NG DNG K THUT HT NHN TRONG CHN TO GING LA CHT LNG CAO

Trn Duy Qu, Bi Huy Thy, Nguyn Vn Bch , o Th Thanh Bng v cs.

NI DUNG CHNH BO CO
1. Lch s ngnh di truyn v chn ging t bin. 2. Cc tc nhn gy t bin v c ch tc ng ca chng
n vt liu di truyn. t bin.

3. Mc tiu, nh hng ca ngnh di truyn v chn ging

4. Nhng kt qa ni bt ca ngnh di truyn v chn ging
la t bin Vit Nam Vit Nam

5. Chin lc pht trin lnh vc di truyn v chn ging la

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Lch s ngnh di truyn v chn ging t bin

A. Trn th gii
T nm 1900 - 1905, nh bc hc ngi o Huygodveri pht
hin ra cy t bin t nhin cy Oenothera lamarkiana. ng nghin cu quy lut pht sinh cc t bin t nhin v hnh thnh nn thuyt t bin. Darwin, Menden.., khng nh cc t bin trong t nhin l ngun bin di phong ph cho qu trnh chn lc t nhin v nhn to hnh thnh nn cc loi mi v ging mi, thch ng tt vi iu kin t nhin v p ng tt nhu cu ca con ngi. n 1/triu, v th lm th no tm ra cc tc nhn nng cao t bin t nhin nn.

Cc nghin ca cc nh sinh hc t th k 19, 20 nh Lamark,

Tuy nhin t bin t nhin xy ra vi tn s rt thp t 1/vn Phng php gy t bin nhn to c bit n t nm
1925 khi Natxon v Philippp pht hin rng tia Roentgen c kh nng gy ra bin d di truyn Nm H ng.
4

A. Trn th gii
Nhng phi sau, cc cng trnh ca Muller, Xapegin,
Stadle.. trn rui dm v ng (1926-1935), Xakharop, Rapoport, Gustafson v cs trn la m, la mch, mch ba gc (1938-1940) thu c rt nhiu t bin c gi tr trong nghin cu di truyn v chn ging. Chnh nhng cng trnh ny l c s cho s ra i ngnh di truyn hc phng x lm nn tng cho s ra i ngnh chn bin t ging phng x. quc t (IAEA) v T chc nng lng th gii (FAO) ti tr m rng hng nghin cu gy t bin ci to nhng ging cy nng nghip v cy cng nghip nhiu nc trn th gii nhm to ra hng lot ging mi nh: la, la m, la mch, to, chanh, ma, chui v nhng loi cy trng khc.

Ngay t nhng nm 1970, C quan nng lng nguyn t

 Cho ti thng 7/2010 (FAO/IAEA Mutant Varieties Database),

A. Trn th gii

trn 2970 ging cy trng c to ra bng gy t bin thc nghim trn phm vi 62 nc. Vic ng dng k thut ht nhn ci tin cy trng mang li hiu qu cc k to ln v kinh t nng nghip. c tnh hng trm t la v hng trm triu hecta gieo trng bng nhng ging cy trng c to ra t t bin.
th gii l mt s ging cy trng nh: c Bermuda, l Nht bn, ging la na ln Remei Nht bn, khoai lang, khoai ty, hoa cc, hoa hng vi mu sc khc nhau v hnh dng cnh hoa a dng cam khng ht ca ran,Vi khng ht A4 ca Trung Quc. c bit Trung quc ging la t bin Zhefu 802 c trng vi din tch ln nht th gii (trn 10,5 triu hecta) v c s dng rng ri trong sn xut trong thi gian kh di trn 10 nm. c trnh by biu di y:

Nhng thnh tu to ln m gy t bin thc nghim em li trn

Mt s thnh tu chn ging t bin trn th gii qua cc nm

6

S lng ging cy trng c to ra nh gy t bin trn th gii (Tnh n 7/2010)

S lng ging

Hn 2970 ging cy trng

Nm

FAO/IAEA Mutant Varieties Database

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B. Vit Nam
Cc nghin cu di truyn v chn to ging vit nam c
tin hnh chm hn nhiu so vi th gii.

Nm 1966, cc nghin cu v nh hng ca ria Gamma,

ngun coban 60, DES, DMS n cc bin d di truyn la, du tm ti b mn Di truyn khoa Sinh i Hc Tng Hp H ni (Trnh B Hu, Phan Phi, L Duy Thnh). T nm 1968 trn u H Lan ca Trn Minh Nam, trn Nigenladamastica ca Phan Phi (1969-1972), trn la Chn Chu ln v Trung Quc 2 ca L Duy Thnh, Trn Duy Qu (1969-1970), tip theo l cc nghin cu trn cy c chua, to, la Vin cy lng thc v thc phm (V Tuyn Hong v cs.. 1975-1980). Nguyn Vn Mng (1971-1974) trn mt s cy trng.

Cc nghin cu nh hng ca tia gama ca Thi Cng Tng,

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B. Vit Nam
Sau gii phng Min Nam cc nghin cu chn to ging t bin pht trin mnh
nhiu vin nghin cu v cc trng i hc: i hc Tng Hp H Ni, i hc s phm H ni I,II,Nng nghip I, Vin Di truyn Nng Nghip, Vin la B sng Cu Long, Vin Khoa Hc k thut Nng Nghip Min Nam, Vin Cy Lng thc v cy thc phm, Vin Nghin cu ht nhn Lt sut sc trong nghin cu di truyn v chn ging t bin cc cy trng khc nhau nh la, ng, u tng, rau hoa: Vin di truyn Nng Nghip (Phan Phi, Trn Duy Qu, Nguyn Hu ng, Mai Quang Vinh v cc cs., 1984- 2009), Vin Khoa hc Nng nghip Min Nam ( Khc Thnh v cc cs., 1995-2009), Vin nghin cu la B sng Cu Long (on Vn Ro, Bi Ch Bu v cc cs., 19902009), Vin Cy Lng thc v thc phm (V Tuyn Hong v cs., 1990-2009), Trng i hoc s pham H ni I (Nguyn Minh Cng v cs 1990-2009),Vin Nghin cu ht nhn Lt (L Xun Thm v cs, 1998-2009). trn cc i tng la, ng, u tng, c chua, du tm, to, hoa cc, hoa hng. tr thnh nc th 4 v chn ging t bin ca chu sau Trung Quc, Nht Bn, n v ng th 8 th gii sau Trung Quc,n ,Nht,Lin X c,H Lan, c, M.

c bit trong nhng nm gn y cc Vin i tin phong v c nhng thnh tu

Cho n nay cc nh khoa hc Vit nam chn to c hn 57 cc ging t bin

Cc tc nhn gy t bin v c ch tc ng ca chng n vt liu di truyn

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Cc phng php gy to t bin

Tc nhn vt l: tia gamma, tia X, cc chm
neutron nhanh, neutron chm

Tc nhn ho hc: EMS, EI, DES, DMS, NMU,

NEU

Kt hp x l tc nhn vt l v tc nhn ho hc
- S dng phng php no tu thuc: i tng, kiu sinh sn ca cy trng, tnh c hiu ca loi tnh trng mong i to ra
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TC NHN VT L
Chiu x tia Cc tm (UV)
Chiu x tia in t trng (electromagnetic radiation)
+ Tia X t my gia tc tia X. + Tia Gamma t ngun Co-60, ngun Cs-137.

Chiu ht in t
+ Chiu tia nhit hoc chiu chm tia ntron chm t l phn ng ht nhn + Chiu chm tia ntron nhanh t l phn ng ht nhn. + Cc tiu phn Beta: Phosphorus 32 hoc Sulfur- 35.

Chm tia ion.

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TC NHN HA HC

Dimethyl Sulfate (DMS) Diethyl Sulfate (DES) Ethyl Methane Sulfonate (EMS) Ethylenimine (EI) N- Nitroso Methyl Urea (NMU) N-Nitroso Ethyl Urea (NEU) Sodium Azide (NaM3) Colchichine

13

C CH TC NG
Nhiu th h cc nh khoa hc trn th gii v Vit nam khi
nghin cu c ch tc ng ca cc tc nhn vt l v ha hc n phn t ADN, NST, t bo, c th, chng gy nn cc phn ng ha hc vi phn t ADN to nn cc dimertimin to nn s thay th cc gc baz nit ca nhm purin bng pyrimidin, hoc to nn cc t on, sai hnh cc NST Ni tm li l to nn nhng thay i v cu trc ca gene, nhng cu trc ny c di truyn cho cc th h sau thng qua cc qu trnh phn bo, nguyn nhim v gim nhim. Cui cng to nn cc c th mi mang nhng c im di truyn khc vi b m. c tnh mong mun nh nng sut cao, kh nng chng chu su bnh, hn, ng to thnh cc ging mi.

Chng ta tin hnh chn lc cc t bin c tao ra theo nhng

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CC PHNG PHP CI TIN GING CY TRNG

Chn lc v bo tn nhng vt liu cy trng sn c trong
t nhin c ci tin mt cch t pht p ng nhng tiu ch ca nh chn ging.

Lai to kt hp chn lc.

Gy to t bin kt hp chn lc.

K thut sinh hc: chuyn gen to ra cc c th c tnh

trng mi GMO (Genetics Modified Organism).

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Mc tiu, nh hng ca ngnh di truyn v chn ging t bin

16

Mc tiu ci tin ging cy trng

To ra ngun gene khi u a dng phc v cc nghin cu l thuyt v di truyn hc v chn to ging cy trng: - Nghin cu bn cht ca gene quy nh cc tnh trng - Lp bn cc gene quy nh cc tnh trng: nng sut, cht lng, tnh chng chu su bnh, kh nng chu ng i vi strees ca mi trng (hn, ng, chua mn, nng, lnh gen bt dc c TGMS..) To c cc ging mi theo yu cu ca ngi tiu dng.

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u im ca chn ging t bin

- To ra s da dng v mt di truyn ca vt liu khi

u nhanh v hiu qu. - To c ging mi trong mt thi gian ngn, c bit i vi loi cy sinh sn v tnh. - Thay i mt hoc mt vi tnh trng m khng lm nh hng ti nhng tnh trng cn li ca cy trng. - Nng cao hiu qu cng tc chn to ging so vi cc phng php chn ging c in v hin i.

Nhng kt qa ni bt ca ngnh di truyn v chn ging la t bin

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 Trong bo co ny, chng ti tp chung trnh by

nhng kt qu ch yu thu c khi x l cc ging la a phng, nhp ni hay cc dng mi lai to c nh cc ging: Tm Thm Thi Bnh, Tm Xun i, D Hng, Chim bu, Cm, Np ci Hoa vng, Np Hi Hu, Mc Tuyn, IR8, C4-63, TB1, cc dng mi lai to nh: ST3, ST5, ST13, ST16, ST19, ST20 bng cc tc nhn vt l v ha hc nh tia Gama, ngun Coban60 vi liu lng 15, 20 v 25 Krad. Cc cht ha hc nh NMU, NEU, EI, DES, DMS vi cc nng 0,01; 0,015; 0,025; 0,030% trong thi gian t 18 -24h ht kh hoc ht ny mm.

 T nm 1980 n nay, chng ti thu c rt nhiu

cc t bin mang nhng c im nng sinh hc qu nh kiu cy cy ci tin, cc yu t cu thnh nng sut cao nh cu trc bng, khi lng 1000 ht tng cao, cht lng go cao, do thm. c bit nhiu t bin chn sm cy c hai v/nm, khng c nhiu su bnh nh o n, bc l, kh vn, ry nu..

Nhiu trong cc s t bin nu trn c trc tip

chn lc qua mt s th h tr thnh ging hoc c lai tch ly vi b m hoc lai gia cc dng t bin vi nhau v thng qua chn lc tr thnh ging phc v sn xut.

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T nm 1985 n nay, chng ti a ra c hng chc ging la t bin c chn lc bng phng php thc nghim nh DT10; DT11; DT13; DT14; DT17; DT33, DT21; DT22; DT37; DT38; A20; DT122; D271; CM1, Tm thm t bin, ST3; MT4; MT2 Nhiu ging n nay vn tn ti trong sn xut. Ngoi cc t bin c li cho cng tc chn to ging cn c cc t bin phc v cho cc nghin cu lp bn di truyn v gen nh gen bt dc c TGMS, gen khng o n, bc l, ry nu, gen mi thm, gen khng mn cm vi quang chu k.. Mt v d in hnh l ging la t bin DT10 ca Vin Di truyn Nng Nghip nhn c t vic chiu x ht kh ca ging C4-63 liu 20Kr. Ging ny c cng nhn tiu chun ging quc gia nm 1990. Khc vi ging b m, ging DT10 c kh nng chu lnh, chu ng, chng chu bnh tt hn b m v cho nng sut kh cao v n nh (nng sut t 5,5 - 7,0 tn /ha). Ging la ny c trng nhiu vng min bc Vit Nam vi din tch khong 1 triu hecta, chim 33% din tch trng la thp k 90. Mt s dng t bin c trin vng c s dng nh ngun vt liu ban u cho cc php lai nh trng hp ca ging la A20, DT22. 22

 Phn ln cc ging t bin c to ra bng chn lc

trc tip t cc dng t bin trin vng t cc th h M3 tr i, chn ra nhng dng c nhng tnh trng tt hn ging gc v mt s c tnh nng hc nh nng sut, cht lng (Hnh 2, Hnh 3, 4).

Mo

AABBCC.. AABBCc AABBCC AaBbCc

M1 T bo khi sinh th 1 T bo khi sinh th 2 T bo khi sinh th 3 M2 (Cc th h con M1) 1st sector 1/3 Plants from: AABBC.3/4 AABBcc.1/4

Cc bin i di truyn trong loci

2nd sector 1/3 AABBCC (Khng phn ly)

aa- Thn ln; bb- mm bch tng; cc: lp phn trn l

Chn lc cc cy thn ln t bin Tn s ca cc cy thn ln M2 Kiu hnh: aaB.CC 1/3 x 3/16=3/48 Kiu gen: aaBBCC 1/3 x 1/16=1/48 (1/16)

3rd sector 1/3 A.B.C. 9/16 A.bbC 3/16 aaB.CC 3/16 aabbC 1/16

Tt c nhng c th c kiu hnh aaB.CC c trng ring bit M3 tip tc chn lc

Hnh 2: S k thut chn t bin truyn thng cy trng sinh sn hu tnh 24 (la, ng, u tng)

Hnh 3: B NST la O.sativa. L

25

Hnh 4: Cc kiu t bin NST hu k nguyn phn

26

 Nhng tnh trng c ci tin thng qua t bin

thng l nng sut, gim chiu cao, chng chu, su, bnh (i vi la: DT10, DT11,...); ph v tnh cm quang (tm thm t bin, DT21); Cht lng ( la CM6); Chu stress ca mi trng ( la CM1), c t bin thay i mu sc ca ht go lm tng tnh thng phm trn th trng nh trng hp ging la Chim bu a phng ht go , nng sut thp, ging t bin CM1 v CM6 ht go chuyn sang mu trng lm tng gi tr thng phm (Bng 1; 2,3); Hnh 5,6).

Hnh 5: Cc kiu t bin tnh trng cy la

Hnh 6: Cc kiu t bin tnh trng cy la

Bng 1. Cc ging la mi c to ra bng phng php gy t bin nhn to

Cc ging t bin
DT10 DT11

Phng php x l t bin

Chiu x (1) 20 Kr i vi ht kh 20 Kr i vi ht kh 20 Kr i vi ht kh 20 Kr i vi ht kh 20 Kr i vi ht kh 20 Kr i vi ht kh Chiu x callus 10-20 Kr vi ht t Ha cht(2)
M2: 0,025% NEU1 M2: 0,025% NEU 0,015% NMU2

Kt hp (1 ) v 2

Kt hp 1 v lai to

+ +

A20
CM1 CM6 DT33 DT36
Tm thm t bin

DT21 DT22
Khang Dn t bin 10-20 Kr vi ht t 25 Kr i vi ht kh

NEU 0,025% NEU 0,025%

+ +

DT38 CL9 PD2

Ht kh
Ht t

+ +
30

Bng 2. Phng php chn lc cc ging la t bin

Teen ging
DT10 DT11 A20 CM1 CM6 DT33 DT17 DT36 DT21 Tm Thm t bin DT22 Khang Dn t bin DT38 CL9 PD2 ST3 + + + + + + + + +

Phng php chn lc Trc tip Gin tip

+
+ + + + + +
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Bng 3: Cc tnh trng c ci tin ging la t bin

Ging STT 1 Ngun gc C4-63 t bin DT10 Tnh trng c ci tin nh t bin Nng sut cao, chu lnh, chu su bnh

Nm c cng nhn tin b khoa hc k thut

1990 (Ging quc gia)

2
3 4

C4-63
DT10/CR203 A8 Chim bu Chim bu

DT11
DT13 A20

Cht lng tt, ngon cm hn DT10

cht lng cm kh hn DT10, nng sut cao Chu hn, chu phn mn, cht lng tt

1995 (Ging quc gia)

1998 (Ging quc gia) 1993 (Ging quc gia)

CM1

Chu phn mn, i mu ht t sang trng, chn 1999 (Ging quc gia) sm Chu phn mn, mu ht, cht lng, mi thm 2000 (Ging tm thi)

CM6

CR203 VN-01/ b BG-902 (F6)

DT33

Nng sut cao hn ging gc 20%, chu c bnh 1994 (Ging quc gia) bc l Nng sut cao, khng su 2002 (Ging tin b bnh KHKT)

DT36

Bng 3: Cc tnh trng c ci tin ging la t bin

9 10 DT10/ OM80 DV2/ np 415 Tm thm Hi hu DT21Dai loan Khang dn DT17 DT21 Tm thm t bin DT22 DT38 Khang dn t bin Bc thm t bin CL9 Ph v tnh cm quang, cht lng tt hn Nng sut cao, cht lng tt Nng sut cao, cht lng tt

Nng sut cao, cht lng tt

Ph v tnh cm quang, nng sut Ph v tnh cm quang, ngn ngy Nng sut cao, cht lng tt Nng sut cao, cng cy, chng Nng sut cao Ph v tnh cm quang, cht lng tt hn Ci thin cht lng go

1999 (Ging tm thi) 2000 (Ging quc gia) 2000 (ging quc gia) 2002 (Ging tm thi) 2007 (Ging tm thi) 2007 (Ging Quc gia) 2006 (Kho nghim quc gia) 2006 (Ging quc gia) 2006 (Ging quc gia) 2006 (gingKN quc gia) c cng nhn ging quc gia nm 1999

11 12 13

14

Khang dn Bc thm Gy t bin con lai IR64/Khang dn 18 Np t bin 415/KT90 Sc Trng (ST) IR64

15

16

17 18 19

PD2 ST3 B VN 95-20

 Ngoi ra, kt hp nghin cu t bin vi nghin cu sinh

hc phn t cng l mt hng i cn c pht huy.

Vic kt hp gia k thut sinh hc phn t vi nghin cu

gy to t bin c thc hin mt cch cht ch, c th cung cp nhng phng php nghin cu chnh xc, hiu qu, nhanh v kinh t hn trong cng tc ci tin cy trng theo hng chn ging t bin v d nh nghin cu lp bn di truyn v gen nh gen bt dc c TGMS, gen khng o n, bc l, ry nu, gen mi thm, gen khng mn cm vi quang chu k (Hnh 7,8).

S dng cc ch th phn t v PCR xc nh t bin mc phn t

TILLING (Targeting Induced Local Lesion in Genome) phng php sng lc nhanh v chnh xc cc dng t bin mc phn t (Till et al. 2003). 35

The TGMS-VN1 gene in a Vietnamese rice variety was mapped. Hnh 7: Bn gene bt dc c nhy cm vi nhit (TGMS-VN1) cy la
36

Marker assisted selection against rice blast

Hnh 8: Bn gen khng o n cy la

37

Chn lc cc ging la t bin nng sut cao, cht lng tt, c sn: Tm Thm t bin

38

Np DT22

39

40

41

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43

Chin lc pht trin lnh vc di truyn v chn ging Vit Nam

44

 Cc kt qu trnh by trn chng t phng php chn to

ging t bin nh k thut ht nhn l rt hiu qa:

- To c nhiu t bin gi tr trong nghin cu l thuyt di

truyn v lp bn gene cy trng:

- To c nhiu vt liu khi u cho cng tc chn to ging:

+ Chn lc trc tip thnh ging mi

+ Lai to gia cc t bin gia cc ging ban u hay gia cc
t bin vi nhau to nn ging mi.

Nng cao hiu qa v rt ngn thi gian to ging mi so vi

cc phng php chn to truyn thng khc hay phng php chuyn gene.

45

 D s dng, gi thnh thp so vi cc phng php chn to

ging hin i khc.

V vy chin lc xy dng v pht trin lnh vc nghin cu di

truyn v chn ging t bin nh s dng k thut ht nhn nc ta theo chng ti phi nh sau: - Nh nc cn xy dng v thc hin chng trnh di hn t nay n 2015 v tm nhn 2030 v ng dng k thut ht nhn trong nng nghip trong c lnh vc gy t bin thc nghim cy trng. Hng nm c nhng ti trng im cp nh nc tp chung vo cc cy trng ch lc nh la, ng, u tng, caf, ch, rau hoa.. gm nh xng, my mc nh my chiu x, hoc xy dng mt trng Gamma l tt nht v c th chiu x nhiu loi cy trng cng mt lc, nhiu giai on khc nhau ca chu trnh pht trin c th ht phn, hp t, tin phi, ht chn sinh l, cy ang sinh trng v pht trin Do s to ra c nhiu vt liu khi u trn nhiu vt liu khc nhau cng mt lc phc v cng tc nghin cu di truyn v chn to ging cy trng nc ta. 46

- Nh nc cn nhanh chng u t mt h thng c s vt cht

 Cn tp chung o to ngun lc cn b k thut v x l chiu x,

vn hnh cc my mc thit b trng Gamma field, cn b nghin cu v di truyn chn to ging . sc a hng nghin c ng dng ht nhn trong nng nghip ngang tm vi cc nc tin tin khu vc v th gii nc, cc doanh nghip trc tip vo mt s khu trong qu trnh nghin cu chn to ging nh k thut ht nhn nh chnh sch lng bng vi cc nh nghin cu, chnh sch t ai thu vi cc doanh ngip u t vo vic trin khai cc kt qa nghin cu ca lnh vc ng dng k thut ht nhn trong nng nghip. vi cc k thut ca CNSH nh nui cy m t bo, k thut marker phn t, k thut tch v chuyn gene nng cao hn na hiu qa ca k thut ht nhn trong nghin cu di truyn v chn ging cy trng nc ta tin ln mt bc mi, p ng c cc mc tiu kinh t, m bo an ninh lng thc v nng cao cht lng ca mt s mt hng nng sn nh la, go, ch, caf.
47

Nh nc cn c nhng chnh sch thu ht ngun lc trong v ngoi

Cn kt hp cht ch giu phng php gy t bin truyn thng

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