11 Cc bnh ph bin trn mt s cy trng quan trng

Trong phn ny v d v nhng bnh ph bin trn nhiu loi rau v mt cy mu c a ra nhm minh ha s a dng ca cc bnh trn cy trng Vit Nam. Cc bnh c lit k trong mi bng cng cung cp mt danh sch kim tra nhm gip vic quan st trn ng rung. Cc tc nhn gy ra nhiu bnh trong s ny ch c th c chn on chnh xc trong phng th nghim. Vic chn on chnh xc tc nhn gy bnh l v cng cn thit trc khi a ra bin php qun l bnh hi tng hp. Chng hn nh thi r do nm c th do nhiu tc nhn gy ra nh Pythium, Phytophthora, Rhizoctonia v Phoma. Vic phng tr cc loi nm ny i hi c cc chin lc qun l bnh hi tng hp khc nhau. S minh ha cho mi cy trng c a ra nhm gip ngi c nhn bit v tr tm cc triu chng ca tng bnh. Hiu bit ton din v nhng bnh ny s gip ngi c trong vic chn on bnh trn nhiu cy trng khc.

11.1

Cc bnh ph bin trn t

Bng 11.1 cung cp danh mc nhng bnh ph bin trn t Vit Nam (cc con s tng ng vi con s trn biu ). Tt c cc bnh c th cng xut hin trong mt v, v mt cy c th b mt hay nhiu bnh gy hi (Hnh 11.1). Thi r Phytophthora, thi gc thn, ho vi khun, sng r do tuyn trng v su c thn u gy ra triu chng ho tng t.

Phn 11. Cc bnh ph bin trn mt s cy trng quan trng

151

Bng 11.1 Cc bnh ph bin trn t

Bnh 1 2 Thi r Phytophthora Thi gc Tc nhn gy bnh Phytophthora capsici Sclerotium rolfsii Du hiu chn on chnh thi r v ho Cc hch nm nh trn mu nu v si nm trng gc thn Dch khun xut hin thn, thn b bin mu nu Vt bnh mu en, lm xung L non ci cc, km pht trin U sng trn r

3 4 5 6

Ho vi khun Thn th Bnh virt Sng r tuyn trng

Ralstonia solanacearum Colletotrichum sp. Vi rt thc vt Meloidogyne sp.

4 3

2 1 6

152

Cm nang chn on bnh cy Vit Nam

Hnh 11.1 Cc bnh trn t: (a) cy t khe (tri) v b ho (phi) c th do mt s bnh gy ra, (b) thn bin mu nu, triu chng in hnh ca bnh ho xanh do vi khun Ralstonia solanacearum gy ra, (c) thi gc mc trng do Sclerotium rolfsii gy ra, (d) thi r Phytophthora do nm Phytophthora capsici gy ra, (e) t b nhim vi rt ho m c chua, (f ) qu t b bnh thn th do nm Colletotrichum sp. gy ra.

Phn 11. Cc bnh ph bin trn mt s cy trng quan trng

153

11.2

Cc bnh ph bin trn c chua

C chua mn cm vi nhiu loi bnh khc nhau. (Bng 11.2). Cn c cc iu tra thm v bnh c chua Vit Nam nhm xc nh tt c cc loi bnh nghim trng hin c. c bit l cn c cc nghin cu chn on v virt v vi khun gy bnh trn c chua. Cc v trng c chua Vit Nam thng b nh hng bi mt s bnh. Mt cy c th b nhim nhiu bnh, lm cho vic chn on tr nn kh khn.
Bng 11.2 Cc bnh ph bin c chua
Bnh 1 2 Ho vi khun Thi gc Tc nhn Ralstonia solanacearum Sclerotium rolfsii Du hiu chn on chnh Ho, dch khun xut hin thn, thn bin mu nu. Hch nm nh mu nu, trn v si nm mu trng xut hin gc thn. Ho, u sng trn r Nm mu xm mc mt di l L vng, ho, thn bin mu nu, m trn qu m hoi trn l L non b bin mu nu cc b, c cc m hoc vng mu ti l gi Ho, mch dn bin mu nu Cc vng trn ng tm mu en trn l Nm mu xm/ta mc mt di l L qun, nh, bin mu vng

3 4 5 6 7

Sng r tuyn trng Mc sng Thi vi khuna m vi khuna Virt ho m c chuaa Ho Fusariuma m vng Mc l Virt vng ngn

Meloidogyne sp. Phytophthora infestans Clavibacter michiganensis Pseudomonas syringae Virt

8 9 10 11

Fusarium oxysporum f. sp. lycopersici Alternaria solani Cladosporium fulvum (Fulvia fulva) Virt

a Cn xc nhn s c mt ca nhng tc nhn gy bnh ny Vit Nam.

154

Cm nang chn on bnh cy Vit Nam

4 7 11

9 10

6 5 1 8

2 3

Hnh 11.2 Cc bnh c chua: (a) triu chng l qun, vng do vi rt gy ra trn nhng chi mi mc. (b) vt lot do vi khun Pseudomonas syringae gy ra trn qu c chua, (c) Sng r tuyn trng do Meloidogyne sp. gy ra, (d) m mc l do Cladosporium fulvum, (e) m vng trn l do Alternaria solani

Phn 11. Cc bnh ph bin trn mt s cy trng quan trng

155

11.3

Cc bnh ph bin trn lc

Lc d b nhim cc bnh trn r, qu, thn v l (Bng 11.3 v Hnh 11.3). Cn nghin cu chn on thm xc nh nguyn nhn chnh gy thi r v v.
Bng 11.3 Cc bnh ph bin trn lc
Bnh 1 Thi r v qu Tc nhn Pythium/Rhizoctonia Du hiu chn on chnh Cht cy con/thi r Cy bin vng v ho Ci cc R bn b bin mu giai on gia v R ci thi vo cui v v thi qu Hch nm trn nh mu nu v si nm mu trng trn gc Cy ci cc v ho Si nm v bo t mu en gc thn v l mm Ho, thi t thn v l, hch nm ln mu en Mn g st mu trn l Vt bnh mu nu s-c-la m Khm, cn chn on trong phng th nghim

2 3

Thi gc mc trng Thi gc mc trng

Sclerotium rolfsii Aspergillus niger

4 5 6 7

Thi thn G st m l Cercospora Virt khm l

Sclerotinia sclerotiorum Puccinia arachidis Cercospora arachidicola Virt

7 6

4 2 3 1

156

Cm nang chn on bnh cy Vit Nam

Hnh 11.3 Cc bnh lc: (a) g st lc do nm Puccinia arachidis gy ra, (b) m l Cercospora (Cercospora arachidicola) v g st, (c) lc b thi r gy triu chng bin vng v ci cc, (d) thi r con v thi qu do Pythium sp., (e) vt bnh trn l mm lc vi rt nhiu bo t ca nm Aspergillus niger, (f ) thi r Pythium cy con, (g) cy khe (tri) v cy b thi r gy ci cc (phi)

Phn 11. Cc bnh ph bin trn mt s cy trng quan trng

157

11.4

Cc bnh nm ph bin trn hnh

Hnh b nhim nhiu bnh nm trn l, thn c v r (Bng 11.4). Hu ht nm gy bnh c th c phn lp kh d dng trn mi trng nhn to. Lu tc nhn gy bnh sng mai l nm k sinh chuyn tnh v khng th nui cy c trn mi trng nhn to. Nhng bnh lit k trong Bng 11.4 c cc triu chng in hnh v thng c th d dng phn bit ngay trn ng rung v sau xc nhn trong phng th nghim. Nm gy thi c c th tip tc ly nhim trong qu trnh bo qun.
Bng 11.4 Cc bnh nm ph bin trn hnh
Bnh 1 2 3 4 5 6 7 8 Chy u l Sng mai m l Stemphylium Thi c r Thi gc mc trng Thi (t) cung l Thi Fusarium Mc en (thi c) Tc nhn Colletotrichum sp. Peronospora sp. Stemphylium sp. Botrytis byssoidea Sclerotium rolfsii Sclerotinia sclerotiorum Fusarium spp. Aspergillus niger Du hiu chn on chnh u l bin mu nu trng, c a cnh. Nm xm mc m l dng ging m vng Nm mu nu xm v cc khi bo t xut hin trn c Si nm mu trng v hch nm mu nu trn gc Si nm mu trng, hch nm to mu en. Si nm c mu trng n tm nht, khng c hch nm Cc m bo t nh bt mu en (cng l bnh gy thi trong qu trnh bo qun ) R mu hng v vy ngoi mu hng Nm mc dy trng nh bng gn vi cc ti bo t en r rt

9 10

Thi r mu hng Thi thn c

Phoma terrestris (Pyrenochaeta terrestris) Rhizopus stolonifer (R. nigricans)

Hnh cng b nhim bnh chy l do vi khun, thi thn c do vi khun, mt s virt thc vt, v mt vi bnh r do tuyn trng (Hnh 11.4). Cc bnh do tuyn trng ch yu gy ci cc v t khi lm cy cht, v vy thng khng c ch n.

158

Cm nang chn on bnh cy Vit Nam

2 3

6 7 9 8 10

Hnh 11.4 Cc bnh hnh: (a) m l Stemphylium, (b) sng mai do nm Peronospora sp., (c) Cc triu chng thi r mu hng do nm Phoma terrestris

Phn 11. Cc bnh ph bin trn mt s cy trng quan trng

159

11.5

Cc bnh nm ph bin ng

Ng c khuyn co trng lun canh vi cy rau phng tr nhiu tc nhn gy bnh tn ti trong t. Ng c kh nng khng bnh ho do vi khun (Ralstonia solanacearum), Sclerotinia sclerotiorum, hu ht cc loi Phytophthora v tuyn trng gy sng r. Tuy nhin, ng kh mn cm vi cc loi Pythium ph bin v tng i mn cm vi Sclerotium rolfsii v Rhizoctonia spp. (Bng 11.5 v Hnh 11.5). Ng cng d b nhim bnh thi thn v bp do mt s loi Fusarium nhng cc loi ny li thng khng nh hng n cy rau. Danh sch chi tit hn v cc bnh trn cy ng c th tm thy trn trang web http://www.cimmyt.org/english/docs/field_guides/maize/diseases.htm.
Bng 11.5 Cc bnh nm ph bin trn ng
Bnh 1 Ung th ng Tc nhn Ustilago maydis Cc du hiu chn on chnh Cc u sng ln mu trng trn ht, cc m bo t en; cng c th gy nhim hoa c v thn. Thi trong thn thng dn n hin tng li thn b ch vn. Thn v bp thi c th c mu hng ti , v c xut hin si nm. Thi trong thn thng dn n hin tng li thn b ch vn. Li thng c mu tm n ta. Si nm trng pht trin trn bp b bnh bn di lp v. Cc mn hoi t di trn l. Gy ra nhng vt bnh ln mu nu nht, loang l trn l v thn. Cc hch nm mu nu hnh dng bt nh xut hin cc v tr b bnh. Cc vt hoi t hnh thnh trn l.

Thi r, bp v thn do Fusarium

Fusarium graminearum

Fusarium verticillioides Fusarium subglutinans Fusarium proliferatum 3 4 G st Kh vn trn r, thn v l Puccinia sorghi Rhizoctonia spp.

Chy l

Bipolaris maydis (Cochliobolus heterostrophus) Exserohilum turcicum

Chy l Turcicum Thi Pythium trn r v thn Sng mai

Vt bnh c kch thc nh hnh bu dc v mng nc trn l sau hnh thnh cc vt hoi t ln hn. Thi t m thn v cc vt bnh mu nu r. Nm mu xm (cnh mang bc bo t) mc mt di l.

7 8

Pythium spp. Peronosclerospora spp. Sclerospora sp. Sclerophthora spp.

160

Cm nang chn on bnh cy Vit Nam

Hnh 11.5 Cc bnh ng: (a) ung th ng do Ustilago maydis, (b) kh vn do Rhizoctonia solani, (c) si nm trng mc trn bp ng b nhim bnh do Fusarium verticillioides

Phn 11. Cc bnh ph bin trn mt s cy trng quan trng

161

12 Nm, ngi v ng vt: cc vn v sc khe

Mt s nm gy bnh trn ngi v cc ng vt khc - nhng bnh ny c gi l cc bnh nm. Chng hn nh Aspergilllus flavus c th xm nhim vo phi ngi, gy ra cc bnh mn tnh v h hp. V vy, phi thn trng khi lm vic vi cc mu nui cy nm A. flavus (xem Phn 12.2.1). Fusarium oxysporum v F. solani lin quan vi cc bnh mt, mng tay v mng chn. Mt s nm gy bnh trn cy cng c kh nng to cc cht chuyn ha bc hai gi l c t nm. c t nm c th ln vo thc phm ca ngi hoc ca ng vt v gy ra hin tng nhim c t nm. Chng hn nh A. flavus to ra cc c t aflatoxin, mt trong nhng nhm c t quan trng nht. Cc afltoxin c trong mt lot cc sn phm nh lc v ng. c t nm c sn sinh ra t cc si nm v ngm vo gi th (nh ht, rm hay tri cy, xem Phn 12.1). c t c th c to ra v ln tp vo trong nng sn trc khi thu hoch hoc trong qu trnh bo qun sau thu hoch. iu quan trng l nn bo qun ht trong iu kin kh gim thiu s pht trin ca nm v tp nhim c t sau thu hoch. Vic sn sinh c t thay i ty theo loi. Chng hn nh Fusarium graminearum sn sinh zearalenone trong ht ng nhng khng c trong ht la m. Aspergillus flavus cn iu kin nng m pht trin v sn sinh aflatoxins trong ng v lc (Hnh 12.2). Ngay c trong mt loi, vic sn sinh c t cng thay i ng k. Trong loi F. graminearum, cc ngun phn lp c th sn sinh deoxynivalenol hoc nivalenol. Nhng khc bit ny rt quan trng, bi v cc c tnh v tc ng ca chng ln cc loi ng vt khc nhau ng k. Mt s nm sn sinh c t trong cc cu trc nm nh hch nm v bo t. Nhng cu trc ny c th ln tp vo ht hay rm r v v vy tc ng n ngi v ng vt n thc n b nhim nm. V d nh hch nm Claviceps purpurea tng i c.

162

Cm nang chn on bnh cy Vit Nam

Nhiu c t chu c iu kin nng, v vy c th tn ti trong thc phm ch bin nh cc sn phm ht ng cc. Mt s c t trong thc n gia sc c th ly sang tht, sa v trng. Con ngi tiu th c t trong thc n t ng cc, cc loi ht, v cc thc phm ch bin khc.

Si nm

c t nm thm vo m ht ng Ht ng b bnh

Hnh 12.1 Ht ng nhim Fusarium graminearum v s minh ha qu trnh c t nm t si nm thm vo m ht.

Hnh 12.2 Aspergillus flavus hnh thnh bo t trn ht lc b nhim bnh trn mi trng phn lp

Phn 12. Nm, ngi v ng vt: cc vn v sc khe

163

12.1 Cc nm c c tnh ch yu Vit Nam

Bng 12.1 a ra danh sch cc nm ch yu c c tnh Vit Nam, cng vi c t do chng sn sinh v i tng cy trng cng nh ng vt b hi.
Bng 12.1 Cc nm c c tnh ch yu Vit Nam
Loi Aspergillus flavus Fusarium verticillioides Fusarium graminearum c t Aflatoxins Fumonisins Deoxynivalenol Nivalenol Zearalenone Penicillium Cyclopiazonic acid Patulin Ochratoxin A Cy trng Lc, ng Ng La m, la mch, ng La m, la mch, ng Ng Ng cc Tri cy Tri cy ng vt Nhiu loi Nga, ln Ln, gia cm Ln, gia cm Ln Xem ti liu Xem ti liu Xem ti liu

c t do nm sn sinh nh hng bi mt s yu t bao gm: gi th nhit m trong gi th dng nm.

164

Cm nang chn on bnh cy Vit Nam

12.2 Cc loi Aspergillus c c tnh

12.2.1 Aspergillus flavus

Ngun Aspergillus flavus thng c trong lc v ng cc vng nhit i, cng c th tm thy trong cc sn phm bo qun trong kho k c gia v. Pht sinh bnh cy Aspergillus flavus tn ti trn cy lc, nhng dng nh khng gy bnh cho cy ang pht trin. A. flavus lin quan ti bnh thi bp ng trong iu kin nng m. c t Aspergillus flavus c th sn sinh aflatoxin v axt cyclopiazonic. Mt s mu phn lp c c tnh rt cao. Aflatoxin c tim nng gy ung th v c th gy ung th gan. Phng nga Loi ny pht trin 37C v c th gy bnh cho ngi, gy sng phi. Bo t v tnh c th cha aflatoxin. Cn phi cn thn khi tip xc vi mu nui cy ca loi ny (Hnh 12.3). Trnh ht phi cc bo t (bo t v tnh).

Hnh 12.3 Aspergillus flavus, ba tn nm trn mi trng Czapek yeast autolysate agar (tri), bo t v tnh mc y trn u cnh bo t phn sinh (gia), bo t v tnh (phi)

Phn 12. Nm, ngi v ng vt: cc vn v sc khe

165

M t Aspergillus flavus to cc tn nm mu xanh-vng, pht trin nhanh, nht l 3037C. Mt s mu phn lp sn sinh cc hch nm c mu nu m n mu en. Cc u (head) Aspergillus mu xanh-vng v c hnh thi ging nh chi lau sn khi quan st di knh lp soi ni. Nhng u ny thng c cu to t 2 lp cung, cung cp 1 v cung cp 2 (t bo sinh bo t), nhng mt s ch c lp cc t bo sinh bo t.

Khng m cc a cy c Aspergillus flavus. Nm ny c th gy bnh phi trm trng cho ngi.

12.2.2 Aspergillus niger

Ngun Aspergillus niger l mt trong nhng loi Aspergillus ph bin nht. N thng c trong lc, v c th c phn lp t hu ht cc sn phm lu c (nh ng cc, cc loi u, gia v) cng nh trong tri cy kh (Hnh 12.4). Pht sinh bnh cy Aspergillus niger gy ra nhiu bnh khc nhau, bao gm thi gc lc, thi v cht cy con, thi mc cy, thi chm nho, thi en hnh ti v mt lot cc bnh thi trn rau qu sau thu hoch. c t Mt s t A. niger c th to ochratoxin A. Loi tng t A. carbonarius l ngun quan trng sn sinh ochratoxin A v c th l ngun ochratoxin ch yu trong cc sn phm nho v c ph.

Hnh 12.4 Aspergillus niger, ba tn nm trn mi trng Czapek yeast autolysate agar (tri), bo t v tnh mc y trn u cnh bo t phn sinh di (gia), bo t v tnh (phi)

166

Cm nang chn on bnh cy Vit Nam

Phng nga Aspergillus niger v cc Aspergilli en pht trin nhanh trong iu kin 37C v c kh nng gy bnh cho ngi. Chng thng c phn lp t tai ngi b nhim bnh. Cn phi cn thn khi lm vic vi mu nui cy ca loi ny. Trnh ht phi bo t (bo t v tnh). M t Cc tn nm A. niger c mu nu s-c-la n mu en v mc nhanh, nht l 30-37C. Phc hp A. niger bao gm tp hp ca mt s loi khc nhau. u cc loi ny thng c mu nu m n mu en sinh ra trn cc cung di v trng ging nh chi lau sn khi nhn di knh lp soi ni. Hu ht cc loi sn sinh cc u c cu to 2 lp cung vi cung cp 1 (metulae) ln.

12.2.3 Aspergillus ochraceus

Ngun Aspergillus ochraceus l loi nm quan trng gy hi trn cc i tng trong qu trnh bo qun. S c mt ca loi ny c bit n trn nhiu loi hng d tr trong kho, nht l nhng vng nhit i. A. ochraceus v cc loi lin quan khc sn sinh ra c t ochratoxin A gy nhim c cho c ph, ca cao, ht ly du v cc loi ht d tr trong kho. Pht sinh bnh cy Khng pht sinh bnh trong cc iu kin thi tit bnh thng. c t Ochratoxin A c tm ra u tin trn mi trng nui cy A. ochraceus. c t ny do mt s loi thuc nhm A. ochraceus to ra. Phng nga C t bo co v vic Aspergillus ochraceus gy bnh cho ngi. Tuy nhin, cng nh tt c cc nm khc, cn phi cn thn trnh ht phi bo t (bo t v tnh). M t Cc tn nm A. ochraceus c mu nu vng nht, v thng c mu nu hng mt di a nui cy. Nhiu dng cng to cc hch nm mu nu hng. C mt s cc loi tng t thuc nhm A. ochraceus (Hnh 12.5). A. ochraceus mc chm hn A. flavus v A. niger, nht l 37C. Mt s loi trong nhm ny khng pht trin 37C. Cc tc gi chn thnh cm n Tin s Ailsa Hocking v nhng ng gp trong vic m t v hnh nh minh ha trong phn ny.

Phn 12. Nm, ngi v ng vt: cc vn v sc khe

167

Hnh 12.5 Aspergillus ochraceus, ba tn nm trn mi trng Czapek yeast autolysate agar (tri), bo t v tnh mc y trn u cnh bo t phn sinh (gia), bo t v tnh (phi)

12.3 Cc loi Fusarium c c tnh

Fusarium verticillioides v F. graminearum l hai loi Fusarium c c tnh ph bin nht trn ng Vit Nam. Nhng loi ny c th xut hin trong cng mt vng. Cc loi Fusarium khc xut hin trn ng, nhng thng t ph bin hn hai loi c bn n y.

12.3.1 Fusarium verticillioides

Ngun Lin quan ch yu n ng nhng i khi cng c phn lp t cc cy khc. Pht sinh bnh cy Gy thi bp, thn v r ng. Ph bin nht trong cc iu kin m, nng, kh, khi cy b thiu nc. Bnh thi bp cng tr nn trm trng hn nhng bp b su b ph hi. Nm ny c th gy nhim khng triu chng thn ng trong cc iu kin thch hp. c t Fusarium verticillioides to nhm c t fumonisin trong ht ng. Fumonisin B1 l cht c nht v ph bin nht. Fumosisin B1 gy ph phi ln v ha lng no nga. Fumonisin B1 cng lin quan n ung th thc qun ngi. C cc quy nh hn ch vic bun bn ng c ln tp Fumonisin B1.

168

Cm nang chn on bnh cy Vit Nam

M t Sn sinh ra cc si nm mu trng trn mi trng PDA v sc t tm trn thch (Hnh 12.6). Trn mi trng thch nc ct c cha l cm chng hoc cc mu thn la xanh tit trng, F. verticillioides sn sinh cc bo t ln di, thon v kh thng tp trung thnh khi trn cc mu thn/l v sn sinh ra cc chui di bo t nh hnh bu dc t cc t bo sinh bo t n. Chng khng to bo t hu.

Hnh 12.6 Thi Fusarium ng do Fusarium verticillioides (tri), v mu nui cy thun trn mi trng PDA (phi)

12.3.2 Fusarium graminearum

Ngun Vit Nam, Fusarium graminearum ph bin trn ng. Cng pht hin trn mt s loi c vng Sapa. Pht sinh bnh cy Gy thi li, r v thn ng trong cc iu kin nhit m. Nm ny cng gy bnh bc ngn la m v k.

Phn 12. Nm, ngi v ng vt: cc vn v sc khe

169

c t Sn sinh ra trichothecenes, nht l deoxynivalenol v nivalenol. C th tm thy chng trong thc phm cho ngi v ng vt lm t ht ng b nhim bnh. Deoxynivalenol (i khi vit ngn DON) cng c bit n l 'c t gy i ma', bi v n khin ln bing n hoc i ma ty theo nng trong thc n. F. graminearum cng to zearalenone, mt c t nm gy ng dc. c t ny gy v sinh, nht l ln, nhng cng c th nh hng n tru b v cc ng vt khc. M t To ra si nm t mu hng n mu ta trn PDA v sc t ta trn thch (Hnh 12.7). Mt s trng hp si nm c mu vng nht. Bo t ln hnh hi cong vi chiu di trung bnh tp trung thnh khi nh trn mi trng CLA hoc trn cc mu thn la xanh trong mi trng thch nc ct. Khng to bo t nh hay bo t hu. To rt nhiu qu th mu en ng tn trn CLA hoc mi trng thch nc ct c cha gi th thc vt thch hp 20-23C trong iu kin chiu sng. Qu th thng khng hnh thnh iu kin trn 25C trn mi trng nhn to. Qu th cng c th hnh thnh trn thn ng v v bp c trong iu kin m v mt.

Hnh 12.7 Thi Fusarium ng do F. graminearum (tri), v mu nui cy thun trn mi trng PDA (phi)

170

Cm nang chn on bnh cy Vit Nam

13 Phng th nghim chn on v nh li

13.1 Phng th nghim chn on

Nhng gi sau da trn nhng phng th nghim chn on c xy dng ti Chi cc Bo v thc vt Qung Nam, Tha Thin Hu, Ngh An v Trng i hc Nng lm Hu thng qua ti tr ca d n ACIAR Chn on, khuyn nng v phng tr bnh hi cy trng ti cc tnh min Trung Vit Nam, CP/2002/115. Nhng phng th nghim ny c xy dng vi mc ch ch yu l h tr cng tc chn on bnh do nm. Tuy nhin, cc phng tin trong phng th nghim cng thch hp cho vic phn lp cc vi khun gy bnh cy ph bin. Khi lm vic ti bt c mt phng th nghim no, cc vn an ton v ri ro nh hng n sc khe u phi c ch . Ph lc 2, sc khe v an ton, phc ha cc ri ro thng gp phi trong mt phng th nghim nghin cu chn on bnh cy. Tuy nhin, cn tham kho kin ph trch phng th nghim trc khi vo mt phng th nghim khng quen thuc.

13.1.1 V tr phng th nghim

Mt phng th nghim chn on cn c b tr trong mt ta nh vi tng bo v trnh ma. Ti nhng vng nhit i, nm thng mc mt bn trong pha tng c ma ht. Nm mc nh vy c th to cc bo t lm ln tp mi trng nui cy. Phng th nghim nn trn tng hai l l tng nht. iu ny gim cc vn v chut v cc su b khc nh kin. Phng th nghim nn c hai phng ln, mt phng chun b v mt phng sch. Nn c mt phng hoc mt khu vc c mi che dng kim tra s b cc mu t ng rung v ra sch t khi mu r. Ti ni ny, cc mu cy nh c chn lc cho vic phn lp tc nhn nm hoc vi khun trong phng sch. Ni ny cng c th c dng tch ly tuyn trng k sinh thc vt t t.

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Hnh 13.1 Sp xp thit b trong mt phng th nghim chn on (phng th nghim ti Chi cc BVTV Ngh An): (a) v (b) hai v tr trong phng sch, (c) v (d) hai v tr trong phng chun b.

13.1.2 Phng chun b

Phng chun b c dng chun b mi trng, bao gm kh trng dng c v vt liu trong ni hp, kh trng a Petri trong t sy, ra v ct gi thy tinh, ha cht v cc vt dng c bn khc. Phng ny cn c mt qut ht hi ht hi nng t ni hp v t sy.

13.1.3 Phng sch

Phng sch dng phn lp nm v vi khun t cc mu cy bnh c lm sch. Phng cng c dng nui cy cc mu vi sinh vt trong iu kin sch. Cc knh hin vi c t trong phng ny nhm kim tra cc mu nui cy v cc cu trc nm.

Khng kim tra cc cy ln trong phng sch. Phn lp t cc mu cy nh c ra sch bi v t bn ngoi trc khi a vo phng th nghim.

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Nu iu kin cho php, phng ny cn c trang b my iu ha khng kh bo v thit b v cc mu nui cy. Phng cng cn c gi cho khi bi v cn trng. Tuy nhin, ng phng qu kn hi, nu khng m s qu cao v nm mc s pht trin trn tng cng nh trn thit b. Nn dng mt my ht m trong phng ny. Khng c mang t vo phng sch bi v t l ngun ca nhn n nm c th lm ln tp cc mu nui cy.

13.2 B tr phng th nghim

Khi thit k mt phng th nghim, c nhiu kha cnh cn xem xt. iu quan trng l cng vic c tin hnh theo mt th t hp l v nhng bc chnh ca qu trnh chn on nn c tch ring khi nhau. Sau y l s b tr ca mt phng th nghim chn on (Hnh 13.2), ch yu l chn on cc tc nhn nm gy bnh thc vt.

T ng mu nui cy

Bung cy v trng

Gi mu cy Phng sch

Bn lm vic T lnh/t

Knh hin vi/knh lp soi ni

My vi tnh Gi sch Phng chun b Bn

Cn T sy Ni hp T bo qun Bn ra dng c thy tinh

Khu vc c mi che

Bn ra mu vt Bn

Hnh 13.2 S phng th nghim chn on, biu th s b tr thit b v bn.

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13.3 Thit b phng th nghim

13.3.1 Thit b cho phng sch

Nhng thit b cn thit cho phng ny c lit k bn di v minh ha trong Hnh 13.2: Mt knh hin vi c gn vt knh 10, 20, 40, v 100 (vt knh du). Mt knh hin vi c bn loi thng l cho hu ht cc cng vic chn on. Nu c iu kin ti chnh, knh hin vi c th gn vi mt vt knh luyn kim 20 vi khong cch lm vic xa. Vt knh ny l tng cho vic kim tra in situ cc cu trc nm trn mi trng nhn to, bi v khong cch t vt knh ti mu quan st kh xa (xem Phn 6.2.2). Mt knh lp soi ni kim tra cc mu cy bnh tm cu trc nm. iu ny c bit quan trng i vi nhiu tc nhn gy bnh trn l m khng th nui cy c trn mi trng nhn to. Knh cng c dng cy n bo t ny mm hoc cy nh sinh trng si nm lm thun mu nui cy, v nghin cu tuyn trng gy bnh cy (xem Phn 6.2.1). Mt t cy v trng dng mi trng v phn lp nm khi m cy. Trong iu kin kh hu nhit i Vit Nam, c nhiu bo t nm trong khng kh. Nhng bo t ny ln tp mi trng khi , cy phn lp hoc cy truyn mu nui cy, tr khi cc thao tc c tin hnh trong mt t cy v trng. Mt gi di c gn n hunh quang bn trn kch thch vic sn sinh bo t v sc t nhiu loi nm trn mi trng nhn to hoc trn mu l m. Nn c mt gi cho cc mu nui cy sch v mt gi ring cho cc mu phn lp. Mt t mu nui cy s thun tin cho vic nui cc a cy cn iu kin bng ti. Vic ny cn thit i vi cc mu nui cy trn mi trng c cht khng sinh b phn hy di tc ng ca nh sng (v d nh mi trng chn lc cho Phytophthora). Mt t lnh d tr cc mi trng trong chai l, a Perti cha mi trng (trong bch ny lng hoc giy nhm trnh cho mi trng khi b kh), cng nh cht khng sinh, mu nui cy v cc mu bnh nh. Cn c mt cn in t vi chnh xc 0,001 g cn lng khng sinh hoc ho cht nh. Cc bn lm vic ln, mt knh hin vi v cn in t, mt cho cng vic phn lp chung v nui cy. Gh ngi thoi mi cc bn lm vic. S ghi chp mu, ghi chi tit mi ln chn on cng nh danh sch cc mu vi sinh vt lu tr.

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 Mt gi sch gm mt lot cc n bn thng tin v bnh: sch gio khoa cm nang trch lc bnh cc bi bo khoa hc t nht mt my vi tnh ni mng internet v mt my in cho: cng vic lu tr c s d liu tm kim thng tin tip cn cc th vin hnh nh lin lc qua th in t. Cc dng c nh cho cng vic phn lp v cy, bao gm: kp nh que cy cn dao m que cy khun li dao m bt nh du dao nh cn tyl que cy truyn nm (u dp) giy lau tht lam knh v lamen giy lc.

Kim tra tng v thit b thng xuyn xem c b nm mc hay khng.

Sn phng sch cn c lau chi thng xuyn loi b bi bn. Cn phi tt qut v ng ca s trong khi nui cy lm gim s di chuyn ca khng kh trong phng. Cng vic quan trng cn c thc hin trong bung cy c lau sch bng cn 70%.

Kh trng b mt khi cn.

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Hnh 13.3 Dng c cn thit cho vic phn lp, cy truyn, lm thun v gim nh cc tc nhn nm v vi khun gy bnh

Kh trng mt bn v ra tay trc khi lm vic vi bt c mu cy sch no gim nguy c ln tp.

13.3.2 Thit b cho phng chun b

Nhng thit b cn thit cho phng chun b c lit k bn di v minh ha trong Hnh 13.1: Mt t sy kh trng a Petri, a Petri cn c gi trong giy bo hoc ti giy. Mt ni hp nh thch hp cho kh trng 1-2 lt mi trng hoc nc trong bnh tam gic hoc chai Schott. Ni hp cng c th c dng kh trng mi trng hoc nc trong ng nghim thy tinh hoc chai McCartney, pipet v cc thy tinh khc gi trong giy hoc giy nhm.

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 Mt ni p sut kh trng lng nh mi trng v nc. C th mua thit b ny hu ht cc ch ln. Mt cn ( chnh xc n 0,1 g) cn ha cht, khoai ty, c rt v nhng nguyn vt liu tng t cho vic chun b mi trng. Mt bp in nu khoai ty v c rt dng cho mi trng. Mt bn chun b mi trng. Mt bn ra a Petri v cc thy tinh khc. Mt t ng.

13.4 Nh li cho vic nghin cu bnh cy

Nh li l mt phn quan trng ca phng th nghim chn on bi v nh li cn cho vic thc hin qu trnh ly bnh nhn to, nh gi thuc tr nm v cc phng php phng tr bnh khc. Vic thit k cn m bo iu kin gip cho cy mc tt v ngn nga ln tp gia cc th nghim ly bnh nhn to v cc th nghim khc (Hnh 13.4). Mt nh li c bn cn c: mi che trong sut sn xi mng dc thot nc tt thong kh tt trong iu kin tri nng (qut cu thng gi rt hiu qu) (Hnh 13.5) thit k chng chut ngun cung cp nc tt cc bn di (Hnh 13.5) mt khu vc chun b bn trong hoc gn nh li. Mi trong sut cho php t nht 75% nh nng xuyn qua. Dng vt liu polycarbonate lm mi rt tt v chng c tia t ngoi, rt bn v d gn vo khung mi st hay g. Tm nha cng c th dng lm mi nhng ch bn 1-2 nm. Mi thy tinh khng thch hp cho nhng vng c bo t hoc ma . Tt nht l mi c gn sao cho hin c kn (trnh bo t). Li che nng c th c dng trong ma h gim nhit trong nh li (Hnh 13.5). Mt sn xi mng dc thot nc tt v c th c gi sch bng vi xt nc.

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Qut thng gi

NHN T CUI
Thng kh trng chu cy Gi chu cy (ct gi) Bn chun b Bn Thng cha Bn ra chn

NHN T BN

Bn tre

Bn tre

Bn tre

Bn tre

NHN T TRN
Hnh 13.4 S minh ha thit k mt nh li thch hp cho vic ly bnh nhn to v cc cng vic th nghim vi tc nhn gy bnh thc vt.

Hnh 13.5 Nh li dng cho nghin cu bnh cy ti Chi cc BVTV Qung Nam: (a) hnh nh tng qut ca nh li vi li chng cn trng, (b) li che nng v mi tn nha polycarbonate phng vi cc b qut cu thng gi

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Cm nang chn on bnh cy Vit Nam

Bn bn nh li c th l tng gch (cao khong 1m). Li st (nh li B40) hoc li st m km (l khong 1 cm ng knh) c th gn gia cc ct, tng gch thp v ko ct mi. Li st gip thng hi tt ng thi ngn chut v chim chc vo nh li. Li chng cn trng tuy tn km, nhng quan trng v c th ngn cn trng xm nhp vo nh li. Cn mt ngun nc tt gi cho sn sch v cung cp nc sch bnh cho cy. ng dn nc phi c mc trn tng sao cho u vi ti khng bao gi chm sn. C in thp sng v vn hnh thit b th rt tin li. Cc bn thp chng g t cc chu cy nn cao khong 1 m v di khong 2-3 m. cao ny gim ti thiu kh nng b ln tp t sn. Nn c bn di loi d di chuyn, c th di di d dng khi cn cho cc cy cao, cy c thn leo ln gin hoc cy n qu cn nh trng trong chu ln. C th dng bn lm bng tre nhng phi c x l vi thuc tr nm ngn mc. Mt cn a 10 kg nn trong nh li cn cc chu trng cy nhm theo di lng nc trong gi th trng.

13.4.1 Khu chun b

Khu chun b c th trong nh li hoc trong khu nh gn . Nn c cc gi ng cao khi mt sn cha cc chu v dng c. Cng nn c ni cha hn hp gi th hoc ct, x da, mn ca mc hoc cc nguyn liu sch bnh khc trng cy phc v th nghim ly bnh nhn to. Mt bn cn dng cho vic chun b cc chu cy, ly bnh vo t v cc cng vic khc. Mt bn cn lm bng vt liu d kh trng, nh thp khng g hoc hoa.

13.4.2 Hn hp gi th
Gi th sch bnh rt cn thit cho qu trnh ly bnh nhn to v nhiu th nghim khc. Gi th sch bnh cng cn cho vic sn xut cy con v gim hom sch bnh trc khi a ra trng rung th nghim. C nhiu loi gi th. Cc c tnh chnh ca hn hp gi th tt l kh nng gi nc tt v d thot nc. C vi loi hn hp gi th c s dng Vit Nam. Vt liu ph bin bao gm mn ca mc, x da, ct, than bn v phn g vin. Mt s thnh phn ny c th cha tc nhn gy bnh; ct c th cha cc tc nhn gy bnh nh Pythium v Phytophthora. X da v mn ca thng sch bnh.

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t rung thng mang nhiu tc nhn gy bnh thc vt. Nhng tc nhn ny cn c dit tr bng xng hi hoc x l nhit (kh trng bng hn hp hi nc/khng kh 60oC trong 30 pht) trc khi t c th c dng cho qu trnh ly bnh nhn to. t t ng rung cha c x l khng c mang vo nh li bi v cc tc nhn trong t c th lm ln tp nh li. Hn hp gi th mn ca c th c lm vi mn ca, ct v phn g vin (70:28:2 theo th tch) v trong 4-6 thng. u tin, gi th trn ny phi nng ti khong 50oC trong thi gian di loi tr bt c tc nhn gy bnh no tn ti trong . Hn hp gi th cn c mc trong cc thng ln. iu quan trng l trnh gi th tip xc trc tip vi t rung hoc cy bnh. X da c th l mt thnh phn l tng cho gi th. Hn hp gi th cng c th c tit trng bng hn hp hi nc/khng kh nu cha c mc. C th trn hn hp gi th trong my trn xi mng sch. Ht phn bn c th c thm vo trong khi trn.

13.4.3 V sinh nh li
Cn c cc quy nh cht ch i vi cn b s dng nh li trnh lm nhim tp cc th nghim ly bnh nhn to hoc cc nghin cu khc vi tc nhn gy bnh trong t rung. Thit b v quy trnh cn tun th bao gm: t mt bn ra chn ni ca ra vo c dp hoc ng cao su ch dng trong nh li khng mang t t ng rung hoc cy bnh vo nh li loi b cy v t dng trong th nghim ngay sau khi th nghim hon tt v t hy cc cy bnh dng nc sch bnh lun gi cho u vi nc khng chm sn nh phun xt ra sn u n cn b khng c vo nh li ngay sau khi thm rung m phi tm ra v thay qun o sch trc khi s dng nh li kh trng tt c cc chu vi thuc kh trng mnh sau khi dng trong th nghim, nh dung dch nc Javen 1% trong 24 gi cc chu kh trng trn gi cao cch xa mt sn x l bn tre vi thuc tr nm cha ng.

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Cm nang chn on bnh cy Vit Nam

13.4.4 Qun l v dinh dng cy

Trng cy trong chu cho qu trnh ly bnh nhn to v cc nghin cu khc i hi vic qun l cn thn vn dinh dng cy. Cc chu nn c l di y thot nc tt. Si nh c th c di y chu gip cho vic thot nc tt. Mc ch l trnh nc ng trong t ni r pht trin. Cn chu thng xuyn duy tr m n nh trong hn hp gi th v trnh nc ng. Ch nn ti nc cho gi th n ngng nng lc ng rung. Cy cn c trng trong hn hp gi th sch bnh. Vic chn la loi hn hp gi th ty thuc loi cy trng, nguyn liu sn c v tnh cht ca th nghim. Cn cung cp dinh dng cho s tng trng bnh thng ca cy. C th cn b sung thm phn bn dng ht vo gi th trc khi trng. Thng th phn bn lng nh dung dch Hoagland's hoc mt sn phm thng mi c dng 1-2 tun mt ln duy tr s tng trng bnh thng ca cy (Hnh 13.6). Cn bn thm phn lng u n c bit nu cy ln c trng trong chu nh trong thi gian di. Phn N-P-K dng lng m c v cc phn vi lng dng lng m c c sn Vit Nam.

Hnh 13.6 Chun b phn bn thng phm dng trong nh li

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C th dng dung dch Hoaglands thay th (xem cng thc trong Khung 13.1). Vic ny c bit c li nu tnh trng dinh dng cn c gim st, hoc cc cht dinh dng no c cn c loi b trong th nghim nghin cu v dinh dng.

Khung 13.1 Dung dch Hoagland's

Dung dch ny c tt c cc cht dinh dng cn thit cho cy pht trin v tng trng. Dung dch Hoagland's c pha t mt s dung dch m pha sn, trn vi nc trc khi dng. Thm vo mi lt nc: 5 mL 5 mL 1 mL 2 mL 1 mL dung dch nitrat kali dung dch nitrat canxi dung dch photphat axit kali dung dch sunphat magie dung dch vi lng m pha sn

10 mL dung dch st-EDDHA pha sn Dung dch m: 1 M 1 M 1 M 1 M KNO3 Ca(NO3)2.4H2O KH2PO4 MgSO4.7H2O nitrate kali nitrate canxi Photphat axit kali sunphate magie (khong 101 g trong 1 L) (khong 236 g trong 1 L) (khong 136 g trong 1 L) (khong 246.5 g trong 1 L).

Dinh dng vi lng lm sn: 0.046 M 0.009 M 0.765mM 0.320mM 0.111mM H3BO3 MnCl2.4H20 ZnSO4.7H2O CuSO4.5H2O H2MoO4.H2O axit boric clorua mangan sunphat km sunphat ng molybdic acid (85%) (khong 2.86 g trong 1 L) (khong 1.81 g trong 1 L) (khong 0.22 g trong 1 L) (khong 0.08 g trong 1 L) (khong 0.02 g trong 1 L)

Dung dch st-EDDHA pha sn 10mM Fe(NO3)3 st-EDDHA (khong 2.45 g trong 1 L)

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Cm nang chn on bnh cy Vit Nam