JS Beckmann and TS Osborn (eds.), Plant Genomes: Methods for genetcis nad physical mapping, pp. 209-237. Kluwer Academic Publishers, Dordrecht, The Netherlands Lander ES, P Green. 1987. Construction of multilocus genetic linkage maps in human. Proc Natl Acad Sci USA 84:2363-2367 Lander ES, D Botstein. 1989. Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121:185-199 Liu BH. 1998. Statistical genomics: Linkage, mapping and QTL analysis. CRC Press, New York, 611pp. Manly KF, EH Cudmore Jr. 1996. New versions of MAP Manager genetic mapping software. Plant Genome IV (Abs.) p105 McCouch SR. 2001. Genomics and synteny. Plant Physiology 125: 152-155 McCouch SR, RW Doerge. 1995. QTL mapping in rice. Plants 11(12): 482-487 Nelson JC. 1987. QGENE: software for marker-based genomic analysis and breeding Tanksley SD, JC Nelson. 1996. Advanced backcross QTL analysis: a method for the simultaneous discovery and transfer of valuable QTLs from unadapted germplasm into elite breeding. Theor Appl Genet 92:191-203
Terwilliger JD, J Ott. 1994. Handbook og human genetic linkage. The Johnson Hopkins University Press, Baltimore van Ooijen, C Maliepaard. 1996. MAPQTL version 3.0: Software for the calculation of QTL position on genetic map. Plant Genome IV (Abs.) p.105 Weller JI. 1986. Maximum likelihood techniques for the mapping and analysis of quntitative trait loci with the aid of genetic markers. Biometrics 42:627-640 Xu Y. 1997. Quantitative trait loci: separating, pyramiding, and cloning. Plant Breeding Reviews 15. Cornell University, NY 14853-1902, USA

Chng 2
C S DI TRUYN TNH CHNG CHU MN

t nhim mn l mt trong nhng yu t chnh lm kh khn trong chin lc pht trin sn lng nng sn, nng sut / ha, v th thch ln trong mc tiu an ton lng thc, trong iu kin kh hu ton cu ang thay i, bng tan hai cc, nc bin s dng ln e da cc vng canh tc t thp ven bin. t mn c th c phn chia lm hai nhm chnh da theo ngun gc pht sinh mn: mn ven bin (coastal salinity), hoc vng ca sng do nc bin xm nhp vo ma kh, c th trng trt bnh thng trong ma ma v mn bn trong t do mao dn t tng di ln (inland salinity) c th do ph rng, khng c tn cy che ph. Trong nhiu nm qua, ngi ta c gng ci tin nhiu ging cy trng c tnh chng chu mn tt. Tuy nhin, chng ta vn cha hiu mt cch y v bn cht, c ch chng chu v kh nng di truyn tnh trng chng chu mn (Mishra v ctv. 1998). Thnh tu t c trong chn to ging chng chu mn rt chm do nhng nguyn nhn nh sau: kin thc v di truyn tnh chng chu cn hn ch tnh cht phc tp ca c ch chng chu mn (Yeo v Flowers 1986) k thut thanh lc cha hon thin hiu qa chn lc thp tng tc gia tnh trng chng chu mn vi mi trng cha c hiu r (Akbar 1986)
i vi cy la, tnh trng chng chu mn l mt tin trnh sinh l rt phc tp, thay i theo cc giai on sinh trng khc nhau ca cy (Akbar v Yabuno 1972, 1975, 1977). Tnh trng bt th trn bng la khi b stress do mn c iu khin bi mt s gen tri, nhng cc gen ny khng tip tc th hin cc th h sau cng. Phn tch diallel v tnh trng chng chu mn, ngi ta ghi nhn c hai hot ng gen cng tnh v khng cng tnh vi h s di truyn thp (19,18%), v nh hng ca mi trng rt ln (Moeljopawirio v Senadhira 1993, Akbar v ctv 1985, Gregorio v Senadhira 1993). Mc c ngha v hot ng gen cng tnh v khng cng tnh c Mishra v ctv ghi nhn (1996). Nng sut v tnh chng chu mn giai on pht dc th hin rt khc nhau gia cc ging la so vi tnh chng chu mn giai on m (Ikehashi v Ponnamperuma 1978, Akbar v ctv. 1985, Mishra v ctv 1990). Hin chng ta c rt t thng tin v kiu hnh chng chu mn giai on trng thnh ca cy la hn l giai on m. Hu ht cc th nghim u c tin hnh trn giai on m, vi qui m qun th hn ch v ch s Na/K thng c dng nh mt gi tr ch th (Mishra v ctv. 1998). Cy la nhim mn c xu hng hp thu Na nhiu hn cy chng chu. Ngc li cy chng chu mn hp thu K nhiu hn cy nhim. Ngng chng chu NaCl ca cy la l EC=4dS/m (Sathish v ctv 1997). Trong qa trnh b nhim mn, nng ion K+ trong t bo c iu tit tng thch vi c ch iu tit p sut thm thu v kh nng tng trng t bo (Ben-Hayyim v ctv 1987). Nhiu loi thc vt thuc nhm halophyte v mt phn ca nhm glycophyte thc hin hot ng iu tit p sut thm thu lm cn tr nh hng gy hi ca mn. Hot ng ny s gip cy duy tr mt lng ln K+ v hn ch hp thu Na+. 2-1. T MN t mn c xem l t c vn rt ph bin trn th gii, lm hn ch nng sut cy trng. Tnh cht vt l v ha hc ca t mn rt a dng. Bin thin ny ty thuc vo ngun gc ca hin tng mn, pH t, hm lng cht hu c trong t, ch thy vn, v nhit (Akbar v Ponnamperuma 1982).
t mn cha mt lng mui ha tan trong nc vng r cy, lm thit hi n hot ng sinh trng ca cy trng. Mc gy hi ca t mn ty thuc vo loi cy trng, ging cy, thi gian sinh trng, cc yu t mi trng i km theo n, v tnh cht ca t. Do , ngi ta rt kh nh ngha t mn mt cch chnh xc v y . Hi Khoa Hc t ca M (SSSA 1979) xc nh t mn l t c dn in (EC) ln hn 2 dS/m, khng k n hai ga tr khc : t l hp thu sodium (SAR) v pH. Tuy nhin, hu ht cc nh ngha khc u chp nhn t mn l t c dn in EC cao hn 4dS/m iu kin nhit 250C, phn trm sodium trao i ESP km hn 15, v pH nh hn 8,5 (US Salinity Laboratory Staff 1954). t mn kh ph bin vng sa mc v cn sa mc. Mui tch t v mao dn ln t mt, chy trn trn mt t theo kiu ra tri. t mn c th pht trin vng nng m hoc cn nng m trn th gii trong iu kin thch hp nh vng ven bin, mn do nc bin xm nhp khi triu cng, l lt, mn do nc thm theo chiu ng hay chiu ngang t thy cp b nhim mn (Bhumbla v Abrol 1978). t mn b nh hng mn chim 7% din tch t ton th gii. t b nh hng mn khng phi u c kh nng canh tc ging nh nhau, m n c chia ra thnh tng nhm khc nhau s dng t hp l. t b nh hng mn i lc thuc Chu u v Bc M rt t c kh nng trng trt. Chu , hn 80% t b nh hng mn c kh nng trng trt, v c khai thc cho sn xut nng nghip. Chu Phi v Nam M, khong 30% t b nhim mn c kh nng trng trt. Hin tng nhim mn l mi e da ln nht n vic gia tng sn lng lng thc cc quc gia Chu (Abrol 1986). 2-2. C CH CHNG CHU MN C ch chng chu mn ca cy la c bit thng qua nhiu cng trnh nghin cu rt ni ting (Akbar v ctv. 1972, Korkor v Abdel-Aal 1974, Maas v Hoffman 1977, Mori v ctv. 1987). Mn nh hng n hot ng sinh trng ca cy la di nhng mc thit hi khc nhau tng giai on sinh trng pht trin khc nhau (Maas v Hoffman 1977) Nhiu nghin cu ghi nhn rng tnh chng chu mn xy ra giai on ht ny mm, sau tr nn rt mn cm trong giai on m (tui l 2-3), ri tr nn chng chu trong giai on tng trng, k n nhim trong thi k th phn v th tinh, cui cng th hin phn ng chng chu trong thi k ht chn (Pearson v ctv. 1966, IRRI 1967). Tuy nhin, mt vi nghin cu ghi nhn giai on la tr, n khng mn cm vi stress do mn (Kaddah v ctv. 1975). Do , ngi ta phi chia ra nhiu giai on nghin cu mt cch y c ch chng chu mn ca cy trng. Thit hi do mn th hin trc ht l gim din tch l. Trong iu kin thit hi nh, trng lng kh c xu hng tng ln trong mt thi gian, sau gim nghim trng do suy gim din tch l. Trong iu kin thit hi nng hn, trng lng kh ca chi v ca r suy gim tng ng vi mc thit hi. giai on m, l gi hn s mt kh nng sng st sm hn l non (Akita 1986) Thit hi do mn c gy ra bi s mt cn bng p sut thm thu v s tch t nhiu ion Cl- (Iwaki v ctv. 1953, Ota v Yasue 1958, Shimose 1963, Tagawa v Ishizaki 1963, Murty v Janardhan 1971). Thit hi do mn cn c ghi nhn bi hin tng hp thu mt lng qa tha sodium, v c tnh ca sodium lm cho clor tr thnh anion tr (neutral), c tc dng bt li vi mt ph rng v nng (Clarkson v Hanson 1980). S mt cn bng Na-K cng l yu t lm hn ch nng sut (Devitt v cvt. 1981). Ion kali c mt vai tr quan trng lm kch hot enzyme v ng m kh khng tng ng vi tnh chng chu mn ca cy trng, thng qua hin tng tch ly lng kali trong chi thn (Ponnamperuma 1984)
Yeo v Flower (1984) tng kt c ch chng chu mn ca cy la theo tng ni dung nh sau: Hin tng ngn chn mui - Cy khng hp thu mt lng mui d tha nh hin tng hp thu c chn lc Hin tng ti hp thu - Cy hp thu mt lng mui tha nhng c ti hp thu trong m libe. Na+ khng chuyn v n chi thn Chuyn v t r n chi Tnh trng chng chu mn c phi hp vi mt mc cao v in phn r la, v mc thp v in phn chi, lm cho s chuyn v Na+ tr nn t hn t r n chi Hin tng ngn cch t l n l - Lng mui d tha c chuyn t l non sang l gi, mui c nh v ti l gi khng c chc nng, khng th chuyn ngc li. Chng chu m Cy hp thu mui v c ngn cch trong cc khng bo (vacuoles) ca l, lm gim nh hng c hi ca mui i vi hot ng sinh trng ca cy nh hng pha long Cy hp thu mui nhng s lm long nng mui nh tng cng tc pht trin nhanh v gia tng hm lng nc trong chi
Tt c nhng c ch ny u nhm h thp nng Na+ trong cc m chc nng, do lm gim t l Na+/K+ trong chi (< 1). T l Na+/K+ trong chi c xem nh l ch tiu chn lc ging la chng chu mn (Gregorio v Senadhira 1993) Mi mt ging la u c mt hoc hai c ch nu trn, khng phi c tt c (Yeo v Flowers 1984). Phn ng ca cy trng i vi tnh chng chu mn v cng phc tp, l hin tng tng hp t nhng yu t ring l. Yeo v Flowers (1984) kt lun rng phn ng tt nht lm gia tng tnh chng chu mn phi gn lin vi vic ti u ha nhiu c im sinh l, c tnh cht c lp tng i vi nhau . Do vy, mc tiu ca chng ta l phi hp tt c nhng c ch sinh l y vo trong ging la ci tin tnh chng chu mn Abscisic acid (ABA) c xem nh mt yu t rt quan trng ca cy trng phn ng vi nhng stress gy ra do mn, do nhit cao (Gupta v ctv. 1998). Do ABA cn c xem nh l gen cm ng (inducible genes) trong c ch chng chu mn ca cy trng 2-3. DI TRUYN TNH CHNG CHU MN 2-3-1. Nghin cu di truyn s lng tnh chng chu mn Nghin cu di truyn s lng cho thy c hai nh hng hot ng ca gen cng tnh v gen khng cng tnh u c ngha trong di truyn tnh chng chu mn (Mishra v ctv. 1990, Gregorio v Senadhira 1993, Lee 1995). Trong giai on m ca cy la, cc tnh trng chiu di chi, hm lng Na v K trong chi, trng lng kh ca chi v r th hin s khc bit c ngha gia ging khng v ging nhim, tnh trng ny ch yu c iu khin do hot ng ca nhm gen cng tnh. H s di truyn tnh chng chu thng qua cc tnh trng nh vy rt thp (Teng 1994). Trong giai on trng thnh ca cy la, tnh trng chiu cao cy, nng sut trong iu kin x l mn c iu khin bi nhm gen cng tnh (Moeljopawiro v Ikehashi 1981, Akbar v ctv. 1986, Mishra v ctv. 1990) Trong phn tch di truyn s lng thng qua lai diallel 6x6, nng sut la th hin tnh hot ng ca nhm gen cng tnh khng c ngha trong iu kin bnh thng, nhng tr nn c ngha trong iu kin x l mn (Narayanan v ctv. 1990). Nng sut la b gim l do nh hng ca mn. Mt ging la c u th hot ng gen cng tnh i vi nng sut s l iu kin thun li cho chn lc ging trong mi trng mn.
Trong phn tch di truyn s lng thng qua lai diallel 9x9, tnh trng chng chu mn c xem xt qua t l thp ca Na / K trong chi, tnh trng ny c kim sot bi hot ng ca c hai nhm gen cng tnh v khng cng tnh. Tnh trng Na / K thp cn th hin nh hng siu tri v c iu khin bi t nht hai nhm gen tri. nh hng ca mi trng rt c ngha v h s di truyn thp (19,18%) (Gregorio v Senadhira 1993). T , cc tc gi ngh qun th con lai phi tht ln, v vic tuyn chn nn c thc hin cc th h sau cng, di iu kin mn c kim sot cht ch, gim thiu thp nht nh hng bin ng ca mi trng. Trong mt nghin cu v di truyn tnh chng chu mn bao gm cc b m c tnh trng tng phn nhau: ging CSR10 v CSR11 c chn lm b (c tnh trng chng chu mn), ging Basmati 370 c chn lm m (khng c gen khng mn) (Mishra v ctv. 1998). Th h F1 c x l mn c EC=10dS/m, iu kin trng trong chu. Th h F2 c trong trong iu kin bnh thng trn ng rung, chn theo phng php trng dn (bulk). Th h F3 c x l mn giai on m (EC=10dS/m). Qun th cy trng ca cc cp lai c chia thnh nhm ty theo phn ng chng chu i vi mn cc im 1, 3, 5, 7, 9. Gi tr trung bnh c tnh theo cng thc = G + I U = G + I ( fU/n) trong G = s nhm I = qung gia cc nhm f = tn sut cy trong mi nhm U = m s ca cy n = tng s cy Phn b chun c ghi nhn trong qun th F3, vi php th Z = (X- ) / x = trung bnh nhm x = lch chun ng dng php th 2 xc nh phn chun ca gi tr qun th, t suy ra h s skewness theo cng thc nh sau b1 = g1 = m3 / (m2 m2) trong m2 = moment th cp = h2 h12 m3 = moment tam cp = h3 3h2 + 2h13 h1 = fU/n h2 = fU2/n h3 = fU3/n ng dng php th Kurtosis theo cng thc sau g2 = b2 3 = (m4 / m22) 3 trong m4 = moment t cp = h4 4h1h3 + 6h12h2 3h14 v h4 = fU4 / n Bng 1: Kt qa th nghim F1 trong mi trng mn (Mishra v ctv. 1998) T hp lai Bas 370 / CSR10 Bas 370 / CSR11 Pak Bas / CSR10 im chng chu mn 5, 5, 7, 7, 5 5, 5, 3, 5, 5 7, 5, 7, 7, 5 Trung bnh ca 5 cy 29/5 = 5,8 23/5 = 4,6 31/5 = 6,2
Bng 2: Th nghim Z v 2 ca t hp Bas 370 / CSR10 (Mishra v ctv. 1998) im 1 3 5 7 9 S cy 68 426 768 276 247 Gi tr Z Z1 = -1,5511 Z2 = -0,5916 Z3 = 0,3679 Z4 = 1,3274 Vng phn b chun (0-Z) 0-Z1 = 0,4397 0-Z2 = 0,2230 0-Z3 = 0,1435 0-Z4 = 0,4073

Vng gia hai gi tr cn (-)Z1= 0,0603 Z1-Z2 = 0,2167 Z2-Z3 = 0,3665 Z3-Z4 = 0,2638 Z4-(+) = 0,0927
X = 5,233 1,000
n= 1785
2 = 159,18 (P = 0,000) df = 5-3 = 2 H s skewness (g1) = 0,2980 (SD = 0,0579) H s Kurtosis (g2) = -0,4789 (SD = 0,1159) F1 ca tt c cc cp lai u nm gn im gia ca phn b hnh chung, cho thy tnh tri khng hon ton i vi phn ng nhim cng nh phn ng chng chu. Nhng nu im chng chu ca F1 l 5,8 (t hp 1) v 4,6 (t hp 2) cho thy nh hng thay th ca cy b (CSR10 hoc CSR11) i vi cy m gn nh ging nhau. T hp 1 v 2 c mc nghing (skewness) trong phn b chun c ngha v thng k trong khi t hp 3, phn b nghing khng c ngha, chng t rng ch c mt vi gen ch lc tc ng cng vi nhiu gen th yu iu khin tnh chng chu mn (bng 1 v 2). Th nghim ny cho thy tnh trng chng chu mn l mt tnh trng di truyn a gen, khng c nh hng ca cy m (Mishra v ctv. 1998) 2-3-2. Nghin cu di truyn phn t tnh chng chu mn Bn QTL (quantitative trait loci) c p dng trong trng hp nhng tnh trng mc tiu do a gen iu khin (th d nh tnh chng chu mn). Di truyn s lng truyn thng khng th pht hin QTL trn nhng loci ring bit gn vi tnh trng s lng ang nghin cu, v tr ca n trn nhim sc th v lin kt ca n vi nhng gen khc. Bn di truyn phn t vi mt cao s lng marker ph trn ton b nhim th trong genome cy trng s cung cp cho chng ta cng c c kh nng nghin cu tnh trng di truyn s lng phc tp, nh v gen trn nhng nhim th, v xc nh cc gen mc tiu lin kt vi gen khc. Bn QTL v phn tch QTL pht trin theo trnh t nh sau: Trn c s marker hnh thi v di truyn t bo, ngi ta tnh ton gi tr khc bit v kiu hnh lin kt vi tnh trng s lng tng loci ring bit trong mt qun th ang phn ly theo l thuyt ca Thoday (1961), v phng php ny kh ph bin trong phn tch QTL vo thp nin 1980 Mt phng php khc c thut ng l phn tch QTL trn c s tnh trng (traitbased QTL analysis), da trn mi tng quan gia marker v tnh trng s lng (Stuber v ctv. 1980). L thuyt ny cn c theo gi nh rng: p lc chn lc s lm thay i mt cch c ngha tn sut gen ca tng QTL ring bit, m nhng QTL ny lin kt kh cht ch vi nhng marker tng ng trong bn . Lebownitz v ctv. (1987) gi phng php phn tch ny l ba loi hnh b tr th nghim (three kinds of experimental designs). Thun li ca phng php ni trn l to iu kin tt cho vic nh du gen v chn ging m marker phn t (MAS) trong chng trnh chn
to ging ci tin, lm gia tng hiu qa chn lc. Mi tng quan gia mt marker v mt QTL c phn tch da trn phng php ANOVA mt chiu Phng php ANOVA mt chiu ch c th pht hin mt QTL nu n c nh v rt gn marker tng ng. Do , mt marker ph trn genome trong qun th phn ly c th b hn ch, c bit trong trng hp marker hnh thi. S pht trin RFLP, v gn y microsatellite marker v cng phong ph, tho mn yu cu ph kn trn genome cy trng. Cc qun th c s dng cho phn tch QTL l n bi kp (Snape 1988), hi giao (Patersons v ctv. 1991), F2 km vi hi giao (Zhang v ctv. 1992) Phng php m phng ti a (maximum livelihood) c pht trin c on tn sut ti t hp gia mt marker v mt QTL, trn c s tnh ton gi tr kiu gen v phng sai (Weller 1986). Phng php ny t ra c hiu qa hn trong trng hp QTL c tnh cht codominant so vi trng hp dominant. Sau Lou v Kearsey (1989, 1991) xut phng php tng t nh phng php Weller, cc tc gi thc hin php tnh tn sut ti t hp theo gi tr d on ca m phng ti a. Nhng phng php ny ch tht s hu dng trong trng hp xc nh tng locus ring bit ng gp vo tnh trng s lng, v trng hp h s di truyn ln hn 10% Ngi ta tip tc xut phng php s dng nhng marker k cn trn c s phng php m phng ti a khc phc cc nhc im nu trn (Lander v Botstein 1989, Jensen 1989, Knapp 1991). Thng tin t hai marker k cn c tnh cht codominant s cho chng ta khi nim v lin kt ca cc khu vc trn nhim sc th. Sau , Knott v Haley (1992) tp hp nhng so snh gia hai phng php marker n v marker k cn, xut cch tnh chnh xc hn v nh hng v v tr QTL ca tnh trng ang nghin cu. Knapp v Bridges (1990) xut m hnh qun th n bi kp (DH), cn giao ti t hp (RI), hi giao (BC), F2, F1 v m hnh cc tnh trng lin quan c phn b chun khi thanh lc vi stress. Phng php ny ch quan tm n hai marker trong mi ln xem xt. Landers v ctv. (1987) thc hin mt phn mm v cng hu ch, l MAPMARKER/QTL s dng trong phn tch QTL. Hu ht cc m hnh thit lp bn QTL u xem xt nh hng cng tnh ca cc gen mc tiu. Carbonell v ctv. (1992) xut mt phng php pht hin nh hng khng cng tnh ca QTL trong qun th F2. Lou v Kearsey (1992) cng xut mt phng php thng k phn tch qun th F2 v chng minh kt qa ca phng php thng qua m hnh ha trn computer. Thng thng ngi ta ghi nhn mt marker lin kt vi mt gen, nhng thc t mt marker lin kt vi nhiu gen th yu nh hng n tnh trng s lng. Do , mt gi nh khc xut: c an kiu gen thng sai lch nn kh c th c on nh hng epistasis trong di truyn s lng. Jensen (1992) xut mt m hnh phi hp khc phc tnh trng ny, xem xt c trng hp tnh trng lin quan khng phn b chun khi thanh lc vi stress, phng php ny c gi l multiple QTLs effect analysis
Teng (1994) s dng qun th cn giao ti t tp (RI) th h F8 bao gm 324 c th thuc t hp lai gia IR29 / Nona Broka nghin cu di truyn tnh chng chu mn ca cy la. Cc dng RI c thanh lc mn trong nh li iu kin EC = 15 dS/m v iu kin ng rung. Phn tch RFLP vi 5 enzyme phn ct hn ch (DraI, EcoRV, HindIII, ScaI, XbaI) cho thy c 266 RFLP marker, trong 117 th hin a hnh (43,98%), ph trn genome cy la vi mt 15cM / qung. RG100 v RZ323 c ghi nhn cho a hnh r nht trong trng hp DNA ca dng chng chu v dng nhim. Mi ba marker nh v gn RG100 v RZ323 trn nhim th s 3 cng c s dng xem xt lin kt gen. Phn tch ANOVA mt chiu chng minh Nona Broka mang alen khng lin kt vi RG100 v RZ323 ti cc loci s lng, vi gi tr R2 l 17,6% v 29,4% (p < 0,0001), theo th t. Phn tch ANOVA hai chiu, tc gi pht hin thm RZ323 (nhim th s 3) v RG333 (trn nhim th
s 8) lin kt vi QTL chng chu mn, vi gi tr R2 l 40,2% (p<0,001), gii thch 40,2% bin thin kiu hnh v tnh trng sng st ca cy m l do QTL ny iu khin. Cc c th ti t hp mang alen t Nona Broka locus RZ323 v t IR29 locus RG333 c kiu hnh sng st lu hn trong mi trng mn so vi nhng t hp khc c cha c hai alen. Kt qa cho thy c s bin thin vt tri (transgressive) i vi tnh trng thi gian sng st ca cy m Bng 3: Phn tch QTL theo phng php cch qung (interval) i vi tnh trng hp thu K, Na v t s Na/Ka chi thn (Teng 1994) Ch tiu Hp thu K 195-209 206-200 3- 93 Hp thu Na 195-209 39-188 88 - 67 27 - 25 T s Na/K 195-209 207- 65 3- 93 Qung gia hai marker P3/M9-8 Saltol RG375 P4/M3-2 P1/M1-3 - P2/M1-3 P3/M9-8 Saltol P1/M5-3 P3/M9-1 P1/M10-6 P1/M710 P1/M3-10 P1/M38 P3/M9-8 Saltol G291 P1/M7-8 P1/M1-3 P2/M1-3 Nhim sc th 1 4 12 1 3 3 10 Gi tr LOD Phng sai kiu hnh c gii thch (%) 80,2 83,5 21,2 64,6 17,1 16,0 35,6
17,23 5,34 3,46 14,54 3,17 3,02 3,96
1 10 12
14,51 3,60 3,14
64,3 86,1 18,5
QTL c khm ph c nh hng iu khin tnh trng hp th K chi, nh v trn nhim sc th s 1, s 4 v s 12 (bng 3), vi phng sai kiu hnh c gii thch l 80,2%, 83,5% v 21,2%, theo th t. QTL c nh hng n hot ng iu khin tnh trng hp thu Na, nh v trn nhim th s 1, 3, v 10. i vi t s Na/K, c 3 QTL nh v trn nhim th s 1, 10 v 12 c gi nh l gen iu khin tnh trng ny, vi bin d kiu hnh c gii thch l 64,3%, 86,1% v 18,5%, theo th t (bng 3). QTL c quan st trn nhim th s 1 i vi 3 tnh trng: Na thp, K cao, t s Na/K thp vi gi nh c lin quan n chng chu mn. Bn QTL (trn c s AFLP v STS marker) cho thy gen ch lc iu khin tnh trng chng chu mn nh v trn nhim th s 1 (saltol). Bn cnh gen ch lc, 3 QTL c ghi nhn c quan h vi tnh trng hp thu cao K, 4 QTL c quan h vi tnh trng hp thu thp Na, v 3 QTL c quan h vi tnh trng t s Na/K thp. Nhng QTL ny nh v trn nhim th s 1, 3, 4, 10 v 12 (Teng 1994) 2-4. S TH HIN GEN CHNG CHU MN Trong nng nghip, thit hi do mn, lnh, v kh hn c nh hng nghim trng nht i vi nng sut cy trng (Boyer 1982). c bit thit hi do mn c th lm thay i hot ng sinh trng, pht trin, nng sut v lm cht cy. Nhiu nghin cu mong mun tm ra c ch chng chu mn mt cch r rng, xy dng mt chng trnh ci tin ging chng chu c hiu qa chn lc cao. Trong lnh vc nghin cu sinh l thc vt, hng lot nh hng stress do mn cho thy rng thc vt t bo v mnh khi nhng thit hi do mn gy ra theo m hnh phn ng oxygen (Kawasaki v ctv. 2001), trnh thiu ht nc, tng cng hp th ion trong chu trnh quang hp (Noctor v Foyer 1998, Dat v ctv. 2000).
S th hin gen chng chu mn xt v lnh vc sinh hc phn t l mt khm ph v cng th v. Tn hiu c truyn vo t bo, cc gen c chc nng chuyn mn c khi ng v hng lot cc qa trnh chuyn m, gii m xy ra. Kawasaki v ctv. (2001) s dng phng tin microarray theo di s th hin ca phn t transcript v tng qa trnh th hin gen iu khin tnh chng chu mn trong cy la. Trn c s th vin cDNA ly trch t r la v b marker EST (expressed sequence tags) ca genome cy la chng chu mn ni ting Pokkali, ngi ta p dng k thut microarray kim sot nhng transcript trong vic so snh vi nghim thc khng x l mn, thi gian thay i t 15 pht n 1 tun l trong iu kin gy mn nhn to. Vt liu c s dng l ging la Pokkali (chun khng) v ging la IR29 (chun nhim). Nhm tc gi ny tp trung xem xt phn ng i vi stress do mn trong qun th lai c 1728 transcript dn sut t r la ca cy b x l mn (NaCl nng 150mM). Kt qa ny cho thy mt tin trnh iu tit gen chc nng vi nhiu mc khc nhau ca transcript. Trong giai on u tin, p ng ca IR29 chm hn Pokkali. Sau 3-6 gi x l mn, mc phong ph ca transcript thay i nhanh trong Pokkali, nhng IR29 c mt s suy gim trong vng 3 gi u tin, dn n ci cht ca ging nhim mn ny ngay sau . Hot ng quang hp, s lu thng qua kh khng, v h hp c ghi nhn sau khi x l 150mM NaCl. Quang hp gim trong vng 20 pht v n nh pht th 30 (Hot ng quang hp c o theo gi tr mol photons / m2 / giy, hot ng ny ch cn khong 1/10 so vi bnh thng) trong ging chun khng Pokkali. Trong iu kin x l mn lu hn, Pokkali tip tc pht trin vi tc quang hp thp, 7 ngy sau khi x l mn, tng lng cht kh tng gp i. Pokkali duy tr lng nc trong chi trong 6 ngy b stress. Ngc li, IR29 th hin phn ng chm hn i vi stress, v tt c cy la b cht kh trong vng 24 gi. iu ny cho thy tnh chng chu ca Pokkali l mt c ch th hin nhanh sau khi c tn hiu mn, gip n chng chu thit hi do mn tt hn ging nhim IR29 (Kawasaki v ctv. 2001) 2-4-1. Ph th hin transcript (transcript abundance profile) So snh s th hin transcript trong r la Pokkali trong iu kin bnh thng v trong iu kin b stress do mn, ngi ta ghi nhn s khc bit v kh nng chng chu ca kiu hnh ny (Kawasaki v ctv. 2001). Thng tin v ph th hin transcript ca cy trng c thu thp thng qua so snh gia hai nghim thc: bnh thng v c x l mn, vi cc chui m l d liu thu thp (bng 4). Trong th vin cDNA ca r la, nhng EST c xp loi theo ph th hin nh sau: th vin OC, khng c stress: 1106 EST, th vin OD, OE v OF, c stress: 1418 EST. Khong 41% chui m ca th vin OC c xem l nhng protein khng c xp hng (unclassified proteins), bao gm c nhm c thut ng chuyn mn l no hits, trong iu kin b x l mn. Trong xp hng kiu no hits, c 324 EST c tm thy vi d liu khng c tnh cht phong ph. Tnh cht ng dng c tm thy trong nhiu marker EST ny, nhng 154 EST ca nhng cy b thit hi do mn khng th hin mt cch c ngha tnh cht ng dng vi EST ca d liu. S khc bit chnh ca transcript thuc nghim thc bnh thng v nghim thc b stress do mn ch: hot ng tng hp protein gim, xt v mt chc nng. Tri li, chng ta quan st thy transcript th hin rt nhiu trong hot ng lm d dng s lu thng trong t bo v hot ng t v ca t bo (bng 4)
Bng 4: Chc nng ca nhng transcript trong ging la Pokkali, c trng cho th vin cDNA (Kawasaki v ctv. 2001) Chc nng ch yu Bnh thng a S lng % 354 32,0 253 22,9 109 9,9 94 8,5 59 5,3 59 5,3 45 4,1 29 2,6 29 2,6 24 2,2 22 2,0 13 1,2 0,9 10 0,5 6 100 1106 X l NaCl b S lng % 41,6 590 10,7 152 10,0 142 16,2 230 2,5 36 3,6 51 2,9 41 2,8 39 1,2 17 3,1 44 2,8 40 1,3 18 0,8 12 0,4 6 100 1418
Protein khng xp loi Tng hp protein Hot ng bin dng No hit T chc t bo nh v protein Truyn tn hiu Nng lng Chuyn m To iu kin chuyn dch Cu sng, bo v, v pht trin t bo Tng trng v phn bo Pht sinh t bo (biogenesis) Di chuyn gia cc t bo Tng cng a th vin OC (khng c stress) b phi hp th vin OD, OE, v OF (c stress)
C 4 th vin cDNA t r la Pokkali: th vin OC s dng RNA ca r la 10 ngy tui, khng x l mn, th vin OD t r la 30 pht sau khi thu hoch cy m 12 gi tui, th vin OE t r la ca cy m 24-72 gi tui, v th vin OF t r la ca cy m 1 tun tui 2-4-2. Phn tch microarray Nhng phn t DNA ca cc clone c chn lc, kch thc chn vo ln hn 500 bp ca th vin OC, OD, v OE c khuch i vi cp mi T3/T7 (Kawasaki v ctv. 2001). Nhng amplicon di hn 400 bp c in ra. Kt qa microarray c 1728 transcript c xc nhn vi 3 ln nhn (1728 x 3 = 5184 nguyn t trn mi slide). Hin nay, chng ta c th s dng web site: www.stress-genimocs.org tham kho bng kt qa cp nht v tt c EST trong tng array. Sau khi lai v ra mu, ngi ta tin hnh phn tch s liu microarray nh nhng phn mm hin c thng mi ha. Cc tn hiu t nhng m lp li ba ln c tnh theo gi tr trung bnh. Cng tn hiu Cy3 / Cy5 c iu chnh vi s tr gip ca cc gen iu khin bn ngoi c cho thm vo microarray slide (theo qui trnh). Cc t l c tnh ton theo tn hiu Cy3 tng s trn tt c nhng m ca slide tng ng vi tn hiu Cy5 tng s. Theo kim tra ban u, nhng microarray ny ny c lai vi nhng probe mc tiu c nh du bng hunh quang t nhng mRNA ca m l hoc m r khng b stress do mn. Kt qa cho thy, hu ht cc transcript trn array c ngun gc t th vin ca r, th hin mc cao hn so vi transcript c ngun gc t l. Phn tch RNA theo phng php Northern Blot, khng nh tnh cht chuyn bit ca m r theo s liu lai array (hnh 2-1). S thay i cng tn hiu c biu th bng t s th hin, tnh theo log 10 (LR) (LR [Cy3/Cy5]). Kt qa so snh lch chun so vi trung bnh mu ca cc m lai vi RNA cho thy 91% tn hiu cng LR 0,1 tng ng vi lch chun l 1,25 fold. Trong nhm cng LR 0,15 ( 1,4 fold), c 98% tn hiu c ghi nhn. Trong nhm cng LR 0,2 ( 1,6 fold), c 0,18% tn hiu c ghi nhn. Microarray c kh nng pht hin s th hin gen a dng cng tn hiu ln hn 0,2 LR, tng ng vi ngng c nghin cu trc y (Maleck v ctv. 2000, Kawasaki v ctv. 2001). Mc bin thin cao c quan st trong mt th nghim tng t vi RNA ca r sau mt nm trong iu kin ging nh trn. Trong nhm cng LR 0,1, c 76% tn hiu c ghi nhn. Trong nhm cng LR 0,15, c 92% tn hiu c ghi nhn. Ch c 2,7%
tn hiu trong trng hp LR 0,2 v 47 marker ca 1728 EST marker c ghi nhn, s bin thin ny th hin mt cch ngu nhin. 2-4-3. c im th transcript ca ging la chng chu mn trong iu kin b stress Trong tt c nhng th nghim, ngi ta so snh cng Cy3 v Cy5 c nh du (PK nonstress) nhm bnh thng ha hin tng bin thin. Trong nghim thc i chng ny, 95% cc m c ghi nhn cng tn hiu 0,07 LR. Phn t RNA khng b stress (3 gi) (thu thp 6 gi trong iu kin c nh sng) lai vi RNA (khng b stress, thu thp 3 gi trong iu kin c nh sng) cho kt qa ph th hin rt ging vi nghim thc i chng (98% cc m c ghi nhn cng tn hiu 0,01 LR). Nh vy s thay i v m c mt tc dng nht nh (Kawasaki v ctv. 2001) Chc nng iu tit s th hin gen thay i, sau khi stress do mn, c quan st thng qua th nghim so snh nhng probe ti thi im i chng vi nhng probe ti 6 thi im khc nhau, lc thu thp RNA, sau khi x l cho cy b stress. Ch sau 15 pht, Pokkali phn ng, thc hin chuyn m. Sau 15 pht b stress do mn, ch c 2% transcript c iu tit theo kiu UP, hoc theo kiu DOWN vi cng tn hiu 0,2 LR. Sau 14% transcript c iu tit vi cng 0,1 LR (8% ln hn + 0,1 LR v 6% t hn 0,1 LR). Sau 1 gi, 33% ca tt c transcript c bin i cho hot ng chuyn m, vi cng 0,1 LR (16% ln hn +0,1 LR v 17% t hn 0,1 LR), v 10% c iu tit vi cng ln hn 0,2 LR (4% ln hn + 0,2 LR v 6% nh hn 0,2 LR) (Kawasaki v ctv. 2001) Trong cy Arabidopsis thaliana, gen chng chu mn c iu tit vi s th hin gen cao nht vo lc 8 n 10 gi sau khi mt tri mc ln (Fowler v ctv. 1999, Park v ctv. 1999). Cng truyn tn hiu cao ca th transcript GIGANTEA trong cy la Pokkali trong iu kin c mn v khng c mn c ghi nhn l 9 gi sau khi c nh sng, v 10% transcript trong Pokkali iu tit dng UP v DOWN trong vng 1 gi khi b stress do mn (Kawasaki v ctv. 2001). Sau mt tun l, tnh cht iu tit theo kiu UP (aquaporins) dng hi phc. Sinh tng hp protein gia tng giai on u, theo sau l s kch thch nhng transcript c tnh cht p ng vi stress trong vng mt vi gi, v kch thch nhng transcript m nhim chc nng bo v c lin quan Cc tc gi tng kt s khc bit gia ging chng chu mn v ging nhim nh sau: Transcript ca ging Pokkali chng chu mn th hin gen vi cng l mt hng s (0,1 LR) trong tt c cc pha ca stress do mn S th hin ca nhng transcript cn thit cho hot ng t bo, cung cp nng lng cho h hp chu k C3, hot ng chuyn m (mRNA), hot ng chuyn dch (ATPase), sinh tng hp t bo (c bit thnh t bo), sinh tng hp DNA khng b nh hng bi iu kin mn Phn tch microarray cho thy ph th hin ca Pokkali c ghi nhn trong vng 15 pht, v ca ging nhim IR29 c ghi nhn sau 1 gi Phn ng u tin trong iu kin mn l iu tit theo kiu UP ca transcript thuc ging Pokkali (protein ca rib th, CDPK, v nhiu EST cha r chc nng), trong khi nhng hin tng iu tit ny khng c trong ging nhim IR29
2-4-4. Vai tr ca abscisic acid, jasmonate, proline Abscisic acid (ABA) v jasmonate c ghi nhn trong phn ng ca cy trng i vi stress do thiu nc v khi cy b thng. Moon v ctv. (1997) so snh nh hng sinh l hc mc phn t ca jasmonic acid (JA) ( 10M) v ABA i vi stress do mn gy ra trong r la. Chng ta bit rng ABA v JA l mt trong nhng cht iu tit sinh trng (regulator) c th lm thay i s th hin ca gen. Cht iu tit sinh trng JA v hp cht methyl ester ca n (methyl jasmonate = MeJA) xut hin trong cy trng v truyn tn hiu
ca cy khi phn ng vi vt thng, cng nh khi cy b pathogen tn cng (Mueller v ctv. 1993). Jasmonate tch t kh nhanh v c tnh cht chuyn v khi cy b thng hoc c x l vi mt elicitor (Creelman v ctv. 1992, Gundlach v ctv. 1992). C hai JA v MeJA u lm kch thch s th hin gen m ha nhng protein c chc nng khi cy b thng v b ngun nm bnh, vi khun tn cng, th d nh cc loi hnh khc nhau c ch proteinase (Hidman v ctv. 1992), th d nh thionins (Andersen v ctv. 1992), protein vch t bo giu proline (Creelman v ctv. 1992), nhng enzyme trong hn ng sinh tng hp phytoalexin, phenylalanine ammonialyase (Gundlach v ctv. 1992), enzyme chalcone synthase (Lee v ctv. 1996), v hng lot nhng protein khc nh PR protein (pathogenesisrelated) (Schweizer v ctv. 1997).
L R R 1/5 R 1/10
metallothionein-like protein OsMT-1 (transcript ID #1272) osrt1 (transcript ID #1719)
-tubulin (transcript ID #1320) rRNA
Hnh 2-1: S chuyn bit ca m trong hot ng lai microarray Phn tch RNA gel blot i vi nhng clone c chn lc vi 10g RNA / ct, RNA c ly trch t r la v l la ca nghim thc i chng. Lai RNA c hon tt trong cng iu kin s dng microarray. Hnh nh c ghi nhn trn phim X-quang, c phn tch bi phn mm GelExpert (version 3.5), Nucleotech, San Carlos. Gi tr log-10 ratio i vi cc tn hiu ca r v l trong RNA gel blot and microarray nh sau: OsMT-1, 1,10/0,98; ars1, -0,4/-0,4; v -tubulin, 0,02 /0,01 (Kawasaki v ctv. 2001).
Jasmonate cn c tm thy trong hot ng kch thch protein bt hot rib th ca l kiu mch (Reinbothe v ctv. 1994) v nhiu dng khc ca nhm ng dng lipoxygenase (LOX) ca thc vt (Bell v ctv. 1995). Abscisic acid l hormone thc vt c xem nh mt tn hiu quan trng trong phn ng phn t v sinh l thc vt khi cy thiu nc, cy b nh hng ca mn, hoc nhit lnh (Zeevarrt v Creelman 1988, Moon v ctv. 1997). ABA v s thiu nc lm kch thch s th hin ca hng lot gen c nhim v ci tin tnh chng chu kh hn, t , cc lp khc nhau ca protein LEA (late-embryogenesis abundant) hnh thnh nn mt s cn xng sinh hc, gip cy chng chu stress. Trn c s cu trc nh vy, nhng phn t protein LEA c nhim v bo v thnh t bo, lm lun chuyn nc, hoc lc cc ion. Stress gy ra do thiu nc s kch hot s chuyn m ca gen LEA thng qua l trnh ABA-l thuc v ABA-c lp c nghin cu rt k (Ingram v Bartels 1996, Shinozaki v YamaguchiShinozaki 1997). Tuy nhin, hin chng ta c rt t hiu bit v nhng nh hng giai on hu chuyn m c kim sot bi ABA (Moon v ctv. 1997). Abscisic acid c chng minh l hot ng ca gen trong phn ng ca cy khi b thng tn. Mc ABA ni sinh ca thc vt sau khi b tn hi gia tng ti ch v pht trin theo h thng ni hp. Cc nh nghin cu s dng t bin thiu ABA theo di mi tng
quan gia s gia tng ABA sau khi b thng tn vi s biu hin ca nhng phn t c ch proteinase (Hidmann v ctv. 1992). Phn ng ABA ca gen pin2 c chc nng tng hp protein trong dch bo khi t bo b thng. Gen LOX ca Arabidopsis c tm thy do ABA kch thch lm gen ny hot ng, chng minh s hp lc ca c hai cht iu ha sinh trng (Melan v ctv. 1993). Moon v ctv. (1997) tng hp nh sau: JA kch hot mt peroxidase ca cation, hai protein mi c kch thc 32 v 28 kD, nhng protein PR c tnh cht acid nh PR-1 v PR-10, v protein SalT phn ng vi stress do mn r la. Hu ht nhng protein trong phn ng ca JA trong r c tch t li khi cy la b stress do mn gy ra JA khng kch thch protein LEA nhm s 3 c tnh cht p ng vi ABA Trong iu kin mn, ABA kch thch hin tng tch t transcript ca gen oslea 3. JA, ABA, v stress do mn kch thch s tch t transcript ca gen salT v gen osdrr, chng mang mt m ca protein PR-10 Mc ABA ni sinh trong r v mc methyl jasmonate gia tng rt khc nhau v lng v v thi gian b stress do mn ko di
Trong mt nghin cu khc v th hin ABA vi binding protein (BP) trong c ch chng chu mn ca cy la, Gupta v ctv. (1998) ghi nhn nhng im chnh nh sau ABA kch thch nhng gen c nhim v quan trng trong iu khin tnh chng chu mn S th hin yu t chuyn m ghi nhn nguyn t ABRE (vit tt t ch abscisic acid response element) nh mt hot ng iu ha khi cy b stress do mn Trong iu kin b stress do mn trong 72 gi, hai transcript c tch t trong r la Pokkali (chun khng) c pht hin l 2,0 kb (r2.0) v 1,5 kb (r1.5). C hai transcript ny c pht hin sau 6 gi b stress do mn, hoc c x l ABA. Giai on th hin hot ng iu tit gen vi nguyn t ABRE-binding protein trong phn ng vi mn xy ra vo lc 26 gi sau khi x l 200mM NaCl S xut hin ca ABRE-binding protein th bt hot v tin-sinh tng hp ca cy i chng v hot ng ca GTP c th l mt l trnh iu tit s th hin ca nhng gen chng chu mn Mi quan h gia hot ng truyn tn hiu v s iu ha gen chng chu mn rt c quan tm nghin cu. Thc vt c kh nng chng chu c nhng th thch ca mi trng nh kh hn, nhit lnh, mn bng c ch iu tit (adjustment) nhiu hn c ch that (escaping). Nhng phn ng nh vy c s h tr c lc ca ABA, vi nhng knh truyn tn hiu hin nay vn cha c bit r rng. Mt tn hiu c ghi nhn l Ca++, v chnh ABA lm tng hm lng Ca++ trong t bo bo v dn n s ng m kh khng khi cn thit (Wu v ctv. 1997). C 3 cht lm nhim v iu tit Ca++ l: inositol (1, 4, 5)triphosphate (IP3), cyclic adenosine 5-diphosphate ribose (cADPR), v nicotinic acid adenine dinucleotide phosphate (NAADP+). Receptor ca IP3 c bit kh r nhng receptor ca cADPR v NAADP+ cha c bit nhiu lm. Receptor c tnh gi nh ca cADPR l ryanodine (RyR) (Wu v ctv. 1997). Cc xt nghim sinh hc trn cy Arabidopsis thaliana cho thy hm lng cADPR gia tng theo ABA Cng trnh nghin cu ca Igarashi v ctv. (1997) nh tnh gen i vi :pyrroline-5-carboxylase synthetase (cOsP5CS) v xem xt mi tng quan gia th hin gen ny vi tnh chng chu mn ca cy la. Ngi ta phn lp mt cDNA i vi cOsP5CS, mt enzyme dng trong sinh tng hp proline. Sau , ngi ta tin hnh xc nh tnh cht th vin cDNA ly trch t cy m 14 ngy tui ca ging la Akibare. Chui amino acid b phn gii ca protein P5CS th hin 74,2% dng tng ng (homology) i vi P5CS ca Arabidopsis thaliana, v 75,5% dng tng ng i vi P5CS ca Vigna aconitifolia. Phn
tch Northern blot cho thy phn t RNA ca gen ny b kch thch trong iu kin x l nng mn cao, hoc iu kin c ABA. S tch ly proline cng c quan st nh mt kt qa th hin gen trong iu kin cy la b x l mn. Proline l mt yu t c th kch thch s iu tit gen i vi hin tng co nguyn sinh trong cy la khi b x l mn hoc kh hn (Strizhov v ctv. 1997, Iyer v Caplan 1998) 2-5. NGHIN CU CHUYN CI TIN GING LA CHNG CHU MN NG BNG SNG CU LONG (Lang v ctv. 2001a, 2001b, 2001c) Vng trng la b nhim mn ng Bng Sng Cu Long (BSCL) c khong 700.000 ha. Mn xm nhp t thng 12 n thng 5. Nng dn y thng ch ma trng la. Tuy nhin do lng ma tht thng, cy la vn c th b mn gy hi giai on m, hoc giai on tr n chn. Vic xc nh tiu chun chn ging chng chu mn, xc nh cc tnh trng cn thit, c ch khng mn giai on m, v giai on pht dc l mc tiu ca nhiu chng trnh chn ging. Tnh trng c quan tm nhiu l mc tn thng trn l giai on m, t l ht bt th giai on pht dc, t s Na+/K+ ca chi thn, trong iu kin mi trng mn. Anh hng gy hi do mn trn cy la rt phc tp, chng ta khng ch quan st tnh trng hnh thi, m cn tnh trng sinh l, sinh ha, tng tc vi mi trng. Do , vic chn lc c th chng chu mn khng th cn c trn mt tnh trng ring bit no (Akbar v ctv. 1986). Ging chng chu mn ni ting Nona Bokra c ghi nhn tt giai on m v giai on tng tng, nhng ging chun khng Pokkali c ghi nht tt giai on pht dc (Senadhira 1987). Ging c , v c Phng c nh gi nh ngun cho gen khng BSCL (Bu v ctv. 1995). Chng trnh chn ging la chng chu mn tp trung khai thc ngun ti nguyn di truyn a phng, tnh thch nghi vi mi trng v tp qun canh tc. K thut di truyn phn t hin c ng dng to ging la nng sut cao v chng chu iu kin bt li chnh. Chen v ctv. (1991) ghi nhn s c mt ca nhm alen M-20 gia hai loci RG711 and RG4 trn nhim th s 7. Qun th F2 ca cp lai gia M-20 vi ging 77-170 cho thy: gen khng ch yu nh v trn nhim th s 7, lin kt vi marker RG711 vi khong cch di truyn 7.02.9 cM (Zhang v ctv 1995). Nhm marker phn t "microsatellite"c xem nh rt ph hp trong trng hp xc nh gen chng chu mn, vi th a hnh (polymorphism) phong ph (KunShen v ctv 1993). Mc tiu nghin cu: (1) xc nh vt liu cho gen chng chu mn, (2) tm hiu mt vi c s sinh l tnh chng chu mn, (3) pht trin ging la chng chu mn BSCL. 2-5-1. VT LIU & PHNG PHP Vt liu bao gm cc ging la a phng c truyn, ging la ci tin trong chng trnh lai, vi ging i chng khng quc t l Pokkali v i chng khng a phng l A69-1, ging chun nhim l IR28. S dng qun th F2 ca t hp lai IR28 / c Phng. Thu hoch hai ht trn mi bng ca mi c th F2 theo di s phn ly F3. Dung dch mn c to ra bng cch khuy u mui n trong nc ct (5 g trong 1 lt nc), iu chnh sao cho dn in EC = 10 dS/m. Thang im nh gi c trnh by trong bng 1, trn c s quan st bng mt. Vic nh gi bt u t tun l th ba sau khi x l mn.
Bng 5: Tiu chun nh gi (SES) giai on tng trng v pht trin. im 1 3 5 7 9 Quan st Tng trng bnh thng, khng c vt chy l Gn nh bnh thng, nhng u l hoc vi l c vt trng, l hi cun li Tng trng chm li; hu ht l b cun, ch c vi l cn c th mc di ra Tng trng b ngng li hon ton; hu ht l b kh; mt vi chi b cht Tt c cy b cht hoc kh

Mc chng chu Chng chu tt Chng chu Chng chu trung bnh Nhim Rt nhim
Microsatellite: Sn phm PCR c chun b trong 10mM Tris-HCl (pH 8), 50mM KCl, 1.5mM MgCl2, 1 unit ca TAKARA Taq, 4 nmol dNTP, 10pmol primer v 50ng DNA. S dng thermal cycler 9600 (Perkin-Elmer), chu k nhn gen c thit k nh sau: tch dy n 95oC trong 5 pht, tip theo l 35 chu k: 94oC trong 60 giy, 55oC trong 30 giy, 72oC trong 60 giy. Ln ko di phn ng cui l 72oC trong 5 pht. Sau khi thc hin PCR, chng ti thm vo 13l dung dch m (98% formamide, 10mm EDTA, 0.025% bromophenol blue, 0.025% xylene cyanol. Mc a hnh ca sn phm PCR c pht hin nh ethidium bromide sau khi in di trn 5% agarose gel. Microsatellite marker c ghi nhn s c bng khng v nhim theo s 1 (chng chu mn) v 2 (nhim). Trong nghin cu c ch tnh chng chu mn, vic x l mn c thc hin trn cy m 3-tun- tui, trong mi trng dinh dng Yoshida. Thm vo 50ml NaCl sao cho nng t 0.5% v 1.0% mi mt tun. Ch tiu quan st c ghi nhn vo la 1 v 2 tun sau khi x l mn. Sodium v potassium c o bng quang ph k ngn la. Hai ging chng chu l CSR10 v CRS13 ca n c so snh vi hai ging nhim l IR20 v IR28 trn c s cc ch tiu sinh l, sinh ha. 2-5-2. KT QA & THO LUN 2-5-2-1. Xc nh vt liu lai to Nghin cu di truyn s lng gen chng chu mn c nghin cu trn c s tnh trng hnh thi hc (Bu v To 1993). u th lai c ghi nhn i vi nng sut ht, s bng trn bi, v chiu cao, do hot ng ca gen tri v tng tc khng alen. Trong iu kin mn, ht chc trn bng c gi tr ng gp trc tip ln nht n nng sut (Bu v Trng 1988). S ht chc trn bng v t l ht lp l hai thng s ng tin cy nht ca ch s chn lc trong iu kin mn. nh hng ca gen khng cng tnh (non-additive) i vi cc yu t cu thnh nng sut c ghi nhn, tr trng lng 1000 ht (To v ctv. 1992). C t nht 5 nhm gen iu khin tnh trng chiu cao cy ca ging la nc su trng vng ven bin b ngp mn Gi tr kh nng phi hp chung cao i vi tnh trng sc cha c ghi nhn trn ging la a phng vng mn l La Giu, Ba xe giai, v Bng Hng (Lang 1994). Chin lc to chn ging chng chng chu mn c xem nh l cch lm kinh t v c hiu qa nht gia tng sn lng thc vng b nhim mn. Chng ta c gng tm kim tnh trng t ch tiu chn lc n gin nht cho nh chn ging. Nhng thng s c cng b v ngh cn quan tm l mc thit hi trn l giai on m, ht hu th giai on pht dc, t s Na+/K+ ca chi trong iu kin mn (Bu v ctv. 1995). Hiu qa chn lc ging la chng chu mn vn cn rt thp bi v tnh cht phc tp ca vng t kho st, v nhng yu t mi trng khc c lin quan nhng khng kim sot c. C hai hoc nhiu gen s lng iu khin tnh chng chu mn, lin kt kh cht ch vi mi trng c ghi nhn, nh phng tin marker phn t RFLP (Kuraka v ctv. 1994, McCouch v ctv. 1998).
Vin La BSCL thanh lc 418 mu ging la a phng trong iu kin mn 612dS/m, c 44 mu ging chng chu tt l Nng Co , Sc Nu (Bu v ctv. 1995), v gn y l c , c Phng, Tri My, C ung Trng (bng 6) 2-5-2-2. Nghin cu c ch chng chu mn Cc ging la nhp ni IR37109-61, BW298-2, CSR9, CSR10, CSR13, v ging la ca Vin OM723-7, OM723-11 c kh nng phc hi tt v cho nng sut cao trn cc vng mc tiu, thng qua h thng kho nghim ging hng nm Hai ging la chng chu mn ca n l CSR10 (M40-431-24-114 / Jaya) and CRS13 (CSR1 / Basmati // CSR5) c s dng so snh vi hai ging nhim IR20 v IR8 (By 1993). mn lm gia tng hm lng sodium trong c hai cy la khng v nhim mn. mn lm gim ng k hm lng potassium cy nhim, nhng lm tng potassium cy chng chu, c hai giai on tng trng v pht dc (bng 7) Ging la chng chu mn c hm lng Na+ gim do n duy tr c mt lng cao K+ trong thn (Krishnamurty v ctv. 1987, Prat v Fathi 1990). Tnh chng chu mn Salinity tolerance (vi tiu chun chn lc Na+/K+ ca chi thn) c iu khin bi hot ng ca c hai nhm gen cng v tri (Gregorio v Senadhira 1993). Tnh trng chng chu mn c iu khin bi t nht hai nhm gen c tnh siu tri. Nhm th nht c th iu khin s loi thi Na+ v nhm th hai iu khin s hp thu K+. T s Na+/K+ trong chi thn ca cc ging chng chu mn thng nh hn so vi ging nhim khi b x l mn. T s Na+/Ca++ ln trong ging nhim so vi ging chng chu (Subbarao et al. 1990). Ging la chng chu mn c th cn nh c ch tch t mt lng cao spermidine v spermine so vi nhng cy m cha x l mn c hai giai on. Hm lng putrescine hi tng ln trong cy m b x l mn so vi cy bnh thng (By 1993). Ging nhim mn thng duy tr mt lng cao putrescine, kt hp vi hm lng thp ca spermidine v spermine trong cy m b x l mn c hai giai on (bng 8). Katiyar v Dubey (1990) ghi nhn rng mn lm cho hm lng polyamine gia tng ng k trong cy m; mn 14 dS/m putrescine tng gp i trong ging nhim. Hp cht polyamines bao gm putrescine, c bit vi chc nng kch thch s tng trng ca cy trng, cho thy rng nng ca nhng amines ni sinh ny c th l mc gii hn ca s tng trng (Smith 1982, 1984). Spermidine v spermine c bit trong c ch ngn nga hot ng ca fructose-1,6biphosphatase trong lc lp (Costa v Bagni 1983, Costa v ctv. 1984). S c mt ca spermidine synthase trong lc lp cho thy rng spermidine c th l mt "modulator" rt quan trng ca fructose-1,6-biphosphatase in vivo (Sindhu v Cohen 1983). Putrescine tch t trong cy gip cy thch nghi vi cc iu kin gy stress t bn ngoi, c bit trng hp thiu K+ v Mg++, hp thu NH4+ trong iu kin b acid ha, v trng hp b nhim mn (Altman v ctv. 1982, Young v Galston 1983, Flores v ctv. 1984, Smith 1984). Nng putrescine cng s gia tng nu c yu t gy sc lm co nguyn sinh hoc yu t mt nc ca t bo (Flores v Galston 1984). S cn bng in gia cation / anion cng chng minh c hin tng gia tng putrescine khi co nguyn sinh, nhng s tch t proline trong iu kin kh hn v mn c th c xem xt t mt c ch ging nhau, putrescine tch t nh mt sn phm d tr trong nhng iu kin b stress rt a dng (Smith 1984) (Bng 8). Bng 6: Thanh lc ging la chng chu mn giai on m, trong mi trng Yoshida + NaCl Ging c c Phng im chng chu 6dS/m 12dS/m 3 5 3 5
Tri My 3 7 C ung trng 3 7 La Trng Ct 5 7 Nng Thit 5 7 Nng Tt 5 7 C ung 5 7 C ung Phn 5 7 Phng Ln Lem 5 7 Nng Thm Ht Lu 5 7 Np Trng Ct 5 9 Tm 5 9 La Nng Thm 9 9 Trng Chm 5 9 Nh Thc C 5 9 Nng Qut 5 9 Nng R 5 9 Chiu 3 9 Nng Hng 5 9 Nng Qut 5 9 Ba L 9 9 Trng Ln 9 9 Nh Thm Ln 3 9 Np M 3 9 c Trng 3 9 Nanh Chn 3 9 Np Mng chim 3 9 Tiu Chm 3 9 Trng Tp 3 9 Mt Bi 9 9 Nng Keo 7 9 NpThan 9 9 IR28 (chun nhim) 9 9 Pokkali (chun khng) 3 5 (1: chng chu tt, 3: chng chu, 5: chng chu trung bnh, 7: nhim, 9: rt nhim)
Bng 7: nh hng mn i vi s tch ly cht kh, hm lng Na, K ca cy la (By 1993) Ging Nghim thc Cht kh (g / cy) 1TSX 0.58 0.49 0.45 0.61 0.47 0.37 0.51 0.32 0.28 0.63 2TSX 1.38 1.01 0.90 1.17 0.77 0.68 1.22 0.64 0.43 1.42 Sodium (meq /g TL kh) 1TSX 2TSX 0.74 0.64 1.30 0.82 1.60 0.88 0.96 0.90 1.80 1.16 2.42 1.26 0.82 0.94 1.26 2.22 2.14 3.00 0.60 0.70 Potassium (meq /g TL kh) 1TSX 2TSX 1.12 0.92 2.60 1.34 3.20 1.64 1.38 1.10 2.72 1.32 3.00 1.58 1.20 1.40 0.85 0.96 0.54 0.46 1.00 1.36
CSR10 CSR13 IR20 IR8
i chng 0,5% NaCl 1,0% NaCl i chng 0,5% NaCl 1,0% NaCl i chng 0,5% NaCl 1,0% NaCl i chng

0,5% NaCl 1,0% NaCl LSD 0.05 (ging) LSD 0.01 LSD 0.05 (ging x nng mui) LSD 0.01 TSX: tun sau khi x l mn 0.41 0.36 0.06 0.09 0.06 0.09 0.84 0.71 0.10 0.15 0.11 0.17 0.98 1.38 0.20 0.30 0.21 0.40 1.56 2.14 0.47 ns 0.55 ns 0.82 0.58 0.22 0.34 0.26 0.39
0.94 0.68 0.33 0.49 0.39 0.57
Bng 8: Hm lng polyamine tng s, putrescine, spermidine v spermine trong chi thn ca ging chng chu mn v ging nhim (mo/100g trng lng ti) (By 1993) Polyamine Putrescine Spermidine Spermine Nghim thc 1 Tsx 2 Tsx 1 Tsx 2 Tsx 1 Tsx 2 Tsx 1 Tsx 2 Tsx 35 17 67 57 73 60 134 175 CSR10 C 51 31 107 80 80 63 0.5%NaCl 174 238 104 57 155 106 83 65 1.0%NaCl 228 342 41 22 74 63 83 64 150 198 CSR13 C 57 39 92 72 95 68 0.5%NaCl 179 244 102 55 145 99 99 73 1.0%NaCl 227 346 IR20 C 167 228 74 99 71 88 22 41 0.5%NaCl 205 298 103 156 77 95 25 47 1.0%NaCl 240 338 103 184 82 101 28 53 45 23 86 66 97 68 157 228 IR8 C 55 26 95 68 141 98 0.5%NaCl 192 291 62 28 102 73 166 1.0%NaCl 223 330 122 LSD0.05 (ging) 9 12 3 6 3 9 6 2 LSD0.01 13 18 4 9 5 ns 10 3 (Gng x%NaCl) 4 8 11 5 8 3 19 10 LSD0.05 5 11 16 7 11 5 27 15 LSD0.01 Tsx: tun sau khi x l mn Trong iu kin x l mn, hm lng spermine v spermidine tng rt nhiu trong ging chng chu mn so vi ging nhim. Ging 2-5-2-3. Phn tch bn di truyn QTL tnh chng chu mn Qun th cn giao ti t hp bao gm 108 dng t t hp lai Tenasai 2 / CB c s dng trong phn tch bn QTL tnh chng chu mn ca cy la. Tenasai 2 l ging cho gen chng chu mn ca Trung Quc v CB l ging nhim c ngun gc t M. Nhng tnh trng c quan st trong iu kin mn 12 dS/m l : s ngy cy m sng st (SD), trng lng kh ca r, trng lng kh ca chi, hm lng Na+, K+ v t s Na+/K+ trong chi. Marker phn t c s dng trong phn tch bn di truyn l RFLP v microsatellite vi 108 marker, ph trn 12 nhim sc th ca cy la. Bn c ph vi tng s 2.340,5 cM, trung bnh 21,68 cM / qung gia marker v loci. Nhng marker phn t lin kt vi loci th hin tnh chng chu mn phn ln nh v trn nhim th s 1, 2, 3, 9, 11 v 12. Bn di truyn QTL c phn tch xc nh nh hng gia QTL v cc tnh trng lin quan n kh nng chu mn ca cy la. Nhng xt nghim c p dng b sung trong phn tch QTL l php th chi bnh phng, phn tch tng marker ring (SMA), phn tch mapping tng qung (IM). Kt qa gia cc phng php ny cho nhng gi tr thng nht kt lun. Tnh trng c lin quan n kh nng chng chu mn: SD do bn QTL iu khin, trng lng kh ca chi do mt QTL, trng lng kh ca r do hai QTL, s hp thu Na+ do mt QTL, s hp thu K+ do mt QTL, v t s Na+/K+ do bn QTL (hnh 1). Thng qua phn tch QTL, s bin thin kiu hnh ca SD thay i t 5,2 n 11,6%, ca nhng tnh trng cn li
l 4,80 n 14,38%. Nhng QTL iu khin chung tnh trng RD v trng lng kh ca r, RD v Na+/K+ nh v trn nhim th 3, v 9. Nhng QTL iu khin chung tnh trng Na+/K+ v K+ nh v trn nhim th s 12. Kt qa ny gii thch hin tng bin thin vt tri ca con lai so vi b m i vi cc tnh trng c lin quan n kh nng chu mn (Lang v ctv. 2000) Bng 9: Xc nh QTL tnh trng s ngy sng st sau khi x l mn (SD) bng phng php SMA (single marker analysis) ca 108 dng RIL t t hp Tenasai 2 / CB (Lang v ctv. 2001b) Marker Nhim Ngun Gi tr trung bnh ca Gi tr P R2 sc th alen (1) alen (2) (%)3 C711 11.5 9 T 28.790.26 0.0080 C 25.640.77 C1454 11.6 9 T 28.790.36 0.0235 C 23.220.00 R1751 6.9 9 T 28.920.28 0.0277 C 27.630.60 R3156 8.5 3 T 28.300.28 0.0250 C 30.901.21 C560 6.6 2 T 28.380.25 0.0298 C 31.601.56 0.0402 C747 6.1 2 T 28.220.30 C 29.600.60 0.0450 R26 5.2 2 T 28.460.25 C 31.061.60 0.0500 C178 5.8 1 T 28.840.26 C 26.661.61
(1) alen ca Tenasai k hiu T, alen ca CB k hiu C (2) Xc sut ca trung bnh nhm khc bit do ngu nhin (3) Phn trm bin thin kiu hnh ca tnh trng s ngy sng st (SD)
a hnh c ghi nhn trn 74 RFLP gia b v m, v y l nhng marker rt c ch xc nh tnh r rng ca nhng mu d on trong nghin cu. Trong 91 cp mi ca microsatellite c 51 marker cho kt qa a hnh. ln phn t ca nhng SSR ny bin thin t 95 bp (RM227) n 220 bp (RM219, RM242). iu tra 74 RFLP, c 42 marker phn ly mt cch c ngha theo t l 1:1. Tt c marker (tr mt marker) th hin lch v pha kiu gen ca Tenasai 2 (ging chng chu mn). Bng 9 cho thy 8 marker nh v gn nhng QTL iu khin tnh trng s ngy sng st (SD) trong iu kin mn. Bn marker th hin QTL iu khin gia tng tnh chng chu mn t kiu gen ca b m, nhng hai marker th hin nh hng ln nht thuc v kiu gen ca ging Tenasai 2. Nhng QTL mc tiu nh vy nh v trn nhim sc th s 1, 2, 3, v 9. Tt c 8 marker ny ng gp mt cch c ngha v lch vi t l phn ly 1:1 gia nhm kiu gen chng chu v nhm kiu gen nhim. Bng 10 cho thy gi tr trung bnh ca trng lng chi thn (SW) ca cc dng cn giao c cha alen ca Tenasai 2, c khuch i bi primer RM209, ln hn gi tr trung bnh c ngha so vi nhm c alen ca ging CB. Locus ny nh v trn nhim th s 11, ni y khng c QTL marker ca cc tnh trng khc lin quan n hin tng chng chu mn nh v. QTL i vi tnh trng trng lng r (RW) lin kt vi nhng marker nh v trn nhim th s 3 v 9, trng vi tnh trng SD, nhng ch c mt marker R3156 c ch nh chung cho c hai tnh trng RW v SD
Hm lng Na v K b nh hng bi cc loci nh v trn nhim th s 6, 7 v 12, cng th hin trn nhim th s 1 v 2.
Hnh 2-2: Ngun A t Tenasai 2 - ging chng chu mn, v ngun B t CB- ging nhim mn. Cc nh biu th vng di truyn ca ging nhim CB, trong khi cc lm biu th vng di truyn ca ging chng chu Tenasai 2, gip gia tng tnh trng mc tiu (Lang v ctv. 2001b)
Hnh 2-3: Bn QTL ca nhng tnh trng mc tiu lin quan n hin tng chng chu mn trn qun th F8 (RIL) ca t hp lai Tenasai 2 / CB (Lang v ctv. 2001b)
Bng 10: Xc nh QTL ca cc tnh trng trng lng r (RW), trng lng chi thn (SW), hm lng Na trong chi, hm lng K trong chi, t l Na/K, bng phng php SMA ca 108 dng RIL t t hp Tenasai 2 / CB (Lang v ctv. 2001b) Tnh trng St.wt Marker Nhim Kiu Trung bnh alen Gi tr Gi tr P th gen F RM209 11 R2 (%) DPE
T 9.24 0.0036 14.3 T 284.56.53 C 8 220.08.54 R3156 3 T 5.87 0.018 9.34 C 72.422.04 C 92.0 4.98 C563 3 T 5.23 0.025 6.95 C 73.041.86 Rt.wt C 87.53 5.34 C397 9 T 4.00 0.050 6.35 T 77.28 2.04 C 61.81 2.60 RM240 2 T 5.21 0.025 7.22 C 0.43 0.01 C 0.48 0.01 Na+ R1167 6 T 3.68 0.050 4.35 C 0.440.01 C 0.550.01 G24 12 T 10.57 0.002 17.4 C 0.730.02 K+ C 5 0.970.11 C86 1 T 5.03 0.029 9.14 C 0.570.02 C 0.730.04 G24 12 T 4.83 0.032 8.81 T 0.610.02 C 0.450.07 Na+/ K+ RM214 7 T 3.86 0.050 5.86 C 0.590.02 C 0.810.06 C747 2 T 3.71 0.050 5.25 T 0.620.02 C 0.540.04 alen ca Tenasai k hiu T, alen ca CB k hiu C DPE: Direction of phenotypic effect: xu hng kiu hnh theo kiu alen ca T: Tesanai 2; v C : CB Phng php nhy cm hn trong pht hin QTL l phng php phn tch cng mt lc nhng nh hng ca marker lin kt trong cng mt qung ca nhim sc th khi nghin cu v di truyn tnh chng chu mn, c tm tt trong bng 11. nh hng ca nhng vng khc nhau trn mi nhim sc th i vi 6 tnh trng mc tiu (tiu chun chn lc) lin quan n hin tng chng chu mn c th hin trong hnh 2-2. Cc nh biu th vng di truyn ca ging nhim CB, trong khi cc lm biu th vng di truyn ca ging chng chu Tenasai 2, gip gia tng tnh trng mc tiu. Trong bng 11, c 7 on thuc 4 nhim sc th iu khin tnh trng SD, v mt vi on iu khin cc tnh trng mc tiu cn li. Tnh trng SD c chung nhng on nh v trn nhim sc th s 2 v tnh trng RW c chung nhng on nh v trn nhim sc th s 3 Bn vng: C560 - C747, R3156 C397, C1454 C397, v G24 R1684 c nh hng rt ngha vi hai tnh trng mc tiu (bng 11). Nhng nh hng ca cc vng khc nhau nh vy trn tng nhim sc th i vi 6 tnh trng mc tiu c tm tt trong hnh 22. Hu ht cc tnh trng mc tiu u th hin nh hng mt cch ngha trn cc vng c xc nh ca genome (bng 11), nhng tnh trng trng lng chi, hm lng K+ th hin nh hng ch trong mt qung ca bn di truyn m thi.
Bng 11: QTL ca cc tnh trng c xc nh bng phng php phn tch qung (interval analysis) trn bn di truyn (hnh 2-3) (Lang v ctv. 2001a) Index (6-7) (18-19) (19-20) (37-38) (80-81) (79-80) (81-82 (91-92) (36-37) (37-38) (80-81) (104-105) 15-16) (1-2) (19-20) (61-62) (104-105) Qung gia hai marker C178-R210 R26 - C560 C560- C747 R3156- C563 C1454 -C397 C711-C1454 R1751-C397 Nhim th Gi tr P 0.010 0.000 0.000 0.008 0.000 0.008 0.000 0.003 0.000 0.00001 0.006 0.010 0.002 0.020 0.000 0.0001 0.0001 centiMorgan 7.7 15.7 11.9 33.1 16.1 19.5 8.4 34.5 5.3 33.1 16.1 12.3 45.3 33.0 11.9 37.7 12.3
S ngy sng st (SD) 1 2 2 3 9 9 9 TL chi thn (SW) RM209-RM206 11 TL r (RW) C563 -C63 3 R3156 63 3 C397-C1454 9 + Hm lng K G24-R1684 12 Hm lng Na+ RM240-RM213 2 T l Na+/K+ C86-RM212 1 C747-C560 2 RM214- R1789 7 G24- R1684 12
Hnh 2-3 cho thy bn lin kt gen v marker vi 74 RFLP v 31 SSR th hin a hnh, vi chiu di tng cng l 2.340,5 cM. Khong cch trung bnh gia hai marker l 21,68 cM. Nhng qung c gi tr nh nht trong nhim sc th s 9 (16,20 cM) v ln nht trong nhim sc th s 4 (30,75 cM). V cn li mt t cc qung thiu khong 50 cM. Vic xc nh c th v tr ca QTL iu khin hin tng chng chu mn cn c thc hin trn cc t hp lai khc nhau (bng 12, Lang v ctv. 2001c)
Bng 12: Phn ng ca cc dng con lai BC2F2 ca 12 t hp lai ( giai on m, EC=36 dS/m) T hp lai IR64 / Cheng-Hui 448 IR64 / OM1706 IR64 / FR13A IR64 / Type3 Teqing / OM1706 Teqing / Amol3 IR68552-55-3-2 / Cheng-Hui448 IR68552-55-3-2 / OM1706 IR68552-55-3-2 / FR13A IR68552-55-3-2 / Madhuhar IR68552-55-3-2 / Type3 IR68552-55-3-2 / Almol3 Pokali (chun khng) PSBRc88 (chun khng) IR29( chun nhim) IR64 (chun nhim) Phn ng vi mn 5 5 5 3 5 5 7 3 3 5 3 5 3 3 9 5
Bng 13: Kt qa thanh lc ccdng BC2F2 trong iu kin mn EC = 18 dS/m v nhng marker tng ng ca n (Lang v ctv. 2001c) BC2F2 ca cc t hp lai Code S cy Phn ng vi mn EC = 18dS/m R R R R R Marker xcnhn
IR64/Cheng-Hui448 32005 11 RM292,RM242 IR64/OM1706 32058 11 RM276,RM315 IR64/FR13A 32035 11 RM315 Teqing/OM1706 32134 8 IR68552-55-3-2 / 32146 11 Chenghui448 IR68552-55-3-2 / 32197 11 R RM223 OM1706 IR68552-55-3-2 / 32175 5 R FR13A IR68552-55-3-2 / 32177 24 R OSR2 Madhukar IR64/Type3 32044 6 R NTP/Type3 32184 13 R Teqing/Almol3 32088 10 R IR68552-55-3-2 / 32160 11 R Almol3 Qun th hi giao ci tin BC2F2 bao gm 12 t hp lai, vi cc vt liu cho gen chng chu mn dng lm b l OM1706, FR13A, Cheng Hui 448, Almol 3, Type 3, v Madhadar, cc vt liu c dng lm m (ti tc) l IR64, IR68552-55-3-2, Teqing (Lang v ctv. 2001 c).

tch.
S cp primer tng ng vi 150 microsatellite markers c s dng trong phn
S th hin a hnh gia b m, s truyn vo ca gen mc tiu (introgression) thng qua biu hin ca con lai BC2F2 c ghi nhn trn nhim th s 1, 6, 8 v 9. nh gi kiu hnh giai on m, trong iu kin thanh lc mn mi trng dinh dng c khuyn co EC = 18dS/m. Chin lc chn ging chng chu nh marker phn t (MAS) nn tp trung trn nhim th s 1 v s 8, c bit l nhim th s 1, vi marker RM315 Marker OSR1 rt hu ch c khuyn co cho iu tra s khi tnh a hnh ca b m 2-5-2-4. Chn ging chng chu mn nh marker phn t (MAS) T kt qa ny, chng trnh ci tin ging la chng chu mn ti Vin La BSCL c thc hin theo hng xc nh mt locus chnh c quan h vi tnh chng chu mn c hai giai on tng trng v pht dc, trn c s l thuyt "chn ging nh marker phn t" (MAS). Qun th F2 ca IR28 / cPhng c th nghim, sau chng ti chn ra 257 con lai F3 ly trch DNA, v ng dng microsatellite marker tm kim th a hnh trong qun th phn ly ny. Cy m ca tng c th c trng trong dung dch dinh dng Yoshida (Yoshida v ctv. 1976) nh gi kiu hnh. iu chnh nng mn vi dn in EC=6 dS/m. Ba ngy sau , nng mn c gia tng ln 12dS/m bng cch cho thm NaCl vo dung dch dinh dng. Dung dch ny c thay i nh k sau 8 ngy, v duy tr pH lun lun l 5,0 trong ban ngy. Cy la thng chng chu c trong thi gian ny mm, nhng s tr nn nhy cm vi mn vo giai on m c 3 l, ri tr nn chng chu trong giai on tng trng, sau li mn cm trong lc th phn v th tinh (IRRI 1967). C mt vi nghin cu cho rng la khng nhy cm vi mn trong lc tr bng (Kaddah v ctv. 1975). V vy, mun bit phn ng ca cy la i vi stress do mn gy ra mt cch y , chng ta phi xem xt chng mi giai on sinh trng khc nhau. im chng chu mn th hin khc nhau giai on tng trng v giai on pht dc. Chng ti ghi nhn c s phn ly vt tri ca con lai so vi b m v tnh cng chu mn. iu ny chng t hot ng ca mt b alen ng nht trn cc loci khc nhau iu khin tnh chng chu (Zhu et al.1999).
Bng 14: Chui k t ca microsatellite markers cho th a hnh trong qun th cn giao ti t hp ca 108 c th F9 thuc t hp lai Tenasai 2 / CB, v IR28 / c Phng (Lang v ctv. 2001a) Marker Nhim th Chui k t primer c Phng CAAATCCCGACTGCTGTCC TGGGAAGAGAGCACTACAGC CCAGATTATTTCCTGAGGTC CACTTGCATGGTTCTGCATTG CCGGTATCCTTCGATATTGC CCGACTTTTCCTCCTGACG TGGTCAAACCAAGGTCCTTC GACATACATTCTACCCCCGG CTGATGATAGAAACCTCTTCTC AAGAACAGCTGACTTCACAA GAGTGAGCTTGGGCTGAAAC GAAGGCAAGTCTTGGCACTG CTCGTTTATTACCTACAGTACC CTACCTCCTTTCTAGACCGATA CAGATTGGAGATGAAGTCCTCC CCAGCAAGCATGTCAATGTA CCCATGCGTTTAACTATTCT CGTTCCATCGATCCGTATGG AGAAGCTAGGGCTAACGAAC TCACCTGGTCAGCCTCTTTC 126 170 140 130 150 120 140 150 120 90 ln phn t (bp) IR28 Tenasai 2 136 126
CP 106 160
RM237 RM231 RM232 RM218 RM214 RM223 RM201 RM202 RM206 RM235 *Dominant
1 3 3 3 7 8 9 11 11 12
190 190 160 120 114 118 163 160 150 150 180 180 * 200 mono mono
150 140 140 160 110 100
135 130
Theo kt qa nh gi kiu hnh, c Phng c im 3, v IR28 im 9 giai on tng trng. C khong 19.4 % c th F3 th hin tnh chng chu ging nh c Phng, v 23.3 % nhim mn gia on m. Quan st trong giai on pht dc, c 13 % c th F3 chng chu mn nh c Phng, v 33.3 % nhim mn. Theo kt qa nh gi kiu gen nh microsatellite (SSR), nhiu marker c xc nh cho kt qa a hnh r rng gia c Phng (type 1) v IR28 (type 2). So vi Pokkali, c Phng th hin tnh chng chu tt hn giai on m, v chng chu tng ng giai on pht dc S primer c th nghim thnh cng l 30 trong trng hp khuch i locus ca c Phng v IR28. Phn t DNA ca b m c nh gi c mc a hnh cao vi 10 marker nh sau: RM202, RM223, RM231, RM235, RM237, RM218, RM214, RM201, RM232, RM206. Trong , marker RM206 ch c khuch i trong c Phng, n th hin tnh cht "dominant", phn ln cc SSR cn li u c tnh cht "codominant" (bng 14). Marker RM218 v RM232 th hin n hnh trong trng hp cp lai Tesanai 2 / CB, nhng th hin a hnh rt tt trong qun th con lai ca IR28 / c Phng. Kt qa cho thy th a hnh DNA gia b m t c 33.3 % thng qua microsatellite marker (bng 14). Bng 15: Kim nh li chnh xc gia nh gi kiu gen v kiu hnh trong MAS, vi qun th con lai F3 ca IR28 / c Phng (Lang v ctv. 2001a)

Kiu hnh S cy RR Giai on m Chng chu 25 23 01 19 00 0 2 4 2 02 22 00 20 92 88 82 90 RM223 RS SS Gi tr chnh xc (%)
25 Nhim Giai on pht dc Chng chu Nhim 23 22
Nhng marker cho kt qa a hnh trn b m c dng phn tch qun th con lai (hnh 2-4). C 9 marker cho thy khng c lin kt vi gen chng chu mn trong qun th con lai ca IR28 / c Phng. Ch c RM223 lin kt r rng vi gen chng chu mn vi khong cch di truyn l 6.3cM trn nhim th s 8 giai on m. on phn t th hin trn bng c kch thc 90-190 bp. Tnh cht a hnh nh vy c th c khai thc trong chin lc chn ging nh marker phn t (MAS) pht hin con lai c gen chng chu mn. Phn tch kiu gen cho thy, alen khng ca c Phng c kch thc phn t 120 bp, v alen nhim ca IR28 c kch thc phn t 160 bp. Qun th F3 c thanh lc mn trong iu kin EC =10 dS/m giai on pht dc. Trong 23 c th c kiu gen ng hp t khng, trn c s lin kt vi marker RM223, 19 c th thuc kiu gen RR, 4 thuc kiu gen Rr, vi mc chnh xc 82% (bng 10). Trong 22 c th c kiu gen ng hp t nhim, trn c s lin kt vi marker RM223, 20 c th thuc kiu gen SS v 2 thuc kiu gen Rs, vi mc chnh xc 90% (bng 15). So snh nh gi kiu gen vi kiu hnh ca tnh trng chng chu mn c hai giai on m v pht dc, marker RM223 t kt qa tng ng 76%. Tuy nhin, nh gi c nh hng ca mi trng, tt c gi tr v nh gi kiu hnh ca qun th u phi c thc hin, v DNA phi c ly trch t qun th trng dn ny. Nhiu tc gi nhn xt do cc gi tr ghi nhn c b nh hng bi mi trng qa ln (Lander and Bostein 1989), cho nn s dng qun th cn giao ti t hp RI (recombinant inbred lines), qun th n bi kp DH (double haploid lines) v qun th F3 trng dn s lm gia tng hiu qa ca k thut "QTL mapping". Tng tc gia RM223 v im nh gi kiu hnh s gii thch lm th no c th ny chng chu tt hn cc th kia, hoc so vi b m. Kt qa nghin cu ny cho thy, c th cc loci iu khin tnh chng chu mn ca c Phng cha ng nhng alen gn RM223, phi hp vi tnh chng chu NaCl giai on m v giai on pht dc.
Hnh 2-4: Ghi nhn th a hnh sn phm PCR ca ging chng chu c Phng (1) v ging nhim IR28 (2), vi cc microsatellite marker nh sau A: RM202, B: RM223, C: RM231, D: RM235, E: RM237 (Lang v ctv. 2001a)
IR64 / ChengHui IR64 / OM1706
IR64 / FR13A
NTP/Mahduka
Hnh 2-5 a: So snh QTL iu khin tnh trng chng chu mn thng qua qun th BC2F2 ca cct hp lai khc nhau, trn nhim sc th s 1 (Lang v ctv. 2001c). Ch locus RM315 trong IR64 / Cheng Hui, IR64 / OM1706, v IR64 / FR13A, locus OSR2 trong NTP / Mahduka
Hnh 2-5b: Bn lin kt gen vi SSR trn nhim sc th s 8, ca qun th BC2F2 thuc t hp lai IR64 / FR13A ti locus RM223
Microsatellite c th c khai thc trong trng hp tnh trng s lng c kim sot bi QTL trong chin lc chn ging nh marker phn t (MAS). Cng vic tip tc l thc hin k thut "fine mapping" c nhng marker lin kt vi gen mc tiu c khong cch gn hn, s chn lc s chnh xc hn. 2-5-2-5. Pht trin ging la trin vng cho vng b nhim mn Bng 16: Thanh lc mn giai on m trong dung dch dinh dng Yoshida + NaCl, cn c trn thit hi l la. Ging AS996 OM1490 AS1007 OM1838 OM2031 OM2401 OM1346 OM1348 OM1849-5 A69-1 OM723-7 IR28 (chun nhim) Pokkali (chun khng) c (a phng) EC = 6dS/m 3 3 5 3 3 3 3 3 7 3 3 5 1 1 EC = 12dS/m 9 5 7 7 9 9 7 9 9 7 7 9 3 1
Trong vng ven bin BSCL, ging la IR42 c pht trin vi qui m rng v n nh t nhng nm u ca thp nin 1980 cho n nay, nht l Bn o C Mau. Bn cnh , ging la A69-1 cng c trin khai mt vi nm, nhng sau khng pht trin c v phm cht go km. Ging nhp ni IR29723-143-3-2 c nng dn chp nhn, mc d n rt nhim bnh bc l. Mt vi ging la trin vng c pht trin trong nhng nm qua nh sau: Nhm trung ma (125-140 ngy) OM344 Mahsuri / IR42 OM723 A69-1 / NN6A OM861 Ba Thiet / IR42 OM916 BG380 / A69-1 OM924 IR29723-143-3-2 / OM80 OM1571 A69-1 / OM87-9 Gn y, nhm c thi gian sinh trng sm hn c khuyn co v pht trin tt Hm Tru (OM576) Hungari / IR48 115-120 ngy OM1314 OM80 / OM576 105-110 OM1490 OM606 / IR44592-62-1-3-3 90-105 OM2031 Thi Lan / Bng Hng 90-105 Nhm trung ma vn c khuyn khch v c nhiu ging mi c nng dn v cn b khuyn nng ch nh sau: OM1346 IR42 / OM739-7-2-2-1 120-130 ngy OM1348 IR42 / OM736-8-1-1 125-130 OM1849 OM723-11 / KSB54 130-135 Tp Hnh t bin Tp Hnh (c truyn) 130-135
Kt qa thanh lc mn c trnh by bng 7 cho thy phn ng chng chu ca cc ging trin vng so vi i chng Pokkali v c . Mt vi vng trng la ven bin cn chu nh hng ngp mn do thy triu, do vic thanh ngp tnh chng chu ngp hon ton cn phi xem xt. Nghin cu cho thy khng c tnh i khng gia chng chu mn v nng sut, do chn ging chng chu mn l chng trnh c tnh kh thi rt tt (Shannon v Akbar 1978). Cc qun th la hoang Oryza rufipogon v Oryza officinalis c su tp vng ngp mn s c khai thc lm vt liu lai, ging nh vic khai thc cc ging la c truyn c , c Phng v Sc Nu. Tn sut ti t hp s c gia tng nu chng ta thc hin chng trnh hi giao ci tin chn c kiu hnh mong mun trong qun th con lai. TI LIU THAM KHO Abrol IP. 1986. Salt-affected soils: problems and prospects in developing countries. In: Global Aspects of Food production. Oxford, 1986, pp. 283-305 Akbar M, GS Khush, D HilleRisLambers. 1985. Genetics of salt tolerance. In: Rice Genetics, IRRI Philippines, pp. 399-409 Akbar M, IE Gunawardena, FN Ponnamperuma. 1986. Breeding for soil stress. Pages 263272 in Progress in rainfed lowland rice. International Rice Research Institute, Los Banos, Philippines. Akbar M, T Yabuno. 1972. Breeding for saline resistant varieties of rice. I. Variability for salt tolerance among some rice varieties. Jpn J Breed 22: 277-284 Akbar M, T Yabuno. 1975. Breeding for saline resistant varieties of rice. III. Response of F1 hybrids to salinity in reciprocal crosses between Jhona 349 and Magnolia. Jpn J Breed 25: 215-220 Akbar M, T Yabuno. 1977. Breeding for saline resistant varieties of rice. IV. Inheritnace of delayed type panicle sterility induced by salinity. Jpn J Breed 27: 237-240 Akbar M. 1986. Breeding for salinity tolerance in rice. In: Prospects for Biosaline ResearchProceedings of US-Pakistan Biosaline Research Workshop. Karachi, Pakistan, pp. 3854 Akbar M, FN Ponnamperuma. 1982. Saline soils of South and Southeast Asia as potential land. IRRI, Los Banos, Philipines. Akita S. 1986. Physiological bases of differential response to salinity in rice cultivars. Paper presented in Project Design Workshop for Developing a Collaborative Research Program for the Improvement of Rice Yields in Problem Soils. IRRI, Los Banos, Philippines Andresen I, W Becker, K Schluter, J Burges, B Parthier, K Apel. 1992. The identification of leaf thionin as one of the main jasmonate-induced proteins of barley (Hordeum vulgare). Plant Mol Biol 19:193-204 Atman A, R Friedman, D Amir, N Levin. 1982. Polyamine effects and metabolism in plants under stress conditions. Pages 483-494 in Plant growth Substances. London, Academic Press. Bay ND. 1993. Biochemical changes in salt tolerant and susceptible varieties of rice (Oryza sativa L.) under saline conditions. MSc. Thesis. GB Pant University of Agriculture and Technology, Pantnagar, India.
Bell E, RA Creelman, JE Mullet. 1995. A chloroplast lipoxygenase is required for woundinduced jasmonic acid accumulation in Arabidopsis. Proc Natl Acad Sci USA 92:8675-8679 Ben-Hayyin G, P Spiegel-Roy, H Neumann. 1985. Relation between ion accumulationof saltsensitive and isolated salt-tolerant cell lines of Citrus aurantium. Plant Physiol 78:144-148 Bhumbla DR and IP Abrol. 1978. Saline and sodic soils. In: Soils and Rice. IRRI, Los Banos, Philippines, pp. 719-734 Boyer JS. 1982. Plant productivity and environment. Science 218:443-448 Buu BC, DX Truong. 1988. Path analysis of rice grain yield under saline conditions. IRRN 13(6): 20-21 Buu BC, NT Lang, PB Tao, ND Bay. 1995. Rice breeding research strategy in the Mekong Delta. Pages 739-755 in Proc. of the Int. Rice Res. Conf. Fragile Lives in Fragile Ecosystems, IRRI, Philippines. Buu BC, PB Tao. 1993. Genetic nature of some agronomic traits in rice (Oryza sativa L.) through triple test cross analysis. OMonRice 3:5-10 Carbonnell EA, TM Gerig, E Balansard, MJ Asins. 1992. Interval mapping in analysis of nonadditive quantitative trait loci. Biometrics 48:305-315 Chen SY, LH Zhu, J Hong, SL Chen. 1991. Molecular biological identification of rice salt tolerant line. Acta Bot Snia 33:569-573. Clarkson DT, JB Hanson. 1980. The mineral nutrition of higher plant. Ann Rev Plant Physiol 31:239 Costa G, N Bagni. 1983. Effects of polyamines on fruit set of apple. Hort. Sci. 18:59-61 Costa G, R Bagni, N Bagni. 1984. Influence of putrescine on fruit set of apple. Acta Gortic. 149:189-195 Creelman RA, ML Tierney, JE Mullet. 1992. Jasmonic acid/methyl jasmonate accumulate in wounded soybean hypocotyls and modulate wound gene expression. Proc Natl Acad Sci USA 89:4938-4941 Dat J, S Vandenabeele, E Vranova, M Van Montagu, D Inze, F Van Breusegem. 2000. Dual action of the active oxygen species during plant stress responses. Cell Mol Life Sci 57:779-795 Devitt D, WM Jarreli, KL Stevens. 1981. Sodium-potassium ratios in soil solution and plant response under saline conditions. Soil Sci Soc Amer J 45:80-86 Flores RE, AW Galston. 1984. Osmotic induced polyamine accumulation in cereal leaves. I. Physilogical parameter of the response. Plant Physiol. 75:102-109 Flores RE, ND Young, AM Galston. 1984. Polyamine metabolism and plant stress. JK Key and T Kosuge, eds. Vol. 22. UCLA Symp. Mol. Cell Biol. Fowlers S, K Lee, H Onouchi, A Samach, K Richardson, B Morris, G Coupland, J Putterill. 1999. GIGANTEA: A circadian clock-controlled gene that regulates photoperiodic floweringin Arabidopsis and encodes a protein with several possible membranespanning domains. EMBO J 17:4679-4688 Gregrio GB and D Senadhira .1993 Genetic analysis of salinity tolerance in rice. Theor.Appl.Gen. 86:333-338
Gundlach H, MJ Muller, TM Kutchan, MH Zenk. 1992. Jasmonic acid is a signal transducer in elicitor-inducer plant cell cultures. Proc Natl Acad Sci USA 89:2389-2393 Gupta S, MK Chattopadhyay, P Chatterjee, B Ghosh, DN SenGupta. 1998. Expression of abscisic acid-responsive element-binding protein in salt tolerant indica rice (Oryza sativa L. cv. Pokkali). Plant Molecular Biology 37:629-637 Hildmann T, M Ebneth, H Pena-Corte, JJ Sanchez-Serrano, L Willmitzer, S Prat. 1992. General roles of abscisic acid and jasmonic acids in gene activation as a result of mechanical wounding. Plant Cell 4:1157-1170 Igarachi Y, Y Yoshiba, Y Sanada, K Yamaguchi-Shinozaki, K Wada, K Shinozaki. 1997. Characterization of the :-pyrroline-5-carboxylase synthetase and correlation between the expression of the gene and salt tolerance in Oryza sativa L. Plant Mol Biol 33:857-865 Ikehashi H, FN Ponnaperuma. 1978. Varietal tolerance of rice to adverse soils. In: Soils and Rice. IRRI, Philippines, pp. 801-803 Ingram J, D Bartels. 1996. The molecular basis of dehydration tolerance in plants. Annu Rev Plant Physiol Plant Mol Biol 47:377-403 IRRI (International Rice Reseaerch Institute). 1967. Annual report for 1967. IRRI, Los Banos, Philippines Iwaki S, K Ota, T Ogo. 1953. Studies on the salt injury in rice plant. IV. The effects on growth, heading and ripening of rice plant under varying concentration of sodium chloride. Proc Crop Sci Jpn 22:13-14 Iyer S, A Caplan. 1998. Products of proline catabolism can induce osmotically regulated genes in rice. Plant Physiol 116:173-181 Jensen J. 1989. Estimation of recombination parameters between a quantitative trait locus and two marker gene loci. Theor Appl Genet 84:49-56 Jensen J. 1992. Interval mapping of multiple quantitative trait loci. Genetics 135:205-211 Kaddah MT, WF Lehman, BD Meek, FE Robinson. 1975. Salinity effects on rice after the boot stage. Agron J 67:436-439 Katiyar S, RS Dubey. 1990. Changes in polyamine titer in rice seedlings following sodium chloride salinity stress. J. Agron. Crop Sci. 165:19-27 Kawasaki S, C Borchert, M Deyholos, H Wang, S Brazille, K Kawai, D Galbraith, HJ Bohnert. 2001. Gene expression profiles during the initial phase of salt stress in rice. The Plant Cell 13:889-905 Knapp SJ, and WC Bridges. 1990. Using molecular markers to estimate quantitative trait locus parameters powers and genetic variances for unreplicated progeny. Genetics 126:769-777 Knapp SJ. 1991. Using molecular markers to map multiple quantitative trait loci: models for backcross, recombinant inbred, and double haploid progeny. Theor Appl Genet 81:333-338 Knott SA, and CS Harley. 1992. Maximum likelihood mapping of quantitative trait loci using full-sib families. Genetics 132:1211-1222 Korkor SA, and RM Abdel-Aal. 1974. Effect of total salinity and type of salts on rice crop. Agric Res Rev 52(5):73-78
Krishnamurty R, V Anbazhagan, KA Bhagwat. 1987. Accumulation of free amino acid and distribution of sodium chloride and potassium in rice varieties exposed to sodium chloride stress. Indian J. Plant Physiol. 30:183-188 Kun-Sheng Wu, Seteven D Tanksley. 1993. Abundance, polymorphism and genetic mapping of microsatellite in rice . Mol gen genet . 241:225-235. Kurata N, Y Nagamura, K Yamamoto, Y Harushima, N Sue, J Wu, BA Antonio, A Shomura, T Kiriha, K Hayasaka, A Miyamo, A Monna, HS Zhong. Y Tamura, ZY Wang, T Momma, Y Umehara, M Yano, T Sasaki, Y Minobe. 1994. A 300 kilobase interval genetic map of rice including 883 expressed sequences. Nat. Genet 8: 365-372 Landers ES and D Botstein. 1989. Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121:185-199 Landers ES, P Green, J Abrahamson, A Barlow, MJ Daly, SE Lincoln, L Newburg. 1987. Mapmarker: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174-181 Lang NT, S Yanagihara, BC Buu. 2001a. A microsatellite marker for a gene conferring salt tolerance on rice at the vegetative and reproductive stages. SABRAO 33(1):1-10 Lang NT, S Yanagihara, BC Buu. 2001b. QTL analysis of salt tolerance in rice (Oryza sativa L.). SABRAO 33(1):11-20 Lang NT, Z Li, BC Buu. 2001 c. Microsatellite markers linked to salt tolerance in rice. OMonRice 9: 9-21 Lang NT. 1994. Studies on heterosis of some physiological traits and grain yield of rice (Oryza sativa L.). PhD Thesis. National Institute of Agricutural Sciences, Hanoi, Vietnam Lang NT. 2000. Quantitative trait loci for salt tolerance in rice via molecular markers. OMonRice 8:37-48. Lebonitz RJ, M Soller, JS Beckmann. 1987. Trait-based analysis for detection of linkage between marker loci and quantitative trait loci in crosses between inbred lines. Theor Appl Genet 73:556-562 Lee J, B Hause, J Lehmann, A Graner, B Pathier, C Gernack. 1996. Isolation, characterization and expression DNA coding for a jasmonate-inducible protein of 37 kDa in leaves. Plant Cell Environ 19:700-707 Lee KS. 1995. Variability and Genetics of salt tolerance in japonica rice. PhD Thesis, University of the Philippines, Los Banos, Philippines, pp. 108 Lou ZW and MJ Kearsey. 1992. Intermapping of quantitative trait loci in an F2 population. Heridity 69:236-242 Maas EV, GJ Hoffman. 1977. Crop salt tolerance current assessment. ASCE J Irrig and Drainage Div 103:115-134 Maleck K, A Levine, T Eulgem, A Morgan, J Schmid, KA Lawton, JL Dang, RA Dietrich. 2000. The transcriptome of Arabidopsis thaliana during systemic acquired resistance. Nat Genet 26:403-410 McCouch SR, G Kochert, ZH Yu, ZY Wang, GS Khush, WR Coffman, SD Tanksley.1998. Molecular mapping of rice chromosomes. Theor,Appl, Genet.76:815-829. Melan MA, X Dong, ME Endara, KR Davis, FM Ausubel, TL Peterman. 1993. An Arabidopsis thaliana lipoxygenase can be induced by pathogens, abscisic acid, and methyl jasmonate. Plant Pysiol 101:441-450
Mishra B, M Akbar, DV Seshu, D Senadhira. 1996. Genetics of salinity tolerance and ion uptake in rice. IRRN 21:38-39 Mishra B, M Akbar, DV Seshu. 1990. Genetic studies on salinity tolerance in rice towards better productivity in salt-affected soils. Rice Research Seminar, July 12, 1990, IRRI, Phillippines, 25 p. Mishra B, RK Singh, V Jetly. 1998. Inheritance pattern of salinity tolerance in rice. J Genet & Breed 52: 325-331 Moeljopawiro S, H Ikehashi. 1981. Inheritance of salt tolerance in rice. Euphytica 30: 291300 Moon A, E Prinsen, G Bauw, M van Montegu. 1997. Antagonistic effects of abscisic acid and jasmonates on salt stress-inducible transcripts in rice roots. The Plant Cell 9:22432259 Mori IK, T Kinoshita. 1987. Salt tolerance of rice callus clones. Rice Genetics Newsl 4:112113 Mueller MJ, W Brodscheim, E Spannagl, MH Zenk. 1993. Signaling in the elicitation process is mediated through the octadecanoid pathway leading to jasmonic acid. Proc Natl Acad Sci USA 90:7490-7494 Murty KS, KV Janardhan. 1971. Physiological consideration for selection and breeding of varieties for saline and alkaline tracts. Oryza 8 [Supp. 2]: 85-100 Narayanan KK, S Krishnaraj, SR Sree Rangaswamy. 1990. Genetic analysis for salt tolerance in rice. Paper presented during the Second International Rice Genetic Symposium, May 14-18, 1990. IRRI, Manila, Philippines Noctor G, CH Foyer. 1998. Arscobate and glutathione: Keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249-279 Ota K, T Yasue. 1958. Studies on salt injury in crops. XII. The effect of sodium chloride solution on the germination capacity of paddy seed. Proc Crop Sci Soc Jpn 27(2):223225 Park DH, DE Somers, YS Kim, YH Choy, HK Lim, MS Soh, HJ Kim, SA Kay, HG Nam. 1999. Control of circadian rhythms and photopediodic flowering by the Arabidopsis GIGANTEA gene. Science 285:1579-1582 Paterson AH, S Damon, JD Hewitt, D Zamie, HD Rabinowitch, SE Lincoln, EC Lander, SD Tanksley. 1991. Mendelian factors underlying quantitative traits in tomato: Comparision across species, generation, and environments. Genetics 127:181-197 Pearson GA, SD Ayers, DL Eberhard. 1966. Relative salt tolerance of rice during germination and early seedling development. Soil Sci 102:151-156 Ponnamperuma FN. 1984. Role of cultivar tolerance in increasing rice production in saline lands. Strategies for crop improvement. John Wiley and Sons, New York, 443 p. Prat D, RAE Fathi. 1990. Variation in organic acid and mineral compartments in young Eucalyptus seedlings under saline stress. Plant Physiol. 79:479-486 Reinbothes S, B Mollenhauer, C Reinbothe. 1994. PR and RIPs: the regulation of plant gene expression by jasmonate in response to environmental cues and pathogens. Plant Cell 1197-1209 Sathish P, OL Gamborg, MW Nabors. 1997. Establishment of stable NaCl-resistant rice plant lines from anther culture: distribution pattern of K+/Na+ in callus and plant cells. Theor Appl Genet 95:1203-1209
Scheizer P, A Buchala, P Silverman, M Seskar, I Raskin, JP Metraux. 1997. Jasmonateinducible genes are activated in rice by pathogen attack without a concomitant increase in endogenous jasmonic acid levels. Plant Physiol 114:79-88 Senadhira D. 1987. Salinity as a contraint to increasing rice production in Asia. Required worshop on maintenance of life support species in Asia Pacific Region 4-7 April ,1987. Shimose N. 1963. Physiology of salt injury in crops. I. Effect of iso-osmotic pressure due to sodium chloride and sodium sulfateon the growth and absorption of mineral element by rice plants. Jpn Soil Sci. Tokyo 34:107-111 Shinozaki K, K Yamaguchi-Shinozaki. 1997. Gene expression and siganl transduction in water-stress response. Plant Physiol 115:327-334 Sindhu RK, SS Cohen. 1983. Some properties of the spermidine synthase of Chinese cabbage. Pages 371- 380 in Advances in polyamine research. Vol. 4, New York Raven Smith TA. 1982. The function and metabolism of polyamines in higher plants. Pages 463-472 in Plant Growth Substances. London/New York, Academic Press. Smith TA. 1984. Putrescine and iorganic ions. Adv. Phytochem. 18:7-54 Snape JW. 1998. The detection and estimation of linkage using double haploid or single seed descent populations. Theor Appl genet 76:125-128 SSSA (Soil Science Society of America). 1979. Glossary of soil science terms. Madison, Wisconsin, 36 pp. Strizhov N, E Abraham, L Okresz, S Blickling, A Zilberstein, J Schell, C Koncz, L Szabados. 1997. Differential expression of two P5CS gene controlling proline accumulation during salt-stress requires ABA and is regulated by ABA 1, ABA 1 and AXR 2 in Arabidopsis. Plant Journal 12(3):557-569 Stuber CW, RH Moll, MM Goodman, HE Schaffer, BS Weir. 1980. Allozyme frequency changes associated with selection for increased grain yield in maize. Genetics 95:225236 Subbarao GV, C Johansen, MK Jana, JVDK Rao. 1990. Effect of sodium/calcium ratio in modifying salinity response of pigeon pea (Cajanus cajan). J. Plant Physiol. 136:439443 Tagawa T, N Ishizaki. 1963. Physiological studies on the tolerance of rice plants to salinity. Proc Crop Sci Soc Japan 31(3):249-252 Tao PB, DX Truong, BC Buu. 1992. Assessment breeding materials from salt tolerant deep water rices in the Mekong Delta . OMonRice 2: 56-63 Teng S. 1994. Gene tagging for salt tolerance in rice (Oryza sativa L.). PhD Thesis. The University of the Philippines, Los Banos, Laguna, Philippines, 118 p. Thoday JM. 1961. Location of polygenes. Nature 191:368-370 US Laboratory Staff. 1954. Diagnosis and improvement of saline and alkali soils. Agric Hard B 60. United States Department of Agriculture, Washington DC Weller JI. 1986. Maximum likelihood techniques for mapping and analysis of quantitative trait loci with the aid of genetic markers. Biometrics 42:L627-641 Wu Y, J Kuzma, E Marechal, R Graeff, HC Lee, R Foster, NH Chua. 1997. Abscisic acid signaling through cyclic ADP-ribose in plants. Science 278:2126-2130
Yeo AR, TJ Flowers. 1984. Mechanism of salinity resistance in rice and their role as physiological criteria in plant breeding. In: Salinity tolerance in plants. WileyInterscience, New York, pp. 151-170 Yeo AR, TJ Flowers. 1986. Salinity resistance in rice and a pyramyding approach to breeding varieties for saline soils. In: Plant Growth, Drought, and Salinity. Ed. By NC Turner and JB Passioura. CSIRO, Melbourn, Australia. pp 161-173 Yoshida S, F Fornoda, JH Cock, KA Gomez. 1976. Laboratory manual for physiological studies of rice . International Rice Research Institute, P.O. Box 933, Manila, Philippines. Young ND, AW Galston. 1983. Putrescine and acid stress. Plant Physiol. 71:767-771 Zeevaart JAD, RA Creelman. 1988. Metabolism and physiology of abscisic acid. Annu Rev Plant Physiol Plant Mol Biol 39:439-473 Zhang GY, Y Guo, SL Chen, SY Chen. 1995. RFLP tagging of a salt tolerant gene in rice . Plant Sci 110:227-234. Zhang XF, JA Masjidis, ZL Hu. 1992. Methods for detection and estimation of linkage between a marker locus and quantitative trait loci. Plant Breeding 109:35-39 Zhu HL, GP Hayes, A Kleinhofs, D Kudrna, Z Liu, L Pron,.T Bsteffeson, H Toojinda, H Vivar.1999. Does function follow form? Principal QTLs for fusarium head blight(FHB) resistance are coincident with QTLs for inflorescence traits and plant height in a doubled -haploid population of Barley . Theo. Apple Genet : 1221-1232.

Chng 3
C S DI TRUYN TNH CHNG CHU KH HN

Kh hn v mn l hai yu t c bn lm hn ch s tng trng v nng sut cy trng. Mc d chng ta c kh nhiu cng trnh nghin cu tnh trng chng chu kh hn ca cy trng, trong sut hai thp nin 1980 v 1990, nhng thnh tu c th v chn to ging chng chu kh hn vn cha mang li ngha ln lao (Reddy v ctv. 1999). Phng php tip cn th nht l phng php phn tch s ng gp ca cc tnh trng c lin quan, vi m hnh QTL (quantitative trait loci). Phng php tip cn ny ph hp vi hu ht cc loi cy trng ch yu nh cy la, nh bn di truyn vi nhiu marker phn t DNA ph kn trn nhim sc th. Hu ht cc nghin cu v tnh trng chng chu kh hn u quan tm nh gi cc tnh trng c lin quan vi n s sng v pht trin ca. cy trng trong iu kin kh hn. Phng php phn tch di truyn phn t gip cho ni dung y t hiu qa cao hn, trong khi thc hin trn tng tnh trng, trc khi chng ta xem xt s sng v pht trin ca cy. Vic nh gi kiu hnh ca nhng tnh trng ring bit y l v cng quan trng, ging nh vic nh gi kiu hnh ca s p ng cy trng trong iu kin c kim sot nh li hay phng th nghim, trc khi chng ta nh gi ngoi ng rung. Mt s lin kt v cng cht ch c chng minh, l thc hin cc bc trong qui trnh MAS (chn ging nh marker phn t) Phng php tip cn th hai l sng to ra mt kiu bin d mi c chc nng c hiu bit cn k trong phn ng ca cy trng khi b stress do kh hn, thng qua k thut chuyn np gen. Phng php chuyn np gen cho thy l phng tin nhiu tim nng gn vo cy mc tiu mt ph rt rng ca cc gen vi kh nng iu tit mt cch linh hot v tr up hoc down trong qu trnh bin dng kt hp vi hin tng p ng khi cy b stress do kh hn. Nhng phng php chuyn np vi mt gen hay mt nhm gen trong mt l trnh c bit no c th khng thch ng trong iu kin chng chu kh hn, bi v cc sn phm di truyn iu khin tnh chng chu kh hn xut pht t rt nhiu l trnh khc nhau (Ingram v Bartels 1996). Thng thng, nhiu protein hoc nhng phn t osmolyte c trng lng phn t thp, ng vai tr iu tit p sut thm thu, s c tch t trong khi b stress do kh hn. Ngi ta cha hiu r: yu t no chi phi hin tng thay i rt nhiu v ng gp tch cc vo tnh chng chu kh hn. Phng php chuyn np gen c xem nh rt c ch nhng n vn cha cho chng ta bit cc thng tin ng gi Phng php tip cn th ba l xc nh cc gen ng c vin (candidate genes) i vi tnh chng chu stress do kh hn, hoc mn, vi nhng pht trin khng ngng ca k thut chuyn np gen, bn cnh nhng thnh tu v cng ngh phn lp gen (gene isolation) v thao tc gen (gene manipulation) 3-1. XC NH TNH TRNG THNH PHN TRONG CHNG CHU HN S th hin tnh chng chu kh hn c quan st thng qua nhng tnh trng c th nh hnh thi r cy, l, chi thn, phn ng co nguyn sinh, bao phn, qa trnh tr bng, v.v.. Nhng tnh trng nh vy c gi l tnh trng thnh phn (component traits). Bng 1 cho chng ta khi nim v tnh trng thnh phn v cc loci tnh trng s lng (QTL) c lin quan n tnh chng chu kh hn ca mt vi cy trng. Ngi ta nhn thy c s ging nhau v c bn v mt vi khc bit nh v c ch chng chu kh hn, s cu thnh nng sut ca nhiu loi cy trng khc nhau. S ging nhau v c ch chng chu hn mc phn t rt r rng so vi sinh l hc v hnh thi hc. Chng ta c th nghin cu t mt cy ri suy din i vi nhiu cy khc (Reddy v ctv. 1999)
Hu ht cc nghin cu v marker phn t u quan tm n nhng thnh phn rt c bit trong s kin chng chu kh hn, l kh nng ca r cy pht trin su xung tng t bn di, tnh trng phun ru v tung phn vi thi gian cch qung c xc nh (ASI = c vit tt t ch anthesis to silking interval) s iu tit p sut thm thu (OA = c vit tt t ch osmotic adjustment) hin tng bin dng ABA (abscisic acid) hin tng nng hc WUE (water use efficiency c ngha l hiu qa s dng nc)
Nhiu nghin cu khc cn cp n nng sut v thnh phn cu to nng sut. Do , ngi ta c gng xc nh v tr cc QTL cho c tnh trng c lin quan n chng chu kh hn v nng sut, cng nh thnh phn nng sut. Qun th c dng trong phn tch bn QTL thay i vi nhiu th h khc nhau. Rt him khi, QTL c pht hin trong mt qun th ca nhng cp lai khc nhau (Reddy v ctv. 1999). Ngi ta thng gp nhiu kh khn khi so snh kt qa ghi nhn c t qun th n bi kp (DH) vi qun th cn giao ti t hp (RILs), hoc qun th chn ging no . Ngi ta cng gp kh khn khi so snh kt qa ca nhng qun th c ln khc nhau (s dng con lai c s dng khc nhau, t vi chc dng n hng trm dng con lai) S QTL c tm thy i vi tnh trng chng chu kh hn thng thay i t 1 n 4 i vi mt tnh trng thuc thnh phn trong s kin chng chu hn, v nhng QTL ny thng tri rng trn ton b genome vi nhiu nhm lin kt gen. Th d, tnh trng WUE (hiu qa s dng nc) c tm thy vi rt t QTL, t 4 n 5 trong genome cy u nnh (Milan v ctv 1998). Trong mt vi trng hp, c nhng QTL nh v trn cng mt nhm lin kt gen, iu khin nhiu tnh trng quan trng nh kh nng iu tit p sut thm thu (OA), kh nng chng chu s thy phn, chng lin kt vi tnh trng hnh thi hc ca r la (Lilley v ctv. 1996). Bin thin kiu hnh i vi tng tnh trng xt trn mi v tr QTL xoay quanh gi tr 10%. Trong trng hp ngoi l, QTL i vi tnh trng chiu di r giai on 28 ngy tui bin thin 30% (Price v Tomas 1997). Mi quan h gia nng sut v QTL thng cho kt qa m tnh trong vi trng hp, nhng i vi tnh trng ASI, kt qa chng chu hn khng i khng vi nng sut (Ribaut v ctv. 1996). Nhng QTL i vi tnh trng ASI c tnh n nh nhiu nm, trong iu kin mc stress khc nhau, s c xem nh l nhng ng c vin trong chin lc p dng MAS. Bng 1: Cc loci tnh trng s lng (QTL) lin quan n tnh chng chu kh hn ca mt s loi cy trng (Reddy v ctv. 1999)
Tnh trng c tnh ca r 1.Hnh thi 2.Phn b r c tnh ca l v chi 1.L v s dn truyn qua kh khng 2.Cng lc m Cy trng La La La La Qun th lp bn QTL Tc gi
203 RILs ca cp lai Co39 x Moroberekan qun th DH: 105 dng 178 dng F2s 118 dng F2s v b m l japonica
Champoux v ctv. (1995) Yadav v ctv (1997) Price v Tomas (1997) Redona v Mackill (1996)
c tnh sinh l 1.Hm lng nc v kh nng iu tit nguyn sinh cht 2.c im tung phn, phun ru 3.Mc thit hi trc khi tr 4.Mc thit hi sau khi tr 5.S tch t ABA

La mch Bp Cao lng Cao lng Bp 187 dng RILs 272 dng F2s 98 dng RILs 98 dng RILs 81 dng F2s Teulat v ctv. (1997, 1998) Ribaut v ctv.(1996) Tuinstra v ctv. (1996) Tuinstra v ctv. (1998) Quarrie v ctv (1997)
Hiu qa s dng nc (WUE) 1.WUE 2.WUE, 13C phn bit nhm
La m C chua
150 dng DH F3s v cc t hp BC1F1, 11 t hp c WUE cao v 8 thp
Quarrie v ctv.(1995) Martine v ctv.(1989)
Nng sut, thnh phn NS, tng tc GxE 1.Nng sut & thnh phn NS 2.Tng tc GxE Phi hp cc tnh trng c gi tr kinh t 1.u th lai 2.Hiu qa s dng N Bp Bp 264 t hp hi giao 100 dng RILs Stuber (1997) Bertin v ctv.(1997) Bp Bp 150 dng F2:F3 272 dng F2s Weldboom v Lee (1994) Ribaut v ctv.(1997)
3-2. NHNG MARKER PHN T DNA & BN QTL Nhng marker phn t DNA c nh ngha nh nhng ct mc trn ng thng, chng c xem xt mc phn t DNA, hoc nhim sc th, m nhng khc nhau v di truyn c th c tm thy nh nhiu cng c xt nghim c tnh cht phn t (Li 1999). Nhng loi marker phn t chnh thng c s dng l: RFLP l marker c tnh cht ng tri (co-dominant) v rt ng tin cy. K thut ny s dng cDNA hoc th probe, c nh du bng phng x, v s phn ct hn ch ca enzyme RAPD l mt dng marker DNA c sn sinh nh k thut PCR. Mc khuch i cao t mt cp mi n gin, ngn, c thit k ngu nhin (10 mer). y l marker c tnh tri (dominant) v mc tin cy thp STS l marker c thit k t chui k t ca RFLP (20-24 mer), c tnh cht ng tri, c sn sinh nh k thut PCR, i khi n cng th hin tnh tri. y l phng php c th phi s dng enzyme phn ct trong trng hp s a hnh khng th hin r. Kt qa c tin cy tt. Microsatellite hoc SSR l marker c thit k t nhng v tinh c chui k t gin n, lp li nhiu ln. Cp mi c thit k t trc v sau nhng v tinh nh vy. marker c tnh cht ng tri, mc tin cy cao, kh nng d tm a hnh kh nhy. Do , n nhanh chng thay th RFLP v RAPD vi cc m tp di-nucleotide hoc tri-nucleotide (GA, GT, CAT, CTT) trong genome cy la AFLP l mt dng trong nhng marker da trn c s PCR, theo cch chn lc ca hai adaptor (l nhng enzyme phn ct hn ch). N phi hp c hai tnh cht ca
RFLP v RAPD, nn c kh nng pht hin a hnh rt tt. Nhng n thng c tnh cht tri (dominant) nn gp hn ch khi th hin alen ln
Nhng marker phn t ni trn u c th c p dng trong bn QTL nghin cu tnh chng chu kh hn ca genome cy la. Hng u tin hin nay c ngi ta quan tm l s dng microsatellite (SSR). Mc tiu c bn ca bn QTL l tm hiu c s di truyn ca nhng tnh trng s lng bng cch xc nh s lng, cc v tr, nhng nh hng ca gen, v hot ng ca nhng loci bao gm tng tc gen (epistasis) v tng tc QTL x E (mi trng). Mt mc ch khc ca bn QTL l xc nh nhng marker mang tnh chn on i vi nhng kiu hnh c th no , sao cho vic p dng MAS tr nn c hiu qa, phc v yu cu chn dng (ging) chng chu kh hn, chng chu mn, v.v.. Mc tiu lu di ca th nghim v bn QTL l cloning cc gen iu khin tnh trng s lng v cng phc tp, thng qua tip cn k thut map-based cloning (Li 1999). Nguyn tc lp bn QTL vi marker phn t DNA l pht hin cho c nhng kt hp gia marker v tnh trng trn c s lin kt gen, thng qua cc phng php b tr th nghim v phng php phn tch thng k chnh xc. Qun th phc v cho nhng yu cu nh vy thng l: F2, hi giao (BC), n bi kp (DH), hoc cn giao ti t hp (RIL) t mt t hp lai gia hai dng cn giao, phn ly nhng tnh trng s lng mc tiu v nhiu marker i km theo. Qun th ny phi c nh gi kiu hnh rt y , v c nh gi kiu gen thng qua bn lin kt gen vi mt marker ph trn nhim th dy c. Cc phng php phn tch thng k phi p ng yu cu chnh xc, ng tin cy, thng quan phn tch tng marker n, phn tch bn cch qung, v m hnh tuyn tnh. Cho n nay, ngi ta bit rng QTL nh hng nhiu tnh trng nng hc quan trng. Nhng QTL ny ng vai tr nh hng chnh (tnh cng v/hoc tnh tri), chng c tm thy thng qua m hnh main-effect QTL c m t bi Lander v Botstein (1989), Zeng (1994), Li (1999). M hnh main-effect QTL c nhng c im c bn nh sau: Nhng gen ch lc phi c nh hng n tnh trng di truyn cao, chng c xem nh nhng QTL c nh hng ln (gi tr LOD score phi ln) v c tnh cht xc nh r rng thng qua qun th v mi trng Nhng QTL mu phi c nh hng chnh kh nh v i din cho hn 80% loci c nhn dng trong genome cy trng Nhng th nghim MAS i vi ni dung chuyn cc QTL mong mun ci tin tnh trng s lng phi c thc hin vi s thn trng nht nh. Bi v tng tc gia QTL vi mi trng, hin tng epistasis cn phi c gii thch
Hin tng epistasis v s kim sot bin d di truyn c cp trong cc m hnh ci tin. Bn cnh tng tc QTL x E cng uc tho lun. 3-3. BN QTL I VI TNH TRNG R LA H thng r pht trin tt l mt tnh trng v cng quan trng gip cy trng chng chu kh hn (c ch thot hn = drought avoidance mechanism). Ngi ta s dng qun th n bi kp (DH) ca cp lai IR64 x Azucena ti Vin La Quc T (IRRI). Sau Shen v ctv. (1999) pht trin qun th gn nh ng gen (NIL) ca IR64 c du nhp vi nhng QTL ch lc. Cc tc gi ghi nhn bn on trn nhim sc th s 1, 2, 7, v 9 l ni nh v cc QTL ch lc trong phn tch chn lc tng QTL mc tiu (hnh 3-1, Bng 2). C 4 dng DH vi hin tng t hp alen ring r ti 4 on nhim sc th ny, trong c t
hn 50% alen ca Azucena. Bn dng ny c chn lm donor (vt liu cho gen chng chu). Nhng dng nh vy c lai lui vi IR64 cho n khi thu hoch qun th BC3F1, ri sau cho chng t th bnh thng. p dng MAS i vi 4 on nhim sc th nh vy hon thin qui trnh chn lc t BC1F1 n BC3F2. Tt c con lai ca BC3F2 mang nhng alen ca donor, nh v ti t nht mt trong bn khu vc mc tiu. Nhng cy mang hai on mc tiu tr ln c xc nh. Bin thin kiu hnh i vi nhiu tnh trng khng phi l mc tiu cng c quan st BC3F2, ngi ta kt lun rng c s th hin v di truyn t th h trc sang th h sau trong qa trnh du nhp gen (introgression) (Shen v ctv. 1999). iu tra c bn phn cn li ca genome cho thy nhng cy ca BC3F2 c mt phn rt nh ca nhng alen thuc Azucena nh v trn nhng vng khng phi mc tiu, gn vi gi tr l thuyt # 3%. Ngi ta thc hin nhiu cp lai gia cc cy BC3F2 mang nhng on mc tiu ca vt liu cho gen iu khin r la pht trin tt, loi tr nh hng di truyn theo kiu genetic drag v nh hng cc QTL mc tiu khc nhau chng lp theo hnh thp (Shen v ctv. 1999) Bng 2: Cc dng n bi kp (DH) c chn lc thc hin hi giao (Shen v ctv. 1999) Mc tiu ban u Mc tiu ln hai Marker Marker Nhim Tn sut alen Nhim th di ca Azucena th ca trong vng qung khng phi (cM) mc tiu 33,8 1 RZ19-RG690-RZ730 61,4 2 RM29-CDO686 38,9 2 RG437-RG17199,2 1 Khng RG157-RZ318 42,4 2 W1-RG331 44,9 7 RM234-CDO418RZ978-CDO38RG351-RM248 30,8 7 Est9-RG351 37,0 9 RZ228-RM242RZ12-RM201RG667
Dng DH P0055 P0035 P0295 P0475
Bng 3: Nhng QTL i vi hnh thi ca r la (Shen v ctv. 1999) Tnh trng THK TRW TRW DR/S DRW DR/T MRL MRL MRL DR/S DR/S DRW DRW Nhim th 1 1 1 1 1 1 2 2 7 7 7 7 7 Qung (interval) RM34-RG345 RG690-RZ730 RZ730-RZ801 RG690-RZ730 RG690-RZ730 RZ730-RZ801 RG171-RG157 RZ318-Pal1 CD0418-RZ978 CD0418-RZ978 RZ978-CD038 RG773-RZ488 CD0418-Z978 Khong cch markerQTL 0.0 12.0 12.0 14.0 10.0 0.0 14.0 0.0 0.0 0.0 2.0 8.0 0.0 V tr QTL 126.3 187.9 202.2 189.9 185.9 190.2 103.0 146.8 128.4 128.4 140.1 8.0 128.4 LOD score 4.7 4.6 5.1 3.1 4.7 5.6 6.3 3.4 7.4 8.1 5.5 3.4 7.0 R2 nh hng cng -0.042 -0.125 -0.158 -0.003 -0.035 -3.758 -6.160 +4.565 -5.078 -0.005 -0.004 -0.035 -0.039
21.4 21.1 35.2 13.3 23.0 22.9 33.4 14.8 28.8 31.2 23.0 21.5 27.7

DRW DR/T MRL TRW 7 7 9 9 RZ978-CD038 CD0418-RZ978 RZ12-RM201 RZ206-RZ422 4.0 0.0 0.0 4.0 142.1 128.4 101.3 34.0 5.3 5.7 4.2 3.6 23.4 24.4 16.8 18.4
-0.036 -3.713 -3.561 -0.112
DWR: trng lng r la di <30cm, DR/S: trng lng r di / thn chi, DR/T: trng lng r di / s chi, MRL: chiu di r ti a, THK: dy ca r, TRW: trng lng r tng s. Khong cch marker-QTL: khong cch tnh bng cM t marker bn tri ca qung. V tr QTL: khong cch tnh bng cM t nh ca nhim sc th. R2: phn trm bin thin kiu hnh ng gp bi QTL gi nh. Gi tr m ca nh hng cng (additive effect) biu th nh hng dng tnh ca kiu hnh t cc alen ca Azucena.
Hnh 3-1: Bn di truyn QTL trn c s qun th n bi kp (DH) ca IR64 / Azucena v tnh trng r la mc su (Shen v ctv. 1999) 3-4. BN QTL I VI TNH TRNG IU TIT P SUT THM THU (OA) Thnh tu ca cng ngh sinh hc trong vic thc hin xy dng bn di truyn phn t cho php chng ta xem xt nhng tnh trng di truyn phc tp nh tnh chng chu kh hn v nng sut (Zhang v ctv. 1999). Ti i hc Texas Tech, nhm tc gi ny thc hin bn di truyn QTL i vi 2 tnh trng quan trng lin quan n s kin chng chu kh hn, l: kh nng iu tit p sut thm thu (OA), v nhng tnh trng hnh thi ca r la.
Hin nay, ngi ta bit rng: nghin cu sinh l hc tm ra 3 hp phn chnh ng gp vo s kin chng chu kh hn ca cy la: (1) kh nng n su ca r xung tng t pha di, (2) kh nng iu tit p sut thm thu (c ch chng chu), gip cy bo v sinh m khng b tn hi do mt nc (3) kh nng kim sot s mt nc bn ngoi kh khng ca l (Nguyn v ctv. 1997) Hin tng iu tit p sut thm thu (OA) l mt hp phn quan trng trong s kin chng chu kh hn. Kh nng OA ca cy gip n tch ly cht ho tan mt cch ch ng trong t bo, khi cy b kh hn. Tnh trng OA kt hp vi tnh trng nng sut n nh trong iu kin b stress do kh hn c quan st la m v cao lng, nhng hin tng ny khng c ghi nhn trn cy la (Zhang v ctv. 1999). Qun th la c s dng trc y lp bn QTL tnh trng OA l: Qun th cn giao ti t hp (RIL) ca t hp lai CO39 / Moroberekan, vi 1 QTL (Lilley v ctv. 1996) Qun th n bi kp (DH) ca t hp lai CT9993 / IR62266, vi 4 QTL (Zhang v ctv. 1999) Qun th la c s dng lp bn QTL tnh trng chng chu s thy phn (vit tt DT t ch dehydration tolerance) l: Qun th cn giao ti t hp (RIL) ca t hp lai CO39 / Moroberekan, vi 5 QTL (Lilley v ctv. (1996) Qun th la c s dng lp bn QTL tnh trng tch ly ABA l: Qun th F2 ca t hp lai IR20 / 63-83, vi 10 QTL (Quarrie v ctv. 1997)
Hnh 3-2: Bn QTL trn c s qun th DH ca t hp lai CT9993-5-10-1-M / IR62266-42-62. ICGY: nng sut ti Israel, iu kin bnh thng. ISGY: nng sut ti Israel, iu kin kh hn. OA: iu tit p sut thm thu. RD: su ca r. RPF: lc nh r. RPI: ch s r pht trin chiu su. YRI: ch s nng sut, TCGY : nng sut ti Thi Lan. TH: dy ca r (Zhang v ctv. 1999)

Mt yu t v cng quan trng lm hn ch vic thc hin bn QTL ca tnh trng OA l thiu nhng qui trnh nh gi kiu hnh nhanh, vi mt s lng ln cc vt liu th nghim. Gn y, c 3 qui trnh thng c s dng xc nh OA so snh vi phng php ti to hin tng hp th nc c xut trong trng hp cy la (Babu v ctv 1999). Qun th n bi kp (DH) ca t hp lai CT9993 / IR62266, vi 154 dng con lai c chn. Nhm tc gi s dng 145 RFLP, 153 AFLP v 17 microsatellite thit lp bn QTL. Tng s l 315 marker. nh gi kiu gen cn c thc hin trn qun th RIL ca t hp lai IR58821-23-B2-1 / IR52561-UBN-1-12, vi 166 dng con lai c chn. Tng s 383 AFLP v 106 RFL c s dng thit lp bn QTL (Zhang v ctv. 1999). Mt QTL c xc nh trn nhim sc th s 8, nh v gia 2 marker RG1 v RG978 (hnh 3-2), vi tn sut bin thin kiu hnh 32-34%. QTL ny nm trong vng ng dng vi nhim sc th s 7 ca la m 3-5. BN QTL TNH TRNG BIU HIN MU XANH CA CAO LNG Trong cy cao lng, kh hn xy ra trc v sau khi tr bng gy ra hin tng chn sm trong tnh trng mc ha gi ca l (senescence). Cy mt mu xanh nhanh chng, ng, v tht thu nng sut. Hn 80% cao lng u th lai c trng ti M di iu kin khng c nc ti, nhng hu ht cc ging ny u c kh nng khng hn giai on trc khi tr bng, hin tng nh vy c gi vi thut ng l stay-green trait (tm dch l tnh trng biu hin mu xanh, tnh trng ko di thi gian ha gi, vit tt l SGT). Tnh trng nh vy l mt c ch khng hn gip cho cy cao lng chng li s lo ha sm di iu kin khc nghit ca m t hn ch trong sut qa trnh sau khi tr. Subudhi v ctv. (1999) xc nh 4 QTL i vi tnh trng SGT ny, trong qun th cn giao ti t hp ca t hp lai B35 / Tx7000, vi 98 dng c chn F7. Ba QTL ch lc ca tnh trng SGT c xc nh trn nhng th nghim ngoi ng, ng gp 46% bin thin kiu hnh. Bn QTL ca tnh trng SGT nh v trn 3 nhm lin kt gen. Ba QTL: Sgt 1, Sgt 2, v Sgt 3 ng gp vo tn sut bin thin kiu hnh l 20%, 30% v 16%, theo th t. QTL th t l Sgt 4 c xc nh trong nm 1994, gii thch 13% s bin thin kiu hnh. i vi tt c QTL ny, nhng alen SGT u do ging m B35 cung cp. Cc tc gi d nh s thit lp bn vt l cc ni nh v ca nhng QTL nh vy c nhng hiu bit r rng hn v di truyn tnh chng chu hn ca cao lng. Hin kt qa nghin cu p dng MAS xc nh s du nhp cc gen Sgt vo cc dng con lai c trin vng (Subhudi v ctv. 1999). Mt cp lai khc B35 / Tx430 cng c thnh lp qun th RIL, vi 96 dng c chn. By QTL ca tnh trng SGT c xc nh, trong c 3 QTL ch lc Sgt A, Sgt D v Sgt G. Nhng QTL ny nh v trn 3 nhm lin kt gen, ng gp 42% bin thin kiu hnh. Bn QTL th yu (Sgt B, Sgt I-1, Sgt I-2, Sgt J) ng gp 25% bin thin kiu hnh ca tnh trng SGT (Subudhi v ctv. 1999). 3-6. BN QTL CC TNH TRNG HNH THI QUAN TRNG Cc tnh trng s lng ca cy la giai on m: tng s r, th tch r, s chi, t l r / chi, chiu di r ti a, im chng chu kh hn (1-5), b dy ca r, chiu cao cy m, trng lng r kh, cun li ca l la, c thit lp bn QTL (Shashidhar v ctv. 1999). Ngi ta s dng qun th n bi kp t cp lai IR64 / Azucena, vi 56 dng c chn. Tng s RFLP marker biu hin a hnh trong iu tra b m, c s dng l 175 marker, bao ph 2.005 cM, vi khong cch trung bnh l 11,5 cM gia hai marker. QTL i vi tnh trng chiu cao cy nh v gia marker RZ730-RZ801 trn nhim sc th s 1. V tr ny tng ng vi gen sd-1 iu khin tnh trng na ln (semidwarf). Hnh 3-3, v bng 4 cho thy: 15 QTL khc c tm thy c hai giai on pht trin, trong c 4 QTL th hin kh ph bin
Bng 4: Nhng QTL gi nh trong 2 giai on tng trng ca cy la: 65 ngy sau khi gieo (DAS) c nc ti bnh thng, v 85 DAS di iu kin b kh hn t ngy 65 tr i (Shashidhar v ctv. 1999).
Tnh trng Ngy ly mu Marker QTL Nhim th Phng sai nh hng cng LOD
Cao cy (cm) S chi

65DAS 85DAS 85DASe RZ730-RZ801 RZ448-RZ337A RZ730-RZ801 RZ519-RZ448 RG958-CD0344 RZ730-RZ801 RZ519-RZ448 RZ-143-RG20 RG810-RG331 RZ448-RZ519 RG449-RG788 A10K250-AG8Aro AC5-RG4188 RG171-RG157 RZ329-RG348 RG91-RG449 RZ617-RG978 RG214-RG143 RG163-RZ590 RG381-RZ19 RG171-RG157 RG472-RG246 W1-RG173 RG477-PGMS0.7 RG134-RZ500 PGMS0.7-CD059 RG171-RG157 Pai1-RZ58 RZ123-RG520 RG403-RZ556 RZ67-RZ70 RZ337B-CD0497 RG171-RG157 RG104-RG348 RG157-RZ318 RG313-RZ556 AC5-RG418B RG157-RZ318 Pal1-RZ58 RG171-RG157 RG348-RG104 Sd1 qPHT3-1 Sd1 qPH3-2 qPHT12-1 Sd1 qPHT3-2 qNOT8-1 qNOT1-1 qNOT3-2 qNOT4-2 qNOT-8-1 qNOT8-4 qNOT2-1 qNOT3-1 qNOT4-1 qNOT8-3 qRTL4-2 qRTL4-1 qRTL1-1 qRTL2-1 qTRN1-1 qTRN1-2 qTRN7-1 qTRN10-1 qTRN7-1 qTRN2-1 qTRN2-2 qTRN2-3 qRTV5-1 qRTV5-3 qRTV7-1 qRTV2-1 qRTV3-1 QRTT2-1 QRTT5-1 QRTT8-1 QRTT2-2 QRTT2-3 qRTW2-1 qRTW3-1 qRS1-1 qDRS4-1 qDRS12-1 qLFR7-1 1 3 1 3 12 1 3 8 1 3 4 8 8 2 3 4 8 4 4 1 2 1 1 7 10 7 2 2 2 5 5 7 2 3 2 5 8 2 2 2 3 1 4 12 7 22.1 27.0 29.2 27.0 16.9 24.9 16.1 15.3 15.8 15.8 19.8 21.3 14.3 15.5 14.6 12.4 25.7 12.7 16.0 13.7 15.4 14.1 24.5 15.7 13.4 15.3 17.4 25.1 17.7 13.5 15.1 14.3 15.6 21.4 26.7 19.1 21.6 16.7 12.4 13.3 20.8 4.32 4.74 4.96 4.66 3.75 5.19 4.04 0.62 -0.96 -0.97 -1.15 1.16 -0.99 -0.99 -0.82 -0.77 1.07 -1.74 -3.04 3.25 -4.01 -6.88 -10.93 -7.25 -6.79 -9.88 -9.96 -11.02 -8.15 -1.51 -1.70 -2.01 -2.07 -1.99 0.05 0.04 0.04 0.05 0.04 -0.42 -0.38 -0.31 -0.27 0.27 0.36 2.53 3.03 3.63 3.42 2.04 2.79 1.92 1.94 2.10 1.95 2.51 2.80 1.73 1.63 1.81 1.61 3.54 1.65 2.07 1.64 1.62 1.81 3.00 2.06 1.75 1.58 1.80 2.69 2.26 1.60 1.82 1.74 1.74 2.89 3.21 2.41 2.94 1.66 1.59 1.58 2.80
S chi
65DAS 85DAS
85DAS
Chiu di r (cm) Tng s r
65 DAS 85DAS 85DAS 65 DAS
85DAS 85DAS Th tch r (cm3) 65 DAS 85DAS 85DAS dy ca r 65 DAS 85DAS TL kh ca r (g) T l r/chi im chng chu hn (1-5) cun li ca l (1-5) 85DAS 85DAS 85 DAS 85DAS 85DAS
RG173-Amy1B RG908-RG91 RG181-RG958 PGM0.7-CD059
12.7 14.4 16.1 11.9
1.63 1.80 2.03 1.55
Cc tc gi cn ghi nhn 21 QTL i vi nhng tnh trng khc trong iu kin b stress do kh hn. Mc d c 2 QTL i vi tnh trng chiu cao nh v nhim th s 1 v 3, kh ph bin, nhng khng c mt loci no i vi tnh trng hnh thi hc ca r c ghi nhn ph bin gia hai giai on pht trin. S khng c mt ca nhng QTL ph bin i vi tnh trng hnh thi ca r gia hai giai on pht trin (65 ngy u c nc, v t ngy 65- n ngy 85: kh hn) cho thy s hin din ca nhng c ch di truyn cng tn ti song song, cng khi ng cc giai on tng trng khc nhau, di iu kin nc ti khc nhau (Shashidhar v ctv. 1999). Th tch r v tng s r trn mt cy gim c ngha v mt thng k di iu kin b stress do kh hn, trong khi chiu di r ti a v chiu cao cy khc bit khng c ngha.
3-7. CHUYN NP GEN MC TIU
Da trn c ch chng chu kh hn ca cy trng, ngi ta c gng thit k nhng clone ca gen mc tiu phc v cho hot ng chuyn np gen v to ra nhng cy trng bin i gen, chng chu kh hn. Bng 5 cho thy danh sch cc gen mc tiu v sn phm ca n c cng b Bng 5: S th hin ca cc gen mc tiu lin quan n tnh chng chu hn, mn trong cy la chuyn gen (Reddy v ctv. 1999)
Gen HVA1 P5CS BetA CodA ADC Enzyme/protein Nhm LEA pyrroline-5-carboxylate synthetase Choline dehydrogenase / betaine aldehyde dehydrogenase Choline oxidase Arginine decarboxylase Ngun La mch (barley) u mothbean E. coli A. globiformis Kiu mch (oat) Phn ng chng chu Kh hn, mn Kh hn, mn Kh hn, mn, nhit Kh hn, mn Kh hn Tc gi Xu v ctv. 1996 Zhu v ctv. 1998 Takabe v ctv. 1998 Murata v ctv. 1998 Capell v ctv. 1998
(LEA: late embryogenesis abundant protein) Gen iu khin s iu tit cht nguyn sinh (OA) lin quan n chng chu kh hn trong cy thucl, cy Arabidopsis thaliana, c chuyn np gen thnh cng l - Nhm mannitol-1-phosphate dehydrogenase vi gen mc tiu l mtlD - Nhm proline bao gm pyrroline-5-carboxylate synthetase, vi gen P5CS. - Nhm glycine betaine trong vi khun l CodA (bacterial choline oxidase) c phn lp t vi khun Arthrobacter globiformis, hoc t Synechoccus sp. - Nhm trehalose t men lm bnh m l gentrehalose-6-phosphate synthetase (TPS1). - Nhm trehalose t vi khun l cc gen: otsA v otsB. - Nhm fructans chnh l nhng phn t polyfructose c phn lp t vi khun Baccilus subtilis, vi gen iu khin tnh chng chu hn l sacB - Nhm D-ononitol th hin gen m ha t cDNA ca myo-inositol Omethyltransferase (IMTI) - Nhm polyamine - Nhm gen c lin quan n stress do oxid ha Protein chc
Protein
Protein mng t bo (transporters, protein knh trao i nc) Yu t bo v ca cc phn t ln (LEA protein, chaperon) Enzyme gii c (GST, sEM) Enzyme chnh trong tng hp th osmolyte (proline, betaine, ng) Proteinase (thiol proteases, Clp protease, v ubiquitine)
Yu t chuyn m (MYC, MYB, bZIP, STRESS do kh hn

Protein kinase (MAPK, MAPKKK, CDPK, S6K) PI turnover (phospholipase C, PIPK, DGK, PAP)
Hnh 3-4:Nhng gen cm Tai lieu nhm: proteinng vi vai tr do khnng vsn phmnggen c th c chia seng stresswWw.SinhHoc.edu.vn tai: chc hn. protein ca vai tr iu tit chia thnh 2 cc tn hiu di truyn v s th hin gen khi cy phn ng vi stress (YamaguchiShinozaki v Shinozaki 1999)
3-8. C CH TRUYN TN HIU
C t nht 4 l trnh truyn tn hiu, khi cy trng b stress do kh hn, mn v lnh (hnh 3-5). l 2 l trnh c tnh cht c lp ABA, k hiu III, IV v 2 l trnh ABA c tnh cht l thuc, k hiu I, II. S tng hp protein v cng cn thit trong trng hp l trnh ABA c tnh cht l thuc (I). Sn phm ABRE cng c ghi nhn trong l trnh ABA c tnh cht l thuc (II). DRE l sn phm c trong l trnh ABA c lp do hin tng iu tit ca gen, khng nhng xy ra trong trng hp kh hn m cn xy ra trong trng hp mn v lnh (IV) (hnh 3-5) Mt l trnh ABA c lp ch kim sot tnh chng chu hn v mn, m khng kim sot tnh chng chu lnh, k hiu III (hnh 3-5).
Kh hn, mn Lnh
nhn tn hiu ABA Tng hp protein (MYC, MYB) I II MYCR2, MYBRS (rd22) th hin gen bZIP ABRE (rd29B) th hin gen ? l trnh c lp ABA DREBs III ? (erd1) (AP2/ERBP) IV (rd29A) th hin gen
Phn ng vi stress v chng chu
Hnh 3-5: L trnh truyn tn hiu v s th hin gen (Yamaguchi-Shinozaki v Shinozaki 1999)
Lnh ABA c lp ABA chuyn m Kh hn, Mn ABA c lp
chuyn m ci bin bZIP
Trans-elements rd29A promoter cis- elements
DREB1/CBF1
DREB2
DRE
ABRE
TATA
Hnh 3-6: M hnh cm ng ca gen rd29A v hot ng ca cc nhn t hot ng c tnh cht cis v trans trong hin tng th hin gen chng chu vi stress (Yamaguchi-Shinozaki v Shinozaki 1999)

Trong hnh 3-6, hai nhn t hot ng theo kiu cis l DRE trong h thng ABA c lp v ABRE trong h thng ABA l thuc, tham gia ng gp vo hin tng cm ng gen rd29A. C hai loi binding protein khc nhau l DREB1 v DREB2 tham gia vo 2 l trnh truyn tn hiu khc nhau, mt p ng vi stress do lnh, mt p ng vi stress do kh hn. DREB1/CBF1 c iu tit trong khi chuyn m, trong khi DREB2 c kim sot c hai giai on: trong qa trnh sau khi chuyn m v trong qa trnh chuyn m. Protein ABRE m ha nhng nhn t chuyn m bZIP. Promoter rd29A lm gim thiu nhng nh hng khng c li cho s sinh trng ca cy, trong khi 35ScaMV lm chm tng trng rt mnh (Yamaguchi-Shinozaki v Shinozaki 1999). 3-9. GEN & S KHM PH L TRNH THNG QUA GENOME HC CHC NNG Mt trong nhng mc tiu chnh ca genome hc v chc nng (functional genomics) l khm ph ra nhng gen ch lc v QTL tng ng vi tnh trng nghin cu. S khm ph cc gen mc tiu hin c pht trin thng qua k thut phi hp c trnh t genome, phn tch microarray, v proteomics (ngnh protein hc), trn c s ngun d liu ca sinh tin hc (bioinformatics) v di truyn phn t (Salekdeh v ctv. 2002). Nhng gen ng c vin c gi vi thut ng short-listed candidate genes c khm ph v lu tr. Chng c th nghim mt cch trc tip trn cy chuyn np gen v hiu qa ca tnh trng chng chu kh hn. Ngnh protein hc (proteomics) c th gip chng ta xc nh nhng gen ng c vin th hin r chc nng v v tr trong genome. Hnh 3-7 s cung cp cho chng ta thng tin v s ng gp ca ngnh hc mi l protein hc v transcript hc (transcriptomics) trong chin lc MAS v chn ging cy trng.
B m chng chu v nhim
Transcriptomics & Proteomics
Gen ng c vin & protein
Phn ly alen
Loci ca gen ng c vin TQL Marker QTL PQL chuyn np gen MAS Chn ging phn t
Transcriptomics Qun th mapping nh gi kiu hnh Proteomics
nh gi kiu gen
Bn di truyn
Hnh 3-7: ng gp ca ngnh protein hc v transcript hc vo chin lc MAS v chn ging cy trng (Salekdeh v ctv. 2002). Gen ng c vin c so snh ng vi tnh cht proteomic hay transcriptomic ca nhng dng chng chu v dng nhim stress do kh hn. TQL: transcript quantity locus, QTL: quantitative trait locus, PQL: proteomic quantity locus
Phn tch proteomic cung cp cho chng ta nhng nhn nhn mi, rng hn v hin tng phn ng ca cy trng i vi stress mc protein. Trong nhng nm gn y, phng php ny pht trin trong khi tm hiu mc nhy cm ca thc vt, v ngi ta xem n nh cng c c kh nng rt mnh trong ci tin k thut in di polyacrylamide hai chiu (2DE). Thut ng 2DE c vit tt t ch two-dimentional polyacrylamide gel electrophoresis. Phng php proteomic cn l cng c pht hin protein, nh lng protein, k thut fingerprinting (in du trong xc nh quan h huyt thng), k thut c trnh t protein tng phn (partial sequencing) bng sc k (mass spectrometry = MS), k thut tin sinh hc (bioinformatics), v cc phng php phn lp gen mc tiu (gene isolation). K thut 2DE cung cp cho chng ta nhng thng tin v nhng thay i trong ng thi pht trin ca protein, phn nh nhng hin tng ci bin xy ra sau khi chuyn m th d nh hin tng phosphoryl ha. Salekdeh v ctv. (2002) p dng phng php ny nghin cu v hin tng p ng ca cy la i vi stress do kh hn v mn. Cc tc gi khm ph trn 2000 protein trong l la b x l kh hn v l la c ti nc y . Vi hn 1000 protein c nh lng mt cch ng tin cy, c 42 protein th hin s thay i v mc a dng v v tr trong genome. Nhm tc gi ny phn lp c nhiu protein trn l la m mc a dng ca n gia tng mt cch c ngha trong sut thi gian b x l kh hn, v trong thi gian ngun nc cung cp gim dn. Ba thay i c n tng nht l (1) hot ng ca actin depolymerase, (2) mt phn t ng dng ca S-like ribonuclease (3) hot ng ca chloroplastic glutathione-dependent dehydroascorbate reductase Nhng so snh mc proteomic gia ging la chng chu mn v ging la nhim mn cho thy c s khc bit protein trong r la trong c hai khi nim protein cu trc (constitutive) v protein cm ng (inducible). Mt enzyme tng hp lignin l caffeoylCoA - O-methyltransferase l kt qa ca s so snh khc nhau ny gia hai ging chng chu v ging nhim. Mc a dng ca ascorbate peroxidase th hin cao hn rt nhiu trong ging la chng chu mn Pokkali so vi ging nhim IR29, trong iu kin cha b x l stress (Salekdeh v ctv. 2002).
Bng 6: Phn lp protein cy la p ng vi stress do kh hn v mn trn c s phn tch protein-fingerprinting (MALDI-TOF MS), hoc phng php c trnh t protein tng phn (ESI-Q-TOF MS/MS) (Salekdeh v ctv. 2002). Spot a TL Mc phn ph t MALDI(kDa) TOF (%) b LD1 46 ESI-Q-TOF Sequence Phn lp Mc AA (%)c 100 Ko di s dch m EF-Tu precursor, chloroplast RUBISCO activase RUBISCO activase Fructose-1,6biphosphate aldolase, chloroplast Triose phosphate isomerase, cytosolic S mud
GITINTATVEYE
AAF15312
LD2 LD3 LD4
43 43 38
37 33 25
P93431 P93431 Q40677
LD5 LD6
28 29
57
P48494
YGYPSEDFFVK e MCFTFDSSENTAVVR S-like nuclease LD7 27 LALTELQALEEHLK 100 GSH-dependent BAA90672 LPEDLTNVHAYTEA DHA reductase LD8 21 LFSR DE[L/I]TASLPADG 80 ADF S30934 RS1 31 VVEVVESSPPEIK 100 ESTs AU091660 BAA96588 SGTTPLSPAIAFILEK 100 RS2 31 46 Caffeoyl-CoA 3- BAA81774 Omethyltransferase RS3 T[L/I]NDT[L/I] Cha c kt qa RS4 28 60 Ascorbate P93404 peroxidase RS5 22 39 ABA- v protein AAB96681 cm ng Asr1 a LDS = protein trong l la, nghim thc x l kh hn, RS = protein trong r la, x l mn b Phn trm amino acid trong protein c ph bi cu ni peptide tng ng trong phn tch MALDI-TOF c Phn trm amino acid trong chui m ESI-Q-TOF c tm thy protein mc tiu d S mu trong NCBI, SWISS-Prot, hoc PIR e Phn lp bng trnh t tng BLASTing ca cDNA tng ng RUBISCO activase: Cy la c xem nh c hai isoform chnh v h thng RUBISCO activase, c kch thc t 43 n 47 kDa. Nhm tc gi ni trn tm thy trong l la trn c s k thut 2DE l: c t nht 4 dng khc nhau ca activase vi kch thc thay i t 42 n 43 kDa. Hai trong s activase biu hin mc a dng cao hn trong iu kin b x l kh hn, so vi iu kin ti nc bnh thng (bng 6).
(1) Nhim v chnh ca RUBISCO activase l phng thch gc phosphate ng c tnh cht c ch, th d nh ribulose-1,5-biphosphate (RuBP) t nhng v tr hot ng ca RUBISCO sao cho nng CO2 trong l c th kch hot ezyme thng qua hin tng carbamyl ha. Kh hn kch thch s ng li ca kh khng to ra nng CO2 cao trong l, lm suy gim hot ng carbamyl ha, v lm khi ng hin tng kt gn c tnh cht dead-end ca RuBP ti v tr hot ng ca RUBISCO. (2) Nhim v gia tng mc phong ph ca activase gia tng c th c mt chc nng c bit lm cho RUBISCO hot ng tr li trong hin tng c nh CO2 thng qua carboxylase, hoc hin tng phn tn nng lng c tnh cht bo v nh phn ng oxygenase trong quang h hp
(3) Nhim v th ba ca RUBISCO activase l bo v sinh tng hp protein lc lp, khi cy b stress do kh hn, hoc do nhit Hu ht cy trng u ch c mt gen RUBISCO activase. N c hnh thnh mt cch tch cc, trong cc intron ca phn t transcript u tin thng b ct ri thnh hai isoform ca activase, ch khc nhau duy nht gc carbon cui cng. Lm th no 3-4 isoform hp li vi nhau thnh mt gen n ? Ngi ta hon ton cha bit r qu trnh nh vy. Guenzi v ctv. (2003) ti i Hc Oklohama thnh cng trong chuyn np gen mannitol-1-phosphate dehydrogenase (gen mtlD) trong cy la m chng chu kh hn v mn. Gen ny c tnh cht b sung b gen vn c trong t nhin, nhm kch hot cy trng chng chu tt hn kh hn (ngun truy cp t http://www.isaaa.org/kc). Chng ta c th tham kho t sch ca Abebe v ctv. (2003) Trong l cy b kh hn v trong r cy b stress do mn, nhng k thut theo di s thay i v mc a dng ca protein, thay i v v tr ca protein c bit v cha c bit, l mt phng php rt ng ch . Nhng protein l sn phm ca qa trnh quang tng hp, s ko di t bo, qa trnh bin dng hp cht chng oxi ha (antioxidant) v qu trnh lignin ha c quan tm trong nghin cu. Do , ngnh hc mi c thut ng proteomics s l cng c hu hiu, gip cho nh chn ging tm kim nhng kiu gen thch hp cho mc tiu ci tin tnh trng chng chu kh hn.
TI LIU THAM KHO Abebe T, AC Guenzi, B Martin, JC Cushman. 2003. Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity. April 11, 2003. Issue of Plant Physiology 131:1748-1755 Babu RC, MS Pathan, A Blum, HT Nguyen. 1999. Comparision of measurement methods of osmotic adjustment in rice cultivars. Crop Sci 39:150-158 Bertin P, A Chacosset, A Gallais. 1997. Physiological and genetic basis of nitrogen use efficiency in maize. In: Tsaftasis A, editor. Genetics, biotechnology and breeding of maize and sorghum. Cambridge (UK): The Royal Society of Chemistry. P 59-64 Botstein D, RL White, M Skolnick, RW Davis. 1989. Construction of a genetic linkage map in man using restriction fragment length polymorphism. Ann J Genet 32:314 Capell T, C Escobar, H Liu, D Burtin, O Lepri, P Christou. 1998. Overexpression ofthe oat arginine decarboxylase cDNA in transgenic rice (Oryza sativa L.) affects normal development patterns in vitro and results in putrescine accumulation in transgenic plants. Theor Appl Genet 97:246-254
Champoux MC, G Wang, S Sarkarung, DJ Mackill, JC OToole, N Huang, SR McCouch. 1995. Locating genes associated with root morphology and drought avoidance in rice via linkage to molecular markers. Theor Appl Genet 90:969-981 Ingram J, D Bartels. 1996. The molecular basis of dehydration tolerance in plants. Plant Cell Rep. 15:963-968 Li Z. 1999. DNA markers and QTL maping in rice. In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 157-172. Lilley JM, MM Ludlow, SR McCouch, JC OToole. 1996. Locating QTL for osmotic adjustment and dehydration tolerance in rice. J Exp Bot 47:1427-1436 Martin B, J Nienhuis, G King, A Shaefer. 1989. Restriction fragment length polymorphisms associated with water use efficiency in tomato. Science 243:1725-1728 Milan MAR, DA Aswey, HR Boerma. 1998. An additinal QTL for water-use-efficiency in soybean. Crop Sci. 38:390-393 Murata N, H Hayashi, A Sakamoto, Alia. 1998. Genetic enhancement of tolerance to multiple stress by metabolic engineering of biosynthesis of glycine betaine. In: proceeding of International workshop on breeding and biotechnology for environmental stress in rice, 26-29 Oct. 1998, Japan. Nguyen HT, RC Babu, A Blum. 1997. Breeding for drought tolerance in rice (Oryza sativa): physiology and molecular genetic considerations. Crop Sci 37:1426-1434 Price AH, Thomas. 1997. Quantitative trait loci associated with stomatal conductance, leaf rolling and heading date mapped in upland rice (Oryza sativa L.). New Phytol 137:8391 Quarries S, A Steed, A Semikhodski, C Lebreton, C Calestani, DA Clarkson, R Tuberosa, MC Sanguineti, R Melchiorre, JL Prioul. 1995. Identification of quantitative trait loci regulating water-nitrogen-use efficiency in wheat. In: II STRESSNET Conference, 2123 Sept. 1995, Salsomaggiore, Italy. European Commission, Rep FII.3-MOR/OCO2. Quarries S, V Lazic-Jancic, M Ivanovic, C Pekic, A Heyl, P Landi, C Lebreton, A Steed. 1997. Molecular marker methods to dissect drought tolerance in maize. In: Tsaftaris A, editor. Genetics, biotechnology and breeding of maize and sorghum. Cambridge (UK): The Royal Society of Chemistry. P. 52-58 Reddy AR, V Ramanathan, N Seetharama, S Bajaj, R Wu. 1999. Genetic improvement of rice for drought and salt tolerance: molecular breeding and transgenic strategies. In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 135-155 Redona ED, DJ Mackill. 1996. Molecular mapping of quantitative trait loci in japonica rice. Genome 39:395-403 Ribaut JM, C Jang, D Gonzales de Leon, GO Edmeades, DA Hoisington. 1997. Identification of quantitative trait loci under drought conditions in tropical maize. 2. Yield components and marker-assisted selection strategies. Theor Appl Genet 94:887-896 Ribaut JM, DA Hoisington, JA Deutsch, C Jang, D Gonzales de Leon. 1996. Identification of quantitative trait loci under drought conditions in tropical maize. I. Flowering parameters and anthesis-silking interval. Theor Appl Genet 92:905-914 Salekdeh GhS, J Sioponco, LJ Wade, B Ghareyazie, J Bennett. 2002. A proteomic approach to analyzing drought- and salt-responsiveness in rice. Elsevier, Field Crops Research 76:199-219
Shashidhar HE, GS Hemamalini, S Hittalmani. 1999. Molecular marker-assisted tagging of morphological and physiological traits at the peak vegetative stage: two constrasting moisture regimes. In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 239-256 Shen L, B Courtois, K McNally, SR McCouch, Z Li. 1999. Developing nera-isogenic lines of IR64 introgressed with QTLs for deeper and thicker roots through marker-aided selection. In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 275-289 Stuber C. 1997. Years of searching and manipulating QTLs in maize. In: Tsaftasis A, editor. Genetics, biotechnology and breeding of maize and sorghum. Cambridge (UK): The Royal Society of Chemistry. P 43-58 Subudhi PK, GB Magpantay, DT Rosenow, HT Nguyen. 1999. Mapping and marker-assisted selection to improve the stay-green trait for drought tolerance in sorghum. In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 183-191 Takabe T, Y Hayashi, T Tanaka, S Kishitanu. 1998. Evaluation of glycine betaine accumulation for stress tolerance in transgenic rice plants. In: proceeding of International workshop on breeding and biotechnology for environmental stress in rice, 26-29 Oct. 1998, Japan. Teulat B, D Tjis, M Khairallah, C Borries, C Ragot, P Sourdille, P Leroy, P Monneveux, A Charrier. 1998. Several QTL involved in osmotic-adjustment trait variation in barley (Hordeum vulgare L.). Theor Appl Genet 96:688-698 Teulat B, P Monneveux, J Wery, C Borries, I Souyris, A Charrier, D This. 1997. Relationships between relative water content and growth parameters under water stressin barley: a QTL study. Nwe Phytol 137:99-107 Tuinstra MR, EM Grote, PB Goldsbrough, G Ejeta. 1996. Identification of quantitative trait loci associated with pre-flowering drought tolerance in sorghum. Crop Sci 36:13371344 Tuinstra MR, G Ejeta, P Goldsbrough. 1998. Evaluation of near-isogenic sorghum lines contrasting for QTL markers associated with drought tolerance. Crop Sci 38: 835-842 Velboom LR, M Lee. 1994. Molecular-marker-facilitated studies of morphological traits in maize. II. Determination of QTLs for grain yield and yield components. Theor Appl Genet 89:451-458 Xu D, X Duan, B Wang, B Hong, THD Ho, R Wu. 1996. Expression of a late embryogenesis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice. Plant Physiol 110:249-257 Yadav R, B Courtois, N Huang, G McLaren. 1997. Mapping genes controlling root morphology and root distribution in a double haploid population of rice. Theor Appl Genet 94:619-632 Yamaguchi-Shinozaki K, K Shinozaki. 1999. Improving drought, salt, and freezing stress tolerance using a single gene for a stress-inducible trascription factor in transgenic plants. In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 173-182 Zeng ZB. 1994. Precision mapping of quantitative trait loci. Genetics 136:1457-1468 Zhang J, HG Zheng, ML Ali, JN Triparthu, A Aarti, MS Pathan, AK Sarial, S Robin, Thuy Thanh Nguyen, RC Babu, Bay duy Nguyen, S Sarkarung, A Blum, Henry T Nguyen. 1999. Progress on the molecular mapping of osmotic adjustment and root traits in rice.
In: Genetic Improvement of Rice for Water-Limited Environments. (Eds.) O Ito, JC OToole, and B Hardy. IRRI, Philippines. P. 307-317
Zhu B, J Su, MC Chang, DPS Verma, YL Fan, R Wu. 1998. Overexpression ofa pyrroline5-carboxylate synthetase gene and analysis of tolerance to water and salt stress in transgenic rice. Plant Sci. 139:41-48
Tai lieu chia se tai: wWw.SinhHoc.edu.vn Chng 4

C S DI TRUYN TNH CHNG CHU NGP NG
Trong thp nin 1980, c khong 30 n 40% din tch trng la Chu thuc loi hnh canh tc b ngp lt hng nm vo v ma, su mc nc trung bnh t 0,5 n 1,0 m n , Bangladesh, Min in, Thi Lan v Vit Nam; nhng vng khc c mc nc rt su (hn 1m) c gi l vng la ni, tp trung Nam v ng Nam Chu (Jackson v ctv 1982, Vergara v ctv 1976 HilleRisLambers v Seshu 1982). Vic iu tra v phn loi vng trng la gp nhiu kh khn v tiu chun nh danh cha thng nht. T nm 1979 n nm 1984, thut ng v vng sinh thi trng la c thay i nhiu ln bi cc nh khoa hc IRRI v ngi ta phi thnh lp mt Uy ban quc t phn loi v nh danh vng sinh thi la, nhm thng nht cch gi tn, gip ch cho vic xc nh mc tiu nghin cu r rng hn (Garrity 1984). Cn c vo su mc nc, Khush (1984) ngh gi nh sau: 0 Nc cn 25 Nc su trung bnh 50 Nc su 100 cm Nc rt su (la ni)
p dng h thng nh danh ny, chng ta c th chia tp on la ma nc su a phng BSCL lm hai nhm: la nc su (mc nc trn rung di 100 cm) v la ni (trn 100 cm).
Cng trnh nghin cu v la nc su c cng b t nm 1934 ti trm la ni Habiganj (Bangladesh) (Jackson v Vergara 1979). Cng trnh tp trung vo vic nghin cu tuyn chn ging ci tin, nh hng ca tui m i vi s chu ngp, iu kin t ai, thi v v phng php gieo s. Trm th nghim la ni Huntra (Thai Lan) c xy dng sau trm Habiganj 7 nm. y, cng trnh tp trung vo vic ci to phm cht ht v nng sut bng con ng tuyn chn t cc ging la a phng. Trong nhng nm 1950, vi cng trnh nghin cu v la nc su c thc hin Nht v Bangladesh (Yamada 1959, Yamada v Ota 1956, Yamada v ctv 1954, Alim v Sen 1955, Alim v Zaman 1985), nhng vn cha c ci tin g v cng tc ging. La nc su bt u c nghin cu IRRI t nhng nm 1960, xem nh mt nhim v quan trng ca Vin. Thut ng floating dwarf c s dng rng ri trong cc chng trnh lai to gia ging la ln v la ni ti IRRI trong thi gian ny. Kh nng phng thch ra cc dng hnh na ln, c th vn lng trong iu kin nc su trung bnh c chng minh vi cc dng trin vng ca t hp lai IR442 (IR95-314 / Leb Mue Nahng 111). Tuy nhin, ngi ta yu cu phi xc nh r v su ca mc nc: su bao nhiu th c gi l nc su ? l cu hi c t ra t nhng chng trnh hp tc quc t v la nc su IRRI. Cc nh chn ging cn c s thng nht v nh ngha vng nc su, bao gm mc v nhng tr ngi chnh ca hin trng c c mc tiu lai to r rng (Jackson v ctv 1982). Hi ngh quc t la nc su ln th nht c t chc ti Bangladesh (1974) vi nhiu bo co nghin cu v ging, di truyn, hnh thi hc, nhng thit hi chnh cc vng la nc su. Hi ngh tho lun vic tng cng s hp tc quc t. Sau vo nm 1975 cc nh khoa hc hp ti IRRI, thng nht mc tiu lai to ging: (1) khng hn, (2) kh nng vn lng, (3) cng lc m, v (4) chng chu ngp. V canh tc, h yu cu nghin cu tp trung: (1) phng tr c di, (2) thi v gieo, (3) mt s, (4) phn bn, (5) phng php canh tc, (6) sa son t. Chng trnh kho nghim la nc su quc t (IRDWON) c t chc t 1976 dn dn p ng phn no yu cu v cng tc ging. Hi ngh quc t la nc su ln th hai c t chc ti Thi Lan (1976) thng nht v c bn vic s dng thut ng ging la nc su v tiu chun ha phng php cho im trong nghin cu tnh chng chu:
Tai lieu chia la nc su lnwWw.SinhHoc.edu.vn trnh se tai: th ba c t chc ti n (1978) xut chng Hi ngh quc t ging
hp tc quc t v k thut RGA (rapid generation advance), pht trin nhanh chng cc dng lai, chng trnh thanh lc chng chu ngp v chu kh hn quc t. Hi ngh quc t la nc su ln th t c t chc ti Thi Lan (1982) nhn mnh v tnh chng chu ngp, xut mc tiu lai to vi tnh chng chu su bnh ring bit i vi tng vng (tuyn trng thn c xem l mt trong nhng i tng chnh): tnh khng hn giai on m, tnh chu ngp cho nhng vng b l lt t ngt. Hi ngh nu r yu cu v ging cho vng la nc su tr nn v cng cn thit, v tm quan trng cho chng trnh lai to. Hi ngh rt quan tm n nhng nghin cu c bn v di truyn - nht l tnh trng chng chu ngp v kh nng vn lng. Hi tho v la nc su (ACIAR workshop) c t chc vo thng 6-1985 ti Australia tp trung tho lun v tnh chu ngp ca cy la (sinh l hc) v nhng kt qu mi nht v c ch chng chu 4-1. TNG QUAN NGHIN CU TRC Y Nm 1941, Ramiah v Ramaswami cng b mt chng trnh nghin cu v di truyn tnh trng ni ca cy la (trn c s dng hnh thn r). c tnh tng trng theo kiu ng rp (b) ca la ni do hai gen ln, lp on iu khin, mt gen lin kt vi gen tri ca tnh chn mun. Nhng pht hin ch yu da trn vic nghin cu qun th F2 c trng trong iu kin nc cn ca cc cp lai, gia la nc su vi ging khng phi nuc su ca n , Bangladesh, Pakistan. Tc gi khng cho rng c kh nng vn lng ca nhng cy ang phn ly. Tc gi ngh mt chng trnh lai cho nhng cy thn ln vi kh nng chi gn v chng chu c nc su. iu ny c th lm c v khng c s tng quan c tnh di truyn gia c tnh chu nc su v chiu cao cy, hoc gia c tnh chu nc su v s chi gn (Choudhury 1975). Cc tc gi iu ngh kiu hnh chi gn c xem nh mt c tnh ci tin ca ging la ni (Jackson v Vergara 1979). Nm 1965, Chang xc nh thm v hai gen ln dw1 v dw2 iu khin tnh trng ni. Nht, Kihara v ctv (1962) nghin cu c tnh tng trng ca cc ging la nc su su tp t Assam (n ), Thi Lan, v Chu Phi. T , ngi ta ngh c tnh ni c th c nghin cu trn c s ca 3 yu t: S pht trin chiu cao v s thch nghi nh vn lng vi iu kin khng c s ngp bt ng lc u (iu kin bnh thng). b) Kh nng vn lng thch nghi vi tc gia tng mc nc trung bnh. c) Kh nng vn lng thch nghi vi tc gia tng mc nc ti a. Tc gi cha cp n vic xc nh gen cho mi yu t, hoc c on s gen ca tng yu t. Theo kt qu ny, cc ging la nc su ni ting u c cha yu t b v c. C s khc bit gia cc ging thuc nhm b v c. Cc gen ni c th c truyn t ging la nc su sang ging la ln, chi khe, dng gn (Kihara v ctv 1962). Yu cu v mt ngun gen phong ph cho chng trnh lai to ging la nc su cng c cc nh khoa hc xut. C hai Oryza sativa L. v Oryza glaberrima Steud u c nhng ging la trng (cultivar) thch nghi vi iu kin ngp l, nhng cc loi c ngun gc Chu cung cp c nhiu dng hnh phong ph hn (Chang v ctv 1977). Nhng ging la thuc Oryza glaberrima Steud c c tnh tng trng kiu b v kh nng g u dy (kneeing) trong khi trng nc cn u c th xem nh dng hnh la nc su (Chang 1979). Tc gi cho bit hin c 646 mu ging la nc su ca Chu v 680 mu ging ca Chu Phi c su tp ti Ngn hng gen ca Vin la Quc t. S thch nghi rng ca cc ging la trng cho thy c ngun gc a h (polyphyletic) ca Oryza sativa L. v lnh vc nc su c ghi nhn nh sau: a)
O. perennis
O. fi O.
O. O.
O O.
Cc ging la nc su c t qu trnh tin ha ca Oryza perennis Moench (Chandraratna 1964, Choudhury 1975). Tt c cc ging la ni c bit hin nay thuc loi hnh Indica, cha thy ging la ni loi hnh japonica, cng nh cha truyn c gen ni ca indica vo japonica (Oka 1975). Tc gi lu : c vi ging la khng th hin tnh vt nc nhng vn c th c gen ni bi c t tin ca n l ging hoang di, ni.
Morishima v ctv (1962) cng b rng la ni dng hoang di khng nhng c kh nng ni m cn khng hn. Tc gi cho rng trong cc ging la trng thuc Oryza sativa L. c s khc bit mt cch ng ch v kh nng ni: t trung bnh n mnh. Kh nng ni l yu t thuc dng hnh hoang di v na hoang di cy la thch nghi vi ch nc thay i bt thng. Ti Vin nghin cu di truyn quc gia Nht bn, 10 ging la hoang v la trng c th nghim theo s phn loi ca Kihara v ctv (1962) c nhng kt qu rt khc nhau i vi yu t a, b v c, cho thy c tnh ni th hin ra vi nhiu hnh khc nhau (Morishima 1975).
c tnh c r ph v chi mt lng ca la ni v la nc su, cng vi c tnh vn lng c dng phn bit gia la ni v la khng ni. S dng cc d kin thu t cc th nghim trong h vi 56 dng la trng v la hoang, Morishima (1975) thit lp cng thc theo phng php phn tch phng trnh bit thc (discriminant function) lm ti a ha nhng khc bit dng hnh ni v khng ni. C s phi hp tuyn tnh ca 4 yu t:
A: S lng c vn di ra (hn 3 cm) B: Chiu di tng cng ca cc lng c vn di. C: S lng c r ph trn thn la D: S chi mt lng trn thn. Phng trnh bit thc XF = 1.00A + 0.47B + 0.50C + 0.24D. Vi phng trnh ny, tc gi iu tra s bin thin v kh nng ni ca mt s dng la trng v la hoang. i vi la trng thuc O. sativa L. v O. glaberrima Steud. Cc ging la ni v khng ni rt d dng phn bit. i vi cc dng hoang di thuc O. perennis Moench v O. brevigulata A. Cheval et Roehr, c s bin thin lin tc v kh nng ni mt mc nht nh. Qun th F2 ca cc cp lai gia la ni v la khng ni cng cho mt bin thin lin tc v XF. iu ny cho thy, c th c nhiu gen iu khin tnh trng ni (Morishima 1975). Cy F2 c trc nghim trong iu kin nc su cho thy s lng v chiu di c tng quan nghch (r bin thin t 0.39 n 0.52). Trong 56 dng c th, hai yu t ny tng quan khng c ngha. iu ny cho thy, s lng v chiu di lng trong nc su c iu khin bi nhiu gen khc nhau (Morishima 1975). Ngoi ra tc gi cn ghi nhn rng, s lng trong iu kin trng nc cn (5cm) c th c dng nh mt ch tiu c on kh nng ni (r= 0.78) trong iu kin ngn ngy v mc nc cn (5cm). Khi nghin cu v cc kiu gen (genotypes) ca cc t hp lai gia la ni v cc dng trung gian, Mukherji v Roy (1977) nhn thy cng thc tnh kh nng ni ca Morishima (1975) cn c thay i, v s lng c r ph v s chi mt lng khng tng quan vi kh nng ni. Yu t chn c lin quan n n l chiu cao cy cui cng. Tc gi ngh mt cng thc mi: FD = 1.00A + 0.50B + 0.20C + 1.00D 0.25E. (FD): kh nng ni, (A): s lng di hn 3cm, (B): chiu cao cy cui cng trong iu kin nc su, (C): chiu di tng cng lng thn c tng thm, (D) chiu di tng cng lng thn c tng thm cho n ngy th 81 k t sau khi gieo, (E): t l vn lng gia iu kin nc su so vi iu kin bnh thng (nc su 5 cm). Ti Huntra (Thi Lan), nghin cu cc t hp lai gia la ln v la ni trn c s theo di s phn ly ca F2, F3, F4 cc su mc nc 90, 110, 140 v 150 cm cho thy: (1) kh nng vn lng ch c th c ghi nhn mc nc su hn 90 cm, (2) vic tuyn chn c tnh vn lng trn c s tng th h ring bit v tng v s khng t kt qu tt, (3) s phn b tn sut ca cc dng F3, F4 chng minh rng c s chuyn bin mnh m t s
vn lng km n tt gia cc cy F3 v dng con chu F4 ca cy y (Supapoj v ctv 1977). iu ny cho thy vic tuyn chn kh nng vn lng s c thc hin khi c s ng hp t. Cng t th nghim ny, kh nng vn lng trong iu kin nc su hn 150 cm c quan h mt thit n chiu cao, cng lc m, gc l, thi gian tr (HilleRisLambers v ctv. 1977). S phn b v tnh vn lng F2 l mt ng biu din lin tc ch c mt nh, c tnh ny c iu khin bi a gen vi tnh tri khng han ton (partial dominance) khng hon ton v ch c mt phn nh c dng hnh ni tht s (Hamamura v Kupkanchanakul 1979, Nasiruddin v ctv. 1982, Supapoj v ctv. 1977), cn li phn ln c dng hnh trung gian, khng c hin tng phn ly vt tri (transgressive segregation). Cc nh nghin cu Gagraghat (n ) cho rng c s phn ly ri rc (discrete segregation) ca dng hnh ni v khng ni bi hot ng ca gen b sung (Nasiruddin v ctv. 1982). Tc gi cn ghi nhn: 1. Trung bnh v chiu cao ca qun th F2 thp hn F1 cho thy s gim st v tnh cht lai i sau (inbreeding depression). 2. C hot ng tng tc khng alen xy ra cho thy phi c nhiu hn mt locus iu khin s vn lng. 3. C hai s vn lng v vn di cy la u c iu khin bi mt c ch di truyn thng nht. S vn di ca cy la thng l s kt hp nh hng ca s vn lng, vn di ca b l, ca phin l (Nasiruddin v ctv. 1977). S vn di b l cn mt lng carbohydrate v oxy cao hn so vi yu cu vn lng (Gomosta v Vergara 1985). Tc gi cho rng c th l th (n-2) l trung tm hot ng, n iu khin s vn lng ca lng th (n-2), bng cch cung cp dng cht, carbohydrate v cc hormones. c tnh to lng thn sm khin cho cy la ni c c im tng trng b, l mt tnh trng di truyn. S phn ha to lng thn sm do mt gen tri iu khin k hiu Nd (Tripathi v Rao 1985). Tc gi cng ghi nhn s iu khin ca hai gen tri Nr 1 v Nr 2, hot ng b sung, i vi s to ra r ph. Cc tnh trng c lin quan khc c cng b nh: Hai gen tri, lp on An 1 v An 2 iu khin tnh trng ru ht, gen tri Gd iu khin tnh ng ngh ca ht. C s lin kt gen gia c tnh tng trng b v tnh trng ru ht, gia tnh trng go vi ng ngh ca ht (dormancy) trong loi hnh cc ging la ni. S vn di lng thn ca ging la nc su a phng xy ra cy m 4 tun tui, trong khi i vi loi hnh ci tin (na ln): 10 tun tui (Keith v ctv. 1986). C s khc bit v ngun gc ca cc nhm la ni v mt a l, khi nghin cu v cc kiu gen vi hm lng esterase khc nhau (Inouye v ctv. 1985). Trong nghin cu ny, cho thy cc ging la ni vt nc gii nht c xu hng tr mun nht, thch nghi vi iu kin ngp lt, nc rt kh chm. Tc gi cn ghi nhn: la ni khc vi la thng l s ko di lng th 5 v 6 khi cho phn ng vi GA (gibberellic acid). Ngoi ra ngi ta cn ch n phng php phn loi nhanh chng cc dng lai c tnh chng chu nc su giai on m, cy la ni c th phn bit bi chiu di ca trc mang l mm (mesocotyl) khi cho ny mm trong ti v gieo su, chiu di ca l th hai (ngn hn), chiu di ca lng th t, v nm khi cho lp t cy m 10 ngy tui, s khc bit v hm lng acid abscisic v acid gibberellic (Hamamura v Saengpetch 1977, Inouye v Xun 1973). V lnh vc nghin cu tnh chng chu s ngp t ngt, cho n nay c rt t thng tin v di truyn tnh chng chu ny (Mohanty v ctv. 1982).
Tnh khng ngp l mt tnh trng quan trng cho la nc cn cng nh cho la ni. La nc cn cn c kh nng sng st sau khi ngp hon ton sut mt ngy hoc mt tun. La ni cng c kh nng chu ngp trong trng hp s vn lng khng theo kp tc nc dng qu nhanh trong ma l bt thng (Vergara v Mazaredo 1975). Kt hp tnh khng ngp vo cc ging la ni s l mt bc ci tin rt ln v kh nng thch nghi ca n. Kh nng vn lng v tnh chng chu ngp l hai tnh trng hon ton khc nhau, s vn di lng thn c xem nh c ch trnh ngp (avoidance mechanism), iu quan trng l c ch phc hi sau khi b ngp hon ton (Riccharia v Misro 1960, De Datta v Banerjee 1972). Tnh khng ngp l kh nng ca cy la sng st sau khi b ngp hon ton, l mt tnh trng di truyn (Mazaredo v Veragra 1982).
Phn tch di truyn v kh nng phi hp cho thy:
1. S chng chu ngp hon ton tri so vi nhim. 2. C s khng i xng gen v cc alen ln thng th hin nhiu hn khi phn ly. Cc alen tri tp trung trong cc t hp c nhng b m khng ngp rt tt nh FR 13A, Kurkaruppan, FR 43 B, Goda Heenati. 3. Hot ng cng hng v khng cng hng u c ghi nhn. Gi tr di truyn c th t n 0,71 cho thy tnh cht quan trng ca hiu qu cng hng. 4. Phn tch diallel cho thy c t nht mt gen ch lc cng tn ti bn cnh nhiu gen khc. 5. Bn cnh tnh tri cn c hin tng epistasis (Mohanty v ctv. 1982).
V lnh vc chng chu kh hn giai on m: cng tc lai to ging khng hn c thc hin t u nhng nm 1970 v c xem nh mt trong nhng yu t chnh cn c nh gi trong chng trnh GEU (nh gi v s dng ngun gen) ca IRRI (Chang v Ctv 1985).
C ch sinh l hc v tnh khng bao gm: 1. S thot khi kh hn (escape) ch yu nh vo chu k sinh trng ngn. 2. S trnh kh hn (avoidance) nh h thng r dy, su v nhy v s cun li, m ra ca phin l. 3. Tnh chng chu (tolerance) nh s iu chnh sinh l hc ca m, t bo v p sut thm thu c ch ny t c bit. 4. S phc hi (recovery) c tng quan n cng lc sinh trng ca ging la. Cc thut ng v c ch chng chu ny c xc nh v tho lun bi nhiu tc gi (Chang v ctv. 1974, Loresto v ctv. 1976). Nghin cu v di truyn tnh khng hn ca cy la IRRI c tp trung vo c ch 1 v 2, ngun gen khng hn rt a dng ca cc ging la trng thuc Oryza sativa L. v Oryza glaberrima Steud. (Chang v ctv 1985). Tc gi thy rng: 1. C ch trnh hn c tng quan nghch vi c ch phc hi. 2. Tnh chn sm gip thot khi kh hn c iu khin bi nhng gen khng u c d phn (anisomeric genes). 3. Kh nng chi km hoc trung bnh ca la ry l tnh tri. 4. a gen iu khin s r la c to ra, s r t l tnh tri, tnh trng r di ca cc ging la ry l tnh tri, trong khi tnh trng r dy b tnh trng r mng ln p. 5. C s kt hp cht ch: dng cy, s chi, chiu di r v b dy r, trong cng mt loi hnh cy la.
Nhng kt qu trong cng tc chn ging:
Cc trm nghin cu la nc su u bt u thc hin vic ci to ging bng cch chn lc cc ging a phng trn rung nng dn, th d nh Leb Mue Nahng 111 Thi Lan, Chenab Sel 64 117 n ... Cng trnh lai to mi c thc hin sau ny ti Bihar, Uttar Pradesh, West Bengal, Assam (n ), (Padmanabhan 1975, Duara 1975, Maurya 1975), ti Indonesia vo nm 1968 (Noorsyami v Hidayat 1975), v ti Thi Lan nm 1968 ni phng thch sm nht ging la nc su qua chng trnh lai to (Prechachat v Jackson 1975). Kh nng sn xut ra cc dng hnh na ln, c th vn lng trong iu kin nc ngp trung bnh c chng minh. Dng la nc su dng ci thin IR442-2-58 c phng thch, tr thnh ging chun khng kh hn sau ny v dng T 442-57 c dng lm dng hnh tiu chun cho nhng nghin cu so snh kh nng vn lng (Jackson v Vergara 1979). Tuy nhin c tnh khng mn cm vi nh sng ngy ngn v tnh d nhim bnh o n v bc l ca n khin cho nhng ging ny khng pht trin c. Li dng c tnh mn cm vi nh sng ngy ngn, Ikehashi (1977) xut p dng k thut RGA (rapid generation advance) p dng trong cng tc chn ging la nc su vi t nht 3 thun li nh sau: 1. To c s n nh nhanh. 2. Phc hi cc kiu hnh do gen ln iu khin. 3. Thc y c s ti t hp di truyn, cha k thun li c bn ca n l rt ngn c thi gian nghin cu to ging rt ln, v ngi ta c th trng c 2-3 v/ nm cy F2, F3, F4 bng k thut che sng (8-10 gi sng/ ngy) v iu chnh nhit cng nh mt cy.
cp n chin lc chn ging la nc su (Swaminathan 1979), chng ta cn c nhiu cng trnh nghin cu c bn v tnh a dng ca ging i vi cc yu cu sau: 1. 2. 3. 4. 5. To ra s lng lng nhiu hn trn thn la vi s vn di lng km cng c. Hot ng bin dng r la phi tht mnh m. S phn ha to lng thn sm hn. S ht v trng lng ht la cao hn cng nh ch s kinh t phi ln hn. Hot ng quang hp l phi hu hiu hn, ngay c trong trng hp b ngp.
Tc gi ngh chng trnh lai to ging vi nhiu mc khc nhau v mc tiu thch nghi: la nc su trung bnh, la chu ngp, la ni nc rt su... iu rt quan trng l mc tiu chn ging phi ht sc r rng nh dng hnh, kh nng vn lng tnh mn cm vi nh sng... (Jackson v Vergara 1979). Bn loi hnh la nc su c ghi nhn nh sau (HilleRisLambers v Seshu 1982, Vergara 1981). 1. 2. 3. 4. La ni: nc dng t t, thi gian ngp ko di La ni: nc dng nhanh, thi gian ngp ko di La b ngp t ngt, thi gian ngp l ngn. La b ngp do nh hng ca thy triu, thi gian ngp xen k. Mi loi hnh c yu cu v dng cy, kh nng chng chu khc nhau. Chng trnh kho nghim la nc su quc t (IRDWON) bt u t nm 1976, cc ging c dng kiu hnh chp nhn c nhiu im v nhiu nm lin tip l: RD 19, BKN7022-6-4, BKN 6986 108-3, PRRS 43/3, SPR 7292-151-2-1-B-B (Seshu 1982). Cc ging c xut lm vt liu lai cho chng trnh la nc su l RD 19, BKNFR 76045-35-1, DWCT 156-1B-B, SPR 7282-2-0-7, BKNPR 76035-106-1, IR8234-OT-9-2, BKNFR 76016-16-0-1 (Jackson v ctv. 1982). Cc ging la a phong c s dng nhiu trong cc chng trnh lai to ging l : Leb Mue Nahng 111, Pingaew 56, Chamara, Khama, Baisbish v kh nng vt nc.

FR 13A, Goda Heenati, Kurkaruppan, Thavalu, Saran Kraham v kh nng chu ngp. Habiganj Aman 1, LMN 111, Sitpwa v kh nng gi u dy. Ngoi ra nhng ging cung cp tnh trng khng bnh bc l, o n, ry nu, tungro, ... cng c thng bo (Seshu 1982, Hille RisLambers v Seshu 1982, Jackson v ctv 1982, Vergara v Mazaredo 1975).
4-2. HNH THI HC CA CY LA THCH NGHI VI VNG B L LT Vng trng la b nh hng bi l lt c gi bng thut ng quc t l flood prone rice. Chng c th c chia thnh 4 nhm nh sau: - Nhm la chng chu ngp hon ton trong vng 10 ngy, sau chng c th phc hi sau khi nc rt. l nhng vng b l qut, hay vng b ngp thnh lnh. Ting Anh gi l flash flood, v tnh chng chu trong iu kin nh vy c gi l complete submergence - Nhm la c kh nng vt nc 5-10cm / ngy, hoc nhiu hn trong vng l lt ko di 3-4 thng / nm. l vng la ni ng bng Sng Cu Long trc y. Loi hnh l lt nh vy c gi vi thut ng quc t l stagnant flood, nc dng t t, ko di nhiu thng. Tnh chng chu trong iu kin nh vy c gi l kh nng vn lng (elongation ability) - Nhm la c kh nng thch nghi vng m ly ven bin, ni thy triu ln xung trong ngy lm cy la b ngp lc triu cng. l vng b ngp xen k (mixed flood) - Nhm la khng c kh nng vt nc, nhng thch nghi tt trong vng nc ngp su, l lt ko di 2-3 thng, thut ng chung gi l deep water rice (la ncsu), la c tnh cm quang, thi gian tr thng xy ra khi nc rt. 4-3. KH NNG VN LNG Kh nng vn lng l tnh trng quan trng ca ging la ni, lm gia tng chiu cao cy la nh c tnh vn di lng thn, vn di b l, v l la, hoc phi hp tt c nhng tnh trng ny cng mt lc. Hin tng vn lng thng xy ra giai on tng trng, v t thy giai on sinh sn. B l v phin l non c th vn di ra rt nhanh trong iu kin cy la b ngp hon ton trong nc. Gomosta (1985) ghi nhn hin tng vn di ca l la nh ngun nng lng ca sn phm quang hp c tch ly dng carbohydrate. Qu trnh tiu th nng lng v tng trng t ra t nhy cm hn khi b ngp hon ton, trong khi kh nng vn lng rt cn nng lng tch ly. Chnh kh nng vn lng c nhiu tc gi cho rng l tnh trng quan trng nht ca la ni gip n sng st v pht trin (Vergara v Mazaredo 1979). Nasiruddin v ctv. (1977) ghi nhn rng s vn di ca cy l kt qa ko di ca t bo. Nhng Sugawara v Horika (1971) cho rng c ch ch yu trong s vn lng l s gia tng s lng t bo, n xy ra mt cch tch cc ti m phn sinh cui lng thn r. iu ny thng nht vi kin ca Inouye (1983). Raskin v Kende (1984) pht hin nng O2 kh thp trong lng thn b ngp nc (khong 3%). Nng O2 kh thp nh vy lm gia tng hin tng tng hp ethylene (C2H4) trong lng thn ca cy la nc su. Ethylene tch ly trong thn r b ngp hon ton, s lm gia tng s tng trng ca lng thn v c ch s tng trng ca l la. nh hng ng thi ca C2H4 v tng trng lng thn lm gia tng nng CO2 trong lng thn (khong 6%). S thch nghi ca cy la nc su vi iu kin ngp nc nh vy l phn ng gim O2, tng CO2 v C2H4 trong lng thn b ngp nc Trong iu kin cy la b ngp nc, tc phn bo xy ra nhanh hn lm gia tng chiu di vng m phn sinh (Metraux v Kende 1984). Khi t bo gi, lng thn khng th vn lng v thiu hin tng phn bo tch cc. Bleecker v ctv. (1987) pht hin kh nng vn lng do s gia tng mc phn bo vng sinh m, trong c s tc ng h tng gia kch thch t sinh trng ethylene v
gibberellic acid, m nhng mc ny thay i ty theo tnh cht ngp khc nhau. S ngp nc lm gim O2 trong lng thn, nng O2 thp nh vy kch hot s tng hp ethylene. Ethylen s tch t nhiu trong lng thn b ngp. Nng cao ca ethylene lm gia tng mc mn cm ca m i vi gibberellic acic (GA), hoc lm gia tng nng gibberellin hot ng, dn n s p ng sinh trng ca cy trong iu kin b stress do ngp nc. Setter v ctv. (1988) cho rng nng ethylene thay i ty theo nhm ging la nc su, c bit nhm ging la c ngun gc Thi Lan. Mt khc, Khan v ctv. (1987) ghi nhn cy m ca ging la nc su trong iu kin b ngp hon ton s sn sinh ra mt lng ethylene ln hn nhiu ln so vi iu kin b ngp khng han ton. Sauter v Kende (1992) quan st thy GA kch thch t bo u tin ko di ra trong sinh m, tip theo l hot ng phn bo tch cc. Sauter v ctv. (1993) ghi nhn gibberellin khng khi ng hin tng ko di ca lng thn, khi sinh m ca n b tch ra trong mt nghim thc ca th nghim. Cch thc hot ng ca cc vi si cellulose (cellulose microfibrils = CMF) mi chnh l yu t ch yu trong tng trng ca t bo. S ko di lng thn s xy ra khi CMF sp xp theo hng tng trng thun chiu. S ko di gp tr ngi khi CMF sp xp theo chiu nghing, khng thun li. S pht hin ca Sauter v ctv. (1995) v cht iu ha sinh trng trong hot ng phn bo sinh m cho thy rng: c s th hin ca hai gen cyclin tng ng: cycOs1 v cycOs2. Hai gen ny l cDNA c clone ha t cy la v l phn t ng dng (homologs) ca cdc2/CDC28, chng c kch hot bi GA. 4-3-1. Di truyn v kh nng vn lng Nghin cu c tnh cht h thng ti Vin La QucT, bi Thakur v HilleRisLamber (1988) cho thy tnh trng kh nng vn lng c iu khin bi hai gen, hot ng b sung. Phn ly trong F2 ca t hp lai gia Leb Mue Nahng 111 / Baisbish cho thy gen iu khin tnh trng vn lng trong hai ging khc nhau, khng alen vi nhau, trong , mi gen u c th iu khin s vn lng, v s hin din ca c hai to ra kh nng vn lng mnh hn. Trong t hp lai gia ging c kh nng vn lng v ging la thp cy, khng vn lng cho thy: t l phn ly 9:7 c ghi nhn trong qun th F2. Phn b lin tc trong qun th F2 vi mt nh, dng phn b chun (Thakur v HilleRisLambers 1989) c ghi nhn, thng nht vi kt qa ca Suge (1988), cc tc gi kt lun rng tnh trng vn lng thn c kim sot bi hai gen tri khng hon ton, vi nh hng c tnh cht b sung, mt gen kim sot sn phm GA, v mt gen kim sot tnh cht p ng ca ethylene Kh nng vn lng tri so vi tnh trng khng vn lng, vi t l phn ly F2 l 13:1 v 15:1 (Saha Ray 1987, Saha Ray v ctv. 1994). iu ny cho thy tnh cht di truyn digenic vi hin tng epistasis tri trong mt trng hp. nh hng epistasis tin-tri (predominant) th hin mnh hn nh hng epistasis cng (additive) v nh hng epistasis tri (dominance), vi vai tr quan trng ca loi hnh lp on trong epistasis. Thch (1994) nghin cu mi quan h gia di truyn kh nng vn lng v tnh trng chng chu ngp t ngun vt liu FR13A cho thy, gen ca FR13A tng tc alen vi mt trong hai gen c tnh cht b sung kim sot kh nng vn lng, gen iu khin vn lng tri so vi gen iu khin tnh chng chu ngp. Trong khi quan st tnh cht siu tri ca tnh trng vn lng F1 ca nhiu t hp lai gia la ni ca Nam v ng Nam , Mishra (1995) ghi nhn hin tng vt tri v chiu cao cy la, t tc gi xut gi thuyt (1) mt vi QTL c chc nng iu khin kh nng vn lng, (2) ging la nc su ca Nam c kh nng vt nc nhanh hn nhm ging la nc su ng Nam , (3) gen vt nc trong c hai nhm ging la ny c th khng ging nhau, (4) t hp lai gia hai nhm ging u c th to ra con lai vt nc tt hn b m. Cp cho im trong nghin cu kh nng vn lng (Mazaredo v Vergara 1977) 1 lng thn di 60cm hoc ln hn

3 5 7 9 lng thn di 40-50cm lng thn di 20-39cm lng thn di 1-19cm khng vn lng, cy cht 4-3-2. Nghin cu la ni BSCL (Bu 1987) Danh sch ccging la ni ng bng sng Cu Long c nghin cu trong bng 1 (Bu 1987)
Bng 1: Phn nhm la ni v khng ni theo ch s XF ca (Morishima 1975). (Bu 1987)
Ging Nng tri (/c) Nng ty m ht ln Ba bng Nng ty ln Nng cht tri Leb Mue Nahng 111 Nng ty nt Nng ty nt Ty nu Ty nt La rung Nng ty m chn lc Nng tay dm (/C) Np nng cht Nng ty rn Nng ty bng da Ba bng RD 19 (/C) Nng ty nh CP15 Cht ct C l Nng cht ct Np chm Nng m to Trng chm (/C) Nng tr di Np rn Np sng ln Np nng cht ui Nng ty C C l La thng 10 LSD 0.05 = 5.014
S mu 0197 0083 0167 0084 0287 0040 0102 0085 0243 0093 0273 0082 0030 0019 0015 0207 0206 0032 0246 0017 0370 0323 0022 0292 0188 0195 0117 0067 0029 0189 0320 0045
A 14 11 13 12 10 12 11 13 13 12 10 13 10 13 11 13 12 10 9 9 10 8 11 9 10 8 8 9 8 10 7 7
B 214 207 201 198 196 182 184 173 171 173 172 163 168 166 164 155 157 157 160 152 147 158 151 150 147 150 141 135 132 126 136 124
C 10.0 8.0 7.0 9.0 7.0 9.0 7.0 9.0 9.0 9.0 6.0 8.5 8.7 5.3 8.0 11.0 8.0 7.0 6.0 10.9 10.0 5.0 4.3 6.0 3.0 4.0 5.8 6.5 7.4 7.0 3.0 5.0
D 0.5 0.7 0.7 0.3 0.7 2.3 0.8 1.5 0.9 0.7 5.0 2.9 1.2 2.4 1.2 2.2 1.1 0.8 1.0 2.5 606 0.0 2.5 1.7 2.4 2.7 2.0 2.1 0.2 1.2 1.6 6.8
XF 129.700 112.458 111.038 109.632 105.788 102.592 102.582 99.170 98.086 97.978 95.040 95.026 93.598 93.246 93.368 91.878 90.054 87.482 87.440 86.490 85.530 84.760 84.720 82.838 81.166 81.148 77.650 76.008 73.788 73.008 72.804 69.292
S tng quan cht ch gia ch s ni XF v t l cy ngoi ln mt nc su 120cm r = 0,816** y = 0,48x + 7,44
c tnh to lng thn rt sm giai on tng trng u tin ca cy la ni c ghi nhn khng nhng iu kin nc su m cn thy iu kin trng cn, to cho n mt dng hnh b (postrate) nh cc loi c ho tho. Ramiah v Ramaswami (1941) s dng c tnh ny nh mt ch tiu ca kh nng ni.
Trong iu kin thanh lc ging ti h nc su ca IRRI (1985) v mc nc dng 5 cm/ngy s dng ging c tui m 30 ngy, cho thy: (bng 4). - Cc ging la ni Ba so, Nng cht ct, Nng m nh, Nng m to, Nng m trng, Nng phc, Nng rng, Nng son, Nng ty, Nng ty C, Nng Ty m, Nng tri, Np c Ba, u vt nc im 1 (IRRI 1980). - Cc ging la nc su nh Trng chm, Trng la, Tu nt, La Thc vn pht trin bnh thng su 70 cm, nhng su 120cm, tt c u b thit hi.
S khc bit v tui m v tc mc nc dng cao mi ngy cho thy c s khc nhau mt t gia 2 th nghim IRRI v Vin La BSCL. Cc ging Nng m to, Nng ty C th hin mc vt nc trung bnh hoc yu trong iu kin th nghim ti Mn, nhng t ra vt nc tt trong iu kin IRRI. Kt qu quan st s phn ly gia dng ng v dng b giai on m ca cc t hp lai (Bng 5) cho thy t l phn ly F2 l 15: 1 v 3: 1 chng minh tnh cht ln, lp on ca gen iu khin tnh trng ny (Ramiah v Ramaswami 1941, Chang 1965).
Bng 2: So snh mt vi c tnh vt nc ca 4 nhm la nc su BSCL
Nhm ging la
Ch s ni XF
I. Nng tri Ba bng Nng ty ln Nng ty nt Trung bnh II. La rung Nng ty m Nng ty rn Nng ty bng da Trung bnh III. HD 19 Nng ty nh CF15 Cht ct Trung bnh IV. Trng chm Nng ch di Np sng ln Ba thit Trung bnh LSD 0.05
129.7 111.1 109.6 102.6 95.0 93.6 92.4 91.9 87.5 87.4 84.8 81.2 81.2 76.2 5.014
Trc nghim giai on m Chiu di (cm) T l cy S lng ngoi ln di hn 3 Thn la Lng di mt nc cm nht (arcsin) 60.00ab 155ab 42 2 60.00ab 164a 27 2 45.00b 147bc 32 1 63.44a 164a 52 3 56.79 158 43 2 42.13bc 144a 20 1 50.77b 154b 33 2 53.73ab 149bc 43 3 42.13bc 145bc 46 2 47.29 148 33 2 18.44d 140cd 33 1 30.00c 135d 48 1 0.57c 110f 0 0 28.66 133 28 1 0.57e 133e 0 0 0.57e 125e 0 0 0.57e 121ef 0 0 0.57e 112r 0 0 0.57 118 0 0 10.42
Nhng s theo cng mt mu t khng khc bit c ngha mc 0.05.
Bng 3. Trc nghim kh nng vn lng ti IRRI 1985 (m 30 ngy tui, tc nc dng 5 cm/ngy)

S ng k Acc.32084 07819 26242 32185 24209 26247 26248 32193 32196 32212 26237 26239 00231 32168 26241 32191 32192 05967 16777 16781 07258 00196 16783 32273 07091 07217 Ging Ba so Leb Mue Nahng 111 Nng phc Nng rng Nng son Nng ty m Nng ty nt Nng ty to Nng tri Np C Ba Pingaew 56 Nng cht ct Nng m nh Nng m to Nng m trng Nng ln Nng ty C Nng ty nu Nng ty nh CF15 Trng chm Trng la Trng nh La thc Trng tp Trng ln Nng cht ct Nng ch RD 19 (C) IR42 (C) 45 cm 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 3 5 5 9 Kh nng vn lng 60 cm 70 cm 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 3 1 1 1 3 1 3 1 1 1 1 1 3 1 3 1 5 3 5 3 5 5 7 7 9 9 9
120 cm 1 1 1 1 1 1 1 1 1 1 1 3 3 3 3 3 3 3 3 9 9 9 9 9 9 9 9 9 9
Bng 4: S to to lng thn v mc vn lng ca cc ging la ni giai on m - tc nc dng 5 cm/ngy, ti h nc su IRRI
Ging Ba so Leb Mue Nahng 111 (C)
S lng thn di hn 3 cm 5.5 6.0
Tng chiu di ca cc lng (cm) 63.4 68.6
Chiu di trung bnh ca lng (cm) 11.74 11.43

Nng cht ct Nng m nh Nng m to Nng m trng Nng ln Nng phc Nng son Nng ty A Nng ty C Nng ty m (C) Nng ty nu Nng ty nh CF 15 Nng ty to Nng tri Np C Ba Pin gaew 56 IR 11141-6-1-4 (C) LSD 0.05 5.0 5.6 5.0 5.2 5.4 5.4 5.0 5.6 5.4 4.8 4.8 5.0 6.2 5.2 4.4 5.4 7.2 NS 64.9 61.0 78.2 65.2 61.5 64.8 73.7 66.9 63.2 67.3 63.2 63.5 77.0 66.0 99.7 69.5 54.5 NS 12.98 14.46 15.64 12.54 11.39 12.00 15.74 11.94 11.70 14.02 13.60 12.70 12.41 12.69 13.56 12.87 7.54 0.662
Bng 5: T l cy ngoi ln mt nc su 120cm, vi tc nc dng 6cm/ngy
Leb Mue Nahng 111/IR48 Nng ty m/IR48 Ba Bng/IR48 Nng ty bng da/IR48 Nng ty m (C) RD 19 (C) IR48 (C)
T l cy ngoi ln mt nc % Su 90 cm Su 110 cm Su 120 cm 54 44 15 48 33 8 80 76 14 80 79 14 100 100 100 100 100 0 0 0 0
T hp lai Ba bng/IR48, Nng ty bng da/IR48 c t l cy ngoi ln mt nc cao nht su 90 v 110 cm .
su 120cm v 7 ngy sau , RD19, b thit hi hon ton, t l cy ngoi ln mt nc ca 4 t hp lai bin thin t 8 - 15%. T hp lai Nng ty m/IR48 c t l vt nc thp nht.
Qun th F2 ca 4 t hp u c tnh trng vt nc nm trong khong gii hn ca b m, ngha l c mt t c th c kh nng vn lng nh cy m (la ni) cn li phn ln c dng hnh trung gian. Nghin cu v vn ny, cc nh chn to ging gi l hin tng khng c phn ly vt tri (transgressive segregation) (Nasiruddin v ctv 1982, Supapoj v ctv 1977). Khng c c th no th hin kh nng vt nc hn hn Nng Ty m, mc d ngoi theo kp, nhng pht trin km.
Tng quan gia s lng v chiu di lng trung bnh l tng quan nghch, rt cht ch r = -0.699* (n = 17 ging la ni).
S lng ca cc ging la ni v la nc su c xu hng tng theo s gia tng ch s ni XF (r = 0.677**). iu ny ph hp xut ca Morishima (1975): s lng trong iu kin nc cn c th dng c on kh nng vt nc. Tc gi cn ghi nhn: s lng v chiu di lng c th c iu khin bi nhiu gen khc nhau. Trong th nghim thanh lc ging ti h nc su ca IRRI (1985), vi mc nc ng 5cm/ngy, s dng tui m 30 ngy cho thy : Ging la ni c kh nng to lng thn rt sm (ghi nhn 60 ngy sau khi gieo) s lng thn bin thin t 4 n 6/thn la. S khc bit v s lng gia cc ging la ni Vit nam khng c ngha (F=1,169 ns), so snh vi cc ging la ni khc nhau Baisbish, IR11141-6-1-4, cc ging la ni Vit Nam c s lng thn t hn v c xu hng pht trin chiu di lng nhiu hn.
Chiu di lng thn trung bnh ca nhng ging la ni Vit Nam khc bit c ngha (LSD0.05= 0,662) cc ging Nng son, Nng m nh, Nng m to, Nng ty m c chu di trung bnh lng ln nht 4-4. DI TRUYN TNH CHNG CHU NGP HON TON
Tnh chng chu ngp hon ton l mt c tnh quan trng cho la nc cn cng nh la ni. La nc cn cn c kh nng sng st sau khi ngp hon ton 1 ngy hoc 1 tun. La ni cng cn c tnh trng ny trong trng hp s vn lng khng theo kp tc d nc dng qu nhanh vo u v cho ch l bt thng gy ra (Vergera v Mazaredo 1975) Kt hp c c hai c tnh vn lng v chu ngp trong chng trnh lai to ging l yu cu c tnh chin lc (Mohanty v ctv 1982). C rt nhiu phng php thanh lc nh gi cc vt liu khi u dng lm b m cho nhng chng trnh lai to ging chng chu ca 18 tc gi bng 20 phng php ring ca h (HilleRisLambers v Vergara 1982, Vergara v Dikshit 1982). p dng phng php thanh lc ngp ca Vergara v Mazaredo (1975), ngp vi su 30 cm, s dng m 10 ngy tui. Kt qu ghi nhn nh sau: 4-4-1. Dng h thang im ca Suprihato (1980) vi 54 ging dng c th nghim. Nhm la ni vng 5,6 (bng 6) chu ngp km, tr ging Nng m to, Nng ty m. Nhm la canh tc vng m ly ven bin, b nh hng ca thu triu ln xung hng ngy, chu ngp kh nht: La thc, Trng tp, Trng ln, Lem ln, Nng ty , Nng ngut.
Tuy nhin s dng h thang im ny, ch cn c vo s l xanh v tnh trng sinh trng ca chng sau khi ngp cho nn kt qu t r rng, v tu thuc vo iu kin xung quanh (nhit , s kho lo khi em mu ra khi h v.v. .) do chnh xc khng cao. 4-4-2. Th nghim c thc hin li vi 100 ging v dng trin vng ti h nc ca IRRI (1985) hai ln lp li kim sot c s ng u v nhit , ngp su hon ton, c ca nc, ng u v s ny mm, s dng h thang im tnh theo t l cy sng st so vi ging i chng khng FR13A (%CS), cho thy:
%CS =
im 1 3 5 6 9
%S
% sng st ca FR13A
x 100
Tnh chng chu ngp (%CS) 100% hoc ln hn 95 99 75 94 50 74 0 49
(CS: c vit tt t ch comparative survival) V phng php chm im: trong iu kin thanh lc nh li, ging chun khng FR13A c t l cy sng st bin thin t 30-95%, v ging chun nhim IR42 t 0-10%. T l sng st trung bnh ca ging chun khng: 60%, cho php kt lun: s liu th nghim chp nhn c. Nh vy s dng % CS chnh xc hn tnh t l cy sng st n thun, lm gim yu t sai bit do ngoi cnh. Cc ging Vit Nam c t l cy sng st thp t 5-10% l Nng m to, Tu nt, Trng nh, Nng ty m, Nng m nh, Nng son, Nng ty. Tt c cc ging la ni v la nc su cn li u b thit hi hon ton. Cc ging FR13A, Chenab sel. 64-117 (n ) v Murkaruppan (Sri. Lanka) t ra chng chu kh nht, c xut lm ngun lai to tnh khng ngp hon ton (Jackson v ctv 1982, HilleRisLambers v Seshu 1982, Mohanty v ctv 1982, Mazaredo v Vergara 1982). Cc dng lai c trin vng: IR33277-1-507-1-1-3 v IR38683-35-2-9-12
C ngun gc t FR13A v Chenab Sel.64-117 (bng 12) c phn ng chng chu ngp kh nht; tnh khng ngp l mt tnh trng di truyn (Mazaredo v Vergara 1982), v tnh chu ngp hon ton tri so vi s nhim, c t nht mt gen ch lc cng tn ti bn cnh nhiu gen khc (Mohanty v ctv. 1982). 4-4-3. Ngun cung cp, tnh khng ngp hon ton t cc ging la a phng BSCL, c th c rt t donors, nhng trong h s GEU ca Trung tm qu gen ca IRRI, ch c 2 ging la ca Vit nam c tnh khng
ngp hon ton cp 1 l ging Mng chim (Acc. 29588) v Np Vn (Acc. 29873). Hai ging ny c thu thp ti Thun hi v ng Nai- trn vng canh tc la nc tri, thng b ngp t ngt trong ma l, thi gian ngp t 7-10 ngy. Do , cn nghin cu nh gi tnh trng ny t cc ging c ngun gc vng canh tc la nc tri thuc min ng Nam b v nhng vng c iu kin t nhin tng t. Tuy c khc bit v c tnh sinh l gia kh nng vn lng v kh nng chu ngp, nhng c hai c tnh ny cn thit cho la ni (Vergara 1976). Bi v la nc su cn c kh nng sng st trong iu kin b ngp hon ton vo giai on m, lc y cy la cha th vn lng c. Mc cy m sng st sau khi ngp hon ton tu thuc vo: thi gian ngp, su mc nc, nhit v c ca nc, lng phn m c bn v cng nh sng. S ngp lm ngn cn s thnh lp sc t l mc d cy khng thiu nh sng, do Suprihatno (1980) xut phng php chm im theo s l xanh v tnh trng sinh trng ca chng. Cn c nhng kin thc c bn v sinh l hc pht trin k thut thanh lc hp l hn (Mazaredo va Vergara 1982). Khi cy la b ngp, lng O2 trong thn gim do s cung cp khng y , s h hp k kh tng ln v s tiu th cc c cht h hp cng tng ln, do Yamada (1959) cho rng: tnh khng ngp ca cy la c xc nh bng s lng c cht h hp c trong cy trc khi b ngp. Cc ging khng c s gia tng v trng lng cht kh rt cao v phc hi nhanh hn sau khi ngp (Mazarado v Vergara 1982). Khc vi kt lun ca Yamada (1959) v hm lng carbohydrate ca ging khng rt cao trc khi ngp: Mazaredo v Vergara (1982) thy rng s sng st v kh nng phc hi nhanh chng tu thuc vo s lng carbohydrate sau khi b ngp. Bng 6: Kt qu th nghim chng chu ngp hon ton (s dng h thang im Suprihatno 1980) ca cc ging la ma ch lc cc vng canh tc khc nhau v nhng dng lai trin vng (Bu 1987).
Ging La thc Trng tp Nng m to Lem ln Trng ln Nng ngut Nng ty m La nc mn Nng keo c rn C ung l Ba thit Ch bin Trng chm Kiu la Tt n Trng phc Cht ct Np c ba
(1= khng 9= nhim)
S ng k 0294 0283 0292 0228 0204 0106 0030 0092 0060 0114 0079 0142 0166 0188 0227 0247 0254 0017 0147
im chng chu 1 2 3 3 3 4 4 4 4 4 4 4 4 4 4 4 4 5 5
Ging Sa mo chm Thn nng Chm rut la Tri my Trng nh La Phi Nng tri Tu binh Ba bng Ty nu Np sng ln Nng ty ln Trung hng Nng ty bng da 342 (c phng/ IR5853) 522 (Tri my ct/424) 375 (Tu binh/NN5A) 523 (IR8608/Trung hng)
S ng k 0182 0050 0073 0086 0252 0255 0197 0176 0167 0243 0067 0084 0334 0207
im chng chu 5 5 5 5 5 5 5 7 7 7 9 9 9 9 9 9 9 9

XF 110 90
70
r = 0.816 ** = 0.48 8+ 77.44
50 30 10 20 30 40 50 60 70
Hnh 4-1a: Tng quan gia ch s ni XFv t l cy vt nc giai on m
Hnh 4-1b: Phn ng ca ging chun khng FR 13A v ging chun nhim IR42 trong th nghim tnh chng chu ngp (Bu 1987) Bng 7: Kt qu th nghim chng chu ngp hon ton theo t l cy sng st so vi i chng khng ti h nc su IRRI (Bu 1987)
Ging
S mu ging
S (%)
CS (%)
im 1-9
Ba so Chenab sel. 64-117 Kurkaruppa La thc Nng cht ct Nng ch Nng m to Nng ln Nng phc Nng ty Nng ty m Nng ty nt Nng tri Tu nt Trng chm Trng la Trng ln Trng nh Trng tp IR33277-1-507-1-1-3 (FR13A/IR9129-209-2-1//IR9129-117-2-3) IR38683-137-5- (Chenab sel. 64-117//IR4744-295-2-3) i chng FR13A (khng) IR42 (nhim)

Acc.32084 0.00 0.00 Acc.846483 48.48 80.80 Acc.15449 57.74 96.23 Acc.00196 0.00 0.00 Acc.26237 0.00 0.00 Acc.07217 2.94 4.90 Acc.00231 9.95 16.59 Acc.26241 0.00 0.00 Acc.26242 3.45 5.75 Acc.24209 3.85 6.42 Acc.26247 6.25 10.43 Acc.26248 3.84 6.41 Acc.32196 0.00 0.00 Acc.10271 0.00 0.00 Acc.16777 0.00 0.00 Acc.16781 2.94 4.90 Acc.32273 4.34 7.25 Acc.07258 6.06 10.10 Acc.16783 2.70 4.50 67.87 112.96 59.50 60.00 0.00 99.17 100.0 0.00
9 5 3 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 1 1 1 9
(1 = khng; 9= nhim) Kh nng chng chu ngp hon ton (submergence tolerance) l mt tnh trng di truyn gip cy la phc hi sau khi b ngp hon ton trong nc (10-14 ngy). Cy c th sng m khng cn c kh nng vn lng. Rt t ging la c truyn c ghi nhn c kh nng ny. Nhng vt liu c cng b l FR13A, FR43B c ngun gc t n , ging Kurkaruppan, Thavalu, Goda Heenati c ngun gc t Sri Lanka (IRRI 1978) Ging la chng chu ngp c kh nng tng hp v tch t nhanh hn v nhiu hn carbohydrate so vi ging khng chng chu ngp, trc khi l bt ng p ti (Emes v ctv. 1988, Setter v ctv. 1997)
Di truyn v tnh chng chu ngp thc s vn cha c hiu r. Mt nghin cu kt lun rng tnh trng chng chu ngp l tnh tri so vi tnh khng chu ngp, vi t nht 3 gen tri, trong 2 gen lp on, v gen th ba hot ng b sung vi 2 gen cn li (Suprihatno v Coffman 1981). Mt vi nghin cu sau ghi nhn tnh trng chng chu ngp c h s di truyn cao v nh hng gen cng tnh vi t nht c mt gen ch lc th hin tnh tri hon ton hoc tri tng phn (Mohanty v ctv. 1982, Mohanty v Khush 1985, Haque v ctv. 1989). C tc gi kt lun y l tnh trng c iu khin bi c gen ch lc v cc gen th yu (Mohanty v Khush 1985), hay c iu khin bi mt gen tri trong FR13A, Kurkaruppan v Thavalu (Thch 1994), hay c iu khin bi ch mt gen tri (Mishra 1995).
Saha Ray v ctv (1993) cho rng tnh chng chu ngp v kh nng vn lng c th c kt hp trong cng mt ging khi chng ta s dng b m l nhng ging c kh nng chng chu ngp rt mnh. Thch (1994) cho rng gen iu khin tnh chng chu ngp ca ging FR13A tng tc alen vi mt trong hai gen hot ng b sung iu khin kh nng vn lng ca ging la ni Jalmagna, do , vic kt hp tnh chng chu ngp ca FR13A v kh nng vn lng ca Jalmagna l khng c tnh kh thi.
Nhng gen mc tiu iu khin tnh chng chu ngp c phn lp bi cng trnh nghin cu ca Xu v Mackill (1995). Cc tc gi ny tm thy gen ch lc iu khin tnh trng chng chu ngp t ging la IR40931-26-3-3-5, nh v trn nhim sc th s 9, lin kt vi RFLP marker (C1232), thng qua , c 69% bin thin kiu hnh v tnh trng ny c gii thch.
Mt nghin cu khcca Vanavichit (1996) cho thy cc gen chng chu ngp ca ging la IR49830-7-1-2-2, lin kt vi RFLP marker (RZ698) v hai RAPD marker (AO2c v HO3b), nh v trn nhim sc th s 9, vi gi tr lin kt khong 4 cM. Phn tch QTL cho thy gen iu khin tnh trng ny lin kt vi RZ70, nh v trn nhim sc th s 5. V tr ca gen ch lc c nghin cu mt cch h thng ti IRRI vi k thut AFLP trn qun th cn giao ti t hp F8 ca t hp lai IR74 / FR13A, nhng gen th yu c cng b nh v trn nhim sc th s 6, 7, 11 v 12 (Nandi v ctv. 1997) 4-5. C CH CHNG CHU NGP V SINH L HC 4-5-1. Dip lc t Ging nhy cm vi stress do ngp ng thng c hm lng dip lc suy gim nhanh theo thi gian cy b ngp. Hin tng quang hp di nc vn c th hot ng, nhng tc quang hp gim do s gim oxygen v thiu nh sng. Hot ng quang hp lin quan rt cht vi hot ng ca RuBP carboxylase trong lc lp, v s ngp hon ton nh vy lm gim i hot ng ca RuBP carboxylase (Rai v Murty 1979). Do , cc yu t nh sng gim, oxygen gim, dip tc t gim, chng phi hp vi nhau lm cho hot ng quang hp gim trong sut thi gian b ngp nc. Nhng ging chng chu c kh nng sng st nh hm lng dip lc gim rt chm, v kh nng duy tr hot ng quang hp trong iu kin nh vy 4-5-2. Carbohydrate Trong sut qa trnh b ngp hon ton, s cung cp oxygen b hn ch. S h hp k kh gia tng (Yamada 1959, Rai v Murty 1976, Setter v ctv. 1987). iu ny to ra mt s tiu th nhanh chng lng carbohydrate v c ch thy phn protein. Hm lng cht bt gim nhanh, ethylene gia tng trong chi thn b ngp, gia tng chiu di l (Setter v ctv. 1988). Trng hp cy la ch b ngp su 30-50cm, khng c s thay i ng k v carbohydrate, tinh bt, ng trong cy la (Setter v cvt. 1987) 4-5-3. Hm lng nitrogen Hm lng nitrogen tng s gim trong cy m b x l ngp hon ton (Mazaredo v Vergara 1982), nhng cy la thuc ging khng chng chu ngp, mc gim ny nhiu hn so vi cy chng chu. 4-5-4. Hot ng ca peroxidase Peroxidase trong qa trnh tng trng v phn ha chc nng c nhiu bo co ghi nhn trong iu kin b ngp nc hon ton (Kadam v ctv. 1973, Murty v Nada 1974), nhng vai tr chnh xc ca n nh th no vn cha c hiu r. C th mi quan h tng tc gia cht kch thch sinh trng IAA v peroxidase gi mt vai tr quan trng ca cy, c bit trong r nhm mc ch gip cy chu ng trong thi gian di hn i vi stress do ngp (Krishnayya v ctv. 1990). Hot ng ca peroxidase b l la gia tng t trc khi b ngp cho n sau khi b ngp trong tt c ccging th nghim. S gia tng ny cng mnh trong ging chng chu. Nhng hot ng peroxidase trong l v r ca ging khng chng chu t ra suy gim trong qa trnh b ngp, trong khi , peroxidase gia tng mt t trong ging chng chu 4-6. NGHIN CU BN DI TRUYN TNH TRNG VN LNG
Ti Vin La QucT, ngi ta s dng k thut AFLP phn tch bn di truyn trn c s qun th cn giao ti t hp (RIL), qun th F8 ca t hp lai IR74 x Jalmagna (Sripongpankul 1998). Tc gi s dng 247 dng lai nh gi kiu hnh, vi hai nghim thc mc nc gia tng chm v mc nc gia tng nhanh. Sau , tc gi chn li 41 dng c kh nng vn lng v 41 dng khng c kh nng vn lng, tng cng 82 dng RIL. K thut AFLP c p dng vi 201 marker phn b trn 12 nhim sc th cy la, vi s phi hp ca 20 cp mi ca Pst1 v Mse1. K thut RFLP cng c p dng vi 7 marker v gen adh c dng kim nh bn . Bin thin kiu hnh ca tnh trng vn lng thn, vn cao cy, v vn di l c trc nghim theo phng php PROC GLM trong phn mm SAS, trn c s ANOVA mt chiu, c hiu chnh trn tng marker ring bit, mi phi hp gia hai marker c ghi nhn c tnh cht gi nh (putatively) v tnh trng kh nng vn lng. Gi tr lin kt c p dng trong MAPMARKER (Lander v ctv. 1987). Khong cch di truyn c tnh trn c s hm Kosambi (Kosambi 1944). Nhng gen ch lc v gen th yu c phn tch theo phng php MAPMARKER/QTL (Lander v Botstein 1989). Theo nguyn tc, tt c marker u c th nghim vi t l phn ly 1:1 trong MAPMARKER/QTL Sripongpankul (1998) thit lp bn AFLP trn qun th RIL ca t hp lai IR74 / Jalmagna vi LOD score l 4,0, bn cnh , bn RFLP cng c thit lp tng thm mc tin cy ca kt qa, vi s tham gia ca 11 enzyme phn ct hn ch. nh gi kiu hnh cho kt qa phn lp 41 dng c kh nng vt nc ging nh Jalmagna v 41 dng khng c kh nng vt nc ging nh IR74. Kh nng vt nc k hiu ELON, kh nng vn lng thn: INTN, v kh nng vn l: LE. Bng 8: Phn b marker trn tng nhim sc th (Sripongpankul 1998) Nhim sc th S Chiu di (cM) 354.73 1 204.87 2 145.64 3 154.68 4 207.93 5 81.91 6 85.82 7 96.40 8 198.79 9 75.73 10 350.61 11 246.44 12 Tng 2203.55 cng Marker 36 16 12 15 16 10 7 14 13 7 43 23 212 Khong cch trung bnh (cM) 9.85 12.80 12.14 10.31 13.00 8.19 12.26 6.89 15.29 10.82 8.15 10.71 10.87
Hnh 4-2a, b cho thy: nhng gen chnh iu khin tnh trng vn cao (ELON) v tnh trng vn lng thn (INTN) c nh v trn bn AFLP, nm gia hai marker k cn P2/M6-7 v RG109 trn nhim sc th s 1, vi gi tr LOD l 6,9 v 10,6 theo th t. Khong cch di truyn gia P2/M6-7 vi hai tnh trng l 14,6 cM, gia RG109 vi hai tnh trng l 9,6cM. Tnh trng vn di ca l ( LE ) c nh v trn bn AFLP, on u ca nhim sc th s 4, vi gi tr LOD l 3,9, lin kt vi marker P3/M7-10 trong khong cch 19,4cM.
Chn QTL phi hp vi tnh trng vn cao thn nh v nhim sc th s 1, 4, 5, 6, 10 v 12 Su QTL phi hp vi tnh trng vn lng thn nh v trn nhim sc th s 1, 4, 5, 6, v 10 Ba QTL phi hp vi tnh trng vn di ca l nh v trn nhim sc th s 1, 4, v 12 Phn tch ANOVA cho thy: mt gen ch lc lin quan n tnh trng ELON c LOD = 34,9, nh v trn nhim sc th s 1, vi bin thin kiu hnh c gii thch l 83,9%. Bn cnh , 2 QTL th yu nh v trn nhim sc th s 1, s 4, mt QTL trn nhim scth s 5, 6, 10 v 12
Von cao cy (%): ELON Vn lng thn (cm): INTN Vn di l (%): LE
Hnh 4-2a: QTL iu khin tnh trng vn lng trn bn AFLP ca qun th RIL thuc t hp lai IR74 / Jalmagna (Sripongpankul 1998)
i vi tnh trng INTN, mt gen ch lc vi LOD = 31,9, nh v trn nhim sc th s 1, vi 86,7% bin thin kiu hnh c gii thch. Bn cnh , mt QTL th yu nh v trn nhim scth s 4, 5, 6 v 10 i vi tnh trng LE, mt gen ch lc vi LOD = 11,4, nh v trn nhim sc th s 4, vi 47,3% bin thin kiu hnh c gii thch. Bn cnh , mt QTL th yu nh v trn nhim scth s 1 v 12 Hai QTL ph bin kt hp c 3 tnh trng mc tiu c tm thy trn nhim sc th s 1 v 12. Cc QTL th yu c tm thy trn nhim sc th s 9, nh v gn gen iu khin tnh chng chu ngp hon ton (Sripongpankul 1998)
Hnh 4-2b: chia se tnh trng vn lng trn bn AFLP ca qun Tai lieuQTL iu khintai: wWw.SinhHoc.edu.vn th RIL thuc t hp lai IR74 / Jalmagna (tip theo)
4-7. CLONE HA GEN OsGAPDH IU KHIN TNH CHNG CHU NGP Ngi ta phn lp rt nhiu clone iu khin tnh chng chu ngp cy la. Mt trong nhng clone nh vy c tn l OsGAPDH (Arumugam Pillai v ctv. 2002). N th hin mc cao trong sut 12 gi b ngp. Mt tng ng trong GenBank Database cho thy OsGAPDH c chui k t di truyn tng ng vi glyceraldehyde-3-phosphate dehydrogenase khng c kh nng o chiu ca genome cy bp. Chui k t ca OsGAPDH bao gm 1772 bp vi khung c m ORF di nht, m ha 499 amino acid v ln phn t ca n l 54,2 kDa. Phn tch Southern blot cho thy c mt hoc hai bn sao ca OsGAPDH biu hin trong genome ging la Yukihikari (Nht). Ngi ta phn tch RFLP v xc nh v tr ca gen OsGAPDH nh v trn nhim sc th s 8. S th hin ca gen ny cc m t bo c bit cho thy rng mc cao ca phn t mRNA thng c tm thy trn bng la (Arumugam Pillai v ctv. 2002). Nhng cy la biu hin tnh chng chu kh hn, chu ngp v phn ng vi nghim thc x l ABA u cho thy c s gia tng mc tch t th transcript ca gen OsGAPDH. Hin tng kch thch t bo ca Escherichia coli cha plasmid ca pGST-OsGAPDH s to ra mt s tch t mt s lng ln protein c tnh cht ti t hp (recombinant protein), vi kch thc phn t 83,2 kDa (Arumugam Pillai v ctv. 2002). Enzyme GAPDH thun khit c hot tnh iu kin pH 8,5, nhit 50oC, enzyme ny b c ch bi ATP v ADP. Tnh chng chu ngp trong cy la lin quan n kh nng duy tr nng lng cung cp cho qa trnh ln men ru vi mc nhanh, v yu cu rt cao v carbohydrate. Nhng nghin cu v th hin ca cc gen c sn phm protein trong thc vt bc cao trong iu khin him kh v ho kh c thc hin trong rt nhiu nm v c nhiu bo co ghi nhn tm quan trng ca c ch nng lng trong hin tng chng chu ngp (iu kin him kh), nhiu protein sn sinh trong iu kin him kh c tn :anaerobic protein vit tt l ANP bao gm nhng enzyme lm nhim v thy phn v nhim v ln men (Vartapetian v Jackson 1997). Hin tng thy phn trong iu kin him kh v hin tng ln men v cng quan trng trong vic cung cp nng lng cho cy trng. S th hin ca nhng gen iu khin tin trnh ny (th d glyceraldehyde-3-phosphate dehydrogenase, enolase, alcohol dehydrogenase v pyruvate decarboxylase) u c kch hot bi iu kin him kh (anaerobiosis) (Umeda v Uchimiya 1994, Sachs v ctv. 1996). S kch hot nh vy rt cn thit cho hin tng chng chu stress ca cy khi phn ng vi iu kin ym kh. T bo s cht thng qua cc s kin sau y: (1) b ng c ethanol do qa trnh ln men ru, (2) t bo cht b acid ha (hin tng acidosis), (3) khng pht ra nng lng (ATP) duy tr hot ng ca t bo, (4) cht do bin dng ri lon bi oxy ti hp thu sau mt thi gian khng c oxy. Tin trnh lm chm s ln men ru hoc lm sai lch s hnh thnh ethanol s gp phn lm gim mc thit hi di dng hp cht t gy hi nh malate. y l kin ca ca Vartapetian v Jackson (1997) da trn c s sinh ha gip cho cy thch nghi v sng st. Protein ANP c nghin cu rt su trong trng hp r cy bp, vi 20 protein chim khong 70% tng s protein c sn sinh trong vng 5 gi trong iu kin him kh, v ng vai tr hot ng xc tc chnh trong phn ng thy phn hoc hot ng bin dng phosphate ng (Sachs v ctv. 1996). Sn lng ANP tip tc tng cho n khi r bp cht sau 72 gi khng c oxy (Sachs v ctv. 1980). y l kt qa ca hin tng chuyn m c kch hot v hin tng gii m c chn lc trn c s hot ng phosphoryl ha sinh tng hp protein nh l nhng yu t khi ng (Webster v ctv. 1991). Nhng cng trnh u tin v c ch kch hat do iu kin ym kh, pht sinh ra nhng th transcript v s tch t mRNA, th d nh adh1 trong genome cy bp, do Rowland v Strommer (1986) cng b. Ging la japonica ph bin Yukihikari l ging la c nng sut cao c s dng trong nghin cu. Arumugam Pillai v ctv (2002) cho ny mm 50 ht trong l tam gic 300-ml, nhit 28oC. M t bo c thu thp vo giai on cy la 6 tun tui. Cy la c trng trong iu kin 16 gi sng, 6 gi ti / ngy, trong t nh n (growth
chamber) nhit 28oC. Cy la 6 tun tui c trng ring trong iu kin ti hon ton 28oC trong 2 ngy. Phn t RNA c ly trch t ton thn cy la ri phn tch Northern blot vi nghim thc i chng, nghim thc kh hn, nghim thc mn, nghim thc b ngp hon ton. i vi nghim thc x l kch thch t, cy m c phun mannitol (0,5 M), elephon (10-2 M), v ABA (10-5 M). Mu DNA c ly trch v c phn ct hn ch bi BamHI, BglII, EcoRI, HindIII. in di c thc hin trn agarose gel 0,8%. Bn RFLP c thc hin trn qun th n bi kp (DH) bao gm 127 dng DH ca cp lai ZYQ8 (indica) x JX17 (japonica). V tr trn nhim sc th ca gen OsGAPDH c tm thy trn bn bn lin kt gen do He v ctv. (1999) thc hin. Genome ca qun th DH c phn ct hn ch bi ScaI v in di trn agarose gel 0,8% v c chuyn n mng nylon. Thc hin lai DNA vi probe l OsGAPDH cDNA. Phn tch bn di truyn theo MAPMARKER/EXP version 3.0 (Lander v ctv. 1987). Phn t RNA tng s c ly trch t cy m 6 tun tui bng phng php SDSphenol ca Bachem v ctv. (1996). tt c mu c bo qun trong nitrogen lng (-80oC). Phn t RNA c chit tch rin bit trong 1,2% (w/v) agarose gel (3 Vcm-1) cha 2,2 M formaldehyde v c chuyn n Highbond-N+ nylon membrane (Amersham) s dng 20x SSC (Sambrook v ctv. 1989). 4-7-1. Phn tch Southern Hnh 4-3 biu th nucleotide v chui m amino acid ca cDNA clone gen mc tiu OsGAPDH, v hnh 4-4 biu th cch sp xp ca gen OsGAPDH trong genome cy la so vi nhng chui m ca genome cy khc (bp, u, thuc l,..) Hnh 4-5 cho thy kt qa phn tch Southern vi 4 enzyme phn ct hn ch. Bng c ln phn t 16,0 kb v 5,6 kb c pht hin bi BamHI v EcoRI. Cc bng c ln phn t 2,8 v 3,8 kb c pht hin bi enzyme BglII v 2,0 v 5,0 kb do enzyme HindIII (hnh 4-5). Nhng tn hiu ca Southern blot cho thy c mt hoc hai bn sao ca OsGAPDH xut hin trong genome cy la. Bn lin kt gen ca He v ctv. (1999) cho thy trong tng s 243 RFLP v microsatlelite marker phn b trn 12 nhim sc th, gen OsGAPDH c pht hin trong qun th n bi kp F2, nh v trn nhim sc th s 8, gia hai marker RG1 v CT56 (hnh 4-6) 4-7-2. Phn tch Northern Arumugam Pillai v ctv. (2002) c gng phn bit tng phn ca mRNA gen OsGAPDH tch t nhiu m khc nhau trong genome cy la (bng, phin l, b l, lng thn, mt lng, v r). Cc tc gi ghi nhn mRNA biu hin nhiu nht trong bng la, hin tng chuyn m xy ra mnh m nht trong cc m t bo bng la (hnh 4-7). nh hng vt c tm thy trong b l, lng thn v mt lng. Th transcript nh vy rt kh tm thy l gi v r.
Hnh 4-3: Chui trnh t nucleotide v amino acid ca phn t cDNA thuc gen OsGAPDH (Arumugam Pillai v ctv. 2002)
Hnh 4-4: Cch sp xp ca gen OsGAPDH trong genome cy la so vi nhng chui m ca genome cy khc (Zea mays, Pisum sativum, Nicotiana plumbaginifolia, Apium graveolens) (Arumugam Pillai v ctv. 2002
S th hin gen OsGAPDH
Phn t RNA tng s ca OsGAPDH c xc nh trong cy m vi nhng nghim thc x l khc nhau (hnh 4-8). Cy la b x l trong iu kin kh hn, ngp hon ton v ABA, s gia tng hm lng mRNA tch t. nghim thc x l nhit lnh 5oC, nghim thc mannitol, ethephon, mc tch t mRNA tng i thp. i vi x l kh hn, mc th hin cao nht th transcript tch t xy ra lc 12 gi sau x l. Nhng mc th transcript ny s gim i qua ngy hm sau, ri tng ln ti a vo giai on 3 ngy sau khi x l kh hn. Hin tng p ng rt mnh m do x l ngp hon ton c quan st vo lc 12 gi sau khi x l, sau s th hin ny gim rt nhanh. Kch thch t sinh trng ABA cng biu bin mt s kch hot kh mnh trong vng 1 ngy khi cho x l ABA, sau th transcript ny gim chm trong 2-3 ngy sau (hnh 4-9).
Hnh 4-5: Phn tch Southern blot OsGAPDH
Hnh 4-6: Bn di truyn genome cy la, nhim sc th s 8 ni nh v gen OsGAPDH (Arumugam Pillai v ctv. 2002)
4-7-3. S th hin dung hp GST-OsGAPDH Mun xc nh protein OsGAPDH c hot tnh ca glyceraldehyde-3-phosphate dehydrogenase, nhm tc gi ni trn tm cch th hin protein y OsGAPDH 54, kDa (499 aa) trong t bo E. coli l protein ti t hp GST-OsGAPDH. S tch t ca GST (29
kDa) c quan st khi E. coli c cha pGEX-6P-3. S kch thch ca t bo E. coli cha plasmid pGST-OsGAPDH bi IPTG s to ra hin tng tch t s lng ln protein c tnh cht ti t hp (83,2 kDa) (hnh 4-10). Tnh cht hot ng hay c ch ca mt s h gen khc nhau c th to ra s th hin khc nhau ca mt enzyme nh glyceraldehyde-3-phosphate dehydrogenase v pyruvate decarboxylase ca cy bp v cy Arabidopsis thaliana (Russell v Sachs 1989, Peschke v Sachs 1993, Yang v ctv. 1993). Di truyn tnh chng chu trong nhng trng hp ny l di truyn s lng, do n c th tn ti dng mt vi gen ch lc v nhiu gen th yu (polygenes). Mi n lc nhm xc nh vng mc tiu ca GAPDH v cng cn thit i vi tnh trng chng chu ngp hon ton v kh hn (Arumugam Pillai v ctv. 2002).
Hnh 4-7: Phn tch Northern blot OsGAPDH mRNA ca phn t RNA (30 l / lane) ly trch t bng la, la la, b l la, mt lng, lng thn, v r la (Arumugam Pillai v ctv. 2002)
Hnh 4-8: nh hng ca nhng nghim thc stress khc nhau v kch thch t sinh trng biu th bng mc th hin transcript (Arumugam Pillai v ctv. 2002)
Hnh 4-9: Phn tch Northern s th hin ca gen OsGAPDH thng qua nghim thc kh hn, ngp hon ton v x l ABA (Arumugam Pillai v ctv. 2002)
Hnh 4-10: Th hin protein dung hp GST-OsGAPDH trong E. coli. Lane 1: protein tng s DH5 c cha pGEX-6P-3, khng chn vo -IPTG. Lane 2: protein tng s DH5 c cha pGEX-6P-3, khng chn vo +IPTG. Lane 3: protein tng s DH5 c cha pGSTOsGAPDH tr i IPTG. Lane 4: protein tng s DH5 c cha pGST-OsGAPDH cng IPTG. Lane 5: protein dung hp pGST-OsGAPDH tinh khit bi glutathione-Sepharose 4B trong sc k kh. Lane 6: protein tinh khit pGST-OsGAPDH c phn ct bi PreScission Protease. Lane 7: protein tinh khit OsGAPDH bi tin trnh glutathione-Sepharose ct 4B. Lane 8: nhng marker phn t: phosphorylase b 94 kDa, BSA 67 kDa, ovalbumin 43 kDa, carbonic anhydrase 30 kDa, trypsin inhibitor 20.1 kDa v lysozyme 14.4 kDa. Tt c mu protein in di trn 12% SDS-polyacrylamide gel, nhum mu Coomassive Brilliant Blue R250 (Arumugam Pillai v ctv. 2002)

TI LIU THAM KHO
Alim A, and H Zaman. 1958. Effect of mechanical up rooting on the yield or deepwater aman paddy. Agric. Pakistan 9: 337-339 Alim A, and JL Sen. 1955. Floating paddy in East Pakistan. IRC newsletter. 14 June: 13-14. Arumugam Pillai M, Z Lihuang, T Akiyama. 2002. Molecular cloning, characterization, expression and chrosomal location of OsGAPDH, a submergence responsive gene in rice (Oryza sativa L.). Theor Appl Genet 105:34-42 Bachem CW, RS van der Hoeven, SM De Bruijin, D Vreugdenhil, M Zabeau, RG Visser. 1996. Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. Plant J 9:745-753 Bleecker AB, J Schuette, H Kende. 1987. An evaluation of 2,5-norbornadiene as a reversible inhibitor of ethylene action in deep water rice. Plant Physiol 84: 395-398 Bi ch Bu. 1987. Nghin cu mt vi c tnh quan trng ca cc ging la nc su a phng ng bng sng Cu Long. Lun n PTS Khoa Hc Nng Nghip. Chuyn ngnh Ging 4 01 05. Vin KHKTNN Vit Nam. 112 trang Chandraratna MP. 1964. Genetic and breeding of rice Longmans London. 389 p. Chang TT, JT Armenta-Soto, CX Mao, R Peiris, and C Loresto. 1985. Genetic studies on the components of drought resistance in rice (Oryza sativa L.) Paper presented at the ins. Rice Genetics symposium. 27-31 May 1985. IRRI. Los Banos, Philippines. Chang TT. 1979. Deepwater rice germplasm. Paper 35-37 in Proc. of the 1978 deepwater rice workshop. 17-19 Aug. 1978, Calcutta, India. Chang TT, AP Marciano, and GC Loresto. 1977. Morpho-agronomic various ness and economic potential of Oryza glaberrima and wild species in the genus Oryza. Paper 67-76 in IRATORSTOM meeting of African rice species, Paris, France. Chang TT, GC Loresto, and O Tagumpay. 1974. Sereening rice germplasm for drought resistance. SABRAO 6: 9-16. Chang TT, EA Badernas. 1965. The morphology and varietal characters of the rice plant. Tech. Bull. 4, IRRI, Lobanos, Laguna, Philippines. Choudhury MA. 1975. Genetics and breeding of deepwater rice. Pages 83-86 in . Proceeding of the Int. Seminar on deepwater rice 1974. Bang ladesh. De Datta SK, and B Banerji. 1972. Performance of flood resistance and deepwater rice in relation to growth and yield under different cultural practices. Indian J. Agric. Sci. 42 (8): 664-670. Duara BN. 1975. The deepwater rice of Aman State. Pages 55-60 in Proceeding of the international seminar on deepwater rice. Aug. 21-26, 1974. Dacca, Bangladesh Emes MJ, CP Wilkins, PA Smith, K Kurkanchanakul, K Hawker, WA Charlton, EG Cutter. 1988. Starch utilization by deep water rice during submergence. In; Proceedings of the 1987 International DeepWater Rice Workshop, 20-30 Oct, Bangkok, Thailand. IRRI, Philippines. Pp. 319-326 Garrity DP. 1984. Rice Environments classification: A comparative review. Pages 11-26 in Terminology for rice growing environment. IRRI, Los Banos, Philippines. Gomosta AR, and BS Vergara. 1985. Factor effecting elongation of deepwater rice under submergence. Paper presented at the IRRIs Saturday seminar. Aug. 1985. IRRI 6 pages (mimeo) Gomosta AR. 1985. Factors affecting elongation of deep water rice under submergence. PhD Thesis, University of The Philippines at Los Banos, Laguna, Philippines, 290 pp. Hamanura K, and K Kupkanchanakul. 1979. Inheritance or floating ability in rice. Jpn. J. Breed. 29 (3): 211-216. Hamamura K, and P Saengpetch. 1977. Technique to distinguish floating rice from non-floating types in seedling stage. Pages 93-100 in proceeding of the 1976 int deepwater rice workshop 8-10. Nov. 1976, Bangkok, Thailand. Haque QA, D HilleRisLambers, NM Tepora, QD de la Cruz. 1989. Inheritance of submergence tolerance in rice. Euphytica 41: 247-251 He P, SG Li, Q Qian, YQ Ma, JZ Li, WM Wang, Y Chen, LH Zhu. 1999. Genetic analysis of rice grain quality. Theor Appl Genet 98:502-508
HilleRisLambers D, and DV Seshu. 1982. Some ideas on breeding procedures and requirement for deepwater rice improvement. Pages 29-44 in proceeding of the 1981 int deepwater rice workshop. IRRI, Los Banos, Philippines. HilleRisLambers D, and BS Vergara. 1982. Summary results of an international collaboration on screening method for flood tolerance. Pages 347-354 in proceeding of the 1981 int. deepwater rice workshop, IRRI, Los Banos, Philippines. HilleRisLambers D, C Setabutara, N Supapoj, K Kupkanchanakul and E Shuwisitkul. 1977. segregation for elongating ability in two crosses of floating rice with ordinary lowland rice. Pages 37-44 in proceeding of the 1976 int. deepwater rice workshop 8-10 Nov. 1976, Bangkok, Thailand. Ikehashi H. 1977. New procedure for breeding photoperiod - sensitive deepwater rice with rapid generation advance. Pages 45-54 in proceeding of the 1986 int. deepwater rice workshop. 8-10 Nov. 1976 Bangkok, Thailand. Inouye J, and VT Xuan. 1973. On the growth habits of floating single and double - Transplanting rice plants in South Vietnam. I. Mesocotyl elongation in darkness. Ipn. J. Trop. Agric. 17 (2): 75 - 80. IRRI. 1980. Standard evaluation system. The International Rice Research Institute, Los Banos Philippines. Jackson BR and BS Vergara. 1979. progress in deepwater rice research. Pages 3 - 12 in proceeding of the 1978 int. deepwater rice workshop, Aug. 17-19, 1978. Calentta, India. Jackson BR, D HilleRisLambers, and C Prechachat. 1982. Development of deepwater rice varieties and breeding materials for south and Southeast Asian countries. Pages 19-28 in proceeding of the 1981 int. deepwater rice workshop. IRRI Philippines. Kadam SS, J Singh, SL Mehta. 1973. Changes in isoenzymes in embryo and endosperm of normal and opaque-2 Zea mays during inhibition. Phytochemistry 12: 1221-1225 Khan AA, R Thakur, M Akbar, D HilleRisLambers, DV Seshu. 1987. Relationship of ethylene production to elongation in deepwater rice. Crop Sci 27: 1188-1196 Khush GS. 1984. Terminology for rice growing environments. Pages 5 -10 in Terminology for rice growing environments. IRRI, Philippines. Kihara H, TC Katayama, and K Tsunewaki. 1962. Floating habit of 10 strains of mild and cultivated rice. Jpn. J. Genet. 37: 1-9. Krishnayya GR, RM De, SB Lodh. 1990. Physiological basis for submergence tolerance in rice. Oryza 27:286290 Kosambi DD. 1944. The estimation of map distances from recombination values. Ann. Eugen. 12: 172-175 Lander ES, D Botstein. 1989. Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121: 185-199 Lander ES, P Green, J Barahamson, A Barlow, MJ Daly, SE Lincoln, L Newbury. 1987. Mapmarker: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1: 174-181 Loresto GC, TT Chang, and C Tagumpay. 1976. Field evaluation and breeding for drought tolerance. Philippines J. Crop Sci.1: 36-39. Maurya DM. 1975. Concept of flood and flood resistance in rice. Pages 170-177 in proceeding of the int. seminar on deepwater rice. Aug. 21-26.1974. Dacca, Bangladesh. Mazaredo AM and BS Vergara. 1982. Physiological differences in rice varieties tolerant of and susceptible to complete submargence. Page 327-342 in proceeding of the 1981 int. deepwater rice workshop. IRRI, Los Banos Philippines. Mazaredo AM and BS Vergara. 1977. Screening for rapid elongation ability in small water tank. In; Proceeding of the International Deepwater Rice Workshop, Bangkok, Thailand. IRRI, Los Banos, Philippines. Pp. 109-113 Metraux JP, H Kende. 1984. The cellular basis of the elongation response in submerged deepwater rice. Planta 160:73-77 Mishra SB. 1995. Genetics of submergence tolerance and elongation ability of flood-prone rice. Plant Breeding, Genetics and Biochemistry Division. IRRI, Philippines. 16 pp.
Mohanty HK, GS Khush. 1985. Diallel analysis of submergence tolerance in rice, Oryza sativa L.. Theo Appl Genet 70:467-473 Mohanty HK, B Suprihatno, GS Khush, WR Coffman, and BS Vergara. 1982. Inheritance of submergence tolerance in deepwater rice. In: Proceeding of the 1981 int. deepwater rice workshop. IRRI, Los Banos Philippines. pp121-134 Morishima H. 1975. Floating ability as an adaptive character of rice and its memoring method. Pages 109-114 in proceeding of the int. seminar on deepwater rice. Aug 21-26, 1974, Dacca, Bangladesh. Morishima H, K Hinata, and HI Oka. 1962. Floating ability ang drought resistance in wild and cultivated species of rice. Indian J. Genetic Plant Breed, 22:1-11. Mukherji DK, and SKB Roy. 1977. Genotypic adaptability of rice (Oryza sativa L.) and a suggested formula for measuring adaptation to water depths above 30cm. Pages 55-65 in proceeding of the workshop on deepwater rice, 8-10 Nov. 1976. Bangkok, Thailand. Murty KG, KK Nanda. 1974. Changes in peroxidase isoenzymes of Phaseolus mungo hypocotyl cutting rooting. Phytochemistry 13:1089-1093 Nandi S, PK Subudhi, D Senadhira, NL Manigbas, S sen-Mandi, N Huang. 1997. Mapping QTLs for submergence tolerance in rice by AFLP analysis and selective genotyping. Mol Gen Genet 255: 1-8 Nasiruddin M, A Wahale, and BA Dewan. 1982: Genetic of plant and internode elongation ability in rice. Pages 113-120 in proceeding of the 1981 deepwater rice workshop IRRI, Los Banos Philippines. Nasiruddin M, MS Ahmed, SNH Zanman, and E Haq. 1977. Effects of flooding on morphological characteristic of floating rice varieties of Bangladesh. Pages 67-74 in proceeding of the workshop on deepwater rice.8-10 Nov. 1976. Bangkok Thailand. Oka HI. 1983. Conservation of the heterogeneous rice germplasm observation workshop. IRRI, Los Banos Philippines. Oka HI. 1975. Floating rice, an ecotype adapted to deepwater paddies - Review from the viewpoint of breeding. Pages 277-287. from rice in Asia. National Institute of genetics. Mishima, Japan. Padmanabhan SY. 1975. deepwater rice India and its improvement. Pages 51-54 in proceeding of the int. seminar on deepwater rice. Aug. 21-26, 1974. Dacca, Bangladesh. Peschke VM, MM Sachs. 1993. Multiple pyruvate decarboxylase genes in maize are induced by hypoxia. Mol Gen Genet 240:206-212 Prechachat C, and BR Jackson. 1975. Floating and deepwater rice variation in Thailand. Pages 39-45 in proceeding of the int. seminar on deepwater rice, Aug. 21-26, 1974. Dacca, Bangladesh. Rai RSV, KS Murty. 1976. Effect of submergence on some physiological changes in rice seedlings. Ind. J. Exp. Biol. 14:369-370 Rai RSV, KS Murty. 1979. Note on the effect of complete submergence of RVBP carboxylase activity on rice seedlings. Indian J. Aric. Res. 13: 61-63 Ramiah K, and K Ramaswami. 1941. Floating habit in rice. Indian J. Agric. Sci. 11: 1-8. Raskin I, H Kende. 1984. Regulation ofgrowth in stem sections of deepwater rice. Planta 160: 66-72 Riccharia RH, and B Hisro.1960. Flood and deepwater rices and future plan of their improvement. Indian Agric. 4: 135-143. Rowland LJ, JN Strommer. 1986. Anaerobic treament of maize roots affects transcription of Adh1 and transcript stability. Mol Cell Biol 6:3368-3372 Russell DA, MM Sachs. 1989. Differential expression and sequence analysis of the maize glyceraldehyde-3phosphate dehydrogenase gene family. Plant Cell 1:793-803 Sachs MM, M Freeling, R Okimoto. 1980. The anaerobic proteins of maize. Cell 20:761-767 Sachs MM, CC Subbiah, IN Saab. 1996. Anaerobic gene expression and flooding tolerance in maize. J Exp Bot 47:1-15 Saha Ray PK, D HilleRisLambers, NM Tepora. 1993. Combination of stem elongation ability with submergence tolerance in rice. Euphytica 68:11-16 Saha Ray PK, D HilleRisLambers, NM Tepora. 1994. Genetics od stem elongation ability in rice (Oryza sativa L.). Euphytica 74:137-141
Sambrook J, EF Fritsch, T Maniatis. 1989. Molecular cloning: a laboratory manual. 2nd edn. Cold Spring Harbor Laboratory, Cold Spring harbor, New York Sauter M, K Kende. 1992. Gibberellin-induced growth and regulation of the cell division cycle in deepwater rice. Planta 188:362-368 Sauter M, RW Seagull, H Kende. 1993. Internodal elongation and orientation of celluluose misrofibrils and microtubules in deepwater rice. Planta 190:354-362 Sauter M, SL Mekhedov, H Kende. 1995. Gibberellin promotes histone H1 kinase activity andthe expression of cdc2 and cyclin genes during the induction of rapid growth in deepwater rice internodes. The Plant Journal 7:623-632 Seshu DV. 1982. Multilocation screening for rice varietal traits relevant to deepwater culture. Pages 6-45-61 in proceeding of the 1981 international deepwater rice workshop. IRRI, Los Banos Philippines. Setter TL, MB Jackson, I Waters, I Wallace, H Greenway. 1988. Foodwater carbon dioxide andethylene concentrations as factors in chlorosis development andreduced growth of completely submerged rice. In: Proceedings of the 1987 International Deepwater Rice Workshop, Bangkok, Thailand. IRRI, pp. 301-310 Setter TL, I Waters, H Greenway, BJ Atwell, T Kupkanchanakul. 1987. Carbohydrates status of terrestrial plants during flooding. In: Crawford RMM (Ed.), Plant Life in Aquatic and Amphibious Habitats. British Ecological Society Symposium No. 5. Blackwell Scientific Publications, Oxford pp. 411-433 Setter TL, E Ellis, EV Laureles, ES Ella, D Senadhira, SB Mishra, S Sarkarung, S Datta. 1997. Physiology and genetics of submergence tolerance in rice. Annals of Botany 79:67-77 Sripongpankul K. 1998. Gene mapping andquantitative trait loci analysis of flood tolerance in rice (Oryza sativa L.). PhD Thesis. University of The Philippines, Los Banos (UPLP), IRRI, Philippines. 137 P. Sugawara T, T Horikawa. 1971. Studies on the elongation of internodal elongation and nodal root development in the floating rice. Plant Cell Physiol 26:607-614 Suge H. 1988. Physiological genetics of internode elongation in submerged deepwater rice. In: Proceedings of the International Deepwater Rice Workshop, Bagkok, Thailand . IRRI, Philippines. pp 245-259 Supapoj N, C Setabutara, K Kupkanchanacul, and R Shuwisitkul. 1977. Segregation for elongation ability in two crosses of floating rice with ordinary lowland rice. Pages 29-36 in proceeding of the int. workshop on deepwater ric, 8-10 Nov. 1976. Bangkok Thailand. Suprihatno B. 1980. Inheritance of submergence tolerance in rice (Oryza sativa L. ) Ph.D. thesis, University of Philippines. Los Banos, Laguna, Philippines. Suprihatno B, WR Coffman. 1981. Inheritance of submergence tolerance in rice (Oryza sativa L.). SABRAO 13:98-108 Swaminathan MS. 1979. International seminar on deepwater rice keynote address. In proceeding of the 1978 int. deepwater rcie workshop. 17-19 Aug 1978. Thach TD. 1994. The genetic association between elongation ability and submergence tolerance in rice (Oryza sativa L.). MSc Thesis, Central Luzon State University, Nueva Ecija, Philippines. 65 pp. Thakur R, D HileRisLambers. 1988. Inheritance of elongation and submergencetolerance in rice (Oryza sativa L.). In: Proceedings of the International DeepwaterRice Workshop, Bangkok, Thailand. IRRI pp.269274 Thakur R, D HilleRisLambers. 1989. Inheritance of plant elongation ability at seedling stage in rice. Euphytica 41:227-233 Umeda M, H Uchimiya. 1994. Differential transcript levels of genes associated with glycosis and alcohol fermentation in rice plants (Oryza sativa L.) under submergence stress. Plant Physiol 106:1015-1022 Vartapetian BB, MB Jackson. 1997. Plant adaptations to anaerobic stress. Ann Bot 79(Suppl A): 3-20 Vergara BS, and NN Dishit. 1982. Research strategies for rice areas with excess water. Pages 187-199 in Rice Research strategies for the future. IRRI, Los Banos Philippines. Vergara BS and A Mazaredo. 1979. Effect of presubmergence drought on internode elongation of deepwater rice. Pages 41-44 in proceeding of the 1978 int. deepwater rice workshop. 17-19 Aug. 1978, Calcutta, India.
Vergara BS, B Jackson, and SK De Datta. 1976: Deepwater rice and its response to deepwater stress. Pages 321346 in proceeding of the symposium on climate and rice. IRRI Sep. 24-27, 1974. Los Banos Philippines. Vergara BS and AM Mazaredo. 1975. Screening for resistance Aug. 21-26, 1974. Dacca, Bangladesh. Webster C, RL Gaut, KS Browning, JM Ravel, JKM Roberts. 1991. Hypoxia enhances phosphorylation of eukaryotic initiation factor 4A in maize root tip. J Biol Chem 266:23341-23346 Xu K, DJ Mackill. 1995. RADP and RFLP mapping of a submergence tolerance locus in rice. Rice Genetics Newsletter 12: 244-245 Yamada N. 1959. Physiological basis of resistance of rice plant against overhead flooding [in Japanese, English summary] Bull, National Institute Agricultural Science Ser. No. 9 110 P. Yamada N, and Y Ota. 1956. Varietal difference in resistance of rice plant to submersion under water [in Japanese, English summary]. Proceeding Crop Sci. Soc. Jpn. 24: 151-153. Yamada N, A Osada, and Y Ota. 1954. Changes of metabolism of rice plant under flooding (prelim, rep.). Proc. Crop Sci. Soc. Jpn. 22: 57-58. Yang Y, HB Kwon, HP Peng, MC Shih. 1993. Stress response and metabolic regulation of glyceraldehyde-3phosphate dehydrogenase genes in Arabidopsis. Plant Physiol 101:209-216

Chng 5
C S DI TRUYN TNH CHNG CHU C NHM
Nng sut cy trng trn t acid, t phn b nh hng rt ng k do nhiu yu t: (1) c st, (2) c nhm, (3) pH thp, (4) thiu ln, v.v... Trong chng ny, chng ta cp n tnh trng chng chu c nhm ca cy trng trn t chua (pH thp), hoc t phn (pH thp, nhiu lu hynh, nh ng bng sng Cu Long) c nhm c xem nh yu t hn ch chnh i vi s tng trng ca cy trng (Kochian 1995). Ngi ta cha bit nhiu v c ch chng chu c nhm ca cy trng mc sinh hc phn t. Trong cc loi m cc, cy la mch en (rye) c xem nh l cy c tnh chng chu c nhm tt nht. Aniol v Gustafson (1984) pht hin tnh chng chu ny cy la mch en, c iu khin bi nhng gen ch lc nh v trn nhim sc th 3R v 6R, vi nhng gen khc nh v trn nhim sc th 4R. Hai loci tri: Alt1 trn nhim th s 6R v Alt2 trn nhim th s 4R, c thit lp bn (Gallego v ctv. 1998, Miftahudin v ctv. 2002). i vi la m, tnh trng ny c iu khin bi 2-3 gen tri hoc a gen (Aniol v Gustafson 1984). Ngi ta xc nh c mt gen chnh AtlBH iu khin tnh trng chng chu c nhm, nh v trn nhim sc th 4DL, ng gp 85% bin thin kiu hnh (Riede v Anderson 1996). Tang v ctv. (2000) thit lp bn mt gen iu khin tnh chng chu nhm trn mt nhnh vai ca nhim th 4H ca la mch (barley), lin kt vi marker Xbcd117, khong cch lin kt l 2,1 cM, lin kt vi marker Xwg464 v Xcdo1395, khong cch lin kt l 2,1 cM. i vi cy bp, nhiu phng th nghim cng tin hnh nghin cu tnh trng chng chu c nhm, h ghi nhn tnh trng ny do a gen iu khin (Magnavaca v ctv. 1987). Nghin cu tnh trng ny trong cy la vn cn t bo co. S dng k thut phn t, Wu v ctv.(2000) xc nh c nhiu QTL c lin quan n tnh chng chu nhm, trong mt qun th cn giao ngu nhin ca t hp lai IR1552 / Azucena. Nguyn v ctv.(2001) pht hin 5 QTL iu khin tnh chng chu nhm, nh v ri rc trn 5 nhim sc th, vi mt QTL chnh thc nh v trn nhim th s 1. Nguyn v ctv. (2002) tm thy 10 QTL nh v trn 9 nhim sc th iu khin tnh chng chu nhm, trn c s qun th lng bi kp (DH) ca t hp lai CT9993 / IR62266. Nhng pht hin ny khng nh rng: tnh chng chu c nhm l mt tnh trng phc tp i vi cy la, trc y, ngi ta ngh rng r la trong t acid phi tip xc v b ng c bi ion H+, nhng hin nay ngi ta cho rng chnh ion Al3+ l tc nhn (Coronel 1980, Thaworuwong v van Det 1974). Trong cc th nghim ca tc gi, vi pH t 3.5 - 5.0, khng thy nh hng c hi r la nu khng c s hin din ca nhm. Chnh s pht trin ca r la l ch tiu quan st d nht cc nng gy c hi ca nhm. Thng thng, gy c hi ca nhm khng xut hin pH ln hn 5,5 (Mc Cart v Kaprath 1965), m ch gy thit hi trm trng pH nh hn 5,0. K thut thanh lc tnh chng chu c nhm trong mi trng dinh dng c nhiu thun li hn thanh lc ngay trn t acid, bi v rt kh kim tra c nng nhm mt cch chnh xc trong t (Froy 1974, Howeler v Cadavid 1976, Martinez 1976), cha ni s thay i pH lm nh hng n dng nhm ho tan v mc gy c hi trong t (Moore 1974).
C 3 k thut thanh lc nhm trong mi trng dinh dng: chiu di r tuyt i, s phc hi sinh trng ca r v nhum mu khi phn ng vi hematoxylin. Nhng k thut o chiu di r tuyt i c p dng ph bin nht (Coronel 1980). Sau khi c c chiu di tuyt i (MRL), ngi ta cn dng php tnh chiu di r tng i (RRL) lm gim bt s sai bit v di truyn ca s sinh trng r la (Coronel 1980). Khi nim v chiu di tng i (RRL) c xem nh l t s ca chiu di r 2 mc nhm khc nhau.
RRL=
Chiu di r nng 30 ppm Al Chiu di r nng 0 ppm Al
Hin tng u tin ca cy trng khi b c ca nhm l s ko di ca r b c ch. R b thng tn trong iu kin nng nhm cao, biu hin thng qua cc triu chng nh sau: ngn, dy, sm mu, dn d gy, t m nhnh, chiu di tng s ca r v th tch r cy gim r rt.
c nhm cn c th nh hng n s tng trng ca chi thn, thng qua s gii hn di chuyn cht dinh dng, v nc do hin tng hp thu hoc truyn dn qa km
5-1. GING LA NC SU CHNG CHU C NHM NG BNG SNG CU LONG Hu ht cc vng la nc su BSCL l t phn, nng st v nhm cao, gy c hi cho cy la giai on m trc khi ma ngp, c bit trong iu kin ma tht thng (van Breeman 1978, V Tng Xun v ctv 1982). Cy la chu tc dng ca nhiu yu t cng mt lc, chng hn nh s thiu ln, tha st, nhm v cc hp cht c cha lu hunh (S) trn t phn. Cc biu hin v triu chng gy c ny rt phc tp, v s tc ng c tnh cht tng hp ca n. Tnh chng chu c nhm ca cc ging la nc su ph bin BSCL c phn tch. Th nghim c thc hin ti IRRI, trong phytotron, nhit 29/21oC (ngy/m), m tng i 70%. Hm lng nhm cho vo mi trng dinh dng l 0 ppm v 30 ppm ly t dung dch IM (AlCl3, 6H2O) Sau khi c c chiu di tuyt i (MRL), ngi ta cn dng php tnh chiu di tng i (RRL) lm gim bt s sai bit v di truyn ca s sinh trng r la (Coronel 1980). p dng php th chi bnh phng v s ng nht ca phng sai (Gomez 1984), chng ti xp nhm nhng ging c tnh chng chu c nhm t kh, trung bnh n nhim, so vi ging i khng (bng 1).
Bng 1. Chiu di r tuyt i ca cc ging khng nng 0 v 30ppm Al trong mi trng dinh dng (Buu 1987).
Ging Leb Mue Nahng Tu nt (La ging) Ba so Khao Daeng Habiganj Aman 1
Chiu di r (cm) 0ppm Al 30ppm Al 13.751.65 10.301.34 10.051.41 13.191.42 13.780.95 9.370.61 7.041.57 4.430.97 8.441.10 8.810.39
RRL 0.68 0.68 0.65 0.64 0.64

Nng phc HTAFR 77022-14-157-6-1 Sa quay IR 11185-RRR-88-2 Ba chc IAC3 (C khng) IR45 (C nhim) 12.181.72 10.831.91 10.931.47 11.361.63 9.300.89 20.362.41 14.841.3 7.711.76 6.780.38 6.851.33 7.131.09 5.780.93 16.512.71 6.111.76 0.63 0.63 0.63 0.63 0.62 0.81 0.41
RRL: chiu di r tng i (relative root length)
bng 1, cc ging khng c gi tr RRL ng nht (2=0,416, p=0,50). Ging Leb Mue Nahng 111 (LMN111), Habiganj Aman1, Khao Daeng v Ba so u c c hai tiu chun v chiu di tuyt i v tng i. Cc ging la ni Ba so (ng Thp Mi), Nng Phc (T gic Long Xuyn) u th hin tnh khng c nhm rt tt. Ging Ba chc v Tu nt ph bin vng Ty Sng Hu v T Gic Long Xuyn cng th hin tnh khng nhm. Ging Sa quay thuc nhm la ma sm - ph bin An Bin (Kin Giang), trng trn t nhim mn v than bn, cng uc ghi nhn c tnh khng. Khao Daeng - ging la nc su go ca Thi Lan v ging la ni LMN111 trc y c ghi nhn khng nhm rt kh (HilleRisLambers v Sittiyos 1985), cng th hin kt qu tng t trong th nghim ny. Dng la nc su trin vng IR11185-RRR-8-2 t t hp lai IR2070-41439/LMN111 c ghi nhn c tnh khng ging nh LMN111. 39 dng ging c gi tr RRL ln hn trung bnh mu c lit k bng 3 v c xp nhm theo s ng nht v phng sai. Mc phn ng i vi nng nhm 30 ppm th hin rt r (2=14.37). Khng ghi nhn triu chng c hi trn l gi. Cc ging la ni Nng tri, Nng m nh, Nng m to (ng Thp Mi), Nng Ty, Nng son (T Gic Long Xuyn) c phn ng hi khng. Ging ph bin trn vng Ty Sng Hu l Trng Chm, Trng La cng c phn ng hi khng. Ging Trng ln, La Thc ph bin trn t phn b nhim mn cho thy c phn ng nhim. Thc hin cn trng lng kh bng cch sy mu trong t nh n 150 0F trong 4 ngy. Cn ring mu r la v mu chi thn (Bng 2) Nhn chung trng lng kh ca chi v r u c xu hng gim nng 30 ppm Al, nhng kt qu khng r rng nh quan st chiu di r. p dng khi nim v chiu di r tng i (RRL) l phng php thanh lc nhm n gin v hiu qu hn ht. Trong th nghim ny, tng quan gia RRL v chiu di r tuyt i (MRL) rt cht ch (hnh 5-1) r = 0,798 ** (n=107) y= 0,039 x + 0,215 Hnh 5-2 v 5-3 cho thy, s bin thin ca trng lng cht kh thng ln hn nhiu so vi bin thin v chiu di. Do phng php o chiu di r quan st phn ng chng chu vi c nhm c th cho bit kt qu chnh xc hn cn trng lng kh. Bng 2: Trng lng kh ca r v chi ca cc ging khng 2 nng nhm 0ppm v 30ppm (Buu 1987).

Ging
0ppm Al 30ppm Al 0ppm Al
Trng lng r (mg/cy) Trng lng chi (mg/cy)

30ppm Al
Leb Mue Nahng Tu nt (la ging) Ba so Khao Daeng Habiganj Aman 1 Nng phc HTAFR 77022-14-157-6-1 Sa quay IR 11185-RRR-88-2 Ba chc IAC3 (C khng) IR45 (C khng)
22,2 14,1 22,3 21,6 20,9 17,1 13,0 21,6 16,4 16,8 28,6 30,5
20,2 16,2 21,5 26,8 16,5 10,2 15,3 18,0 18,1 15,0 27,3 17,3
53,2 15,1 53,6 63,6 55,5 35,3 30,4 65,7 38,5 52,2 53,2 56,7
48,5 61,6 50,9 59,8 40,8 26,1 38,0 51,5 43,4 55,1 59,4 39,5
Th nghim nhum hematoxylin r la cng kh so snh v kt lun. Cn tip tc thanh lc v ghi nhn mc ph bin ca tng ging la trn nhng loi t phn khc nhau (Sulfaquepts), kt hp vi s thanh lc c st c nhng kt lun tip theo gip cho s b tr c cu ging hp l v xc nh mc tiu lai to ph hp cho tng vng.
Bng 3a. Danh sch cc dng ging c RRL ln hn trung bnh mu (Bu 1987) Hng 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 Dng ging Leb Mue Nahng 111 Tu nt (la ging) Ba sao Khao Daeng Habigan j Aman 1 Nang phc HTAFR 77022-14-157-6-1 Sa quay IR1185-RRR-88-2 Ba chuc Nng m nh Nng m to BGD4-7-3PE-1-12 Nng ty Habiganj Aman 8 FR13A SPR7411-7-2-1 C l Jhora Trng la Nng ty Nng tri IR33277-1-507-1-13 IRRS 43/3 BKN 6986-1 IR5853-196-1-1 Baisbish HKNPR 76010-108-2-0 SPR 7294-136-2-4-1-0-8-2 IR31086-12-1-3-3-1 Khama Khao Dawk Mali 105 RRL 0.68 0.68 0.65 0.64 0.64 0.63 0.63 0.63 0.63 0.62 0.60 0.60 0.60 0.60 0.58 0.57 0.57 0.56 0.56 0.56 0.56 0.55 0.55 0.54 0.54 0.54 0.53 0.53 0.53 0.53 0.53 0.53 2 0.416
3.361
4.193

33 34 35 36 37 38 39 Tu nt Huyt rng D10 Nng ty C Nng sen IR28932-9-3-3 Trng chm IR11288-B-B-69-1 0.53 0.52 0.52 0.52 0.52 0.51 0.51
x= 0.49** 0.02
MRL ( ) Hnh 5-1: Tng quan gia chiu di tuyt i (MRL) v chiu di r tng i (Buu 1987)
Hnh 5-2: Bin thin se tai: wWw.SinhHoc.edu.vn i v Tai lieu chiachiu di r tngca 2(RRL) i trng lng kh ca r tng i (RRW) nng 30ppm Al ging chng (Bu 1987)
Hnh 5-3: Bin thin chiu di r tuyt i (MRL) v Trng lng r (DRW) nng 30ppm Al ca 2 ging i chng (Bu 1987) Bng 3b: Kt qa thanh lc nhm b ging la c su tp t nhiu quc gia (Khatiwada v ctv. 1996), gi tr RRL c theo th t t cao xung thp.
Ging Siyam Kuning Gudabang Putih Siyam Lemo Khao Deang Siyamhalus Bjm-12 Ketan Seribugangtang Bater Raden Rati Padi Kanji Bjm-13 Batang Pane Bjm-14 C ung Bjm-10 Padi Jambi Gablak Cablak Barito Ngun gc Indonesia Thi Lan Indonesia Vit Nam Indonesia RRL (trung bnh ca 2 ln lp li) 1.159 1.135 1.105 1.088 1.083 1.059 1.058 1.056 1.050 1.047 1.043 1.040 1.039 1.038 1.037 1.035 1.032 0.956 0.944

Engatek Bjm-15 Siyam Kuning Quisidugo La Thc Gudabang Kuning Bjm-17 Kutik Putih Kapuas Baiang 6 Pontianak Nng Ci Bayar Kuning Bjm-11 Trng Ha Bnh Alabio Khao Seetha Gaw Diaw Bow Khao Taeng La Thc C Talang A Mahakam Galambong Ti Nguyn Ketumbar Thm Rn Talang B ui Tru C ung Phn Gogo Ranceh c Phng Nng Go Masirit Kapus Yaca S-1 Atanha Nng Co Thn Nng Pokkali Sc nu Silla S-4 IRAT104 (chun khng) IR1552 (chun nhim) CV % LSD 0,05 Malaysia Indonesia Indonesia Vit Nam Indonesia Vit Nam Indonesia Vit Nam Indonesia Thi Lan Indonesia Thi Lan Vit Nam Indonesia Vit Nam Indonesia Vit nam Indonesia Vit Nam Indonesia Vit Nam Indonesia West Africa Vit Nam India Vit Nam West Africa 0.930 0.928 0.925 0.922 0.918 0.916 0.903 0.902 0.886 0.886 0.853 0.850 0.848 0.845 0.845 0.807 0.807 0.801 0.800 0.794 0.784 0.783 0.773 0.768 0.742 0.738 0.729 0.701 0.700 0.698 0.676 0.671 0.670 0.622 0.661 0.659 0.623 0.623 0.621 0.617 0.587 0.573 0.450 0.828 0.572
12.50 0.213
5-2. XC NH QTL IU KHIN TNH CHNG CHU C NHM By v ctv. (2003) s dng 171 dng F6 (dng cn giao ti t hp = RILs) c chn mt cch ngu nhin t 312 dng ca t hp lai IR64 / Oryza rufipogon (acc. 106424). Mu ging la hoang Oryza rufipogon ny c thu thp t Trm Chim (ng Thp Mi), ni c xc nh l t phn nng. Cc dng RIL c to ra thng qua phng php chn lc SSD (single seed descent) cho n F6 ti IRRI.
IR64 l ging la indica cho nng sut cao v phm cht go tt, pht trin rng khp Chu , nhng rt nhy cm vi c nhm (Khatiwada v ctv. 1996). La hoang Oryza rufipogon l loi hnh a nin, genome AA ging nh b genome ca la trng IR64, chng mc hoang di trong vng m trng, t phn ng Thp Mi. B m v con lai c thanh lc nhm trong dung dch dinh dng Yoshida (Yoshida v ctv 1976). Nghim thc nhm c thm vo l 0 v 40 ppm Al (tng ng 1.48 mM), nhm hot ng Al3+ t AlCl3.6H2O pH 4. Dung dch dinh dng c thay mi ngy, iu chnh pH 5. Khay th nghim v cy m c gi trong iu kin phng th nghim c nhit 27 2 0C, vi 12 gi sng 300 PPFD (photo proton flux density) Th nghim c b tr theo kiu khi hon ton ngu nhin, 4 ln nhc li. Chng trnh MAPMARKER/EXP v.3.0 (Lander v ctv. 1987, Lincoln v ctv. 1992) c s dng phn tch bn lin kt gen, vi hm Kosambi (Kosambi 1944). Chng trnh MAPMARKER/QTL version 1.1 c s dng xcnh cc loci nh hng n nhng tnh trng s lng mc tiu trn c s phn tch bn cch qung (Paterson v ctv. 1988, Lincoln v ctv. 1992). S dng LOD score = 2,4 lm ngng chn lc s hin din QTL trn c s khong cch tng s ca bn , v khong cch trung bnh gia nhng marker (Lander v Botstein 1989). Vi ngng ny, mt QTL c th c pht hin trong genome vi xc sut tin cy 0,05 (Paterson v ctv. 1988, Lander v Bostein 1989). Bin thin kiu hnh c gii thch bi tt c QTL thng qua m hnh phng trnh a tuyn trong MAPMARKER/QTL. Tng tc gia nhng loci trn bn c th hin bng cch s dng QTLMapper version 1.0 (Wang v ctv. 1999) Ba tnh trng mc tiu c xem xt l Chiu di r i chng: Chiu di r b stress: Chiu di r tng i: CRL SRL RRL
Phn tch ANOVA pht hin s khc bit c ngha gia b m vi con lai dng cn giao ti t hp c trnh by bng 4
Bng 4: Thng s ca 3 tnh trng c ghi nhn trn b m v 154 dng con lai RIL, 4 ln lp li (By v ctv. 2003) Tnh trng CRL IR64 O. rufipogon RILs SRL IR64 O. rufipogon RILs RRL IR64 O. rufipogon RILs Min. 4,48 1,76 0,18 Max. 19,28 9,83 0,88 Trung bnh 11,30** 7,10** 9,98** 2,56** 5,50** 3,99 0,20** 0,77** 0,41 CV (%) 11,12 16.18 18,93 LSD0.01 2,02 1,18 14,30
5-2-1. iu tra a hnh trong b m S dng 5 enzyme EcoRI, EcoRV, DraI, HindIII v XbaI phn ct hn ch genome ca b m. S dng 400 RFLP marker ca i Hc Cornell v ca chng trnh RGP (Nht) trong nghin cu a hnh. Kt qa c 274 probe (65,9%) th hin a hnh. Mc a hnh th hin tng ng vi cc enzyme EcoRI (14,0%), EcoRV (12,5%), DraI (19,8%), HindIII (27,7%) v XbaI (19,5%) Trong 168 SSR marker c s dng, c 112 marker (66,7%) th hin a hnh. Tc gi xem xt 13 gen ng c vin iu khin tnh chng chu c nhm theo ngun s liu GenBank, trong c 4 gen (30,8%) c tm thy biu hin a hnh gia b v m 5-2-2. Phn ly v thit lp bn a hnh c xem xt trn c s 238 marker rt r rng trong b m, t chng c s dng iu tra trn qun th con lai. Tc gi thit lp bn vi 151 marker loci v phn tch QTL trong dng con lai cn giao ti t hp (RIL). Hu ht nhng marker ny th hin bng lai n, tr RG788 c hai copy. Marker ny c nh du l suffix A v B. Nhng alen khng r ngun gc, khng phi ca b m (hon ton mt tn hiu) c tm thy trong qun th RIL ny l 7,3%. Trng hp nh vy c x l nh trng hp thiu s liu. Phn ly ca 151 marker trong bn lin kt gen c trc nghim thng qua php th Chi bnh phng. Kt qa cho thy hu ht (122 marker) phn b theo l thuyt Mendel 1:1, chim 80,7% tng s. Hu ht marker (118 marker) thin lch v IR64, 4 marker lch v Oryza rufipogon, v 29 loci c t nht 50% alen thuc v Oryza rufipogon. Mc lch ny (skewness) trong qun th RIL [60% nghing v alen IR64, 40% nghing v alen la hoang] gy nh hng n kh nng thit lp bn marker de novo trn c s s liu phn ly ca nhhng dng RIL (By v ctv. 2003)
Bng 5: Nhng QTL gi nh trong trng hp 3 tnh trng CRL, SRL v RRL (By v ctv. 2003)
Tnh trng QTL Qung Nhim Chiu th di (cM) Khong cch vi marker u tin (cM) 4,0 8,0 0,0 18,0 6,0 0,0 18,0 18,0 8,0 nh hng cng LOD R2 (%) [a]
CRL QAlCr12.1 RG341-RZ397 SRL QAlSr2.1 QAlSr3.1 QAlSr8.1 RG139-RG324 CDO1395-RG391 RG28-RM233
12 2 3 8 1 3 7 8 9
8,9 11,6 0,5 31,0 6,5 0,6 29,8 31,0 10,0
1,834(I) 3,235(O) 1,478(O) 1,915(O) 0,100(O) 0,167(O) 0,126(O) 0,104(O) 0,109(O)
2,8 2,9 6,2 3,1 45,6 2,4 8,3 5,4 2,5 2,6
10,3 26,4 18,7 20,8 9,0 24,9 22,5 20,8 9,9 70,8
M hnh QTL ti ho RG406-RZ252 RRL QAlRr1.1 CDO1395-RG391 QAlRr3.1 RZ629-RG650 QAlRr7.1 RG28-RM223 QAlRr8.1 RM201-WALI7 QAlRr9.1 M hnh QTL ti ho [b]
(I): IR64, (O): Oryza rufipogon [a] bin thin kiu hnh c gii thch bi tng QTL ring bit [b] bin thin kiu hnh c gii thch bi tt c QTL Chin lc thit k mt bn lin kt gen t qun th c tnh cht nghing lch nh vy (skewness) c Wang v ctv. (1994) xut. Trc tin, nhng marker ny cng thuc v nhm lin kt c xc nh trn c s bn c mt cao v lin kt di truyn trong cy la (Causse v ctv. 1994, Harushima v ctv. 1998, Temnykh v ctv. 2001). Sau , t l ti t hp, v tr th t trn nhim sc th c xc nh bng lnh Three Point v First Order trong phn mm chng trnh my tnh, LOD = 3,0 trn c s s liu ca qun th RIL. i vi nhng marker m v tr ca n cha c bit r, chng ta phi dng phng php phn tch hai im vi LOD 10-12 tm thy nhm lin kt ca chng. Cu lnh Try c s dng i vi nhng marker khng lin kt (unlinked). Kt qa phn tch cho thy, 151 marker loci phn ra thnh 13 nhm lin kt (ring nhim th s 1 c hai nhm lin kt). Cc nhim scth c nh hng theo nhnh vai ngn nm pha trn ca v tr tm ng (Singh v ctv. 1996). Th t ca marker trn bn c xp theo nguyn tc thng nht ca i Hc Cornell (Causse v ctv. 1994, Temnykh v ctv. 2001), ca chng trnh genome cy la (RGP) ti Nht (Harushima v ctv. 1998), v bn cy la c pht trin t qun th O. sativa x O. rufipogon (Xiong v ctv. 1997, Xiao v ctv. 1998). Tng chiu di bn l 1.755 cM, khong cch trung bnh gia hai marker l 11,6 cM (hnh 5-4) Nhim sc th s 10 c s marker th hin t nht. Cho d 12 SSR v 20 RFLP nh v trn nhim th ny, nhng ch c 6 marker c thit lp trn bn . S th hin a hnh km trn nhim th s 10 cho thy: mt vi vng ca genome ging la trng v la hoang c th biu hin kh ph bin hin tng suy gim (descent), hoc O. rufipogon ng vai tr dn xut ca con lai (derivative) to ra trng thi gn gi v huyt thng (proximity) ca loi hoang di mc ng rung ca nng dn trng la Chu (th d hin tng la c) 5-2-3. Bn QTL
Tc gi xc nh 9 QTL vi gi tr LOD 2,4 c phn lp i vi 3 tnh trng mc tiu (bng 5). Nhng QTL gi nh ny c ghi nhn vi nhim sc th m n nh v, gi tr LOD, phn trm bin thin kiu hnh c gii thch, nh hng cng (additive effect). S QTL iu khin tnh trng thay i t 1 QTL i vi CRL cho n 5 QTL i vi RRL. Bin thin kiu hnh cng thay i t 9,0% n 26,4%. V tr ca nhng QTL c xem xt trn hnh 5-4 Tnh trng chiu di r i chng (CRL): ch c 1 QTL, l QAlCr12.1, nh v trn nhim th s 12, hai marker k cn ca n l RG341 v RZ397, iu khin tnh trng chiu di r trong iu kin khng c stress do nhm. QTL ny gii thch 10,3% bin thin kiu hnh vi 1 alen (chiu di r di hn) t ging la IR64. S gia tng ca chiu di r do alen ny l 1,83cm Tnh trng chiu di r b stress do nhm (SRL): c 3 QTL nh hng, c tm thy trn nhim th s 2, 3 v 8. l QAlSr2.1 nh v trn nhim th s 2, hai marker k cn l RG139 v RG324, n c gi tr R2 cao nht (26,4%). Tip theo sau l QAlSr8.1, hai market k cn l RG28 v RM223, trn nhim th s 8 (20,8%). V cui cng l QAlSr3.1, hai marker k cn l CDO1395 v RG391, trn nhim th s 3 (18,2%). Nhng QTL ny ng gp 45,5% bin thin kiu hnh. y Oryza rufipogon c nhng alen u th v tnh trng chiu di r trong c 3 QTL ni trn. nh hng allelic thay i t 1,46 cm (QalSr3.1) n 3,24 cm (QalSr2.1 ) Tnh trng chiu di r tng i (RRL): c 4 QTL nh hng, c tm thy trn nhim th s 1, 3, 7 v 8. l QAlRr1.1 nh v trn nhim th s 1, hai marker k cn l RG406 v RZ252, n c gi tr R2 l 9,0%). K n l QAlRr3.1, hai market k cn l CDO1395 v RG391, trn nhim th s 3 (24,9% cao nht). Tip theo sau l QAlRr7.1, hai market k cn l RZ629 v RG650, trn nhim th s 7 (22,5%). QAlRr8.1, hai market k cn l RG28 v RM233, trn nhim th s 8 (20,8%). V cui cng l QAlRr9.1, hai marker k cn l RM201 v WALI7, trn nhim th s 9 (9,9%). Tng s QTL trong m hnh gi nh nh vy gii thch c 70,8% bin thin kiu hnh. nh hng ca alen thay i t 10,0% (QAlRr1.1) n 16,7% (QAlRr3.1) (By v ctv. 2003) 5-2-4. Phn tch epistasis Hin tng epistasis l thnh phn quan trng ca di truyn i vi nhng tnh trng phc tp nh tnh chng chu c nhm. Tng tc gia QTL v nhng loci ci bin c th c xem nh mt dng chnh ca epistasis i vi cc tnh trng s lng (Yu v ctv. 1997). Phn tch epistasis gia cc loci trn bn ca 3 tnh trng chnh c th hin bng 6, nh phn mm QTLMapper version 1.0 (By v ctv. 2003) Bng 6: Phn tch epistasis i vi nhng loci ca gen iu khin tnh trng chiu di r i chng (CRL), chiu di r b x l (SRL), v chiu di r tng i (RRL) trong qun th RIL ca t hp lai IR64 x Oryza rufipogon (By v ctv. 2003)
Tnh Nhim Qung gia 2 trng sc th marker (i) RM233-RM203 CRL 2 S2486-RZ590 4 Rali33-RG172 6 RZ509-RM2 7 SRL RRL 3 1 S1845-CDO1395 RM1-RM283 Nhim sc th 5 12 12 9 6 3 Qung gia 2 marker (j) RM249-RM164 RM17-RG901 RM17-RG901 Wali7-RZ404 R2(%) RG653-RZ828 R2(%) CDO1395-RG391 A(i) A(j) AA(ij) LOD 1,39** 1,19** 1,60** 1,92** 51,21 0,39 0,00 0,033 2,84 3,11 4,16 3,82 5,20 7,73
0,18 0,03 0,29 0,43 0,53 0,18 0,32 0,18 0,00 0,00 1,00** 0,10 22,84 0,00 0,030 0,112

4 9 Rali26-RG788B RM201-Wali7 5 12 RM249-RM164 RG543-RG341 R2(%)
0,024 0,065** 0,006 3,34 0,029 0,036** 0,060 3,35 0,000 36,59 0,000
Du * v ** biu th mc ngha 0,05 v 0,01, theo th t A(i): nh hng alellic site (i) A(j): nh hng allelic site (j) AA(ij): tng tc khng alen gia site (i) v site (j) R2(%): biu th mc kiu hnh c gii thch site (i), site (j) v nh hng epistatic (ij) Khi xem xt tnh trng chiu di r trong iu kin c x l nhm, trn nhim sc th s 3, qung gia hai marker S1845-CDO1395 c mt tng tc vi khu vc c xc nh bi RG653-RZ828 (trn nhim th s 6). nh hng allelic ti site (i) rt c ngha v thng k v c gii thch 22,8% kiu hnh th hin ra. Tuy nhin, nh hng allelic site (j) v nh hng tng tc khng alen gia site (i) v site (j) khng biu th mc ngha r rng, v khng c nh hng n bin thin kiu hnh (bng 6). Chiu di r trong iu kin bnh thng, ch c mt QTL nh v trn nhim th s 12 iu khin trong ging IR64. Tuy nhin, bn cp loci c tnh cht epistatic c ghi nhn vi mc c ngha cao. nh hng chnh ca nhng loci ny khng biu hin bin thin v kiu hnh, nhng n biu hin hin tng tng tc khng alen vi nhau, vi R2=51,2%. Kt qa ny cng c cng b bi Zhang v ctv. (2001). Khi xem xt tnh trng chiu di r tng i, tc gi tm thy 6 khu vc khc nhau trn nhim sc th (1, 3, 4, 5, 9, v 12) biu th tng tc vi nhau. nh hng allelic site (j) c ghi nhn rt c ngha, vi 36,59% kiu hnh c gii thch bi QTL. nh hng allelic site (i) v nh hng epistatasis khng c ngha thng k, v R2 cng bng 0 (bng 6). Khu vc QAlRr3.1 nh v trn nhim th s 3 c tng tc khng alen vi khu vc RM1RM283 trn nhim th s 1, cho d tng tc ny khng c ngha v mt thng k. Khu vc QAlRr9.1 nh v trn nhim th s 9 tng tc vi khu vc RG543-RG341 trn nhim th s 12, cho kt qa khng c ngha v mt thng k. Trong mt qun th mapping khc, Wu v ctv. (2000) tm thy 3 cp loci tng tc theo kiu epistasis, vi bin thin kiu c gii thch khong 20% i vi tnh trng chiu di r tng i, trong khi kt qa ny (IR64 x O. rufipogon) cho thy epistasis khng c ngha (R2=0,00). Phn tch epistasis cho thy tnh chng chu c nhm giai on cy m c iu khin bi u th ca hot ng gen cng tnh, ging nh cy la m (Bona v ctv. 1994) v cy bp (Lima v ctv. 1995). Kt qa phn tch QTL v epistasis cho thy kiu gen c nng sut cao nh IR64 c th kt hp vi ngun cho gen chng chu c nhm ca Oryza rufipogon, lm tng cng tnh chng chu ca IR64 thng qua biu hin kiu hnh ca con lai. Trong vic khai thc chin lc MAS (marker-assisted selection), QTL chnh lin kt vi CDO1395 nh v trn nhim th s 3 c th c xem nh ng c vin p dng vic chn con lai c gen chng chu c nhm (By v ctv. 2003) Chiu di r tng i (RRL) l mt thng tin v cng quan trng, lin quan trc tip n tnh chng chu nhm ca cy la. Do , QTL iu khin tnh trng RRL c s dng so snh vi nhng t liu tham kho c lin quan. C 5 QTL iu khin tnh trng RRL c cng b trong mt qun th mapping khc. Khu vc QAlRr.1 (R2=9,0%) nh v trn nhim th s 1, c tm thy tng t nh vy qun th IR1522 x Azucena (Wu v ctv. 2000), CT9993 x IR62266 (Nguyn v ctv. 2002), OM269 x Chim bu (Nguyn v ctv. 2001) (Hnh 5-5) Khu vc QAlRr9.1 nh v trn nhim th s 9, c tm thy tng t nh vy trn qun th IR1552 x Azucena (Wu v ctv. 2000), CT9993 x IR62266 (Nguyn v ctv. 2002) (Hnh 5-5) Nhm xc nh nhng QTL chung cho la v loi m cc khc, By v ctv. (2003) so snh vi kt qa c cng b trn la m (Riedle v Anderson 1996), trn cy la mch
en rye (Miftahudin v ctv. 2002), cy bp (Sibov v ctv. 1998), v cy la mch (barley) (Tang v ctv. 2000), trn c s s dng bn so snh (Ahn v Tanksley 1993), Gale v Devos 1998) v trn c s s dng mt b RFLP probe c tnh cht so snh ca i Hc Cornell. Phn tch ny cho thy khu vc QAlRr3.1 c nh hng kiu hnh ln nht c gii thch bi QTL (R2=24,9%) trong qun th IR64 x Oryza rufipogon, v chng c nh hng kh thp (R2=9%) trong qun th IR1552 x Azucena (Wu v ctv. 2000), nh v trn nhim th s 3, tng ng vi gen iu khin chng chu c nhm thuc nhm s 4 ca Triticeae (hnh 5-6) 5-2-5. ng dng marker trong chn ging chng chu v gen ng c vin Nhng marker phn t lin kt cht ch vi nhng QTL s v cng hu ch cho nh chn ging trong chin lc MAS, chng ta c th chn lc c th mong mun nhng th h u tin. Nhim sc th s 1 c cc nh khoa hc Nht thc hin k thut c trnh t (sequencing), vi 418 BAC clone, trong 353 clone c c xong vo thng T nm 2002. H s dng bn tng hp (integrated genetic map) v tm ra RG406, mt trong nhng marker lin kt vi QTL nhim th ny (QAlRr1.1), vi khong cch di truyn l 0,3 cM. Marker ny c neo bi BAC clone k hiu l OSJNBa0014K08f trn contig s 20 thuc bn vt l cy la Clemson. Chui k t ca BAC c gi i phn tch gen mc tiu theo chng trnh rice genome ti Nht Bn. Khng c trnh t no lin quan n c ch loi tr acid hu c c tm thy trong BAC. i vi QAlRr9.1, ngi ta tm thy trong CT9993 l ging c tnh chng chu c nhm mnh nht (Nguyn v ctv. 2000). Mt QTL ch lc khc iu khin tnh chng chu nhm c tm thy trong CT9993 x IR62266 (R2=28,7%), nh v trn nhim th s 8 lin kt vi hai marker k cn l C1121 v M53. Mt trong nhng QTL ch lc c xc nh trong nghin cu ca By v ctv. (2003) cng c tm thy trn nhim th s 8 (R2=20,8%), lin kt vi RG28 v RZ650. Theo kt qa bn tng hp ca Japanese Oryzabase, marker C1121 cch khong vi marker RG333 l 27,7 cM, v gia hai marker RG28 vi RG333 l 21,8 cM. iu ny cho thy, nhng QTL ny nh v trn cng mt vng ca nhim sc th. Nh vy QTL ny c th c xem l mt trong hai qun th c nn tng di truyn khc nhau. QTL c nh hng ln nht l QAlRr3.1 (R2=22,9%) trong qun th IR64 x Oryza rufipogon, nhng mt QTL c nh hng nh hn (R2=9,0%) trong qun th IR1552 x Azucena u cng nh v trn nhim sc th s 3. Xt theo hin tng synteny trong chng m u, chng tng ng vi chromosome thuc nhm s 4 ca Triticeae (Gale v Devos 1998). Nhng cng b trc khng nh rng c mt vng genome c bo tn trn vai nhnh di ca nhim th tng ng s 4 iu khin tnh chng chu nhm trong cy la m (AltBH), trong cy rye (Alt3), trong cy la mch (Alp) (Miftahudin v ctv. 2002). Gen iu khin tnh chng chu c nhm trong la m v rye lin kt kh cht vi marker BCD1230, v lin kt khng cht ch lm vi CDO1395, nhng ring cy la mch, gen ny lin kt rt cht vi CDO1395. Nh vy, iu ny gi ra mt tng rng AltBH, Alt3 v Alp l nhng gen c loci tng ng (orthologous loci) v mc cao ca hin tng synteny gia nhng nhim sc th 4DL, 4RL, v 4HL. Chng c th cng chia s mt chc nng chung. Mt trong nhng c ch chng chu trong Triticeae l c ch loi thi nhm (Al exclusion) (Kochian 1995). C ch ny c trung gian bi tin trnh phng thch nhm hot ng t acid hu c (ch yu l malic acid), acid hu c gi cht Al+++ theo dng chelate trong vng r v ngn cn nhm xm nhp vo nh r la. Minh chng v hin tng sinh l hc ny c gii thch rt r rng nh nhng loci tng ng iu khin tnh trng chng chu nhm trong Triticeae. QTL ch lc iu khin tnh chng chu nhm c ghi nhn trn nhim sc th s 3 biu hin s tng ng vi cc gen iu khin tnh chng chu nhm thuc nhm s 4 ca Triticeae. Nhng s biu hin v mt sinh l thc vt ca tnh chng chu ny rt thiu trong
cy la so vi cc cy trong nhm Triticeae. nh gi kiu hnh nhng cy b m cho thy Oryza rufipogon c kh nng chng chu nhm cao nht so vi cc ging thuc Oryza sativa. By v ctv. (2003) c gng tm nhng gen ng c vin lin quan n tnh chng chu nhm trn nhim sc th s 3 trn c s s liu sau khi c trnh t (sequencing). Trong 160 BAC clone phn lp t nhim sc th s 3, ch c 53 clone c xc nh trnh t chui m. iu khng may mn trong nghin cu ny l: hu ht nhng clone ny nh v trn nhnh vai di ca nhim sc th, trong khi gen iu khin tnh chng chu nhm c xc nh trn nhnh vai ngn. Tuy nhin, ngi ta rt cn phi thc hin mt s kt ni rt tt gia bn di truyn v bn vt l tm hiu nhng gen ng c vin nh vy trn nhim sc th s 3. Chui m di truyn ca bn gen cy la c cng b (Goff v ctv. 2002, Yu v ctv. 2002) l iu kin v cng thun li. 5-2-6. To ra clone ca nhng QTL iu khin tnh chng chu nhm t la hoang Nhng QTL c xc nh trong genome ca Oryza rufipogon l nhng ng c vin y tim nng i vi vic pht trin cc dng gn nh ng gen (NILs) cloning cc gen iu khin tnh chng chu nhm trong cy la. Nhng QTL ch lc trn nhim sc th s 3 s c ch trc tin. Chng gii thch khong 25% bin thin kiu hnh v biu hin tng ng vi nhng gen chng chu nhm thuc nhm s 4 ca Triticeae QAlRr1.1 trn nhim sc th s 1 c nh hng rt thp (9,0%) i vi bin thin kiu hnh. Tuy nhin, khu vc ca chng biu hin mt cch n nh vi mt QTL c pht hin t nhng cng trnh nghin cu trc y thuc 3 qun th mapping khc. QAlRr7.1 trn nhim sc th s 7 c nh hng ln ng hng th hai (22,5%) i vi bin thin kiu hnh. Hin n tm thi l QTL duy nht trong qun th mapping ny, cha c bo co trong cc qun th trc . QAlRr8.1 trn nhim sc th s 8 c c nh hng ln (20,8%) i vi bin thin kiu hnh. N biu hin mt QTL trn cng mt khu vc ca nhim sc th trong qun th mapping ca CT9993 x IR62266, trong , ging CT9993 c xc nh l ging c kh nng chng chu nhm tt nht trong Oryza sativa (Nguyn v ctv. 2000) Tuy nhin, v cc vng trong genome cha nhng QTL gi nh (QAlRr7.1, QAlRr8.1) vn cn rt rng (29,8 v 31,0 cM, theo th t), chng ta cn phi nghin cu thm nhng marker b sung v lp y khong trng, nhm tha mn mc tiu cloning chnh xc hn QAlRr9.1 trn nhim sc th s 9 c nh hng thp (9,9%) i vi bin thin kiu hnh. Nhng n biu hin kh n nh trong 3 qun th mapping khc nhau. N lin kt cht ch vi marker WALI7 (2,0 cM), mt cDNA ca genome cy la m Pht trin qun th NIL s gip cho cc nh chn ging nh gi ng n nh hng ca cc alen iu khin tnh chng chu nhm ca la hoang Oryza rufipogon du nhp vo ging la trng. Cc qun th hi giao qui m ln dn xut t qun th NIL c bit, cng c th c khuyn co trong thc hin yu cu fine mapping v cloning nhng QTL mc tiu (By v ctv. 2003) Nghin cu su hn v c ch sinh l cy trng v cc gen iu khin tnh chng chu nhm trong cy la s cho chng ta bit c s tin ha v tnh cht a dng ca hin tng chng chu nhm trong cy la v nhng loi m cc khc c quan h gn gi. 5-3. CHN TO GING LA CHNG CHU NHM Vin La BSCL tin hnh thu thp mu qun th la hoang Oryza rufipogon trong nm 1989-1991 ti Trm Chim, ng Thp Mi, k hiu acc.106412, acc.106423, acc.106424 (Ngn hng gen IRRI), ni c xem l t phn nng. Vin La Quc T thc hin cp lai gia IR64 v nhng qun th ny. Con lai F2 c chuyn v Vin La BSCL thanh lc trong iu kin t phn Tri Ho An, Phng Hip, Cn Th.
Ba dng c trin vng c kho nghim l AS1007 IR64 / Oryza rufipogon (Acc.106412) AS833 IR64 / Oryza rufipogon (Acc.106424) AS996 IR64 / Oryza rufipogon (Acc.106412)
Chng u c thi gian sinh trng ngn (95-100 ngy), dng cy ci tin, nng sut cao, chng chu ry nu v bnh o n, chng chu c c nhm, thch nghi trn t phn BSCL, c bit l ging AS996. T 38064 ha nm 2000, ging AS996 pht trin gn 100 nghn ha vo nm 2002 Bng 7: Kt qa thanh lc nhm (30ppm Al) giai on m
Vt liu Oryza rufipogon (acc.106412) AS996 OM1490 OM1314 C ung (chun chng chu) IR29 (chun nhim) LSD0.05 Chiu di r tng i (RRL) 1.158 1.133 1.058 0.899 0.843 0.514 0.230
Bng 8: Kho nghim nng sut ging la v ng xun 2000 trn vng t c vn
Ging AS996 OM1870 OM2401 IR64 CM16-27 OM2031 T.bnh LSD 0.05 Tn Thnh 7.93 7.87 7.47 7.33 7.13 6.60 7.73 Mc Ha 1 7.15 7.01 6.30 6.94 6.44 7.08 6.82 Mc Ha 2 5.40 4.25 4.34 4.68 3.88 3.89 4.41 Th Th Tn Tr Vnh Tha 1 Tha 2 Hng 8.30 7.20 5.98 5.53 7.13 6.86 5.89 5.87 7.00 7.53 5.88 5.93 7.20 6.40 5.69 6.47 7.67 7.67 5.88 5.73 7.10 6.83 5.57 5.53 7.42 7.08 5.82 5.84 Th X 6.50 6.73 6.26 5.93 6.38 6.26 6.35 Chu Thnh 5.60 5.23 5.53 4.80 4.63 4.47 5.04 Trung bnh 6.62 6.31 6.24 6.16 6.15 5.92 0.435
Bng 9: Phn tch tng tc gia kiu gen v mi trng (ng xun 2000, h thu 2000)
Ging AS996 OM1840 IR64 OM2401 CM16-27 OM2031 Ging OM2401 AS996 OM2395 CM42-94 OM2031 IR64
ng xun 2000 Trung bnh H s gc 6.48 0,84 6.41 1,13 6.30 0,81 6.24 0,89 6.19 1,19 5.93 1,12 H thu 2000 Trung bnh H s gc 4.60 1.85 4.52 0.71 4.49 1.42 4.33 0.68 4.25 1.12 4.14 0.22
SE 0,141 0,082 0,183 0,128 0,108 0,113 SE 0.238 0.251 0.409 0.172 0.235 0.271
Bng 10: Kho nghim AS996 trn vng t ph sa c nc ti, ng xun 2000 (t/ha)
Ging OM2031 AS996 OM1870 CM16-27 OM2401 IR64 Ging OM2031 AS996 OM2395 CM42-94 OM2401 IR64 Tn Chu 5.90 8.67 7.50 6.83 6.51 6.85 An Ph 6.70 7.60 6.70 6.30 6.23 6.80 Chu Ph 7.73 8.25 7.53 7.65 7.45 8.57 Ph Tn Long Xuyn Thoi Sn 7.33 7.73 7.17 7.70 7.37 7.00 Ch Mi 5.30 5.50 4.90 4.80 5.00 4.60 Trung bnh 7.05 7.69 7.35 7.14 7.14 7.27 Trung bnh 5.44 5.88 5.67 5.54 5.44 5.48

ng xun 2000 8.00 6.33 8.14 5.67 7.55 7.00 7.04 6.50 7.68 6.73 7.61 6.33 H thu 2000 Thoi Long Dien Ph Tn Sn 5.80 5.00 4.40 6.10 4.90 5.30 7.06 4.70 5.00 5.90 5.60 5.10 5.30 5.40 5.30 5.80 4.80 5.40
Bng 11: Tng tc gia kiu gen x mi trng trn vng t ph sa c nc ti

Trung bnh 7.06 7.69 7.35 7.14 7.15 7.27 ng xun 2000 H s gc 1.304 1.555 0.334 0.728 0.711 1.367 SE 0.529 0.798 0.169 0.326 0.304 0.365 Trung bnh 5.90 5.49 5.69 5.47 5.55 5.31 H thu 2000 H s gc 1.255 1.043 1.254 0.499 0.679 1.270 SE 0.247 0.209 0.370 0.181 0.204 0.055
AS996 OM2031 OM2395 OM2401 CM42-94 IR64
Ging AS996 c cng nhn ging quc gia vo nm 2002, l ging c lai to thnh cng theo chin lc du nhp gen mc tiu t la hoang vo la trng chng chu c c nhm
Hnh 5-4: Bn QTL cc tnh trng chng chu nhm vi 151 marker loci t 171 dng RIL ca IR64 x Oryza rufipogon (acc 106424). Tnh trng CRL: chiu di r i chng, SRL: chiu di r trong iu kin x l nhm, RRL: chiu di r tng i (By v ctv. 2003)
IR1552 x Azucena (Wu et al 2000)
CT9993x IR62266 (Nguyen et al 2002)
OM269 x Chim bu (Nguyen et al 2001)
IR64 x O. rufipogon
BS125 x O. longistaminata //BS125 (Causse et al 1994)
IR1552 x Azucena (Wu et al, 2000)
CT9993 x IR64 x IR62266 O. rufipogon (Nguyen et al 2002)
Hnh 5-5: So snh QTL iu khin tnh chng chu nhm trn (a) nhim sc th s 1, v (b) nhim sc th s 9 (By v ctv. 2003)
Cy la IR64 x O. rufipogon
Cy la IR1552xAzucena (Wu v ctv. 2000)
Barley (Tang v ctv. 2000)
La m Rye (Riede& (Miftahudin Anderson 1996) v ctv. 2002)
Hnh 5-6: So snh QTL iu khin tnh trng chng chu nhm gia nhng cy thuc nhm m cc (By v ctv. 2003)
TI LIU THAM KHO Ahn S, SD Tanksley. 1993. Comparative linkage maps of rice and maize genomes.Proc Natl Acad Sci USA 90:7980 -7984 Aniol A, IP Gustafson.1984.Chromosome location of genes controlling aluminum tolerance in wheat, rye, and triticale.Can J Genet Cytol 26:701-707 Bona L, BF Carver, RJ Wright, VC Baligar. 1994. Aluminium tolerance of segregating wheat population in acid soil and nutrient solutions.Commu Soil Sci Plant Anal 25:327 339 By ND, DS Brar, BC Buu, NV Tao, PN Luong, HT Nguyen. 2003. Identification and mapping of the QTL for aluminum tolerance introgressed from new source, Oryza rufipogon Griff, into indica rice (Oryza sativa L.). Theor Appl Genet 106:583-593 Bi ch Bu. 1987. Nghin cu mt vi c tnh quan trng ca cc ging la nc su a phng ng Bng Sng Cu long cho cng tc chn ging. Lun n PTS Khoa hc nng nghip. Vin KHKTNNVN, H Ni. 112 pp. Causse MA, TM Fulton, YG Cho, SN Ahn, J Chunwongse, K Xu, J Xian , Z Yu, PC Ronald, SE Harrington, G Second, SR McCouch, SD Tanksley. 1994. Saturated molecular map of the rice genome based on an interspecific backcross population.Genetics 138:1251-1274
Coronel VP. 1980. Response of rice and wheat at seedling stage to aluminium. Peper presented at IRRI. Saturday seminar, Nov. 8, 1980, Los Banos, Philippines. 55 p. Gale MD, KM Devos.1998. Comparative genetics in the grasses. Proc Natl Acad Sci USA 95:1971-1974
Gallego FJ, B Calles, C Benito.1998. Molecular markers linked to the aluminum tolerance gene Alt1 in rye.Theor Appl Genet 97:1104 - 1109 Goff SA, D Ricke D, TH Lan, G Presting G,et al. 2002. A draft sequence of the rice genome (Oryza sativa L.ssp.japonica ). Science 296:92-100 Gomez KA and AA Gomez. 1984. Statistical procedures for Agricultural Research, IRRI, Second edition. 680 pages
Harushima Y, M Yano, A Shormura, M Sato, T Shimano, Y Kuboi, T Yamamoto, SY Lin, BA Antinio, A Parco, H Kajiya, N Huang, K Yamamoto, N Nagamura, N Kurata, GS Khush, T Sasaki. 1998. A high-density rice genetic linkage map with 2,275 markers using a single F2 population.Genetics 148:479 494
HilleRisLambers D, and P Sittiyos. 1985. Rice breeding for deepwater areas. Paper presented at the Int. Rice Research conference, 1-5 June 1985. IRRI. 17p. (mimeo). Howeler RH and LP Cadacid. 1976. Screening of rice cultivars for tolerance to Al - toxicity in nutrient solutions as compares with a field screening method. Agron.J. 68 (4): 551-555.
Khatiwada SP, D Senadhira, AL Carpena, RS Zeigler, PG Fermadez. 1996.Variability and genetics of tolerance for aluminum toxicity in rice (Oryza sativa L.).Theor Appl Genet 93:675 681 Kochian LV. 1995.Cellular mechanisms of aluminum toxicity and resistance in plants.Annu Rev Plant Physiol Plant Mol Biol 46:237-260 Lander ES, D Bostein.1989. Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps.Genetics 121:185-181 Lander ES, P Green, J Abrahamson, A Barlow, DJ Daly, SE Lincoln, L Newburg. 1987. MAPMAKER:an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations.Genomics 1:174-181 Lima M, M Filho, PR Furlani. 1995. Diallel cross among inbred lines of maize differing in aluminum tolerance.Braz J Genet 18:579-584 Magnacava R, CO Gardner, RB Clark.1987. Inheritance of aluminum tolerance in maize. In: Gabelman HW, Loughman BC (eds): Genetic aspects of plant mineral nutrition. Martinus Nijhoff Publishers, Dordrecht, The Netherlands, pp 201-212
Martinez CP. 1976. Aluminum toxicity studies in rice (Oryza Sativa L.) Ph.D. Thesis, Oregon State University, Corvalis, Oregon. 113g.
McCart GD and EJ Kamprath. 1965. Supplying Ca and Mg for cotton on sandy low cation exchange capacity soils. Agronomy. J. 57: 404-406. Miftahudin G, J Scoles, JP Gustafson. 2002. AFLP markers tightly linked to the aluminum tolerance gene Alt3 in rye (Secale cereale L.).Theor Appl Genet 104:626-631 Nguyen VT, BD Nguyen, S Sakarung, C Matinez, AH Paterson, AH Nguyen. 2002. Mapping genes controlling Al tolerance in rice:comparing different genetic backgrounds.Mol Genet Genomics (in press) Nguyen VT, LD Thanh, AH Paterson, LT Binh, HT Henry. 2000. Rapid screening method for aluminum tolerance in rice in nutrient solution (in Vietnamese with English abstract,ISSN: 0866-8566). J Genet Appl 2:5-11 Nguyen VT, MD Burrow, HT Nguyen, BT Le, TD Le, AH Paterson. 2001. Molecular mapping of genes conferring aluminum tolerance in rice (Oryza sativa L.).Theor Appl Genet 102:1002-1010 Panaud O, X Chen, SR McCouch. 1996. Development of microsatellite markers and characterization of simple sequence length polymorphism (SSLP)in rice (Oryza sativa L.). Mol Gen Genet 252:597-607 Paterson AH, ES Lander, JD Hewitt, S Paterson, S Lincoln, SD Tankley. 1988. Resolution of quantitative traits into Mendelian factors by using a complete linkage map of restriction fragment length polymorphisms.Nature 335:721-726 Riede CR, JA Anderson. 1996. Linkage of RFLP markers to an aluminum tolerance in wheat.Crop Sci 36:905-909
Sibov ST, M Gaspar, MJ Silva, LMM Ottoboni, P Arruda, AP Souza. 1998. Two genes control aluminum tolerance in maize:genetic and molecular mapping analyses.Genome 42:475-482 Singh K, T Ishii T, A Parco, N Huang, DS Brar, GS Khush. 1996. Centromere mapping and orientation of the molecular linkage map of rice (Oryza sativa ).Proc Natl Acad Sci USA 93:6163-6168 Tang Y, ME Sorrels, LV Kochian, DF Garvin. 2000. Identification of RFLP markers linked to barley aluminum tolerance gene Alp .Crop Sci 40:778-782 Temnykh S, G DeClark, A Lukashova, L Lipovich, S Cartinhour, SR McCouch. 2001. Computational and experimental analysis of microsatellites in rice (Oryza sativa): frequency, length variation, transposon association,and genetic marker potential.Genome Res 11:1444-1452 Thawornwong N and A van Dibst. 1974. Influence or high acidity and aluminum on the growth of lowland rice. Plant and soil 41: 141-159.
van Breemen N, and LJ Pons. 1978. Acid sulfate soil and rice. Pages 739-761. Soil and rice. IRRI, Los Banos Philippines.
Wang D, J Zhu, Z Li , AH Paterson. 1999. Manual for QTLMapper.Version 1.0. A computer software for mapping QTLs Wang GL, DJ Mackill, JM Bonman, SR McCouch, MC Champoux, RJ Nelson. 1994. RFLP mapping of genes conferring complete and partial resistance to blast in a durably resistant rice cultivar.Genetics 136:1421-1434 Wu P, CD Liao, B Hu, KK Yi, WZ Jin, JJ Ni , C He. 2000. QTLs and epistasis for aluminum tolerance in rice (Oryza sativa L.) at different seedling stages.Theor Appl Genet 100:1295 1203 Xiao J, J Li, S Grandillo, SN Ahn, L Yuan, SD Tanksley, SR McCouch SR. 1998. Identification of trait-improving quantitative trait loci alleles from a wild rice relative,Oryza rufipogon .Genetics 150:899-909 Xiong L, KD Lui, XK Dai, SW Wang, CG Xu, DP Zhang, MAS Maroof, T Sasaki, Q Zhang. 1997.A high density RFLP map based on the F 2 population of a cross between Oryza sativa and O.rufipogon using Cornell and RGP markers.Rice Genet Newslett 14:110115 Xuan VT, LT Duong, and VT Guong. 1982. Deepwater rice trials in farmer field in the Mekong Delta in Vietnam. Pages 271-278 in Proceeding of the 1981 Int. Deepwater rice Workshop IRRI, Los Banos, Philippines. Yoshida S, DA Forno, JA Cock, KA Gomez. 1976. Laboratory manual for plant physiological studies of rice,3rd edn.International Rice Research Institute, Manila, The Philippines Yu J, S Hu, J Wang, GKS Wong et al. 2002. A draft sequence of the rice genome (Oryza sativa L. ssp.indica ). Science 296:79-92 Zhang WP, XY Shen, P Wu, B Hu, CY Liao. 2001. QTLs and epistasis for seminal root length under a different water supply in rice (Oryza sativa L.).Theor Appl Genet 103:118-123

Chng 6
C S DI TRUYN TNH CHNG CHU THIU LN
6-1. GII THIU CHUNG Hin tng thiu ln trong t canh tc rt ph bin trong vng trng la Chu , nht l t phn, t acid min Nam Vit Nam. Hin tng thiu ln do nhiu nguyn nhn khc nhau, ty theo tnh cht ca t trng la: kh nng c nh ln ca t, hm lng ln d tiu v ln tng s qa thp trong t. Chu , din tch t trng la b thiu ln nghim trng c khong 35 triu ha (Ni v ctv. 1998). S khc bit c ngha gia cc ging la chng chu hay khng chng chu vi iu kin thiu ln c ghi nhn (IRRI 1971, 1976, Katyal v ctv. 1980, Gunawardena v ctv. 1978, Ikehashi v Ponnamperuma 1978, Senanayake 1984, Fageria v ctv. 1988), m ra trin vng chn to ging la nng sut cao kt hp vi tnh chng chu thiu ln (phosphorous deficiency). Nhng ging la nh vy c th c chia thnh hai dng hnh Ging chng chu iu kin P trong t cao, nhng kh nng c nh P rt ln Ging chng chu iu kin P trong t rt thp, khng c hin tng c nh
Trong dng hnh th hai, bin php bn thm phn ln c tnh kh thi v hiu qa hn ht, nhng nhng vng la nc tri, ngi ta kh c th thuyt phc nng dn bn phn ln cho n khi h thy c bn ln thc s mang li hiu qa v nng sut, v chng minh cho h thy p ng ca phn ln rt chm v t t cho cy la s dng, so vi hiu qa tc th ca vic bn phn m. Ln l mt nguyn t v cng cn thit i vi sinh vt. N l thnh phn quan trng trong kin trc ca phosphate ng, nucleic acid, coenzyme, v phospholipid. L mt thnh phn ca nucleic acid, n tham gia trong qa trnh phn ct t bo v truyn tn hiu di truyn i vi nhng tnh trng. N cn c chc nng d tr nng lng di dng adenosine triphosphate cng nh vn chuyn nng lng trong sut qa trnh tng hp ng, tinh bt v protein. Di dng nicotamide adenine dinucleotide phosphate, ln ng vai tr vn chuyn ion H+ trong chu trnh Krebs v chu trnh phn gii glucose (glycolysis) Cy trng hp thu ln t trong t. Tt c P trong t bt ngun t microcrystallite apatite ca m magmatic (Chaubey v ctv. 1994) 6-1-1. t thiu ln Thng thng, hin tng thiu dinh dng trong t i km theo pH t. C hai nhm t acid v t kim u c hin tng thiu ln. Tnh cht acid ca t lm gia tng hm lng nhm, mn-gan v st trong dung dch t (Ikehashi v Ponnamperuma 1978, Clark v Brown1974). Ln tc ng vi Al, Mn, v Fe di dng hp cht kh ha tan. Trong t kim, P d tiu ty thuc vo hot tnh ca Ca. Phn P c bn vo b gi li mt ngoi theo c ch hp ph (adsorption) v b kt ta dng calcium phosphate, nu chng duy tr dng ny, cy khng th hp th c, dn dn, ln c chuyn i thnh nhng hp cht apatite kh tan. S thiu ln xy ra kh ph bin trong t trng la Bangladesh, Cambodia, Nht, nepal, Philippines, ng Nam Trung Quc, Sri Lanka, i Loan, Thi Lan v Vit Nam (Ikehashi v Ponnamperuma 1978) 6-1-2. Hin tng thiu ln trn cy la P l mt nguyn t di chuyn trong cy ni chung. Xt trn gc dinh dng, ln di chuyn t l gi sang l non. Hin tng u tin c ghi nhn trn l gi, biu th mu xanh
m. Trong sut thi gian b stress do thiu ln, sc t trn l la cng c th xut hin, cho kt qa pha trn gia hai mu xanh . Cy gim cng lc v nng sut. R cy tr nn mc di ra, mnh khnh, i khi dng si mng nh ch. Trong trng hp cy la, hin tng thiu ln biu hin giai on rt sm, cy thp ln, cn li, chi km. L la tr nn ngn, hp chiu ngang, mc thng v c mu xanh m. Bi v P di chuyn, nn l la non vn cn th hin s khe mnh bnh thng so vi l gi, c th l gi chuyn sang mu nu v cht. 6-1-3. Biu hin ca ging la chng chu thiu ln Mt ging la c gi l chng chu vi stress do thiu P khi ging y sn xut ra mt khi lng cht kh nhiu hn ging nhim khi chng c trng trong iu kin thiu ln (Chaubey v ctv.1994), v chng sn sinh ra lng cht kh tng ng vi cc ging nhim khi c cung cp ln ti ho. u im ca ging chng chu nh vy l chng c th duy tr kh n nh c hai iu kin c ln bnh thng, hoc ln c cung cp vi mt lng t cho cy (Gerloff 1987). Mt ging cy trng va chu c iu kin t ngho ln, va chu c iu kin bn ln thp hay bn bnh thng c nh ngha l ging khng p ng vi P (P-non responsive genotype) (Gerloff 1987) Ngi ta cha th nh tnh mt cch chnh xc nhng thuc tnh ca s kin mc hiu qa ca ln i vi cy trng (P-efficiency). Khi nim P-efficiency l mt tng c nhiu nh khoa hc cp. Do , c nhiu nh ngha khc nhau ca cc nh nghin cu i vi khi nim ny. Clark v Brown (1974 a) cho rng: hiu qa p ng ca P l kh nng ca cy trng mt mc P cho php tch ly hm lng P cao hn. Koyama v ctv. (1973) cho rng: mt ging biu hin tnh chng chu ln phi c nng lng dinh dng cao hn i vi P trong t so vi ging nhim. Ging chng chu thiu ln l ging c s lng cht kh c sn sinh ra trn n v hm lng P c hp thu bi cy trng (Chaubey v ctv. 1994)
Tuy nhin, nhng nh ngha ny trn u thng nht: ch s thu hoch (HI = harvest index) l thuc tnh quan trng ca hiu qa p ng ca P Fox (1978) cho rng hiu qa p ng ca P l s lng cht kh c th thu hoch c trn mt n v thi gian, trong iu kin cy c trng trong t thiu ln Clark (1976) cho rng cy c hiu qa p ng vi P cao pht trin tt hn, sn sinh nhiu cht kh hn, biu hin hin tng thiu ln t hn so vi cy c hiu qa p ng vi P thp, khi chng c trng so snh trong iu kin t thiu ln Vi tc gi khc nh ngha v hiu qa p ng ca P nh l nhng gi tr o lng ca trng lng cht kh trn n v ln c s dng, v trn n v ln hp thu (Chaubey v ctv. 1994) Blair v Cordero (1978) cho rng mt cy c hiu qa p ng vi P khi n biu hin kh nng hp thu P cao trn n v chiu di r, hoc trn n v din tch
Nhng kt qa nghin cu v sinh mi hc c thc hin i vi cc loi hoang di cho thy kh nng hp thu P trn mt n v chiu di r hay trn mt n v din tch v vic s dng P bn trong cy, khng c th c xem nh chc nng thch nghi ca cy trong iu kin b stress do thiu ln (Chapin 1980) Do , mc thch nghi ca cc thng s ny c xem nh nhng tiu chun c tnh cht phn lp ging chng chu v ging nhim, vn cn l cu hi ln cha c gii p. Mt trong nhng tiu chun phi la chn nh gi hiu qa p ng vi P c th l s tch ly P trn n v trng lng cht kh c tch ly trn mt n v thi gian (Chesholm v Blair 1988b)
Tc tng trng chm lm gim stress c tnh cht bin dng ca cy trng trong iu kin t thiu ln, l mt tin trnh rt c hiu qa chn lc cy c hiu qa p ng vi P cao (Chapin 1980). Chesholm v Blair (1988a) cho rng tc tng trng tng i v s phn b cht kh n r v chi thn c th c s dng lm tiu chun chn lc cy chng chu v cy nhim vi stress do thiu ln tt hn so vi phng php thu hoch trng lng cht kh giai on cui cng Trong cy la, nht l cy la dng hnh ci tin, nng sut cao, s thiu ln th hin cc yu t hn ch rt nghim trng cho s tng trng (Gunawardena v Wijeratne 1978). So snh vi cc loi cy trng khc, cy la c sc chng chu thiu ln tt hn ht (Chaubey v ctv. 1994) Ging la chng chu thiu ln s to ra kh nng hp thu ln cao hn ging nhim (Koyama 1973). Ging la c hiu qa p ng vi P c xu hng hp thu mt lng ln P v s dng n mt cch c hiu qa trong iu kin trng chng t ngho ln (Katyal 1984) Tnh chng chu s thiu ln ca ging la c o lng bng gi tr: s chi / cy, nng sut (Gunawardena 1979). Trng lng kh ca chi l tnh trng rt nhy cm vi hin tng thiu ln (Fageria v ctv. 1988), k n l trng lng kh ca r, s chi. 6-2. K THUT THANH LC K thut thanh lc ging la chng chu stress do thiu ln c IRRI xut: Bn ht thc ny mm c chn t mi ging la c t trn mt tm li nylon, c khung l tm xp trng, gip cy ni trong dung dch dinh dng Yoshida c cha 0,5 mg / lt P t trong thau nha c th tch 5 lt (Ponnamperuma 1977). Mt nghim thc khc vi 4 ht ny mm / ging c trng trong mi trng dinh dng i chng vi 10 mg / l P. Ging la IR26 c xem nh ging i chng chng chu (T), v IR36 l i chng nhim (S). Sau mi lot bao gm 50 ging thanh lc l xen gia bi ging i chng T v S. iu chnh pH ca mi trng dinh dng hng ngy pH 6,0, v dung dch dinh dng c thay mi tun mt ln Bn tun sau khi gieo, hiu qa p ng vi P ca mi ging la c quan st v cho im theo phn trm s chi tng i (relative tiller % = RT %) S chi trong dung dch cha 0,5 ppm P RT% = --------------------------------------------------------------- x 100 S chi trong dung dch cha 10,0 ppm P RT% c chuyn i thnh thang im chun trong h thng nh gi (SES) ca Vin La Quc T RT% 80-100 60- 79 40- 59 20- 39 0- 19 Thang im SES 1 3 5 7 9 Hiu qa p ng vi P Cao Hiu qa Trung bnh Khng hiu qa Rt khng hiu qa
Trong iu kin ng rung, IRRI ngh phng php th nghim trn rung nng dn. Phn tch P tng s, P d tiu trc v sau khi ngp nc. Hai l c bn ln v khng bn ln u c cy, xi, nh bn, v san mt rung ging nhau. Bn 50-25-20 kg NPK / ha cho l i chng v 50-0-20 kg NPK / ha cho l x l thiu ln. Cc ging la c cy thnh hng di 5m, khong cch cy 20x20 cm. Ging chun nhim c trng sau 20 dng,
v ging chun chng chu c trng ngu nhin trong cc ging trc nghim. Vo lc 4-8 tun sau khi cy, ngi ta tin hnh o m s chi / cy. Vo lc la chn, chn 20 khm la / ging hng gia thu hoch, ri ghi nhn nng sut. Hot ng ca acid phosphatase (APA) c xt nghim bng cch s dng pnitrophenyl-phosphate lm hot cht pH 4,0 (Clark 1976) Xc nh hm lng ln trong chi v r la tnh bng mg / cy v xc nh hiu qa s dng P chi (SPUE), r (RPUE) trn mt n v ln c hp th (Ni v ctv. 1996) 6-3. C CH CHNG CHU THIU LN Tnh trng chng chu stress do thiu ln ca cy la c th c xem xt trn hai c ch khc nhau (Ni v ctv. 1998): (1) hiu ng bn trong (internal efficiency), v (2) hiu ng bn ngoi (external efficiency). Hiu ng bn trong biu th yu cu ln ca cy mc thp Hiu ng bn ngoi l kh nng bt gi, hp thu ln khng d tiu trong t
C ch hiu ng bn trong c th c nghin cu thng qua phng php thanh lc trong dung dch Yoshida nh m t trn vi 2 nghim thc P = 0,5 mg / l v P = 10 mg/l Ni v ctv. (1996) th hin mt th nghim gii thch c ch ny nh nhau: Chn 6 cy m non / mi ging la x l 2 nghim thc ln (10ppm) v thiu ln (0,5ppm) trong dung dch dinh dng Yoshida. Ly mu vo lc 2 tun, 4 tun sau khi nui cy. Phn tch hm lng ln trong chi (SPC) v trong r (RPC), phn tch hiu qa s dng P trn tng lng cht kh chi (SPUE: shoot-P use efficiency) v r (RPUE: root-P use efficiency), tnh bng mg hm lng ln hp thu. Bin thin kiu gen i vi tnh trng chng chu thiu ln c nh ngha l hiu s gia hai gi tr thu c trong hai iu kin ln v thiu ln (Gouley v ctv. 1993) Bng 1: So snh gi tr trung bnh SPC, RPC (mg / cy) v SPUE, RPUE (g/mg) trong hai nghim thc x l ln v thiu ln, vo lc 2 tun sau khi nui cy (Ni v ctv. 1996)
Ging Tesanai 2 IR55178 IR20 pp2462-11 IR31406 Ging Tesanai 2 IR55178 IR20 pp2462-11 IR31406 0,5 ppm P SPC RPC 0,13a 0,35a 0,09b 0,29b 0,06c 0,19c 0,13a 0,36a 0,09b 0,16c 0,5 ppm P SPUE RPUE 0,36a 0,33a 0,44b 0,37a 0,48b 0,29b 0,34a 0,37a 0,33a 0,36a 10 ppm P SPC RPC 0,28a 1,60a 0,28a 1,54a 0,21b 0,89b 0,32a 2,06c 0,41c 1,82d 10 ppm P SPUE RPUE 0,11a 0,11a 0,10a 0,12a 0,09a 0,10a 0,10a 0,10a 0,09a 0,12a Gim SPC 1,25a 1,25a 0,70b 1,70c 1,66c SPUE 0,22ab 0,25ab 0,19a 0,27b 0,24ab RPC 0,15a 0,19a 0,15a 0,19a 0,25b Tng RPUE 0,25a 0,34b 0,39b 0,24a 0,24a
Nhng s theo sau cng mt ch khng khc bit c ngha mc 0,01 (DMRT) Bin thin kiu gen c ngha i vi SPC, RPC, vi s gim gi tr gia 2 nghim thc quan st 10ppm P v 0,5ppm P. Ging IR20 th hin tnh chng chu cao i vi stress do thiu P, chnh lch gi tr trung bnh thp nht chi v r (bng 1), trong khi IR31406 c xem l ging nhy cm nht vi stress
Bin thin kiu gen ca SPUE v RPUE c ngha nghim thc 0,5ppm P, nhng khng c ngha nghim thc 10ppm P. Khc bit gi tr tng ln ca SPUE v RPUE rt c ngha trong khi gim hm lng ln t 10ppm xung 0,5ppm (bng 1). IR20 th hin s khc bit r nht i vi gi tr SPUE Bng 2: Hot ng ca acid phospatase r (RAPA) v hot ng acid phosphatase tit ra bn ngoi (EAPA) tnh bng mol PNP/g trng lng r ti / gi, trong iu kin thiu ln v ln, giai on 2 v 4 tun sau khi nui cy trong mi trng (Ni v ctv. 1996)
Ging Tesanai 2 IR55178 IR20 pp2462-11 IR31406 Tesanai 2 IR55178 IR20 pp2462-11 IR31406 0,5ppm P 10ppm P RAPA EAPA RAPA EAPA (2 tun sau khi trng) 12,57ac 121,71a 29,73a 172,94a 13,22ab 68,97b 27,37c 98,26b 15,23b 116,68a 37,93b 141,26c 13,35ab 129,90a 25,37c 183,32a 10,66c 96,00b 19,75d 80,84c (4 tun sau khi trng) 15,81a 158,10a 26,55a 179,16a 21,75b 175,26b 33,13b 200,48b 19,07c 144,55c 30,14c 147,81c 14,90d 155,16a 22,65d 176,77a 13,62d 107,87d 21,29d 117,55d P-stress kch hot Psi-RAPA Psi-EAPA 51,23a 29,29b 24,58c 53,42d -15,16e 21,06a 25,22b 3,26c 18,61d 9,68c 17,16a 14,15b 22,02c 12,02d 9,09e 10,74a 11,38a 11,07a 7,75b 7,67b
Nhng s theo sau cng mt ch khng khc bit c ngha mc 0,01 (DMRT) Psi: vit tt t ch P-stress induced Hot ng ca men acid phosphatase trong r (RAPA: root-acid phosphatase acitivities) v acid phosphatase tit ra bn ngoi (EAPA: excreted acid phosphatase acitivities) c xc nh. Sau , ngi ta c on gi tr P-stress kch hot RAPA v EAPA bng cch tnh hiu s chnh lch gia RAPA v EAPA trong 2 nghim thc ln v thiu ln. Khi thiu ln, ging la b kch hot sn sinh ra men acid phosphatase r v bn ngoi vng r. Bin thin kiu gen i vi gi tr RAPA v EAPA rt c ngha mc 0,01 (bng 2) Gi tr Psi-RAPA khng n nh gia cc ging so vi gi tr Psi-EAPA. Sau 2 tun, gi tr Psi-RAPA ca ging pp2462-11 cao nht, v gi tr Psi-EAPA ca IR20 cao nht, sau l ging Tesanai-2. Bin thin kiu gen ca gi tr Psi-EAPA gim theo thi gian sinh trng ca cy (bng 2). Hayashi v Takijima (1953) quan st hot ng phosphatase ngoi bo r la trong iu kin thiu ln s gia tng gp 2-fold so vi r la mc trong iu kin bnh thng. Nhng nghin cu trc y ghi nhn r cy la 12 ngy tui c hot ng phosphatase cao gp 5 ln trong t thiu ln so vi cy la bnh thng, v sau hot ng ny gim xung sau 20 ngy tui (Hirata v ctv. 1982) Psi thay i theo hm lng ln trong r v trong chi. Sau hai tun, Psi-EAPA tng quan cht vi s gim SPC, v sau 4 tun Psi-EAPA tng quan cht vi s gim RPC (Ni v ctv. 1996). Tng quan m cng c ghi nhn gia s gim SPC v s tng RPUE, gia s gim SPC v s gim RPC. Ging la chng chu thiu ln c nh gi trn c s mc suy gim hm lng ln tch ly v mc gia tng hiu qa s dng P trong iu lin hm lng P thp dung dch dinh dng C ch trnh n stress do thiu ln ca cy trng c th hin bng cch iu tit s tng trng chm li trong t ngho ln, th d nh cc loi thuc Agrostis, Eucalyptus, Carex, v.v.. (Chaubey v ctv. 1994) 6-3. DI TRUYN TNH CHNG CHU THIU LN
Ngi ta c gng tm hiu nhng tnh trng di truyn c lin quan n hin tng chng chu s thiu ln, thng qua tnh trng hnh thi hc, sinh l hc, sinh ha hc trong qa trnh hp thu khong cht, c ch hp thu Tnh trng hnh thi hc c lin quan l chiu di r, ng knh r, lng ht, nhng chng thng th hin vi gi tr bin thin cao do nh hng mnh m ca mi trng cha kim sot c trong th nghim Tnh trng hp thu P trong t v tnh trng s dng P c hp thu trong cy l hai yu t khng lin kt vi nhau Nhng ging la IR8, IR20, IR36, IR64, Mahsuri, TCNJ1, H4, Bg90-2 c nh gi l ging c hiu qa p ng vi P cao, v chng c mc thch nghi rng, nn c xem nh vt liu cho nghin cu di truyn v to ging mi Thun li ca k thut sinh hc phn t gip cho cc nh nghin cu c th thc hin bn di truyn d dng hn, nht l i vi tnh trng s lng nh th ny. Majumder v ctv. (1989) xut gi thuyt rng tnh trng chng chu thiu ln ca cy la l mt tnh trng di truyn s lng vi nh hng rt mnh m v u th lai. Trong khi , kt qa nghin cu ca Chaubey v ctv. (1994) cho nhng nhn xt khc hn QTL iu khin tnh chng chu thiu ln c thc hin trn qun th con lai ca Nipponbare x Kasalath (japonica x indica) (Wissuwa v ctv. 1998). Tnh chng chu ny biu hin thng qua s khc bit gia cc kiu gen v mc hp thu P (P-uptake) v mc hiu qa s dng P bn trong. Mt QTL iu khin tnh trng P-uptake c tm thy trn bn vai nhim di thuc nhim sc th s 12, qung 13,2 cM, lin kt vi marker k cn C443-G2140. V tr xc nh 54,5 cM, v cch marker C443 mt khong cch di truyn c gi tr 3,0 cM. Nhng QTL th yu c tm thy trn nhim sc th s 2, 6 v 10 (Wissuwa v ctv. 2002). Cng trnh u tin ca Ni v ctv. (1998) v s c mt ca mt QTL ch lc iu khin tnh chng chu thiu ln c cng b trn nhim sc th s 12. N dng nh cha r rng v nh hng ca QTL i vi mc hp thu P hoc hiu qa s dng P. Mt nghin cu su hn trn qun th dng gn nh ng gen (NIL) c pht trin bi ba ln hi giao vi Nipponbare. Dng NIL ny c tn l NIL-C443 vi 91% nn tng di truyn ca Nipponbare, nhng mang trong n mt on Kasalath 50-cM trn nhim sc th s 12, th hin cp marker thuc qung C443-G2140, ni nh v ca mt QTL gi nh iu khin tnh chng chu thiu ln. nh hng ca QTL ch lc ny mt ln na xc nh c trong hai iu kin th nghim: ng rung v trong chu (Wissuwa v Ae 2001a, 2001b). trong iu kin trng la trn t thiu ln, dng NIL-C443 c kh nng hp thu P gp 3 ln so vi Nipponbare. Trong khi , c hai khng biu hin s khc bit g khi trng chng trong t c ln. nh hng ca QTL C443 rt chuyn bit i vi hin tng thiu ln. i vi yu cu pht trin chin lc MAS (chn ging nh marker phn t), hay yu cu map-based cloning, v tr ca mt QTL cn c bit r vi mc tin cy v chnh xc cao (Paterson v ctv. 1990, Yano v Sasaki 1997, Yano v ctv. 2000) Hin nay, QTL C443 c thc hin bn vi mt qung 13,2 cM, v vic xc nh QTL C443 da trn c s mt dng NIL c cha 50-cM Kasalath segment. iu ny cha cho yu cu thc hin MAS, bi v mt qung 13 cM c th cn cha nhiu gen khng mong mun khc xen vo, nht l i vi ging Kasalath, donor ca QTL C443, mt ging la bn a c c tnh nng hc tng qut rt km. Hn na, c th mt QTL c hnh thnh trn bn c qung gi nh qa rng nh vy tng ng vi mt cluster ca nhng gen, mi gen c nh hng tng i nh. Qun th mapping c pht trin bng phng php hi giao hon ton ph hp vi yu cu thnh lp bn QTL (Yano 2001) v hu ht cc yu t di truyn khng lin quan n QTL ny trong qun th phn ly qa lu
6-3-1. Lp bn QTL bng AFLP
Qun th cn giao ti t hp (RIL) F8 ca cp lai IR20 x IR55178-3B-9-3 bao gm 285 dng c s dng trong nghin cu (Ni v ctv. 1998). IR20 c xem l ging cho gen chng chu. Phn tch AFLP theo phng php ca Vos v ctv. (1995). c hai h thng PstI/MseI v EcoRI/MseI u c s dng pht sinh ra nhng marker a hnh AFLP, vi 58 cp primer. Mt b AFLP marker trn bn ca qun th n bi kp IR64/Azucena v qun th RIL ca IR74/FR13A c s dng nh nhng marker c tnh cht neo i vi mi nhm lin kt gen. tt c nhng AFLP marker c tm thy trn 12 nhm lin kt gen LOD > 3, trong phn tch MAPMARKER (Lander v ctv. 1987). Khong cch trn bn di truyn (cM) c tnh da theo hm Kosambi (Kosambi 1944) iu tra DNA trn b m cho thy: c 26 RFLP marker a hnh. Nh vy 26 RFLP marker ny v 30 AFLP c dng nh nhng anchor (neo) hnh thnh bn di truyn (Ni v ctv. 1998), vi tng s chiu di l 1371,8 cM, trung bnh ca qung l 7,62 cM, vi gi tr nh nht trn nhim sc th s 4 (2,07 cM), v gi tr ln nht trn nhim sc th s 3 (11,49 cM). C 7 on cn trng >30cM, v 2 on >50cM trn nhim th s 1 v 2 (hnh 6-1). Bng 3: QTL iu khin cc tnh trng c lin quan n hin tng chng chu thiu ln: RTA: kh nng nhnh tng i, RSDW: trng lng kh tng i ca chi, RRDW: trng lng kh tng i ca r, theo phng php phn tch SMA (singlemarker analysis) (Ni v ctv. 1998)
Tnh trng RTA QTL Qung gia 2 NST marker 1 1 E4/M1-8E4/M3-86 2 E3/M8-3E1/M7-73 RG9-RG241 12 LOD Trung bnh alen IR20 IR55178 73,7 2,42 E4/M1-8 93,1 99,6 99,8 98,8 61,7 64,5 66,9 64,5 67,2 54,1 70,7 71,5 70,8 52,5 45,4 49,2 48,9 49,8 50,6 104,9 130,2 132,1 131,6 92,9 96,8 Marker Pe 27,0 -34,4 -31,0 -27,8 42,9 36,3 35,2 33,0 29,4 14,3 -22,4 -21,1 -18,1 16,1 31,3 25,1 23,9 20,8 18,7 21,3 -32,3 -31,7 -28,4 47,0 42,1 R2 (%) 9,9 33,6 27,5 23,1 54,0 37,7 35,6 31,7 25,4 10,8 34,0 30,7 22,7 14,9 60,8 40,4 35,7 28,0 23,3 8,9 25,2 23,4 18,9 44,2 37,4
RSDW
1 2 3 4
E4/M1-8-E4/M3-8 E3/M8-3-E1/M7-7 E1/M7-7-75RG451 RG9-RG241
1 6 9 12
7,82 E3/M8-3 65,2 E5/M4-3 68,8 E1/M7-7 71,0 104,6 16,51 RG9 P1/M7-7 100,8 102,1 RG241 E1/M5-7 97,5 E3/M6-4 96,6 2,66 E4/M1-8 68,3 7,90 E3/M8-3 E5/M4-3 E1/M7-7 2,47 E1/M7-5 48,4 50,4 52,7 68,6
RRDW
1 2 3
E4/M1-8~E4/M3-8 E4/M8-3~ ~E1/M7-7 RG9~RG241
1 6 12
76,7 16,98 RG9 74,3 RG241 P1/M7-7 72,8 E3/M6-4 70,6 E1/M5-7 69,3 2,42 E4/M1-8 126,2 5,80 E3/M8-3 E5/M4-3 E1/M7-7 12,05 RG9 RG241 97,9 100,4 103,2 139,9 138,9

E3/M6-4 131,8 P1/M7-7 131,1 E1/M5-7 128,6 98,7 99,3 101,5 33,1 81,8 27,1
NST: nhim sc th, Pe: nh hng kiu hnh, R2: phn trm bin thin kiu hnh c gii thch bi QTL
24,2 24,6 16,8
Nhim sc th s 6
Nhim sc th s 12
Hnh 6-1: Bn QTL tnh trng lin quan hin tng chng chu thiu ln (RTA, RSDW, RRDW), qun th RIL ca t hp lai IR20 / IR55178, trn nhim sc th s 6 v 12, vi AFLP marker (Ni v ctv. 1998)
Gen ch lc iu khin tnh chng chu thiu ln c k hiu l PHO. Gen ny c pht hin nh AFLP marker: P1/M7-7 v E1/M5-7, v RFLP marker RG9 RG241 trn nhim sc th s 12 (hnh 6-1) (Ni v ctv. 1998). QTL iu khin tnh trng kh nng nhnh gii thch c 54% bin thin kiu hnh, tng t, QTL iu khin tnh trng trng lng kh ca chi: 60,8% v QTL iu khin tnh trng trng lng kh ca r: 44,2% (bng 3). QTL c pht hin trn nhim sc th s 6 marker loci E3/M8-3 v E1/M7-7 gii thch c 33,6% bin thin kiu hnh ca tnh trng kh nng nhnh, 34% trng lng kh ca chi, 25,2% trng lng kh ca r (hnh 6-1, bng 3) QTL c pht hin trn nhim sc th s 1 marker loci E4/M1-8 v E4/M3-8 gii thch c 9,9% bin thin kiu hnh ca tnh trng kh nng nhnh, 10,8% trng lng kh ca chi, 8,9% trng lng kh ca r Ch c mt QTL c pht hin trn nhim sc th s 9 marker loci E1/M7-5 v RG451 gii thch c 14,9% bin thin kiu hnh ca tnh trng trng lng kh ca chi. cy bp, trng lng kh trung bnh c s dng nh mt thng s nh gi tnh chng chu thiu ln (Reiter v ctv. 1991). Tuy nhin, kh nng nhnh tng i (RTA) vn c xem nh du chun c mc tin cy cao nht trong nh gi tnh chng chu thiu ln ca cy la (Hung 1985, Chaubey v ctv. 1994). Chng ta va tho lun trn vi 3 tnh trng: kh nng nhnh tng i (RTA), trng lng kh tng i ca chi (RSDW) v trng lng kh tng i ca r (RRDW). Chng tham gia rt tch cc vo hin tng chng chu thiu ln. Trong , RTA c th c xc nh mt cch d dng hn trong mi quan h vi tnh chng chu v c xem nh tiu chun chn lc then cht. 6-3-2. Lp bn QTL bng RFLP Cng trnh nghin cu ca Wissuwa v ctv. (1998) Nht Bn c th c xem nh th nghim kiu mu trong phn tch QTL c lin quan n tnh trng chng chu thiu ln. Nhm tc gi ny cn c trn hai hin tng c bn: kh nng hp thu P v hiu qa s dng P trong cy. Ging la japonica c tn Nipponbare (c ngun gc Nht) c lai vi ging la indica Kasalath (c ngun gc Assam, n ). Con lai F1 c lai lui vi ging ti tc Nipponbare. Chn ngu nhin 98 dng con lai BC1F1, thu hoch mi dng mt n hai ht theo phng php SSD (single seed descent) trong sut 5 th h lin tc (Lin v ctv. 1998). Tc gi s dng l ca qun th BC1F5 ly trch DNA v phn tch RFLP (restriction fragment length polymorphism) Trong khi , tc gi thc hin nh gi kiu hnh qun th BC1F16. Qun th ny cn c gi l BIL (Backcross inbred lines). 6-3-2-1. nh gi kiu hnh Bn cnh , nhm tc gi nh gi kiu hnh ca 14 ging la c kh nng hp thu P khc nhau lm chun gi tr so snh: IR26, IR36 (IRRI, Philippines), Yamadanishiki (Nht), Oryza Sabana-6, IAC47, IAC1246, CT11891-2-2-7 (CIAT, Colombia), Progresso, CNA4143, CNA4128, CNA4291, CNA7013 (Brazil), Morogsol (n ), WAB 99-84 (WARDA, Ty Chu Phi). Nghim thc khng bn ln trn t ngho ln l nghim thc x l. Nghim thc i chng c bn 60kg P2O5 / ha. Sau khi trng 125 ngy, cy la th nghim c m s chi / cy. S chi ny c ct v mang v phng th nghim, sy kh 65oC trong 5 ngy. Mu c nghin sau khi xc nh trng lng kh ca tng mu, cho qua ry 1-mm mesh. Hm lng P trong m c tnh trn 1mg mu bt l, s dng my sc k o P theo phng php Hanson (1950), sau khi x l vi HNO3, HClO4, H2SO4 (3:1:1). Hm
lng P tng s c hp thu c tnh theo trng lng cht kh v P trong m, v hiu qa s dng ln c biu th bng trng lng cht kh (g) / mg P c hp thu Bng 4: nh hng stress do thiu ln (-P) so vi nghim thc c bn ln (+P) trong 14 ging la i chng (Wissuwa v ctv. 1998)
Thng s P hp thu (mg / cy) Hm lng P m (mg P / g) Trng lng kh (g / cy) S chi +P 8,95 0,86 10,45 9,37 -P 3,48 0,67 5,19 5,00 Gim (%) 61,0 22,9 50,4 46,7 Mc gim trong qung (%) 28,9-96,5 6,8-40,4 5,3-94,1 15,4-87,5
+P: th nghim c bn 60kg P2O5 / ha -P: th nghim trn t thiu P, ln d tiu (1,0 mg P. kg-1 t, Truog-P; 4,5 mg P / kg t, Bray2-P) Bng 5: H s tng quan gia cc tnh trng mc tiu trong 98 dng BILs (Wissuwa v ctv. 1998). Tt c h s tng quan u c ngha mc 0,01. Hiu qa s dng P -0,72 Trng lng kh 0,96 -0,60 S chi 0,75 -0,52 0,74
Kh nng hp thu P (mg/cy) Hiu qa s dng P (g trng lng kh / mg P) Trng lng kh (g / cy) 6-3-2-2. Xy dng bn QTL
Bn lin kt gen bao gm 245 RFLP marker c ngun gc t chng trnh RGP (Rice genome program) ca Nht c m t bi Lin v ctv.(1998). S liu v s chi / cy v hm lng ln trong cy c chuyn i thnh gi tr log trc khi phn tch. Phn tch QTL theo phng php CIM (composite interval mapping) ca Zeng (1994), trn c s phn mm PLABQTL (Utz v Melchinger 1996, Haley v Knott 1992) Bi v m hnh chuyn mn phn tch trn qun th BIL cha hon thin, cc tc gi s dng s liu ny nh s liu F2 backcross v marker d hp t xem nh s liu thiu. Bc mt, qui trnh SIM (simple interval mapping) phi c hon tt chn lc cc gi tr cofactor. Chn trn computer cc ngng F-to-enter v F-to-drop mc 6.0 nhm ngn nga trng hp nhiu marker lin kt vi mt QTL nh l nhng cofactor. S dng nhng cofactor ny gim bin thin do yu t cn. Tc gi tm kim QTL gi nh vi phng php CIM thang im LOD > 2,80. RFLP v phn tch lin kt trong trng trng hp dng BIL (backcross inbred lines) c m t kh chi tit bi Lin v ctv. (1998). Gi tr khong cch trung bnh gia 245 RFLP marker trong bn QTL l 4,8 cM. Phn trm d hp (4,6%) khc bit khng c ngha vi gi tr hi vng 3,1%, v t l phn ly ph hp vi nh lut Mendel 3:1 i vi hu ht cc marker. Bn QTL gi nh i vi tnh trng hp thu P c tm thy trn nhim sc th s 2, 6, 10 v 12 (hnh 6-2, bng 6), gii thch c 54,5% bin thin kiu hnh ca tnh trng hp thu P. Mt trong nhng QTL ny lin kt vi C443 trn nhim sc th s 12, c nh hng chnh, v ng gp rt ln n bin thin kiu hnh (Wissuwa v ctv. 1998)
Bng 6: QTL gi nh i vi tnh trng hp thu P, hiu qa s dng P, trng lng kh, s chi trong iu kin stress do thiu ln (Wissuwa v ctv. 1998)
Qung gia hai marker Nhim th V tr (cM) Khong cch (cM) +3 +2 +6 -3 Total +3 -4 -4 Total -1 +1 -3 Total +2 -1 -3 Total LOD R2 (%) nh hng thay th +971 +705 -621 +1938 -0,346 +0,303 -0,465 -992 +1560 +3193 +1,84 +1,54 +1,86 Alen dng tnh K K N K N K N N K K K K K
Hp thu P
G227-C365 C498-R1954 R162-R2447 G2140-C443 G227-C365 C946-R1854 G2140-C443 C1488-C63 C191-C498 G2140-C443 R347-C335 C901-G1406 G2140-C443
2 6 10 12 2 4 12 3 6 12 4 12 12
106 13 32 30 106 86 28 86 10 30 32 2 30
2,82 3,52 4,70 10,74 16,25 5,22 4,35 6,57 11,40 3,08 4,71 10,50 12,89 4,37 3,90 7,87 10,97
5,8 9,8 7,7 27,9 54,5 9,8 9,4 19,1 42,1 6,4 9,7 26,5 45,4 9,8 9,5 20,6 40,3
Hiu qa s dng P
Trng lng cht kh S chi
Nhng marker nm gn QTL nht c gch di ch s trong ct 2 Khong cch tnh t marker gn nht (cM) trong ct 5 nh hng thay th c hai alen Nipponbare v Kasalath trn c s gi tr cha chuyn i thnh logarit Ct cui cng biu th alen nh hng kiu hnh ca K (Kasalath) v N (Nipponbare) Gi tr total ca khong cch bao gm tt c QTL gi nh theo m hnh multiple-QTL
Hnh 6-2: Bn QTL cc tnh trng trng lng kh (dry weight), hp thu P (P uptake), hiu qa s dng P (P-use efficiency), v s chi (tiller number), trn qun th BILs ca t hp lai Nipponbare x Kasalath, vi RFLP marker (Wissuwa v ctv. 1998)
Ba trong s bn QTL ch lc, c mt QTL rt quan trng thuc v ging Kasalath. Trong 3 QTL iu khin tnh trng trng lng kh, c 2 QTL nh v trn nhim th s 6 v 12 cng vi tnh trng hp thu P QTL lin kt vi C443 trn nhim th s 12 c nh hng rt ln i vi c 4 tnh trng mc tiu, gii thch 27,9% bin thin kiu hnh ca hp thu P, 19,1% i vi hiu qa s dng P, 26,5% i vi trng lng cht kh, v 20,6% i vi kh nng nhnh (Wissuwa v ctv. 1998). Kt lun ny tng ng vi nhn xt ca Ni v ctv. (1998). Khng c QTL no lin quan n trng lng cht kh c tm thy trn nhim sc th s 2 v 10, nhng cc nh cng nhng qung i vi tnh trng hp thu P c pht hin trong trng hp qung gia G222-C365 (LOD=2,30) trn nhim th s 2, v qung gia R1629-R2447 (LOD=2,19) trn nhim th s 10. Tng t, QTL i vi tnh trng trng lng cht kh trn nhim th s 3 th hin ng thi vi mt nh thy c vi hp thu P nhng LOD qa thp (2,24) i vi tnh trng hiu qa s dng P, c 3 QTL gi nh gii thch 42,1% bin thin kiu hnh (bng 6). QTL ch lc trn nhim th s 12 v QTL th yu trn nhim th s 2 xut hin ng v tr vi QTL ca tnh trng hp thu P (hnh 6-2). nh hng alen ca Nipponbare lm gia tng hiu qa s dng P, v alen ca Kasalath lm gia tng kh nng hp thu ln, c ghi nhn (Wissuwa v ctv. 1998). QTL gi nh trn nhim th s 4 khng tng ng vi nh trong tt c tnh trng mc tiu i vi tnh trng kh nng nhnh, 3 QTL gi nh gii thch 40,3% bin thin kiu hnh (bng 6), trong C443 gii thch 20,6%. nh hng alen ca Kasalath lm gia tng tnh trng ny (Wissuwa v ctv. 1998), mt tnh trng c xem nh tiu chun chn lc ca nhiu nh chn ging (Ni v ctv. 1998) Nhng ging chng chu stress do thiu ln cn phi phi hp c c hai tnh trng hp thu P v s dng P c hiu qa bn trong cy la. Nh vy c hai QTL chung cho c hai tnh trng ny nh v trn nhim sc th s 2 v 12 vi cng qung gia 2 marker G227C365 v G2140-C443. nh hng alen ca Kasalath trong 2 QTL ny: lm gia tng hp thu P, nhng lm gim hiu qa s dng P. Hin tng nh vy do s xut hin c hai loci gia nhng qung u c ngha, c hai lin kt rt cht ch theo kiu thc y (repulsion, dng trans), hoc ch c mt loci nm bn trong qung vi biu hin nh hng ngc i vi hp thu P v s dng hiu qa P. Chn lc cc alen ca Kasalath dng tnh ci tin tnh trng hp thu ln, ng thi n lm gim hiu qa s dng P, l khng th chp nhn c (Wissuwa v ctv. 1998). Tuy nhin, chnh kh nng hp thu P cao l iu kin chnh gip cy la chng chu hin tng thiu ln, trong khi hiu qa s dng ln c nh hng th yu trong vic gia tng tnh chng chu thiu ln. Nhn li h s tng quan bng 5, chng ta s thy r hn iu . Trong th nghim ny, trng lng kh vo lc 125 ngy sau khi trng l mt gi tr ch th tnh chng chu thiu ln, bi v chng ta khng th dng nng sut ht xc nh trong iu kin cc dng BIL th hin rt khc nhau v thi gian sinh trng, dn n ngy thu hoch rt chnh lch. Trng lng cht kh trong iu kin t thiu ln thay i ty theo kh nng hp thu P ca ging la. Fageria v ctv. (1988) khng tm thy mt s tng quan c ngha gia nng sut ht v kh nng hp thu P, trong iu kin th nghim ca nhm tc gi ny khng qa nghim trng v nh hng stress do thiu ln. Cc tc gi ghi nhn: nng sut ht ch gim khong 14% tng ng vi mc P c bn vo, v hm lng P m vn cn duy tr kh tt, trn 1mg P / gr cht kh, m mc ny, ngi ta nh ngha l ngng thiu ln i vi cy la giai on nhnh.
Mt th nghim khc ti IRRI (1994) cho thy tng quan gia hp thu ln v nng sut ht r = 0,40, v Koyama (1973) ghi nhn rng: nng sut ht ty thuc ch yu vo kh nng hp thu P, vi kh nng hp thu P cng cao, mc chng chu thiu ln cng ln. Trng lng cht kh c tng quan kh cht vi kh nng hp thu ln so vi nng sut ht trong rt nhiu nghin cu trc , do s khc bit v ging, khc bit v kh nng chuyn v P t m sinh trng sang m sinh dc (Wissuwa v ctv. 1998), v do s khc bit v ch s thu hoch (HI). Nhng nh chn ging c gng pht trin chng trnh ci tin ging mi, kt hp kh nng hp thu P vi gia tng ch s HI trong mc tiu gia tng s chng chu thiu ln, hn l tp trung mc tiu ci tin hiu qa s dng ln. Gia tng hiu qa s dng P s thu c nhng nh hng ngc li, bi v hm lng P trong ht thp l vn ca cht lng ht (Hedley v ctv. 1994). Hm lng P trong ht c nh hng rt ln n cy m, r pht trin nhanh trong giai on u, nh hng n kh nng hp thu ln. Kh nng nhnh tuyt i v kh nng nhnh tng i chnh l nhng thng tin gin tip i vi tnh trng chng chu thiu ln ca cy la (Hung 1985, Chaubey v ctv. 1994), c ngh lm gi tr ch th (indicator). N c thun li l d xc nh trong sut giai on tng trng, khng ging nh trng lng kh hoc kh nng hp thu ln. Trong 3 QTL iu khin tnh trng kh nng nhnh, c mt QTL th hin ng v tr vi QTL(C443) i vi kh nng hp thu ln v trng lng kh. Mt trong nhng yu im ca gi tr ch th gin tip (kh nng nhnh) tm kim QTL i vi tnh trng c lin quan (chng chu thiu ln) l: nhng QTL nh vy c th i din cho nhng gi tr khc nhau v kiu gen ch c lin quan n s chi m khng c ngha i vi tnh trng mc tiu sau cng. Kh nng nhnh tng qut v mc nhy cm i vi hin tng thiu ln c mi lin quan kh cao nu chng ta s dng s liu s chi tuyt i, v ngc li nu chng ta s dng s liu s chi tng i (t l gia gi tr s chi b stress do thiu ln trn gi tr s chi l bn ln y ). Nhng QTL i vi s chi trong hnh 6-2 cho thy: chng nh v khu vc 32 cM trn nhim th s 4 v 2 cM trn nhim th s 12, trong qung c mc ngha cho php, nh hng di truyn n tnh trng s chi trong iu kin khng c stress (Ray v ctv. 1996). QTL chnh trn nhim th s 12 (C443) khng kt vi vi c hai tnh trng: s chi (Ray v ctv. 1996) v s bng / bi (Lin v ctv. 1996). S dng s chi (mt tnh trng gin tip) c th cho chng ta xc nh c mt QTL chnh ca tnh trng chng chu thiu ln, nhng Wissuwa v ctv. (1998) tht bi trong vic tm kim nhng QTL gi nh c nh hng ph trong tnh trng hp thu ln, hoc tnh trng trng lng kh. Kh nng hp thu ln trong t ngho ln dng nh cn l thuc vo c im ca r la. Ngi ta tp trung nghin cu nhm gii thch nhng QTL gi nh biu th nh hng ca chng i vi tnh trng chng chu thiu ln. Mt v tr trn nhim sc th s 6 c pht hin (C498, 13-cM) vi mt QTL ph ln khu vc y lin quan n kh nng hp thu ln (Ray v ctv. 1996). QTL ny gii thch 10% bin thin kiu hnh ca tng s r trong mt cp lai indica x japonica, v alen ca indica c nh hng dng tnh. QTL trn nhim sc th s 2, 6, v 12 l nhng ng c vin tt i vi tnh trng kh nng hp thu ln, bi v alen dng tnh c ngun gc t Kasalath, ging chng chu thiu ln rt u vit. Wissuwa v ctv. (1998) ngh thit lp qun th NIL (neraly-isogenic lines) tch bit r rng vng trn nhim sc th c cha QTL gi nh i vi kh nng hp thu ln, v thc hin nhng nghin cu c bn v sinh l hc, tm hiu c ch hp thu P ca cy la (Yano v Sasaki 1997). Khi y, nh chn ging c th p dng chin lc MAS trn c s QTL c xc nh r rng, chuyn kh nng hp thu P t ging bn a Kasalath vo ging ci tin, c ch s HI cao, c nng sut cao, trong iu kin b thiu ln. 6-3-3. Gen Pup-1: mt QTL ch lc lm gia tng kh nng hp thu ln Nghin cu ca tp th tc gi Wissuwa ti Nht c pht trin su hn thng qua bn gen Pup-1 (Wissuwa v ctv. 2002). Mt QTL c ln bn vi s p ng kh rng ca qung gia hai marker bao hm mt cluster ca cc gen, mi gen s c mt nh
hng nht nh so vi mt locus n c. Qun th mapping c pht trin theo dng qun th NIL trn c s hi giao vi b m l ging ti tc (recurrent) hon ton tha mn vi yu cu ca k thut fine mapping i vi QTL. Khi nim ny c p dng thnh cng i vi fine mapping nhng QTL mc tiu trn cy bp (Dorweiler v ctv. 1993), trn cy c chua (Alpert v Tanksley 1996), trn cy la (Yamamoto v ctv. 1998). Nhng kt qa cho thy vi QTL c th c x l ging nh gen n theo di truyn Mendel. Wissuwa v ctv. (2002) hnh thnh qun th lp bn QTL th hai bng cch hi giao NIL-C443 vi Nipponbare. Ti mt on phn t 50-cM marker ca Kasalath, ngi ta cng b rt nhiu kt qa trong chng trnh genome cy la ca Nht. Cc h F2 c chn lc v nh gi ngoi ng c hm lng ln rt ngho trong t. Phn tch QTL tiu chun c hon thin bi m hnh substitution mapping do Paterson v ctv. (1990) xut. NIL-C443 c hnh thnh thng qua hai ln hi giao b sung ca mt dng BC1F2 c tuyn chn trong qun th mapping u tin, vi Nipponbare c dng l ging ti tc. iu tra kiu gen nhm duy tr nhng alen ca Kasalath QTL C443, cng nh chn nhng alen khng phi Kasalath cc loci khng c lin quan, thng qua qui trnh s dng 118 RFLP marker trn genome cy la. B sung thm chi tit on nhim th 12 cha 50-cM Kasalath, NIL-C443 cn cha nhng on ngn Kasalath trn nhim th s 1, 8 v 10, chng khng lin kt vi kh nng hp thu ln trong phn tch QTL s khi (Hnh 6-3). Qun th mapping th cp c dng trong k thut fine map i vi QTL ch lc iu khin tnh trng hp thu P c pht trin thng qua hi giao gia NIL-C443 vi Nipponbare. Tc gi thu thp 150 cy F2 nh gi kiu gen vi 42 RFLP marker, v mt nhm ph gm 50 gia nh F2 c chn lc nh gi kiu hnh. nh gi kiu gen c thc hin vi 42 RFLP marker c chn lc t bn gc, mt cao RFLP, vi 3267 marker, thuc chng trnh genome cy la ca Nht (chng ta c th theo di trn web site http:rgp.dna.affrc.go.jp/Publicdata.html). Trn nhim sc th 12, c 22 marker, trong 16 marker nh v mt on ln ca Kasalath, gia 30,0 v 72,5 cM. Nhng marker cn li ph trn nhim sc th s 1 ca Kasalath (8 marker), nhim sc th s 8 (4 marker), v nhim sc th s 10 (8 marker) (Wissuwa v ctv. 2002). Phn tch QTL c hon thnh bng cch s dng s liu ca 50 h F2 nh gi kiu hnh. Kt qa nh gi kiu hnh cho thy s chi tng i trong iu kin thiu ln so vi iu kin ln c khng nh l mt d on gin tip i vi kh nng hp thu ln (Ni 1998, Wissuwa v ctv. 1998, 2002) Chng ta thc hin mt k thut c gi l substitution mapping (tm dch l lp bn chi tit theo phng php loi tr dn) i vi gen Pup1. 6-3-3-1. Bn QTL ca Pup-1 Bng 7: Bn gen Pup-1 i vi tnh trng kh nng hp thu ln trn nhim th s 12 (Wissuwa v ctv. 2002)
Qung gia marker Bn trn c s F2 (n=50) S14025-S13126 Bn trn c s F3 (n=160) S14025-S13126 Bn QTL u tin ca Wissuwa v ctv. 1998 V tr (cM) 53,6 54,8 LOD R2 nh hng cng 0,56** 0,69** nh hng tri 0,05ns 0,38*
16,6 11,5
78,9 28,1

C443-G2140 54,5 10,7 27,9 -
Trong k thut fine mapping F2, mt QTL nh v trn nhnh vai di ca nhim th s 12 gii thch 78,9% bin thin kiu hnh (bng 7), vi 2 marker k cn S14025 v S13126, n lin kt vi S14025 vi khong cch 1,3 cM. Nhm tc gi s dng k hiu Pup-1 (vit tt t ch phosphorous uptake 1) ch nh QTL ny. Phn tch QTL cho thy Pup-1 l do hot ng ca gen cng tnh, trong khi nh trng gen tri khng c ngha. Bn cnh khu vc rt ln trn nhim th 12 ca Kasalath, NIL-C443 cn c nhng on quan trng khc ca Kasalath trn nhim th s 1, 8, v 10 (hnh 6-3) nh hng n tnh trng kh nng hp thu ln. Trong k thut fine mapping F3, bin thin do kiu hnh c gii thch bi QTL gim xung cn 28,1%. y l kt qa tt, bi v cc s liu t c th c thu thp khng c tnh cht lp li. Pup1 lin kt vi 2 marker k cn S14025 v S13126, v lin kt cht vi S13126 Trong th nghim u tin v bn QTL (Wissuwa v ctv. 1998), Pup1 nm gia hai marker C443-G2140, v lin kt cht vi C443 khong cch di truyn l 3 cM. Trong th nghim ny, nhm tc gi s dng bn lin kt gen ca Kurata v ctv. (1994). V tr ca C443 ct mc tng ng vi v tr 54,5 cM, cng thm khong cch 3 cM ca gen Pup-1 (hnh 6-3) S chi c phn b trong qun th ng hp t F3 theo biu hnh chung (phn b chun), trong khi qun th d hp F3 khng phn b chun Thc hin k thut substitution mapping trn on mc tiu ca nhim sc th s 12 ging Kasalath, Wissuwa v ctv. (2002) chn ra 6 h c cht lng thng tin cao trong s 160 c th F3. Cc h (family) khc bit nhau trn 5 on ca nhim th s 1, 8 v 10 trong genome cy la Kasalath (hnh 6-3), nhng tt c cc cy trong cng mt h u cha ng on ging nhau ca Kasalath. Bin thin kiu gen trong nhng h ch c ghi nhn trn nhim sc th s 12. V tr ca gen Pup-1 c th c xem xt theo qui trnh loi dn tng bc mt (step-wise exclusion). Trong kiu gen ca h 14, gen Pup-1 nh v gia hai marker S10705-C449. Trong kiu gen h 92 v 93, ngi ta c th rt ngn qung cha gen Pup-1 gia hai marker S10704-G2140, bi v n th hin mt bin thin kiu gen nm bn ngoi qung m n khng c lin quan n bin thin ca kiu hnh (s chi). Tt c cc dng ca h s 6 c cha on nhim sc ca Nipponbare thuc qung gia marker R1869-S14025, biu hin s phn ly ca kiu hnh. Nh vy, S14025 c th c xem nh mt marker k cn trong phn tch QTL nh vy. Trong h s 97 v 114, c xu hng loi ra qung S13752-G21410 (-C449). Nh vy, theo phng php loi dn cc yu t, ngi ta c th xc nh rng ch c marker S13126 l mt th ng dng phn ly (co-segregates) vi gen Pup-1 (Wissuwa v ctv. 2002). Gen Pup-1 vn tip tc th hin nh hng ca n trong khi xc nh genome ca ging cho (donor) (Wissuwa v Ae 2001a, 2001b) S dng qun th NIL c th s c nhng ri ro nh on nhim th rt nh ca ging cho tn ti v nh hng tim n ca chng li khng ng gp g (Paterson v ctv. 1990). Kiu tn ti nh vy (relics) s biu hin mt cch ngu nhin trong qun th phn ly. Trong th nghim ca Wissuwa v ctv. (2002) khng c relics trong NIL-C443 gy nh hng n tnh trng kh nng hp thu ln v hn 80% bin thin v s chi c gii thch bi locus Pup-1. Tng tc theo kiu epistasis gia Pup-1 v vng cn li trong genome cng khng c ngha.
Phng php tip cn c th c s dng xc nh chnh xc v tr gen Pup-1: phn tch QTL, t v tr Pup-1 trong qung 3 cM lin kt vi marker S14025 v S13126. Phng php lp bn chi tit bng cch loi tr dn (substitution mapping) cho thy: Pup-1 ch ng phn ly vi marker S13126. Qung nh v nh vy c gii hn bi S14025-S13752, nm bn ngoi qung c cha gen Pup-1 (Wissuwa v ctv. 2002). Hin tng ti t hp trong qung S14025-S13126 ca h s 6, trong qung S13126S13752 ca h s 97, 114 (hnh 6-5) cho thy chnh s ti t hp s c th gip cho gi tr bn cao hn v gii thch s kin. Tuy nhin, nhm tc gi ny xut phi a thm nhiu marker trong cc qung nh vy nh v chnh xc hn Pup-1. Chnh Pup-1 l QTL ch lc gy nh hng chnh n tnh trng mc tiu p ng yu cu th nghim v bn . Yamamoto v ctv. (1998) chng minh rng nhng QTL th yu cng c kh nng cung cp thng tin chnh xc cao. Mt QTL c th c xem nh mt yu t di truyn gen n theo Mendel hoc l kt qa ca mt cluster ca nhng gen th yu? l cu hi c t ra rt sm. Trong trng ny, ch c mt QTL nh v trn nhim sc th s 12, gii thch hn 80% bin thin kiu hnh, c th c xem tng ng vi yu t trong di truyn Mendel. Tuy nhin, vi qung 3 cM cha Pup-1 qa rng ri cha nhiu gen. Nh vy, Pup-1 c gi nh do hu qa ca nhiu gen lin kt rt cht (Wissuwa v ctv. 2002) 6-3-3-2. ng dng trong chn ging ng dng marker phn t trong chn ging (MAS) l mc tiu ca nghin cu di truyn tnh chng chu thiu ln. y, ging Kasalath c xem nh ging cho. Ging Kasalath l ging bn a, c ngun gc bang Assam, n , thn cao, d ng, ht nh, v d rng. Phng php tip cn c a thch nht l chuyn gen Pup-1 vo ging la ci tin theo kiu hi giao, vi s tr gip ca MAS (Melchinger 1990, Hospital v Charcosset 1997). Ngi ta phi chn la marker lin kt cht vi gen Pup-1 biu th mc chnh xc cao khi nh gi so snh kiu gen vi kiu hnh i vi tnh trng hp thu P trong iu kin t thiu P. Vic ng dng MAS s gip nh chn ging tit kim cng sc lao ng v thi gian khi nh gi trn ng rung. Chn lc c th ng hp t i vi alen ca ging ti tc (recurrent) (ging c dng hnh ci tin, nng sut cao) trn mt s lng ln marker loci, ph ht b genome ca cy s lm gim i s th h cn thit phc hi genome ging ti tc nhng loci khng c lin quan (Hospital v Charcosset 1997). Frisch v ctv. (1999) c on ch cn bn th h hi giao c s h tr ca MAS, chng ta c th ng dng trn mt qun th nh c s marker point kh hn ch chuyn mt alen mc tiu vo ging ci tin. iu ny c th gip chng ta rt ngn hn 2 th h so vi phng php hi giao truyn thng. Nu qun th ln v marker point nhiu, chng ta s rt ngn hn 3 th h. Pup-1 c xc nh gia hai marker k cn S14025 v S13126 (S13752) l tin cc nh chn ging p dng MAS hi t gen ny vo con lai c trin vng, nm p ng mc tiu chn to ging la chng chu thiu ln. Vi mt qung 3-4 cM va nh gim thiu nh hng linkage drag (lin kt khng nh hng), s hi t cc tnh trng khng mong mun vi alen mc tiu mc chp nhn c ca cc nh chn ging (Frisch v ctv. 1999). i vi mc tiu map-based clonig, qung 3-4 cM c xem nh qa ln. Nh vy mun clone c gen Pup-1, chng ta cn phi b sung thm nhiu marker lin kt cht ch vi gen ny trong mt bn c tn chuyn mn l high-resolution. 6-3-3-3. S dng dng gn nh ng gen trong nghin cu sinh l hc Qun th NIL khng nhng l cng c c tim nng trong nghin cu di truyn, m cn cho php chng ta xt nghim nhng tin trnh sinh l cy trng lin quan n gen mc
tiu, bi v qun th NIL lm gim bin thin khng c lin quan n tnh trng mc tiu. Nhm tc gi s dng qun th NIL-C443 trong so snh vi Nipponbare v NIL b sung c cha on nhim th ca Kasalath thuc nhim sc th s 1, 8 v 10, thiu on nhim sc th s 12, nh tnh QTL iu khin s hp thu P (Wissuwa v Ae 1999). Nghin cu nh vy cho thy rng: ch c nhim sc th s 12 tham gia vo s khc bit kh nng hp thu ln ca cy la Nhm tc gi cha xc nh c c ch sinh l ca tnh trng hp thu P tng ng vi tng kiu gen khc nhau. Nhng th nghim tip sau khng nh kh nng hp thu ln cao ca NIL-C443 l do s tng trng mnh hn ca r la so vi dng hp thu ln thp, hoc do hin tng hp thu P c hiu qa tt hn trn mt n v r la (tnh theo kch c)(cn c gi l hiu qa ca r = root efficiency). Tc tng trng ca r la trong giai on m ca NIL-C443 c kch thch trong iu kin thiu ln (Wissuwa v Ae 2001a). Dng NIL-C443 duy tr kh nng hp thu ln cao hn trong khi mc khc bit v tng trng ca r gim. iu ny gi ra mt khi nim: gen Pup-1 nh hng c hai tnh trng tng trng ca r v hiu qa ca r. Gi nh rng c mt QTL, mt gen, hoc mt c ch s khng ng. Pup-1 c th cha mt cluster ca nhng gen, mi gen s nh hng n mt c ch khc nhau, hoc iu tit gen khi p ng vi tn hiu stress theo kiu up hoc down hng lot cc gen khc khng nh v cng khu vc vi QTL ny. Kh nng gen Pup-1 ch nh hng c ch c tnh cht nguyn nhn, th d nh hiu qa ca r v tng trng ca r ng vai tr th yu. Dng NIL-C443 c 91% alen ca Nipponbare v 9% alen ca Kasalath. Do nhng nh hng th yu c th l nguyn nhn gy ra s khc bit alen nhng loci bn ngoi qung cha gen Pup-1. Nu mt gen (hay nhiu gen) mt QTL gi nh no c clone, chc nng ca chng s c th c nh hng s liu cn bn ca gen (gene-databases)(Wissuwa v ctv. 2002). Ch c nhng tnh trng ng phn ly vi QTL, vi kh nng hp thu P cao, dng nh c lin quan trc tip n c ch chng chu thiu ln nh vy (Wissuwa v ctv. 2002). Nhng dng NIL mi c th c pht trin thng qua k thut fine mapping. Nhng h c s dng trong k thut substitution mapping (hnh 6-4) s c th nhn c mt im khi ng tuyt ho, v nhng dng NIL khc bit v kch c v v tr ca on nhim th Kasalath c ly trch mt cch trc tip, hoc sau mt th h ni tip. Cc tc gi ngh s dng ton b NIL trong th nghim sinh l hc c nhng gii thch c th hn v c ch chng chu trn c s sinh l v di truyn.
R21 74 S17 78
R665
R244 R2635 R1377
Hnh 6-3: Kiu gen ca NIL-C443 cho thy on nhim th s 1, 8, 10 v 12 ca Kasalath, v bn lin kt gen trn nhim th s 12 trn c s s liu marker ca 150 cy F2 thuc qun th QTL th cp (Wissuwa v ctv. 2002)
Hnh 6-4: Nhng kiu gen biu th trn gin ca on nhim sc th s 12 Kasalath c thu thp t 160 c th chn lc F3 c ngun gc t 6 h F2, vi gi tr trung bnh s chi, lch chun, v n c th (Wissuwa v ctv. 2002)
6-4. PHN TCH DI TRUYN S LNG Nghin cu c thc hin ti Vin La ng bng sng Cu Long trong nhng nm 1990 trn c s phn tch di truyn s lng theo phng php truyn thng, thng qua lai diallel vi 6 ging la: OM723-11, OM850 chng chu tt s thiu ln, IR64, IR50404 chng chu trung bnh, IR59606, OM997 chng chu km (Cn v ctv. 1998) Nn c bn ln p dng cng thc 100-60-0 kg NPK / ha v nn khng c bn ln 100-0-0 kg NPK / ha. C hai nn t tri qua 8 v (4 nm) th nghim di hn vi cng cng thc. Bn tnh trng c ghi nhn theo tiu chun chn lc ca IRRI l: kh nng nhnh, thi gian sinh trng, s ht chc trn bng, v trng lng kh ca r. Xt theo kh nng nhnh, OM723-11, OM850 c gi tr kh nng phi hp chung (gca) cao nht, IR59606 c gi tr gca cao nht i vi thi gian sinh trng, OM997, OM723-11 c gi tr gca cao nht i vi trng lng kh ca r, IR50404 c gi tr gca cao nht i vi tnh trng s ht chc trn bng, trong iu kin thiu ln (Cn v ctv. 1998). Kh nng nhnh c xem nh tiu chun chn lc r nht. i vi cc tnh trng nghin cu, hot ng ca gen khng cng tnh c u th trong phn tch phng sai, ngoi tr tnh trng thi gian sinh trng (bng 8 v 9). Thi gian sinh trng c xu th ko di ra trong iu kin thiu ln, tnh trng ny c iu khin bi u th ca nhm gen cng tnh. Xu th ging chng chu s thiu ln c thi gian sinh trng sm hn c kim nh bng t l +ve/-ve.
Bng 8: ANOVA v kh nng phi hp trong Griffing I, model 1

Ngun GCA SCA REC GCA SCA REC GCA SCA REC GCA SCA REC Tnh trng Thi gian sinh trng S ht chc / bng Trng lng kh ca r Kh nng nhnh Khng ln MS F 84,80 188,40** 130,50 512,50** 1,43 3,17** 137,7 2,1* 1691,0 21,2** 242,9 3,0 ns 8,85 4,58** 34,30 17,70** 1,83 0,94 ns 15,47 4,40** 125,02 53,60** 6,43 1,80 ns

C bn ln MS F 133,90 278,90** 294,90 614,30** 12,30 25,60** 67,6 1,8 ns 796,0 21,9** 84,6 2,3* 11,05 4,31** 31,10 12,10** 1,16 0,45 ns 12,19 3,27** 131,10 35,20** 10,52 2,82*
Bng 9: Thng s di truyn ca nhng tnh trng nghin cu trong iu kin thiu ln Tnh trng (H1/D) Kh nng nhnh Thi gian sinh trng S ht chc / bng Tr. lng kh ca r 1,94 0,98 5,80 0,81
1/2
Mc tnh tri 22gca 22gca +2sca 0,16 0,56 0,01 0,03
H2ns (%) 19,70 33,90 3,10 20,90
Trng lng kh ca r c gi tr KD/KR = 1,6 biu th tn sut alen tri cao hn alen ln trong iu kin thiu ln, ng thi t s H2/4H1 = 0,37 biu th kiu hnh c trng lng kh r cao hn trong ging la chng chu thiu ln. Gi tr d on hot ng ca gen cng tnh c tnh theo h s di truyn H2 v cng thc 22gca / (22gca +2sca) cho thy xu th hot ng ca gen khng cng tnh mnh hn gen cng, ngoi tr thi gian sinh trng TI LIU THAM KHO Alpert KB, SD Tanksley. 1996. High-solution mapping and isolation of a yeast artificial chromosome contig containing fw2.2: a major fruit weight quantitative trait locus in tomato. Proc Natl Acad Sci USA 93:15503-15507 Blair GJ, DC Cordero. 1978. Sulfur nutrition of wet land rice. IRRI Res Paper Series 21 Cn NT, ND By, BC Bu. 1998. Genetic analysis on phosphorous deficiency tolerance in rice (Oryza sativa L.). OMonRice 6:20-28 Chapin DC. 1980. Iron toxic soil. In: Soil and Rice, pp.781-800. IRRI, Los Banos, Philippines Chaubey CN, D Senadhira, GB Gregorio. 1994. Genetic analysis of tolerance for phosphorous deficiency in rice (Oryza sativaI L.). Theor Appl Genet 89:313-317 Chesholm DC, GJ Blair. 1988a. Genetic analysis of salinity tolerance in rice. Theor Appl Genet 86:333-338
Chesholm DC, GJ Blair. 1988b. Sodium-potassium ratio in soil solution and plant response under saline condition. Soil Sci Soc Am J 45:80-86 Clark RB. 1975. Characterization of phosphatase of intact maize roots. J Agric Food Chem 23:458-460 Clark T. 1976. Greenhouse screening techniques for tolerance to phosphorous deficiency. Crop Sci Soc of the Philippines, pp.1-7 Clark T, J Brown. 1974a. Techniques for breeding improved rice varieties for problem soil in genetic evaluation and utilization of rice for problem traing course 1990. Special project report, 10pp. Clark T, J Brown. 1974b. Factors affecting rice growth in acid sulfate soil. Soil Sci Soc Am 153:471-476 Dorweiler J, A Stec, J Kermicle, J Doebley. 1993. Teosinte glume architechture 1: a single locus controlling a key step in maize evolution. Science 262:233-235 Fageria NK, OP Morais, VC Baligar, RJ Wright. 1988. Response of rice cultivars to phosphorous supply on an oxisol. Fert Res 16: 195-206 Fox MD. 1978. The salt tolerance of rice. Inst Rice Commun Newsl 10:1-4 Frisch M, M Bohn, AM Melchinger. 1999. Comparision of selection strategies for markerassisted backcrossing of a gene. Crop Sci 39:1295-1301 Gerloff GC. 1987. Intact plant screening for tolerance to nutrient deficiency stress. Plant Soil 99:3-15 Gunawardena SDIE, HMS Wijeratne. 1978. Screening of rice under phosphorous deficient soil in Sri Lanka. Int Rice Res Conf 1978 (No.34). International Rice Research Institute, LosBanos, Philippines Gunawardena I. 1979. Field screening for tolerance to iron toxicity in plant. Journal of Plant Nutrition 11(3):303-319 Haley CS, SA Knott. 1992. A simple regression method for mapping quantitative trait loci in line crosses using flanking markers. Heredity 69:315-324 Hanson WC. 1950. The photometric determination of phosphorous in fertilizers using the phosphovanado-molybdate complex. J Sci Food Agric 1:172-173 Hedley MJ, GJD Kirk, MB Santos. 1994. Phosphorous efficiency and the forms of soil phosphorous utilized by poland rice cultivars. Plant Soil 158:53-62 Hospital F, A Charcosset. 1997. Marker-assisted introgression of quantitative trait loci. Genetics 147:1469-1485 Hung HH. 1985. Studies on tillering ability of rice under phosphorous stress. PhD thesis, Texas A&M University, Texas, USA Ikehashi H, FN Ponnamperuma. 1978. Varietal tolerance of rice for adverse soils. In: Soil and Rice. Symp IRRI, LosBanos, Philippines, pp 801-823 IRRI. 1971. Annual report for 1970. LosBanos, Philippines IRRI. 1976. Annual report for 1975. LosBanos, Philippines IRRI. 1994. Program report for 1993. LosBanos, Philippines Katyal JC. 1984. Screening of rice for adverse soil tolerance. In: Bassam NEL et al. (eds),Genetic aspects of plant mineral nutrition, Kluver Acad Publisher, the Netherlands, pp.523-531
Katyal JC, BD Sharma. 1980. Commun Soil Sci Plant Anal 15:1065-1081
Kosambi DD. 1944. The estimation of map distances from recombination values. Ann Eugen 12:172-175 Koyama MT. 1973. Phosphorous deficient effects on growth of rice at heading stage of development. Soil Sci. 96:105-111 Koyama MT, C Chammek, P Snitwonge. 1973. Varietal differences of Thai rice in the resistance to phosphorous deficiency. Trop Agric Res Cent Jpn Tech Bull 4 Kurata N, Y Nagamura, K Yamamoto, Y Harusima, N Sue, J Wu, BA Antonio, A Shomura, T Shimizu, S-Y Lin, T Inoue, A Fukuda, T Shimano, Y Kuboki, T Toyama, Y Miyamoto, T Krihara, K Hayasaka, A Miyao, L Monna, HS Zhong, Y Tamura, Z-X Wang, T Momma, Y Umehara, M Yano, T Sasaki, Y Minobe. 1994. A 300 kilobase interval genetic map of rice including 883 expressed sequences. Nature Genet 8:365372 Lander ES, P Green, J Abrahamson, A Barlow, MJ Daly, SE Lincoln, L Newburg. 1987. Mapmarker: an interactive computer package for constructing primary linkage maps of experimental and natural populations. Genomics 1:174-181 Lin HX, HR Qian, JY Zhuang, J Lu, SK Min, ZM Xiong, N Huang, KL Zheng. 1996. RFLP mapping of QTLs for yield and related characters in rice (Oryza sativa L.). Theor Appl Genet 92:920-927 Lin SY, T Sasaki, M Yano. 1998. Mapping quantitative trait loci controlling seed dormancy and heading date in rice (Oryza sativa L.) using backcross inbred lines. Theor Appl genet 96:371-375 Majumder ND, DN Borthakur, SC Rakshit. 1989. heterosis in rice under phosphorous stress. Indian J Genet 49:231-235 Melchinger AE. 1990. Use of molecular markers in breeding for oligogenic disease resistance. Plant Breed 104:1-19 Ni JJ, P Wu, AC Lou, YS Zhang, QN Tao. 1996. Low phosphorous effects on the metabolism of rice seedlings. Commun Soil Sci Plant Anal 27(19-20):3073-3084 Ni JJ, P Wu, D Senadhira, N Huang. 1988. Mapping QTLs for phosphorous deficiency tolerance in rice (Oryza sativa L.). Theor Appl Genet 97:1361-1369 Paterson AH, JW Deverna, B Lanini, SD Tanksley.. 1990. Fine mapping of quantitative trait loci using selected overlapping recombinant chromosomes in an interspecies cross of tomato. Genetics 124:735-742 Ponnaperuma FN. 1977. Screening for rice tolerance to mineral stresses. IRRI Res Paper Series 6, IRRI, LosBanos, Philippines. Ray JD, L Yu, SR McCouch, MC Champoux. 1996. Mapping quantitative trait loci associated with root penetration ability in rice (Oryza sativa L.). Theor Appl Genet 92:627-636 Reiter RS, JG Coors, Sussman, WH Gableman. 1991. Genetic analysis of tolerance to lowphosphorous stress in maize using restriction fragment length polymorphism. Theor Appl genet 82:561-568 Senanayake N. 1984. Varietal tolerance to phosphorous deficiency in wetland rice soils. Trop Agric 140:69-78 Utz HF, AE Melchinger. 1996. PLABQTL: a program for composite interval mapping of QTL. J Quant Trait Loci http://probe,nalusda.gov.8000/otherdocs/jqtl
Vos P, R Hogers, M Bleeker, M Reijans, T van der Lee, M Hornes, A Frijters, J Pot, J Paleman, M Kruiper, M Zabeau. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407-4414 Wissuwa M, J Wegner, N Ae, M Yano. 2002. Substitution mapping of Pup 1: a major QTL increasing phosphorous uptake of rice from a phosphorous-deficient soil. Theor Appl Genet 105:890-897 Wissuwa M, M Yano, N Ae. 1998. Mapping of QTLs for phosphorous-deficiency tolerance in rice (Oryza sativa L.). Theor Appl Genet 97:777-783 Wissuwa M, N Ae. 2001a. Genotypic variation for tolerance to phosphorous deficiency in rice and the potential for its exploitation in rice improvement. Plant Breed 120:43-48 Wissuwa M, N Ae. 2001b. Further characterization of two QTLs that increase phosphorous uptake of rice (Oryza sativa L.) under phosphorous deficiency. Plant Soil 237:275-286 Wissuwa M, N Ae. 1999. Molecular markers associated with phosphorous uptake and internal-phosphorous use efficiency in rice. In: Gissel-Nielsen G, A Gensen (eds) Plant Nutrition Molecular Biology and Genetics. Kluver Academic Publishers, The Netherlands, pp. 433-439 Wissuwa M, Y Kuboki, SY Lin, T Sasaki, M Yano. 1998. Fine mapping of quantitative trait loci Hd-1, Hd-2 and Hd-3, and controlling heading date of rice, as single Mendelian factors. Theor Appl Genet 97:37-44 Yamamoto T, Y Kuboki, SY Lin, T Sasaki, M Yano. 1998. Fine-mapping of quantitative trait loci Hd-1, Hd-2, Hd-3, and controlling heading date of rice, as single Mendelian factors. Theor Appl genet 97:37-44 Yano M, T Sasaki. 1997. Genetic and molecular dissection of quantitative traits in rice. Plant Mol Biol 35:145-153 Yano M, Y katayose, M Ashikari, U Yamanouchi, L Monna, T Fuse, T Baba, K Yamamoto, Y Umehara, Y Nagamura, T Sasaki. 2000. Hdi, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANT. Plant Cell 12L:2473-2483 Yano M. 2001. Genetic and molecular dissection of naturally occurring variation. Curr Opini Plant Biol 4:130-135 Zeng ZB. 1994. Precision mapping of quantitative trait loci. Genetics 136:1457-1468

Chng 7 C S DI TRUYN TNH CHNG CHU C ST
7-1. GII THIU CHUNG c st c th lm gim nghim trng nng sut ht (Genon v ctv. 1994). Trong t phn trng la ng Bng Sng Cu Long, c st c xem l yu t hn ch chnh, cng vi hm lng hu c cao, phn ng vi ion st trong iu kin t ngp nc. Trong iu kin canh tc cy trng cn, th d la c canh tc trn t c iu kin thong kh, rt kh tm hin tng ng c do st (Quijano v Mendoza 1994). Trong cc biu loi t c vn , c st thng gp khi iu kin pH t thp, kh nng trao i cation (CEC) thp, trng thi base thp, cung cp Mn thp, v mc thot nc km (Ponnaperuma 1974, Ponnaperuma v Solivas 1982). c st thng c tm thy trn t phn tr (Sulfaquepts), t ph sa c v t ph sa ct km thot nc (Hydraquents, Fluvaquents) cc thung lng, t acid ph sa v ph sa c Tropaquepts v Tropaquents, t than bn (Quijano v Mendoza 1994). c st trong cy la biu th ra bn ngoi cc triu chng nh sau: nhng m nu nh li ti l th hai sau l trn cng, dn dn cc vt nh ny lan rng lm l c mu nu, tm, vng cam ty theo ging la. C khi l la cun trn li. Trong trng hp ng c st qa nng, l th hai tr xung s tr nn nu sm ri cht. Mc tng trng v kh nng nhnh s b c ch, h thng r tr nn th cng, v c mu nu sm. Nu hin tng ng c st xy ra mun, kh nng tng trng khng b nh hng, nhng nng sut ht s b gim do tnh cht bt th. R la km pht trin, en, thi v c st bao bc bn ngoi (Benckiser v ctv. 1982). Bin i v c st rt rng cho nn nh hng ca n rt a dng. Tanaka v ctv. (1966) ghi nhn hm lng st trong t c th gy c cho cy la bin thin t 10 n 1000 mg/L. Mc bin thin rng nh vy khin cho ngi ta rt kh xc nh tiu chun gy hi ca st, dng hin din ca n, giai on cy la nhy cm nht, ging la, s c mt ca cht c ch h hp, tnh trng dinh dng ca cy, v yu t mi trng khc. Trn t phn, c st thng c pht sinh do Fe ho tan mc rt cao (Moormann v van Breemen 1978). Trong t c mc dinh dng thp, hoc b c ch h hp, hm lng Fe t 20 n 40 mg / L s gy c cho cy la (van Breemen 1978). Trong t ct, c st xy ra khi hm lng st trong t bin thin t 40 n 100 mg/L (van Breemen v Moormann 1978) Trong iu kin t acid, ion SO42- v Cl- gy ra hin tng hp thu ion Fe2+ (van Menswoort v ctv. 1985) c st c th l kt qa tng hp ca hng lot cc stress lin quan n dinh dng, ch khng phi ch n thun do ion Fe d tha. Tng quan thun gia Ca trong dung dch t v tng trng cy la trong t phn c bo co (Attananda v ctv. 1982). Vai tr ca calcium i vi c st c nghin cu bi Benckiser v ctv (1984), Howeler (1973), Ottow v ctv. (1983). c st c quan st trn cy la thiu Ca nu hm lng cation trong dung dch t ch yu l st ho tan (Moore v Patrick 1989). Bode (1990) nghin cu c st trn c s hm lng oxygen vng r v nhng enzyme c lin quan. Nhng ging la chng chu c st th hin hot tnh cao hn v superoxide dismutase v peroxidase trong iu kin b stress do st. Ging la IR9764-45-2 c xem nh mt genotype chng chu c Fe. Ngi ta ly trch trong r la hm lng superoxide dismutase v peroxidase cao gp hai ln so vi ging IR64 (i chng) (Bode 1990). Nu chng ta gia tng hm lng Fe trong mi trng th nghim, hot tnh ca
peroxidase s tng ln rt mnh ging chng chu. Trong khi hot tnh ca catalase hi tng mt cht trong ging nhim v khng p ng vi gia tng hm lng st. K thut x l cy la b ng c do st c th l: bn vi, thot thy, bn phn hu c, bn N, P, K v Zn cn i cy la trng thi dinh dng tt. Trong trng hp t c t l Fe/Mn rt cao, ngi ta bn thm MnO2 c th lm gim thiu c st (Nueu v Singh 1984, Ponnamperuma v Solivas 1982) Ging la biu th rt a dng v tnh chng chu c st. Vic ci tin ging c tnh chng chu st cng c t ra rt sm, nhng ci kh nht l qui trnh thanh lc, bi v n phc tp hn thanh lc nhm rt nhiu, lm sao duy tr c mi trng trng thi Fe++, gim s chuyn ho sang Fe+++. Bin thin trong ging la i vi tnh chng chu nh vy c t liu ha to iu kin cho cc nh chn ging la chn vt liu lai thch hp c ging chng chu Fe ngng cho php (Gunawarkena v ctv. 1982, Lai v Hou 1976). Chn lc ci tin ging chng chu st phi c hai iu kin: (1) thng s chn lc trn c s mt tnh trng indicator ng tin cy, vi mt c ch c bit v tnh chng chu, (2) hiu r cn bn di truyn ca c ch chng chu ny (Wu v ctv. 1997) Phn ng lm nu (bronzing) mu l c xem nh phng php xc nh mc chng chu c bit rt sm (IRRI 1965, Ota 1968), nhng ngi ta bit rt t v c s di truyn ca c ch chng chu tng ng vi mc nu. K thut marker phn t ang gip cc nh nghin cu tm hiu c s di truyn trn c hai phng din: tnh trng n gen v tnh trng a gen, tng tc epistasis,.. (Tanksley 1993). Kh nng p dng bn lin kt gen rt kh thi trong vic phn tch tnh trng ny (Causse v ctv. 1994, Kurata v ctv. 1994) 7-2. K THUT THANH LC Thanh lc c th c thc hin trong nh li hoc trn ng rung. Phng php thanh lc trong dung dch dinh dng, trong ct, trong t c x l thm ion Fe2+ l phng php thng c p dng. Fageria v Robelo (1987) s dng 100 ppm Fe2+ pH 4, trong khi Yamaguchi v Yoshida (1981) s dng 200 ppm Fe2+ pH 5. Trong mt bo co khc, Aranda v Orticio (1978) s dng 500 ppm Fe vi sucrose c dng lm cht anti-oxidant (chng -xi ha). y l bin php nhm ngn nga Fe++ chuyn ha thnh Fe+++. Trong th nghim trong chu: t c c st c mang v l th nghim. Ngi ta tin hnh nghin mu t c sy kh trc , xong bn thm NPK v cho vo nhng chu t. Chu t ny c t trong b nc sao cho ngp khi mt t, vi iu kin nc c kh khong. S dng 4 cy m c 2 tun tui cy vo mi chu. Sau khi cy c 4-6 tun, cy la c nh gi kiu hnh theo SES (IRRI 1988) v mc tng trng. Phng php ny c p dng ti IRRI v c thay i mt t ti IITA v WARDA. Trong th nghim ngoi ng, IRRI thit k kiu th nghim nhm thanh lc cc dng con lai c trin vng. Chn rung th nghim c c st v cy nhng dng th nghim trn rung y, vi tui m l 3 tun, mi dng cy thng 3 hng di 5m / hng. Khong cch cy 20 x 20 cm, vi 2-3 cy m / khm la. Ging chun chng chu c cy xen vo mt cch ngu nhin gia cc dng, v ging chun nhim c cy sau mi 10 dng th nghim. Nu rung khng ng u, c th cy ging chun nhim sau mi 5 dng, trong tng khi ngu nhin. Cho im thit hi theo SES vo giai on 4 tun, v 8 tun sau khi cy (Quijano v Mendoza 1994)

Bng 1: Tiu chun nh gi c st (SES) (IRRI 1988)
im 1 2 3 5 7 9 Triu chng Tng trng v nhnh bnh thng Tng trng v nhnh bnh thng, nhng m nu hoc vng cam xut hin u l gi Tng trng v nhnh bnh thng, l gia c mu nu, tm, hay vng cam Tng trng v nhnh chm li, nhiu l i mu Tng trng v nhnh ngng hn, hu ht cc l u b i mu hoc cht Tt c cy cht kh
Thanh lc qun th con lai qui m ln trn ng rung Nhng ging la chng chu c Fe c gii thiu l:
Ngun gc Tn ging
Bangladesh Colombia India Indonesia Liberia Malaysia Philippines Sri Lanka Thi Lan Vit Nam
BR52-8-2, BR95-75-2 Colombia II, CICA 4 IET 4082, IET 2911, IET 5551, Pokkali Sigadis, Banih Kuning Gissi 27, Suakoko 8 Mahsuri IR20, IR38, IR40, IR44, IR46, IR52, IR54 BW78, BW100, BW247-6 Khao Dwak Mali, Nahng Kiew C ung G Cng
7-3. NGHIN CU DI TRUYN PHN T TNH CHNG CHU C ST 7-3-1. Tnh chng chu c st trong qun th n bi kp IR64 x Azucena Cn c vo iu tra DNA trn b m vo nm 1995 ti IRRI, ngi ta ghi nhn IR64 (indica) l ging nhim, v Azucena (japonica) l ging chng chu tt, do , ngi ta thit lp mt qun th n bi kp (double haploid = DH) vi 123 dng c pht trin t cp lai ny (Wu v ctv. 1997, Guiderdoni v ctv. 1992). Sau , ngi ta pht trin mt qun th BC1F1 t 100 dng DH lai lui vi Azucena, do vy n cn c gi l DHBC1F1. Ht ging ca b m, 123 dng DH v 100 dng DHBC1F1 c cy trong khay ct thch anh (quartz sand). Sau mt tun, mi dng chn 12 cy, mi b m chn 24 cy, ri chuyn chng vo chu nha plastic cha 30 lt dung dch dinh dng (Yoshida v ctv. 1976). X l c st bng cch cho vo dung dch dinh dng 250 mg Fe++ / lt dng Fe2(SO4)3, pH dung dch c iu chnh mi ngy sao cho iu kin th nghm pH 4,5 v ngi ta phi thay mi dung dch dinh dng sau mi 3 ngy. Nhit trong nh knh c iu chnh 20-320C ban ngy v 12-200C ban m. Sau 4 tun l, ngi ta thu hoch mi cy vi im nh gi SES bng mt (IRRI 1988) vi triu chng leaf bronzing, xc nh trng lng kh ca chi (SDW) v hm lng Fe trong cy. Gi tr tng i ca mi tnh trng c tnh bng cch ly hiu s gi tr tuyt i ca 2 nghim thc chia cho gi tr tuyt i ca nghim thc i chng v nhn cho 100. Th d gi tr tng i ca trng lng kh chi thn: (SDW i chng SDR x l) RSDW = ----------------------------------------- x 100 SDR i chng
Wu v ctv. (1997) s dng 175 marker thnh lp bn trn nhim sc th s 12 thng qua qun th DH ca IR64 x Azucena, trong bao gm 76 marker ca genome cy la (RG), 16 marker ca genome cDNA cy oat (CDO), 49 marker ca cDNA clone (RZ), 15 gen v 19 marker khc. Tn sut ca alen cy IR64 i vi hu ht nhng marker s dng biu th mt phn b i xng, minh chng mt s cn bng trong vt liu di truyn ca mi b m truyn sang cho con thng qua k thut nui cy ti phn (Huang v ctv. 1997)
Hnh 7-1: Bn QTL tnh chng chu c st trn qun th DH ca IR64 x Azucena ti nhim sc th s 1 v s 8, thanh mu en biu th QTL ch s chng chu, thanh mu trng biu th QTL gi tr tng i ca trng lng kh r (Wu v ctv. 1997) 175 marker biu hin a hnh c s dng lp bn lin kt gen theo MAPMARKER (Lander 1993). Gi tr LOD > 2,4 v p < 0,001 i vi sai s Type 1 c s dng lm tiu ch xc nh mc ngha ca QTL lin kt vi marker loci. Azucena c khng nh l ging c kh nng chng chu c st v IR64 l ging nhim ngng 250 mg Fe++ / lt trong mi trng dinh dng Yoshida (Bode v ctv. 1995) Hm lng st trong dung dch dinh dng gim khong 50% sau 3 ngy trong khi cy tng trng. Do vy, chng ta phi thay mi mi trng sau mi 3 ngy. Cho d c thay mi, nhng hm lng Fe++ vn c xu hng suy gim gia nhng ln thay i nh vy

Bng 2: Ch s im chng chu v trng lng kh ca chi gim tng i (RSDW) vi nhng marker loci tng ng (p , 0,001) (Wu v ctv. 1997)
Thng s Ch s chng chu RSDW Marker RG345 RG381 RG810 RG345 RG381 RG810 RG331 RG978 Nhim th 1 1 1 1 1 1 1 8 Xc sut 0,001 0,001 0,001 0,001 0,001 0,001 0,001 0,005 R2 0,33 0,29 0,12 0,17 0,15 0,23 0,21 0,11 II Kiu gen IR64 4,17 4,02 2,14 39,74 38,63 27,02 26,96 35,83 JJ Kiu gen Azucena 1,99 1,99 3,37 28,43 28,65 37,55 38,24 26,33
Bng 3: Phn tch QTL theo phng php bn cch qung (Wu v ctv. 1997)
Thng s Ch s chng chu RSDW V tr RG345-RG381 RG810-RG331 RG345-RG381 RG810-RG331 RG978-RG1 Nhim th 1 1 1 1 8 Gi tr tnh cng -1,06 0,63 -5,51 5,66 -4,61 R2 (%) 32,3 12,6 15,5 21,5 10,0 LOD 10,10 3,53 4,73 6,38 2,75
Ch s chng chu v trng lng kh ca chi tng i c phn tch c hai phng php SMA (bng 2) v IMA (bng 3). Bn loci ca marker trn nhim sc th s 1 v mt marker trn nhim sc th s 8 c ghi nhn (hnh 7-1). QTL iu khin ch s chng chu gii thch t 12,6 n 32,2% bin thin kiu hnh, trong phn tch bn cch qung. QTL iu khin RSDW gii thch t 15,5 n 21,5% bin thin kiu hnh trn nhim sc th s 1 v 10% kiu hnh trn nhim sc th s 8. Hai loci ca gen c xc nh bi hai marker k cn RG345 v RG381, v lin kt vi marker 810 trn nhim sc th s 1 i vi c hai tnh trng: ch s chng chu v RSDW. So snh trung bnh kiu gen gia hai marker cho thy alen ca tnh chng chu xut pht t Azucena nh v ti locus th nht trn nhim th s 1 v mt locus khc trn hnim th s 8. Alen t locus th nh trn nhim th s 1 xut pht t IR64. Wu v ctv. (1997) cng ang thc hin k thut substitution mapping trn qun th chn lc t cc dng chng chu vi gim thiu ti a genome ca Azucena trong nghin cu tip theo, nhm p ng yu cu ng dng MAS trong chn ging la chng chu c st. 7-3-2. Tnh chng chu c st trong qun th cn giao c lai lui gia Nipponbare / Kasalath // Nipponbare Wan v ctv. (2002) thc hin mt qun th cn giao backcross (BILs = backcross inbred lines) t Nipponbare (japonica) / Kasalath (indica) // Nipponbare, vi 96 dng BIL phn tch bn QTL tnh chng chu c st ca cy la. nh gi kiu hnh c thc hin trn b m v qun th con lai BIL ny, vi s hin din ca cc ging i chng IR26, IR64, IR74, Mahsuri, v Suakoko 8. t th nghim ng c st thuc Tri th nghim ca i Hc Nng nghip Nanjing Trung Quc. t c c st trong vng trng la ca Trung Quc chim khong 7,5-8,0 triu ha, lm nng sut gim 1020% ty theo t v ty theo ging canh tc (Li v ctv. 1991)
Hnh 7-2: QTL xc nh tnh chng chu c st trong qun th Nipponbare / Kasalath // Nipponbare (BIL), vi tnh trng leaf brozing index (LBI): ch s chng chu da trn l b nu, stem dry weight: trng lng kh ca thn la, tiller number: s chi, root dry weight: trng lng kh ca r la (Wan v ctv. 2002)
Ch s chng chu da trn hin tng l b nu (LBI), trng lng kh ca thn r (SDW), s chi (TN), v trng lng kh ca r la (SDW) c s dng nh nhng tiu chun chn lc tnh chng chu c do st sau 4 tun cy trong mi trng dinh dng Yoshida c x l 250 ppm Fe++ Phn b chun (normal) c ghi nhn trong tnh trng LBI, lch tri theo Kasalath c 3 tnh trng cn li, trong 96 c th ca qun th BIL, v c s khc bit rt ngha gia Nipponbare v Kasalath Bng 4: Phn tch QTL ca 4 tnh trng: ch s chng chu (LBI), trng lng kh ca thn (SDW), s chi (TN), v trng lng kh ca r (RDW) trng giai on m, theo phng php bn cch qung (Wan v ctv. 2002)
Tnh trng LBI SDW TN Qung gia hai marker C955-C885 C955-C885 C25-C515 R1928-C178 C955-C885 Nhim th 1 1 3 1 1 LOD 3,17 7,03 5,85 3,49 6,89 R2 (%) 20,5 36,9 47,9 42,4 43,9 Tnh cng 0,0210 -0,0114 0,0072 0,1300 0,0679

RDW R2329-R210 C955-C885 C25-C515 1 1 3 4,32 6,59 5,35 39,2 6,59 5,35
-0,0055 -0,0095 0,0029
Chng ta cha tm ra mi quan h gia hai kt qa ca hnh 7-1 v hnh 7-2 trn nhim sc th s 1 Trong kt qa phn tch qun th BIL, gi tr LOD u ln hn 3,17 (bng 4), bn RFLP v s liu u c thu thp t chng trnh Rice Genome ca Nht phc v nghin cu QTL. Wan v ctv. (2002) xc nh 8 QTL lin kt vi tnh chng chu c st, s dng computer chng trnh Mapmarker/QTL (Lincoln v ctv. 1993). Mt QTL gi nh c tm thy c lin quan n tnh trng LBI trn nhim th s 1, gii thch 20,5% bin thin kiu hnh trong qun th BILs (bng 4, hnh 7-2). Hai QTL c tm thy c lin quan n tnh trng SDW, mt nh v trn nhim sc th s 1 cng v tr vi tnh trng LBI. Ba QTL c tm thy c lin quan n tnh trng RDW, hai nh v trn nhim sc th s 1 v 1 nh v trn nhim sc th s 3 QTL nh v trn nhim th s 3 va c lin quan n tnh trng trng lng kh ca thn, va c lin quan n tnh trng trng lng kh ca r (hnh 7-2) Tt c nhng QTL gi nh ny u gii thch bin thin kiu hnh ca tnh trng kh ln trong qun th BILs t 20,5% n 47,9% Chng ta ch , qung gia hai marker C955-C885, l ni nh v ca QTL gi nh c lin quan n c 4 tnh trng tiu chun (hnh 7-2). Nh chn ging c th li dng c im ny tm QTL hu ch cho chin lc MAS nhm hi t gen chng chu c st vo ging ci tin c ch s thu hoch cao, gia tng nng sut ging la trng trn t c c st cao.
TI LIU THAM KHO
Attananda T, S Vacharotayan, K Kyuma. 1982. Chemical characteristics and fertility status soils of Thailand. Pages 137-156 in Proc Bangkok Symposium on Acid Sulfate Soils. ILRI, Wageningen, Netherlands Benckiser G, JCG Ottow, S Santiago, I Watanabe. 1982. Physicochemical characterization of iron toxic soil in some Asian countries. IRRI Res Paper Series 85 Benckiser G, S Santiago, HU Nueu, I Watanabe, JCG Ottow. 1984. Effect of fertilization on exudation, dehydrogenase activity, iron-reducing population and Fe++ formation in the rhizosphere of rice (Oryza sativa L.) in ralation to iron toxicity. Plant and Soil 79:305316 Bode K, O Doring, S Luthje, HU Nueu, M Bottger. 1995. The role of active oxygen in iron tolerance of rice (Oryza sativa L.). Protoplasma 184:249-255 Causse M, TM Fulton, YG Cho, SN Ahn, J Chungwongse, K Wu, J Xiao, ZH yu, PC Ronald, SB Harrington, GA Second. 1994. Saturated molecular map of the rice genome, based on an inter-specific backcross population. Genetics 138:1251-1274 Fageria NK, NA Robelo. 1987. Tolerance of rice cultivar to iron toxicity. J Plant Nutr 10(6):653-661
Genon JG, N Hepcee, JE Duffy, B Delvaux, PA Hennebert. 1994. Iron and other chemical soil constrainyts to rice in highland swamps of Burundi, Plant and Soil 166(1):109661 Gunawarkena I, SS Virmani, FJ Sumo.. 1982. Breeding for tolerance to iron toxicity. Oryza 19:5-12
Howeler RH. 1973. Iron induced oranging disease of rice in relation to physicochemical changes in a flooded Oxisol. Soil Sci Soc Am Proc 37:893-903
IRRI. 1965. Annual Report 1964, The International Rice Research Institute, Los Banos, Philippines IRRI. 1988. Standard Evaluation System (SES). IRRI, Los banos, Philippines Kurata N, Y Nagamura, K Yamamoto, Y Harusima, N Sue, J Wu, BA Antonio, A Shomura, T Shimizu, S-Y Lin, T Inoue, A Fukuda, T Shimano, Y Kuboki, T Toyama, Y Miyamoto, T Krihara, K Hayasaka, A Miyao, L Monna, HS Zhong, Y Tamura, Z-X Wang, T Momma, Y Umehara, M Yano, T Sasaki, Y Minobe. 1994. A 300 kilobase interval genetic map of rice including 883 expressed sequences. Nature Genet 8:365372 Lai KL, C Hou. 1976. Studies on the differentiation of physiological characteristics of the roots of different type rice plant (Oryza sativa L.). Taiwan Agric Center, Bull of the Phytotron Oct 10, pp. 17-21 Lincoln SE, MJ Daly, ES Lander. 1993. Mapping genes controlling quantitative traits usingg MAPMARKER/QTL version 1.1: a tutorial and reference manual. 2nd ed. Cambridge Mass: Whitehead Institute for Biometrical Research. Moore PA, WH Jr Patrick. 1989. Iron availability and uptake in acid sulfate soils. Soil Sci Soc Am J 53:471-476 Moormann FR, N van Breemen. 1978. Rice: soil, water, and land. Int Rice Res Inst, Los banos, Philippines Nueu HU, VP Singh. 1984. Management of outland rice and fishponds on problem soils in the tropics. Pages 352-366 in Ecology and management of problem soils in Asia. FFTC Book Series No. 27, Taipei Ota Y. 1968. Studies on the occurrence of the physiological disease called bronzing. Bull Natl Inst Agric Sci (Japan) D 18: 31-104 Ottow JCG, G Benckiser, I Watanabe, S Santiago. 1983. Multiple nutritional stress as the prerequisite for iron toxicity of wte land rice (Oryza sativa L.). Trop Agric (Trinidad) 60:102-106 Ponnamperuma FN, JL Solivas. 1982. Field amelioration of an acid sulfate soil with manganese dioxide and lime. Pages 213-222 in Proc of Bangkok Symp on Acid Sulfate Soils. H Dost and N van Breemen (eds.), IILRM, Wagenigen. Ponnaperuma FN. 1974. Problem rice soils. Proc. IRRI Conf., April 22-25, 1974, LosBanos, Philippines, 11p. Quijano C, R Mendoza. 1994. Mineral Toxicities in rice. Breeding rice for flood-prone and problem soil areas. IRRI 1994. pp 99-116 Tanaka A, SA Navasero. 1966. Interaction between iron and manganese in the rice plant. Soil Sci Plant Nutr 12:29-33 Tanksley SD. 1993. Mapping polygenes. Annu Rev Genet 27:205-233
van Breemen N. 1978. Landscape, hydrology and chemical aspects of some problem soils in the Philippines and in Sri Lanka. A terminal report submitted to IRRI, Los Banos, Philippines pp. 247-282 van Breemen N, FR Moormann. 1978. Iron toxic soils. PP. 781-800 in Soils & Rice, IRRI, LosBanos, Philippines van Menswoort ME, SR Lantin, R Brinkman, N van Breemen. 1985. Toxicities of wetland soils. PP 123-138 in Wetland soils: Characterization, Classification, & Utilization. IRRI, LosBanos, Philippines Wu P, A Luo, J Zhu, J Yang, N Huang, D Senadhira. 1997. Molecular marker linked to genes underlying seedling tolerance for ferrous iron toxicity. In: T Ando et al.(eds.) Plant Nutrition for sustainable food production and environment. Kluwer Acadamic Publishers, pp. 789-792 Yamaguchi M, S Yoshida. 1981. Physiological mechanism of rice tolerance for iron toxicity. IRRI, Saturday Seminar, June 6. Guiderdoni E, E Galinato, J Luistro, G Vergara. 1992. Anther culture of tropical japonica x indica hybrids of rice (Oryza sativa L.). Euphytica 62:219-224 Yoshida S, D Forno, JH Cock, KA Gomez. 1976. Laboratory Manual for Physiological studies of rice. The third edition. IRRI, LosBanos, Philippines Huang N, A Parco, TW Mew, G Magpantay, SR McCouch, E Guiderdoni, J Xu, P Subudhi, R Angeles, GS Khush. 1997. RFMP mapping of isozymes, RAPD and QTLs for grain shape, brown plant hopper resistance in a double haplod rice population. Molecualr Breeding Lander ES.1993. Mapmeraker/Exp 3.0 and Mapmarker/QTL 1. 1. Tutorial and Reference manual. Whitehead Institute. 9 Cambridge Center, Cambridge MA 02141 Li DM, JJ Tang, JL Zhou, SQ Li. 1991. The eco-physiological mechanism of rice tolerance for gleyic soil stress and the breeding varieties tolerance for soil-related stress. Rice Review and Abstracts 10:1-4 Wan JL, HQ Zhai, JM Wan, H Ikehashi. 2002. Detection and analysis of QTLs for ferrous iron toxicity tolerance in rice (Oryza sativa L.). Rice Genetics Newsletter 19:56-58

Chng 8
C S DI TRUYN TNH CHNG CHU NHIT LNH
8-1. GII THIU CHUNG Cy la thuc nhm cy nhy cm vi nhit lnh, nht l i vi nhng ging c ngun gc nhit i v cn nhit i. Nhng tn thng do nhit lnh gy ra c th c quan st nhiu giai on tng trng khc nhau ca cy la: ht khng ny mm, cy m km pht trin, hin tng cy ln, cn ci, hin tng bin i mu sc, hin tng thoi ha nh bng la, gia tng t l ht lp, v hin tng ht chn bt bnh thng (Kaneda v Beachell 1973). pha bc nc Nht, hin tng bt th do nhit lnh c xem nh vn nghim trng nht lm nng sut la gim rt nhiu (Saito v ctv. 2001). i vi kiu gen cy la mn cm vi tnh trng bt th do nhit lnh, giai on tng trng nhy cm nht l giai on lm ng, c bit l giai on vi bo t chuyn t pha tetrad sang pha u tin ca nhim th thu nh li, xt trn c s gin phn nhim th trong di truyn t bo (Satake v Hayase 1970). Nhit thp trong sut qa trnh pht sinh vi bo t (microsporogenesis) s lm cho thoi ha vi bo t v to ra hin tng phnh to t bo ht phn. Mt khc, c quan ca non vn duy tr c kh nng th tinh mt cch bnh thng, nhng tin trnh pht trin microspore hon ton b c ch (Hayase v ctv. 1969). Do , hin tng thoi ha microspore, v/hoc hin tng phnh to t bo ht phn c xem nh phn ng ca kiu gen bt th i vi s tn thng do nhit lnh (Nishiyama 1976). C ch chng chu lnh hin nay vn cha c nghin cu mt cch chi tit, cn nhiu cu hi cha c gii p tha ng. Mun t c mt kt qa thnh cng v th phn, cy la cn c s lng ht phn cn thit trong ti phn. S lng ht phn gim theo nghim thc x l lnh lm cho t l bt th gia tng rt c ngha (Satake 1991). Nhng ging la chng chu lnh thng sn sinh ra s lng ht phn nhiu hn ging nhim (Satake v Shibata 1992). Nh vy, chng ta c th ni rng s lng ht phn l mt yu t quan trng trong c ch chng chu lnh. Ngi ta cn ghi nhn s lng ht phn tng quan thun vi chiu di ti phn (Oka v Morishima 1967, Suzuki 1981), v dng nh chiu di ti phn c tng quan vi hin tng chng chu lnh (Tanno v ctv. 1999). Hin tng chng chu lnh l tnh trng di truyn s lng v cha c mt gen ch lc no c xc nh (Saito v ctv. 2001). Futsuhara v Toriyama (1966) c on rng: s gen chng chu lnh c th l 4 gen hoc nhiu hn 4 gen. Chng lin kt rt cht vi gen Pr (tnh trng v tru mu tm), Rc (tnh trng v la ht go mu nu), d2 (tnh trng ln), gh (tnh trng v tru mu vng), nl (neck leaf), v bc (tnh trng thn r gin, d b gy = brittle) trn nhim sc th s 3, 4, 5 v 7 (Futsuhara v Toriyama 1966, Takahashi v ctv. 1973, Sawada 1978) 8-2. DI TRUYN TNH CHNG CHU LNH 8-2-1. Bn QTL trn nhim sc th s 4 Ging la Norin-PL8 c gen chng chu lnh k tha t ging la Silewah (japonica). Saito v ctv. (2001) quan st trn hai on nhim sc th c hi t gen mc tiu t Silewah, nh v trn nhim th s 3 vai ngn, v trn nhim th s 4 vai di (Saito v ctv. 1995). Cc tc gi ny tin hnh lp bn QTL theo kiufine mapping, tp trung trn nhim th s 4, trn c s qun th hi giao (BC) v NIL (nearly isogenic lines). Norin-PL8 l ging la chng chu lnh c pht trin nh lai hi giao gia ging japonica chng chu lnh Silewah vi mt dng la Nht Hokkai 241, trong dng Hokkai 241 c dng lm dng ti tc (recurrent parent), v Silewah l ging cho gen mc tiu (donor). Saito v ctv. (2001) chn c mt dng Syo6 t t hp lai Kirara 397 / Norin-
PL8 // Kirara 397 (qun th BC1F5). Dng Syo6 c cha on nhim sc th s 3 v 4 hi t gen mc tiu t Norin-PL8. Sau hai ln hi giao, tc gi chn mt cy d hp t i vi hi t gen mc tiu nhim th s 4, nhng khng c gen mc tiu nhim th s 3. Cho cy ny t th phn, ngi ta pht trin qun th phn ly k hiu l BT4, v 114 cy BT4 ny c chn s dng trong phn tch bn cch qung. Mu DNA c ly trch t l la theo phng php CTAB (Murray v Thompson 1980). RFLP marker c s dng c ngun gc t chng trnh genome cy la ca Nht (RGP) k hiu l R v C, c ngun gc t Saito v ctv. (1991) k hiu l XNpb. Phng php nh gi kiu hnh tnh trng chng chu lnh trong nc c nhit lnh (Futsuhara v Toriyama 1964), c xem nh l phng php thng nht trong thanh lc ging chng chu lnh quc t. cy la c x l nhit nc 19oC t giai on tng khi s khi n giai on tr. su mc nc 20 cm th nghim ngoi ng v 24 cm trong nh li. nh gi ht hu th kt lun v thanh lc chng chu lnh. Saito v ctv. (2001) s dng phn mm MAPL97 (Ukai v ctv. 1991, Hayashi v Ukai 1994) phn tch QTL v bn QTL. Mun xc nh chnh xc v tr QTL trn nhim sc th s 4, ngi ta pht trin mt qun th hi giao ci tin, chn Kirara 397 lm ging ti tc. B sung thm 5 RFLP marker (R738, R740, R1427, R2737 v C1016), 2SCAR marker (SCAB1 v SCAM20) phn tch. Nhng marker c th hin trn bn (hnh 8-1B). Saito v ctv. (2001) s dng 117 c th ca qun th BT4 phn ly phn tch theo phng php cch qung i vi tnh trng chng chu lnh. Hnh 8-1A cho thy gi tr LOD trn on hi t gen mc tiu ca nhim th s 4, vi thang im kh cao (LOD>25) t trung tm n im cui ca hi t (introgression). Gi tr ti a ca LOD c ghi nhn ti qung gia R2737 v XNpb102. iu ny cho thy rng QTL phn ln nh v trong qung R2737 v XNpb102. Saito v ctv. (2001) cn pht trin qun th NIL thc hin k thut fine mapping QTL iu khin tnh trng chng chu lnh. Ngi ta chn la 12 dng ti t hp (recombinants) gia R738 v R1427 trong qun th BT4. Con lai ca mi recombinant c pht trin thnh nhng dng n nh (fixed) thng qua phng php MAS (markeraided selection). Kiu gen ca mi dng NIL c th hin trong hnh 8-1B. Kiu hnh chng chu lnh c nh gi thng qua gi tr trung bnh ca ht hu th. Kiu gen ca BT4-9-7 l kiu gen ca Norin-PL8 i vi marker R738. Tnh trng chng chu lnh ca BT4-76-2 v BT4-76-7 l kiu gen ca Norin-PL8 i vi marker R738 v R2737. Kt qa nh vy cho thy rng QTL iu khin tnh chng chu lnh nh v gia marker R738 v XNpb102 Bng 1: Tng quan gia nhng on hi t gen mc tiu trn nhim sc th s 3 v 4 vi tnh trng chiu di ti phn (Saito v ctv. 2001) Nhim sc th 3 4 Marker XNpb345 R2737 Chiu di ti phn Kirara 397 [n] Norin-PL8 [n] 2,42 (22) 2,51 (33) 2,38 (26) 2,55 (26) t 2,048 4,257 P 0,0455 <0,0001
Trc , ngi ta tm thy QTL iu khin tnh trng chng chu lnh nh v trn vai ngn ca nhim th s 3 v trn vai di ca nhim th s 4 (Saito v ctv. 1995). Ngi ta cng xc nh dng chng chu lnh Hokkai-PL4, mt dng cng b m vi Norin-PL8, cng c vng hi t gen mc tiu trn nhim th s 4, nhng thiu vng mc tiu trn nhim th s 3. Chnh QTL chng chu lnh trn nhim th s 4 cng c khm ph trong mt
ging chng chu lnh khc Hokkai-PL5, m tnh trng chng chu ny c hi t t ging chng chu c ngun gc javanica (japonica nhit i) Lambayque 1. Do , QTL trn nhim sc th s 4 c xem nh quan trng hn so vi nhim th s 3 trong vic th hin gen chng chu lnh
Hnh 8-1(A): Bn cch qung QTL tnh trng chng chu lnh trn vai di ca nhim sc th s 4, s dng qun th BT4, vi kiu gen ca cc dng NIL. (B) Dng hnh Norin-PL8 biu th bng thanh ngang t en, dng hnh Kirara 397 biu th bng thanh mu trng. Dng hnh ti t hp (recombination) c biu th bng thanh ngang c chm. Kiu gen Ctb-1 v Ctb-2 c d on bng kiu gen marker v tnh chng chu lnh cng c biu hin (Saito v ctv. 2001)
Hnh 8-2: V tr trn bn ca Ctb-1 v Ctb-2 . Bn bn tri c hnh thnh trn c s ti t hp trong qun th BT4. Bn bn phi c hnh thnh trn c s bn lin kt gen theo chng trnh RGP ca Nht (Harushima v ctv. 1998). YAC clones c nh v trn bn theo kt qa nghin cu ca Koike v ctv. (1997)
Hnh 8-1 cn th hin mt QTL chng chu lnh khc nh v gia R740 v SCAB11. Ngi ta tm thy hai QTL chng chu lnh khc nhau nh v trn nhim sc th s 4, mt th hin tnh cht ngoi bin (distal), k hiu l Ctb-1, mt th hin tnh cht ni bin (proximal), k hiu l Ctb-2. Ba dng NIL (BT4-112-1, BT4-10-6, v BT4-50-1) c c Ctb-1 v Ctb-2, nhng tnh trng chng chu lnh ca chng th hin ging vi cc dng NIL ch c mt Ctb1 hoc Ctb-2 (th d nh BT4-76-2, BT4-70-1). Kt qa ny cho thy tng tc gia Ctb-1 v Ctb-2 khng c tnh cht cng tnh (additive) (Saito v ctv. 2001). 8-2-2. V tr trn bn ca Ctb-1 v Ctb-2 SCAB11 v SCAM20 l nhng marker SCAR c pht trin t phn t DNA a hnh c khuch i mt cch ngu nhin (RAPDs). Ngi ta s dng nhng marker RFLP trn bn theo chng trnh genome cy la (RGP) ca Nht (Kurata v ctv. 1994, Harushima v ctv. 1998), nhng v tr ca SCAB11 v SCAM20 trn bn vn cha c xc nh. Do , ngi ta khng th c on chnh xc khong cch di truyn gia Ctb-1 v Ctb-2. xc nh c nhng v tr ca SCAR marker trn bn di truyn RGP, ngi ta s dng bn vt l trn c s YAC clone (yeast artificial chromosome) ca RGP (Koike v ctv.1997). Marker SCAM20 trong YAC clone Y1676, ph kn mt vng 102,0 cM 102,6 cM trn bn . Marker SCAB11 trong YAC clone Y1955, ph kn mt vng 106,7 cM trn bn (hnh 8-2).

cM. 8-2-3. Tnh chng chu lnh v chiu di ti phn
Ctb-1 nh v ti vng c ln 6,7 cM, qut trn mt on t 106,7 cM n 113,4 Ctb-2 nh v ti vng c ln 5,8 cM, qut trn mt on t 96,2 cM n 102,0 cM. Gi tr khong cch t Ctb-1 n Ctb-2 c khong 4,7 n 17,2 cM (hnh 8-2).
Chiu di ti phn ca ging Norin-PL8 l 2,85mm v Hokkai 241 l 2,13mm (Saito v ctv. 2001). Chiu di ti phn ca cc dng con lai F7 phn b theo biu hnh chung (phn b chun), cho thy khng th khng nh chiu di ti phn do mt gen ch lc quyt nh. S phi hp gia chiu di ti phn v cc on nhim th hi t trn nhim sc th s 3 v 4 c th nghim thng qua phn tch phng sai vi mi marker c xem nh mt nghim thc. Mc d marker R2737 trn nhim sc th s 4 tng quan c ngha vi chiu di ti phn, nhng mc tng quan c ngha ca XNpb345 trn nhim sc th s 3 kh thp (bng 1). Kt qa ny dn n mt suy lun rng QTL iu khin tnh trng chiu di ti phn nh v trn vai di ca nhim sc th s 4 (Saito v ctv. 2001). C hai Ctb-1 v Ctb-2 c kh nng kt hp vi tnh trng chiu di ti phn, v s tng tc ca nhng gen ny khng c tnh cht cng tnh. 8-2-4. Bn QTL tnh trng chng chu lnh giai on lm ng Giai on nhy cm nht i vi nhit lnh l giai on lm ng, vo khong 11 ngy trc khi la tr (Andaya v Mackill 2003). Ging la japonica c ghi nhn c kh nng chng chu lnh tt hn ging la indica (Glaszmann v ctv. 1990, Dilday v ctv. 1990). Ngi ta s dng ging la japonica c ngun gc California l M-202, v ging la indica c ngun gc IRRI l IR50 lm b m cho qun th cn giao ti t hp (RIL) vi 191 dng con lai th h F5 v F6 (Andaya v Mackill 2003). Tc gi s dng 300 microsatellite marker v k thut PCR phn tch QTL trn qun th RIL v tnh trng chng chu lnh. nh gi kiu hnh c thc hin trong phytotron trong iu kin 12oC, 12 gi sng / ngy, m tng i trong phng l 72% Bn lin kt gen c thit k theo phm mm MAPMARKER 2.0 (Lander v ctv. 1987), vi chun gi tr LOD = 5,0. Tng s 175 microsatellite marker trn 181 loci c s dng thit k bn QTL, vi chiu di tng cng trn bn l 1.276,8 cM, chiu di trung bnh gia hai marker l 7,1cM. Tc gi s dng phn mm PLABQTL (Utz v Melchinger 1996) trong phn tch bn cch qung. QTL nh v trn nhim sc th s 1, 2, 3, 5, 6, 7, 9, 12 c xc nh vi tr iu khin tnh trng chng chu lnh giai on lm ng. S marker iu tra tnh cht a hnh trn mi nhim sc th bin thin t 15 n 41, t l a hnh bin thin t 27% (nhim th s 8) n 88% (nhim th s 5). Trung bnh a hnh ghi nhn c trong cp lai gia japonica x indica l 67%. Phn tch lch phn ly (segregation distortion) thng qua php th 2 vi ga nh tn sut alen l 1:1 (indica : japonica). Kt qa tn sut alen ca IR50 l 0,52 v ca M-202 l 0,48. Trong s 181 loci c xc nh trn bn , c 12% nghing v alen ca IR50 v 5% nghing v alen ca M202 (hnh 8-3). Tn sut alen i vi IR50 bin thin t 0,19 n 0,72 vi tt c marker nh v trn nhim sc th s 5, 6, 9, 10 v 11, biu th mt s phn b chun ca alen. Cc on nhim th 1, 3 v 8 th hin s lch lc trong phn ly, xu hng nghing v alen ca ging japonica (Andaya v Mackill 2003). Bin thin kiu hnh v s hin din QTL c thc hin trn c s thanh lc stress do nhit lnh 12oC, thng qua s quan st t l ht bt th SFPa ca M-202 t 70% xung 60%, v SFPa ca IR50 t 30% xung 20% vo lc 5-6 ngy sau khi x l nhit lnh,
trong , kt qa quan st 5 ngy u tin cho thy: IR50 gim t ngt SFPa. Tri li, SFPb ca IR50 biu th rt thp vo lc 6 ngy sau x l, trong khi SFPb ca M-202 vn duy tr gi tr 70% cho n ngy th 7 (Andaya v Mackill 2003). Bng 2: Phn tch QTL tnh trng chng chu lnh theo phng php cch qung, qun th RIL t t hp lai IR50 x M-202 (Andaya v Mackill 2003)
QTL % ht hu th
SFPa
Nhim th
Qung gia 2 marker
Ga tr cM
LOD
R2
Tnh cng
nh hng DFEb M M M M M M I I M
qCTB2a qCTB9
SFPb
2 9 1 2 3 5 6 7 12
RM234-RM301 RM257-RM242 RM151-RM259 RM324-RM301 RM156-RM214 RM26 RM334 RM50 RM173 RM129-RM81b RM292-RM260
2,6 5,1 14,5 2,6 3,1 10,4 14,9 10,1 3,1
6,0** 3,7* 5,1** 3,6* 5,9** 4,3** 4,3** 4,0* 3,9*
qCTB1 qCTB2b qCTB3 % hoa la khng pht trin (USP) qCTB5 qCTB6 qCTB7 qCTB12
(17,5)a 16,8 10,7 (27,7) 14,6 10,5 16,5 (15,9) 12,8 12,6 11,7 11,6
5,83 4,68 5,81 4,02 5,76 -10,63 7,42 5,13 - 9,11
(a): biu th tng cng nh hng QTL trong gii thch bin thin kiu hnh, (b): DPE vit tt t ch direction of phenotypic effect, c ngha l xu hng nh hng kiu hnh thuc v M (ging M-202) v thuc v I (ging IR50). Theo kt qa ny: T l phn trm ht hu th c xu hng nghing v kiu gen ca ging chng chu lnh M-202. T l phn trm hoa la khng pht trin USP (undeveloped spikelet percentage) c xu hng nghing v kiu gen ca c hai ging: ging nhy cm vi nhit lnh IR50, v ging chng chu M-202. SFPa c iu khin bi 2 QTL nh v trn nhim sc th s 2 v 9 (bng 2, hnh 8-3), k hiu l qCTB2a v qCTB9. qCTB2a c xc nh gia hai marker RM234-RM301, gii thch 16,8% bin thin kiu hnh qCTB9c xc nh gia hai marker RM257-RM242, gii thch 10,7% bin thin kiu hnh SFPb c iu khin bi 3 QTL nh v trn nhim sc th s 1,2 v 3, gii thch 27,7% bin thin kiu hnh (bng 2) y l cng trnh u tin phn tch QTL tnh chng chu lnh giai on lm ng vi qun th RIL, nghim thc x l nhit lnh trong phng. Bi v trc , c nhiu cng trnh thanh lc trong nc ng lnh, v nhim sc th s 3 v 4 c xc nh l vng nh v ca QTL iu khin tnh chng chu lnh (Saito v ctv. 1995, 2001). Nhng QTL nh v trn nhim sc th s 3 ca cng trnh Andaya v Mackill (2003) khng ging vi QTL ca Saito v ctv. (2001). Gn y nhng QTL nh v trn nhim sc th s 1, 7 v 11 iu khin tnh chng chu lnh giai on lm ng cng c cng b trong t hp lai gia hai ging japonica (Takeuchi v ctv. 2001)
Hnh 8-3: Bn QTL tnh chng chu lnh giai on lm ng, trn c s 191 dng RIL (IR50 x M-202) (indica x japonica), vi microsatellite marker ph trn 12 nhim sc th (Andaya v Mackill 2003) Nh vy c 8 vng nh v trn 8 nhim sc th c ghi nhn c s th hin ca nhng QTL gi nh iu khin tnh chng chu lnh giai on lm ng. Kt qa ny khuyn co mt phng php mi thay v thanh lc mn trong iu kin x l nc lnh, ngi ta c th dng phng c nhit lnh nh gi kiu hnh. Hng lu di, ngi ta c gng xc nh chin lc s dng microsatellite marker p dng MAS trong chn ging chng chu, nhm chi ph v cng sc trong thanh lc tnh chng chu lnh. 8-3. S TRUYN TN HIU & PHN NG IU TIT P SUT THM THU
Kh hn, mn v lnh l 3 yu t mi trng ph bin nh hng n c ch iu tit cht p sut thm thu trong t bo (OA = osmotic adjustment) (Boyer 1982, Thomashow 1994, Bohnert v ctv. 1995). Ngi ta ngh rng cn phi hiu bit mt cch y c ch nhn v truyn tn hiu ca t bo nh mt cha kha bit c phn ng ca cy v c s di truyn tnh chng chu lnh, t chng ta mi c th ci tin ging cy trng theo mc tiu mong mun. Phn ng i vi stress gy ra s bin i p sut thm thu do thiu nc, do nng mui cao c bo co thng qua s th hin ca nhiu gen lm thay i tnh trng ca cy (xem chng 2 v 3). Phn ng ny cng c quan st trong iu kin nhit lnh, vi thut ng osmotic stress responsive genes, vit tt l gen OR (Nodin v ctv. 1991, Thomashow 1994, Giraudat 1995). C hai loi stress i vi p sut thm thu v i vi nhit lnh u lm gia tng hm lng ABA (abscisic acid) (Chandler v Robertson 1994). S th hin ca nhng gen OR c th c kch hot bi vic gia tng hm lng ABA (Zhu v ctv. 1997). Tuy nhin, trong phn tch s th hin gen OR trong cc dng t bin thiu ABA (ABA-deficient)v dng t bin khng nhy cm ABA (ABA-insensitive), ngi ta thy rng s th hin ca mt vi gen OR t ra kh c lp vi ABA (Gilmour v Thomashow 1991, Nordin v ctv. 1991, Gosti v ctv. 1995). Bn cnh , mt nguyn t iu tit DNA c tnh cht cis- acting, c xem nh l nguyn t phn ng vi triu chng mt nc (DRE = dehydation-responsive element), nguyn t ny ch phn ng vi stress do lnh v stress do tn thng cht nguyn sinh, m khng phn ng i vi ABA (Vasil v ctv. 1995). Nh vy, tin trnh truyn tn hiu c tnh cht c lp vi ABA v tin trnh l thuc ABA u c th xy ra, gen iu khin tnh chng chu lnh th hin v iu tit p sut thm thu, bo v cy gim thit hi do stress (Gosti v ctv. 1995, Shen v Ho 1995, Ishitani v ctv. 1997, Stockinger v ctv. 1997). Ngi ta lm th nghim xem xt tin trnh truyn tn hiu OR trong sinh vt n bo Saccharomyces cerevisiae v ghi nhn nhiu thng tin b ch v s xc nh v th hin cc gen OR (Brewster v ctv. 1993, Posas v Saito 1997, Zhu v ctv. 1997), trong , ngi ta ghi nhn c ch nhy cm trong p sut thm thu vn cn l vn th thch rt ln cho cc nh khoa hc. Ngi ta m t: phospholipase C, nhiu yu t chuyn m v nhng protein kinase mang m di truyn ca gen OR, nhng chc nng nhim v ca chng v tin trnh truyn tn hiu vn cha c g r rng (Hirayama v ctv. 1995, Urao v ctv. 1994, Stockinger v ctv. 1997, Nishihama v ctv. 1995). Ishitani v ctv. (1997) pht trin cc dng t bin ca Arabidopsis thaliana nghin cu tin trnh truyn tn hiu i vi stress do lnh v tn thng cht nguyn sinh, hin tng iu tit gen OR v s th hin gen ny. Ngi ta chuyn np gen th khm, c cha luciferase reporter ca con om m, RD29A-LUC vo cy Arabidopsis, cng thc gen ny c khi ng bi DRE/C-repeat- v ABRE-c cha RD29A promoter (YamaguchiShinozaki v Shinozaki 1994). Cy c chuyn np s th hin tnh cht pht quang sinh hc (bioluminescence) khi phn ng vi stress do lnh, x l ABA. Ht ca cy chuyn np dng ng hp t v transgene, c x l t bin bng ha cht EMS (ethyl methanesulfonate). Cy M2 c thanh lc da trn phn ng pht quang sinh hc. Kiu hnh ca nhiu dng t bin vn khng th gii thch hin tng truyn tn hiu OR. Ishitani v ctv. (1997) tm cch thit lp mt m hnh khc trong nghin cu tin trnh truyn tn hiu ny trn c s hai phng n: tin trnh c lp vi ABA v tin trnh l thuc ABA. 8-3-1. iu tit phn ng pht quang sinh hc ca cy Arabidopsis Cy Arabidopsis thaliana chuyn np gen RD29A-LUC l vt liu th nghim c xem xt ti i hc Arizona, M. Vic chn la mt promoter v mt reporter tng ng l
iu kin cn thit thanh lc cc dng t bin i vi mc tiu nghin cu v iu tit gen v truyn tn hiu. RD29A l mt promoter v LUC l mt reporter c chn thanh lc cc mutants ca gen OR. Chc nng ca RD29A c th c bit vi cor78, hoc Iti78 (Howarth v ctv. 1993, Nordin v ctv. 1993), nhng sn phm ca gen vn cha c bit r. Ngi ta xc nh rng promoter ny c th c kch hot bi stress do nhit lnh v do p sut thm thu (Yamaguchi-Shinozaki v Shinozaki 1994). Bn cnh , phn ng a dng ca ABRE/ABA iu ha ABA, m RD29A cng c cha yu t DRE, yu t ny c kch hot bi stress do lnh, nhng khng b kch hot do ABA. Nh vy, promoter ny s to iu kin chng ta nghin cu c hai tin trnh: c lp vi ABA v l thuc ABA. Reporter LUC c chn trong trng hp ny bi v s th hin ca n trong cy c th c pht hin bng cch s dng hnh nh video c thut ng chuyn mn l low-light video imaging thng qua hin tng pht quang sinh hc ca con om m. Cng thc gen RD29A-LUC c chuyn np vo cy Arabidopsis (ecotype C24) theo phng php chuyn gin tip nh Agrobacterium tumefaciens, chuyn qua r (Valvekens v ctv. 1988). Ngi ta chn c mt cy trong 9 cy chuyn np c tnh cht c lp, th hin cao nht hot tnh ca LUC trong iu kin b stress cht nguyn sinh v b stress do lnh. Hai primer ca promoter ny l: 5-TCGGGATCCGGTGAATTAAGAGGAGAGAGGAGG-3 5-GACAAGCTTTGAGTAAAACAGAGGAGGGTCTCAC-3 nh gi kiu hnh c thc hin trn cy m 1 tun tui trng trong mi trng agar, iu kin nhit 0oC, thi gian th nghim 48 gi, v o gi tr pht quang sinh hc bng my low-light CCD imaging. Kt lun: Stress do lnh kch thch rt mnh hin tng pht quang sinh hc trong cy chuyn np gen RD29A-LUC ((Ishitani v ctv. 1997). 8-3-2. Phn lp dng t bin c mc pht quang sinh hc thay i Mt s lng ln cc dng t bin thuc dng: cos: c vit tt t thut ng: constitutive expression of osmotically responsive genes c ngha l dng th hin c bn ca gen phn ng vi iu tit nguyn sinh cht los: c vit tt t thut ng low expression of osmotically responsive genes c ngha l dng th hin thp ca gen phn ng vi iu tit nguyn sinh cht hos: c vit tt t thut ng high expression of osmotically responsive genes c ngha l dng th hin cao ca gen phn ng vi iu tit nguyn sinh cht
Chng c phn lp trong cc dng mutant nh k thut cao trong h thng luminescence imaging. Nhng dng t bin los v hos c gom vo thnh 14 lp ty thuc vo tnh cht khim khuyt trong phn ng vi mt stress hoc kt hp nhiu stress v tn hiu ABA (Bng 3). Trn c s phn lp nh vy, ngi ta ngh mt m hnh truyn tn hiu trong thc vt bc cao (hnh 8-4) Bng 3: S dng t bin c xp hng vo nhng lp khc nhau (Ishitani v ctv. 1997) Lp cos los Tt c Lnh NaCl 1a 9 34 3 3 2b 16 139 7 15 3c 8 198 23 47

ABA Lnh/NaCl NaCl/ABA Lnh/ABA 0 3 2 0 6 2 4 6 18 17 12 9 10 32 32 16 12 11 14 hos 12 21 Tt c 9 9 Lnh 20 1 NaCl 11 3 ABA 9 1 Lnh/NaCl 10 11 NaCl/ABA 5 3 Lnh/ABA a t bin c kiu hnh biu th pht quang mnh (tng cng 103) b t bin c kiu hnh biu th pht quang trung bnh (tng cng 271) c t bin c kiu hnh biu th pht quang yu (tng cng 459)
Hnh 8-4: M hnh truyn tn hiu trong iu kin b stress do lnh (Ishitani v ctv. 1997) 8-4. S BIN I PROTEIN THEO NHIT , MN & KH HN Ti i hc New Delhi, n , ngi ta s dng ging la Lal nakanda c tnh chng chu kh hn rt tt lm vt liu nghin cu (Pareek v ctv. 1998) vi cc nghim thc x l: lnh, nng, ABA, kh hn, v mn. S thay i protein ca ging la th nghim vi trng lng phn t bin thin t 10,2 kDa (phn ng vi mn, kh hn, lnh v ABA) n 123 kDa (phn ng vi mn, v ABA), thay i ty theo loi hnh b stress, thi gian x l v ty theo ni ly trch thn hoc r (hnh 8-5, v 8-6)
Hnh 8-5: in di protein phn t cao, ly trch t thn cy m, acrylamide SDS-gel 7,5%, thng qua nghim thc x l mn (salinity), kh hn (desiccation), lnh (cold), nng (heat), ABA (Pareek v ctv. 1998)
Hnh 8-6: in di protein phn t thp, ly trch t thn cy m, acrylamid SDS-gel 15-22%, thng qua nghim thc x l mn (salinity), kh hn (desiccation), lnh (cold), nng (heat), ABA (Pareek v ctv. 1998) Nhiu protein th hin cc vch chng lp ln nhau theo mc nhy cm vi stress. Nhng protein c trng lng phn t 102, 100, 87, 85, 55, 44, 43.5, 43, 41.7, 39, 36, 32, 31, 29, 26, 24, 23.8, 23, 21.5, 19, 18.2, 16.8 v 16.2 kDa trong r la phn ng vi nhng stress khc nhau (hnh 8-9, v 8-10)
Hnh 8-7: in di protein phn t cao r la trong iu kin nng acrylamide SDS-gel 7,5%, thng qua nghim thc x l mn (salinity), kh hn (desiccation), lnh (cold), nng (heat), ABA (Pareek v ctv. 1998)
Hnh 8-8: in di protein phn t thp r la trong iu kin nng acrylamide SDS-gel 15-22%, thng qua nghim thc x l mn (salinity), kh hn (desiccation), lnh (cold), nng (heat), ABA (Pareek v ctv. 1998)
Nghim thc x l ABA c tnh cht bt chc trong t nhin trong c ch truyn tn hiu v phn ng chng chu, cng cho ra kt qa a hnh protein nh nhng stress trong th nghim.

Hnh 8-9: Biu Venn cho thy protein tch t trong phn ng vi cc stress khc nhau thn cy m, theo kt qa in di acrylamide SDS-gel 7,5% v 15-22%. Trng lng phn t (kDa) ca tng protein i vi stress c nh du (*) biu th s suy gim phn ng chng chu vi stress, v nhng gi tr cn li biu th s gia tng kh nng phn ng vi stress (Pareek v ctv. 1998).
Hnh 8-10: Biu Venn cho thy protein tch t trong phn ng vi cc stress khc nhau r la, theo kt qa in di acrylamide SDS-gel 7,5% v 1522% (Pareek v ctv. 1998).
Kt qa phn tch protein nh vy s cung cp cho nh nghin cu nhng kin thc v: (1) nhng thay i trong th hin gen lm c s gii thch nhng bin d di truyn trong phn ng i vi stress, (2) gi tr tng i ca nhng thay i s th hin gen c tnh cht cm ng v th hin gen c tnh cht c bn trong iu khin tnh chng chu vi stress, v lm r hn kin thc gia gen v protein (sn phm cui cng ca n).
Mt khc, ngi ta ghi nhn tnh chng chu lnh ca nhiu loi cy trng c lin quan mt thit vi cc mc khc nhau v tnh khng bo ha ca acid bo trong phosphatidyl glycerol (PG) c trong cc lp mng ca lp th (chloroplast). Thc vt c t l acid bo khng bo ha cao, th d nh Arabidopsis thaliana, s c kh nng chng chu rt tt hn. Enzyme glycerol-3-phosphate O-acyltransferase (GPAT) c trong lp th ng vai tr quan trng trong vic xc nh mc khng bo ha ca acid bo PG (Yokoi v ctv. 1998). Ngi ta phn lp mt cDNA i vi enzyme GPAT c tnh chuyn bit i vi oleate ca genome cy Arabidopsis thaliana di s kim sot ca promoter cy bp Ubiquitin, gen ny c chuyn np vo cy la bng phng php gin tip thng qua Agrobacterium tumefaciens. Cng thc gen chuyn np: KmR-Ubiquitin-GPAT-NOS ter-HygR-pBIN19 Cc mc ca acid bo khng bo ha trong phosphatidyl glycerol trong cy la chuyn np gen t gi tr 20% cao hn ging la khng chuyn np, nhit x l 17oC. Nh vy, vic du nhp cDNA ca GPAT trong cy Arabidopsis vo cy la to ra mt s khng bo ha acid bo ln hn v lin quan n tnh chng chu lnh trong qa trnh quang hp ca cy la (Yokoi v ctv. 1998). Bertin v ctv. (1997) s dng hin tng hunh quang ca dip lc t thanh lc cc ging la chng chu rt. Nhm tc gi khai thc s bin d t bo soma ca cy la thanh lc cc tnh trng mc tiu. K hiu Fo biu th gi tr ti thiu hunh quang dip lc khi m phn ng trung tm PSII, Fm: biu th gi tr ti a, Fv: biu th gi tr ca ging (=Fm Fo), Fv/Fm l hiu qa ca PSII photochemistry. nh gi hiu qa ca h thng quang hp II (PSII): photochemistry Fv/Fm nhit lnh i vi cc dng soma, ngi ta o mc hunh quang dip lc t ca tng dng soma trong th nghim ny, so vi ging b m ban u, bng mt cng c xch tay fluorometer (PEA analyser, Hansatech, Anh Quc). Nghim thc lnh trong phytotron c duy tr 10/5oC (ngy/m) trong 72 gi, trc khi x l lnh, cy la iu kin nhit l 28/22oC (ngy/m). Mu l la c ly o gi tr Fv/Fm sau 0, 3, 6, 12, 24, 36, 48, 60, 72, 75, 144, v 147 gi v so snh vi mu i chng c duy tr nghim thc 28/22oC. Nghim thc lnh lm kch hot mt s gim ng k Fv/Fm trong ban ngy, v c b p li vo ban m. Tnh cht ny cn ty thuc vo ging chu chng hay ging nhy cm. L la ca nhng dng soma c tnh chng chu lnh c x l cng iu kin nh trn trong 6 ngy, c thu thp o m. Ngi ta quan st s khc bit rt ln gia cc dng soma vi ging gc ca chng v Fv/Fm. Nh vy, phng php o hunh quang ca cht dip lc trong cc nghim thc x l lnh s gip chng ta pht hin s thay i tnh trng chng chu rt ca cy la (Bertin v ctv. 1997). Kt lun: nghin cu ny cho thy hin tng hunh quang thay i trong dip lc t l mt tiu chun quan trng nghin cu tnh chng chu lnh ca cy la, mt vi c ch nhy cm vi lnh c th xy ra m khng ty thuc vo s thay i ca hunh quang dip lc. Nhng c ch khc thng nh vy c th ghi nhn c trong khai thc bin d t bo soma, v nhng dng soma ny phi c trcnhgim tip tc trn ng rung sau khi c kt qa trong phng th nghim (Bertin v ctv. 1997)

TI LIU THAM KHO Andaya VC, DJ Mackill. 2003. QTLs conferring cold tolerance at the booting stage of rice using recombinant inbred lines from japonica x indica cross. Theor Appl Genet 106:1084-1090 Bertin P, J Bouharmont, JM Kinet. 1997. Somaclonal variation and improvement of chilling tolerance in rice: Changes in chilling-induced chlorophyll fluorescence. Crp Sci 37:1727-1735 Bohnert HJ, DE Nelson, RG Jensen. 1995. Adaptation to environmental stress. Plant Cell 7:1099-1111 Boyer JS. 1982. Plant productivity and environment. Science 218:443-448 Brewster JL, T de Valoir, ND Dwyer, E Winter, MC Gustin. 1993. An osmosensing signal transduction pathway in yeast. Science 259:1760-1763 Dilday RH. 1990. Contribution of ancestral lines in the development of new cultivars of rice. Crop Sci 30:905-911 Futsuhara T, K Toriyama. 1964. Studies on testing methods of cold resistance in rice. Jpn J Breed 14:166-172 Futsuhara Y, K Toriyama. 1966. Genetic studies on cool tolerance in rice. III. Linkage relations between genes controlling cool tolerance and marker genes of Nagao and Takahashi. Jpn J Breed 16: 19-30 Gilmour SJ, MF Thomashow. 1991. Cold acclimation and cold-regulated gene expression in ABA mutants of Arabidopsis thaliana. Plant Mol Biol 17:1233-1240 Giraudat J. 1995. Abscisic acid signaling. Curr Opin Cell Biol 7:232-238 Glaszmann JC, RN Kaw, GS Khush. 1990. Genetic divergence among cold tolerant rices (Oryza sativa L.). Euphytica 45:95-104 Gosti F, N Bertauche, N Vartanian, J Giraudat. 1995. Abscisic acid-dependent and independent regulation of gene expression by progressive drought in Arabidopsis thaliana. Mol Gen Genet 246:10-18 Harushima Y, M Yano, A Shomura, M Sato, T Shimano, Y Kuboki, T Yamamoto, SY Lin, BA Antonio, A Parco, H Kajiya, N Huang, K Yamamoto, Y Nagamura, N Kurata, GS Khush, T Sasaki. 1998. A high-density rice genetic linkage map with 2275 markers using a single F2 population. Genetics 148:479-494 Hayase H, T Sasake, N Nishiyama, N Ito. 1969. Male sterility caused by cooling treatment at the young microspore stage in rice plants. II. The most sensitive stage to cooling and the fertilizing ability of pistils. Proc Crop Sci Soc Japan 38:706-711 Hayashi T, Y Ukai. 1994. Detection of additive and dominance effects of QTLs in interval mapping of F2 RFLP data. Theor Appl Genet 87:1021-1027 Hirayama T, C Ohto, T Mizoguchi, K Shinozaki. 1995. A gene encoding a phosphatidylinositol-specific phosphilopase C is induced by dehydration and salt stress in Arabidopsis thaliana. Proc Natl Acad Sci USA 92:3903-3907 Howarth DP, BK Mclarney, MF Thomashow. 1993. Regulation of Arabidopsis thaliana L. (Heynh) cor78 in response to low temperature. Plant Physiol 103:1047-1053
Ishitani M, L Xiong, B Stevenson, JK Zhu. 1997. Genetic analysis of osmotic and cold stress signal transduction in Arabidopsis: interactions and convergence of abscisic aciddependent and abscisic acid-independent pathways. The Plant Cel 9:1935-1949 Kaneda C, HM Beachell. 1973. Response of japonica indica rice hybrids to low temperatures. SABRAO J 6:17-32 Koike K, K Yoshino, N Sue, Y Umehara, I Ashikawa, N Kurata, T Sasaki.1997. Physical mapping of rice chromosome 4 and 7 using YAC clones. DNA Res 4:27-33 Kurata N, Y Nagamura, K Yamamoto, Y Harusima, N Sue, J Wu, BA Antonio, A Shomura, T Shimizu, S-Y Lin, T Inoue, A Fukuda, T Shimano, Y Kuboki, T Toyama, Y Miyamoto, T Krihara, K Hayasaka, A Miyao, L Monna, HS Zhong, Y Tamura, Z-X Wang, T Momma, Y Umehara, M Yano, T Sasaki, Y Minobe. 1994. A 300 kilobase interval genetic map of rice including 883 expressed sequences. Nature Genet 8:365372 Lander ES, P Green, J Abrahamson, A Barlow, M Daly, SE Lincoln, L Newburg. 1987. Mapmarker: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174-181 Murray MG, WF Thompson. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acids res 8:4321-4325 Nishihama R, H Banno, W Shibata, K Hirano, M Nakashima, S Usami, Y Machida. 1995. Plant homologues of pathways in yeast and animals. Plant Cell Physiol 36:749-757 Nishiyama I. 1976. Male sterility caused by cooling treatment at the young microspore stage in rice plants. XIII. Ultrastructure of tapetal hypertrophy without primary wall. Proc Crop sci Soc japan 45:270-278 Nodin K, P Heino, ET Palva. 1991. Separate signal pathways regulate the expression of a low-temperature-induced gene in Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 17:1233-1240 Nordin K, T Vahala, ET Palva. 1993. Differential expression of two related, low temperatureinduced genes in Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 21:641-653 Oka H, H Morishima. 1967. Variations in the breeding systems of a wild rice, Oryza perennis. Evolution 21:249-258 Pareek A, SN Singla, A Grover. 1998. Protein alterations associated with salinity, dessication, high and low temperature stresses and abscisic acid application in Lal nakanda, a drought-tolerant rice cultivar. Current Sci 75(11):1170-1174 Posas F, H Saito. 1997. Osmotic activation of the HOGMAPK pathway via Ste11p MAPKKK: Scafford role of Pbs2p MAPKK. Science 276:1702-1705 Saito A, M Yano, N Kishimoto, M Nakagahra, A Yoshimura, K Saito, S Kuhara, Y Ukai, M Kawase, T Nagamine, S Yoshimura, O Ideta, R Oshawa, Y Hyano, N Iwata, M Sigiura. 1991. Linkage map of restriction fragment length polymorphism loci in rice. Jpn J Breed 41:665-670 Saito K, K Miura, K Nagano, Y Hayano-Saito, A Saito, H Araki, A Kato. 1995. Chromosomal location of quantitative trait loci for cool tolerance at the booting stage in rice variety Norin-PL8. Breed Sci 45:337-340 Saito K, K Miura, K Nagano, Y Hayano-Saito, H Araki, A Kato. 2001. Identification of two closely linked quantitative trait loci for cold tolerance on chromosome 4 of rice and their association with anther length. Theor Appl Genet 103:862-868
Satake T, H Hayase. 1970. . Male sterility caused by cool treatment at the young microspore stage in rice plants. V. Estimation of pollen developmental stage and the most sensitive stage to coolness. Proc Crop Sci Soc Japan 39:468-473 Satake T, M Shibata. 1992. Male sterility caused by cool treatment at the young microspore stage in rice plants. XXXI. Four components participating in fertilization. Jpn J Crop Sci 61:454-462 Satake T. 1991. Male sterility caused by cool treatment at the young microspore stage in rice plants. XXX. Relation between fertilization and the number of engorged pollen grains among spikelets cooled at different pollen developmental stages. Jpn J Crop Sci 60:523-528 Sawada S. 1978. Study of sterile-type cool injury in rice plants with special reference to the mechanism and inheritance of sterility. Res Bull Obihiro Univ 10:837-883 Shen Q, THD Ho. 1995. Functional dissection of an abscisic acid (ABA)-inducible gene reveals two independent ANA-responsive complexes each containing a G-box and a novel cis-acting element. Plant Cell 7:295-307 Stockinger EJ, SJ Gilmour, MF Thomashow. 1997. Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cisacting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. Proc Natl Acad Sci USA 94:1035-1040 Suzuki S. 1981. Cold tolerance in rice plants with special reference to the floral characters. I. Varietal differences in anther and sigma lengths and the effects of planting densities on these characters. Jpn J Breed 31:57-64 Takahashi M, T Kinoshita, K Takeda. 1973. Character expression of some major genes in rice and their agronomic application. J Facul Agr Hokkaido Univ 57:275-293 Takeuchi Y, H Hayasaka, B Chiba, I Tanaka, T Shimano, M Yamagishi, K Nagano, T Sasaki, M Yano. 2001. Mapping quantitative trait loci controlling cool-temperature tolerance at the booting stage in temperate japonica rice. Breed Sci 58:240-245 Tanno H, J Xiong, L Dai, C Ye. 1999. Some characteristics of cool weather-tolerant rice varieties in Yunnan province, China. Jpn J Crop Sci 68:508-514 Thomashow MF. 1994. Arabidopsis thaliana as a model for studying mechanisms of plant cold tolerance. In Arabidopsis C Somerville and EM Meyerowitz (Eds). Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. Pp. 807-834 Ukai Y, R Oshawa, A Saito. 1991. MAPL: a package of microcomputer programs for RFLP linkage mapping. Rice Genetics Newsl 8:155-158 Urao T, T Katagiri, T Mizoguchi,, K Yamaguchi-Shinozaki, N Hayashiba, K Shinozaki. 1994. Two genes that encode Ca++-dependent protein kinases are induced by drought and high-salt stresses in Arabidpsis thaliana. Mol Gen Genet 244:331-340 Utz HF, AE Melchinger. 1996. PLABQTL: a program for composite interval mapping of QTLs. J Quant Trai Loci 2: (http://probe.nalusda.gov:8000/otherdocs/jqtl/jqtl19968001/utz.html) Valvekens D, M van Montagu, M van Lijsebettens. 1988. Agrobacterium tumefaciensmediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85:5536-5540 Vasil V, WR Marcotte Jr, L Rosenkrans, SM Cocciolone, IK Vasil, RS Quatrano, DR McCarty. 1995. Overlap of Viviparous1 (VP1) and abscisic acid response elements in
the Em promoter: G box elements are sufficient but not necessary for VP1 transactivation. Plant Cell 7:1511-1518
Yamaguchi-Shinozaki K, K Shinozaki. 1994. A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high salt-stress. Plant cell 6:251-264 Yokoi S, SI Higashi, S Kishitani, N Murata, K Toriyama. 1998. Introductionof the cDNA for Arabidopsis glycerol-3-phosphate acyltransferase (GPAT) confers unsaturation of fatty acids and chilling tolerance of photosynthesis on rice. Mol Breed 4:269-275 Zhu JK, PM Hasegawa, RA Bressan. 1997. Molecular aspects of osmotic stress in plants. Crit Rev Plant Sci 16:253-277

CH DN (INDEX)
ABA Abebe abiotic stress ABRE abscisic acid ACIAR acid phosphatase (APA) actin depolymerase ADC adenosine triphosphate adh Adsorption AFLP Agrobacterium tumefaciens Agrotis Ahn Akbar allele mining aluminum toxicity tolerance anaerobic protein Anchor anchor probe Andaya nh hng cng nh hng tri anisomeric genes ANOVA mt chiu ANP (anaerobic protein) Anti-oxidant Apatite Apium graveolens Arabidopsis thaliana Arumugam Pillai AS1007 AS883 AS996 ascorbate peroxydase ASI association genetics Ba bng Ba chc Ba so Babu BAC clone BAC contig backcross inbred lines bn AFLP bn RFLP bn QTL bn QTL ca Pup-1 bn so snh bn vt l Banih Kuning Basmati 370 Basten By BC Beachell Benckiser Bertin BetA bin i protein theo nhit biu Venn BIL binding protein bioinformatics bioluminescence biotic stress Blair Bohnert Bostein Boyer Brachiaria decumbens bronzing Bu C ung C ung G Cng C ung Phn CaMV35S Cn cn giao ti t hp candidate genes carbohydrate Carex Causse Causse CBF1 cDNA CGIAR chalcone synthase Chang Chapin Chaubey chelate Chenab Sel 64-117 Chesholm
bn cch qung bn chi tit theo phng php thay dn bn gen bn lin kt gen chiu di r tng i chiu di ti phn Norin-PL8 chip sinh hc chloroplast chng chu c nhm chng chu c st chng chu kh hn chng chu lnh chng chu mn chng chu ngp chng chu ngp hon ton chng chu nhit lnh chng chu thiu ln chng lp theo hnh thp Choudhury chu trnh Krebs chu trnh phn gii glucose chng trnh genome cy la chuyn m gen OR chuyn np gen CIAT CICA 4 CIMMYT CIP cis-element Clark c ch chng chu in-vitro c ch chng chu ngoi ng co nguyn sinh CO39 / Moroberekan coastal salinity CodA codominant marker comparative mapping comparative survival (CS) composite interval mapping contig cor78 Coronel IR1552 / Azucena cos co-segregates CRL crop replacement CSR10 CSR13

Cht ct ch s XF chiu di r b stress chiu di r i chng DFE (direction of phenotypic effect) dip lc t distal dominant marker dng cn giao ti t hp (RILs) dng ng gen dng n bi kp (DH) dng gn nh ng gen (NILs) dng hi giao (BC) dng hi giao ci tin (ABC) donor DRE (dehydration-responsive element) DREB1A Dubcovsky Dw1 Dw2 i Hc Cornell i Hc New Delhi i Hc Nng Nghip Nanjing nh gi kiu gen nh gi kiu hnh t acid t bc mu t mn t thiu ln in di protein phn t cao in di protein phn t thp iu tit p sut thm thu iu tit nguyn sinh cht iu tra a hnh trong b m c c Phng n bi kp ui Tru EAPA EC Elicitor ELON empirical approach EMS Epistasis Escherichia coli E. coli
Ctb-1 Ctb-2 C l cysOs1 cysOs2

EST ethyl methanesulfonate Ethylene Eucalyptus expressed sequence tags
deep water rice deletion tilling fine mapping fingerprinting flash flood flood prone Flowers fluorometer Fluvaquents Fox FR13A Frisch functional genomics Futsuhara Fv/Fm Gale gen cng tnh gen khng cng tnh Gene isolation Gene machines Gene manipulation genetic drag genetic map genome hc genomics Genon genotyping gi tr d on hot ng ca gen cng tnh giai on lm ng gibberellic acid Gilmour ging cho ging chng chu ging khng p ng vi P ging ti tc Gissi 27 Glaszmann glyceraldehyde-3-phosphate dehydrogenase glycerol-3-phosphate O-acyltransferase glycine betaine Goff Gomosta GPAT
Fageria FAO Hedley hiu qa p ng ca P hiu qa s dng P HilleRisLambers hi giao Hi Nc Quc T hi t gen Hokkai 241 homeologous segment homeologue region homeology Hordeum vulgare hos Howarth Howeler Huang HVA1 Hydraquents hydrophobic barrier ICARDA ICRISAT ideotype breeding approach IET4082 Igarashi IITA Ikehashi indica indicator Ingram inland salinity Inouye interval mapping INTN introgression IR11141-6-1-4 IR20 IR20 x IR55178-3B-9-3 IR26 IR28 IR29 IR36 IR38
Graminae Gregorio Guiderdoni Gunawardena

IR40 IR42 IR442 IR442-2-58 IR46 IR50 x M-202 IR50404 IR52 IR54 IR59606 IR64 IR64 / Oryza rufipogon low-light CCD imagining la mch en la ni la nc su La Thc luciferase luminescence imagining
Hm lng Fe trong cy Hamamura hp thu c chn lc Harushima Hayashi h s Kurtosis h s skewness IR64 x Azucena IR74 x Jalmagna IR74/FR13A IR9764-45-2 IRDWON IRRI Ishitani Iti78 IWRA Jackson japonica jasmonate Jensen Kaneda Kasalath Kasalath segment Katyal Kawasaki kh nng nhnh kh nng phi hp chung (gca) kh nng vn lng Khao Daeng Kh hn Khush Kihara Kihara 397 Kilian Knapp knockout populations Koike Kosambi Koyama Kupkanchanakul Kurata k thut 2DE k thut chng gen k thut cloning lai diallel Lander
Mackill Mahsuri main-effect QTL mannitol-1-phosphate dehydrogenase map-based cloning MAPL97 MAPMARKER MAPMARKER 2.0 MAPMARKER/EXP version 3.0 MAPMARKER/QTL marker-assisted selection Martinez MAS Mazarado McCouch Melchinger Melchinger Mendoza microarray microsatellite microsatellite marker microsporegenesis microsynteny Milan Mishra mixed flood m phng ti a Mohanty Mng chim Monsanto Moon Moore
Lang LE LEA (late-embryogenesis abundant) leaf brozing leaf brozing index (LBI) Leb Mue Nahng Leung Li likelihood Lin Liu LOD logarith likelihood los Nng ty m Nng tri Nelson Np vn Nhm T Vn v Nghin Cu Nng Nghip Quc T Ni Nicotiana plumbaginifolia NIL-C443 Nipponbare Nipponbare x Kasalath Northern blot Nueu O. breviligulata O. longistaminata O. nivara OA Oka OM1314 OM1490 OM269 x Chim bu OM723-11 OM850 OM997 OR Oryza glaberrima Oryza officinalis Oryza rufipogon Oryza sativa OsGAPDH osmolyte osmotic adjustment P5CS Paterson P-efficiency peroxidase Peroxidase

Moormann Morishima mRNA Mse1 MtlD mc hp thu P mc ha gi ca l multiple QTL effect analysis multiple-locus Murray Murty Nahng Kiew Nng Ci Nng son PLABQTL p-nitrophenyl-phosphate Pokkali polyamine Ponnamperuma Porteresia coarctata PR protein Price profile proline promoter promoter rd29A protein cm ng protein cu trc protein SalT proteinase proteomics proximal Pst1 Pup-1 P-uptake putrescine pyruvate decarboxylase QTL qun th cn giao ti t hp qun th n bi kp qun th F2 qun th gn nh ng gen qun th hi giao qun th knockout quantitative trait loci Quijano R2737 Ramaswami Ramiah
phn b chun phn tch epistasis phn tch QTL phn trm sodium trao i (ESP) pht quang sinh hc phenotyping php th 2 PHO phosphatyl glycerol photochemistry phng php CTAB phng php o hunh quang cht dip lc physical map phytotron Pingaew 56 Pisum sativum RIL RM223 Rpg1 RPUE RRL RSDW RUBISCO RUBISCO activase RuBP rye RZ730-RZ801 Sa quay Saccharomyces cerevisiae Saito Salekdeh Sasaki Satake SCAB11 SCAB20 SCAR SD (survival days) Senadhira senescence SES Setter SGT Shinozaki siu tri Sigadis Silewah simple interval mapping single marker analysis single nucleotide polymorphism single seed descent single-QTL

RAPA RAPD RAPDs RD19 RD29A-LUC recombinant recombinant inbred lines recurrent Reddy relative root length relative tiller % reporter
RFLP RGA (rapid generation advance) RGP Ribaut Stuber s th hin gen Suakoko 8 Sub1 submergence tolerance substitution mapping Sulfaquepts superoxide dismutatase Suzuki Swaminathan Syngenta synteny ti hp thu Ti Nguyn Takeuchi tn sut alen Tanksley Tanno Tu binh Tu nt Ty Sng Hu Temnykh Tenasai 2 Terwilliger Thch Thakur th ng dng phn ly th nghim di hn Thinopyrum bessabaricum thoi ha nh bng la thoi ha microspore Thoday Thomashow Thompson
sinh hc phn t skewness SMA SNP Solivas Southern blot spermidine spermine SPUE Sripongpankul SRL SSD SSR stagnant flood stay-green trait step-wise exclusion stress Strizhov STS transgene transgressive transgressive segregation trehalose Triticeae trng lng kh ca chi (SDW) trng lng kh ca r Tropaquents Tropaquepts truyn tn hiu T Gic Long Xuyn tng tc GxE tng tc QTL x E ubiquitin Uchimiya Ukai Umeda USP Utz van Breeman Vergara

Th vin cDNA Th vin DNA t l +ve/-ve t l cy ngoi ln mt nc t l hp thu sodium (SAR) t l ht lp t l Na+/K+ tiu chun nh gi c st TILLING tn hiu Cy3 / Cy5 tnh cht ngoi bin tnh cht ni bin tnh trng thnh phn Tolerance Toriyama Trm Chim Trng chm transcript trans-element Vigna aconitifolia Vos vt tri Wan Wang Weller Wissuwa Wu WUE Xu Xun YAC clone Yamada Yamaguchi-Shinozaki Yano Yeo Yokoi Yoshida Yu Zea mays Zhang Zhu

Co So Di Truyen Cua Tinh Chong Chiu Voi Moi Truong Cua Cay Lua

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Co So Di Truyen Cua Tinh Chong Chiu Voi Moi Truong Cua Cay Lua

Uploaded by

Copyright:

Available Formats

Tai lieu chia se tai: wWw.SinhHoc.edu.

C S DI TRUYN TNH CHNG CHU I VI THIT HI DO MI TRNG CA CY LA

NH XUT BN NNG NGHIP Thnh ph H ch Minh 2003

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

1-12-2-3. Php th t n gin trong qun th hi giao

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Php th F = MSM / MSG(M) 1-12-2-5. M phng trong qun th hi giao

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

aaqq aaQQ AaQQ, Aaqq, aaQq

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn

Tai lieu chia se tai: wWw.SinhHoc.edu.vn