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Chng 1 GII THIEU

1.1 at van e Ca chua la mot trong so nhng cay rau mau quan trong, c phan bo rong rai nhieu nc tren the gii. Tuy nhien viec gieo trong ca chua van gap nhieu kho khan, tr ngai. Mot trong nhng nguyen nhan lam han che s phat trien san xuat va lam giam nang xuat ca chua o la sau va benh hai. Trong o ang chu y hn ca la benh heo ru do vi khuan Ralstonia solanacearum gay ra. ay la loai vi khuan co kha nang gay hai tren 200 loai cay co, ac biet gay hai nang tren cay ho ca nh ca chua, ca tm, khoai tay, thuoc la va cac cay khac ho nh au phung, gng, chuoi, chung co kha nang song rat lau trong at va lay lan rat nhanh, di chuyen t ni nay sang ni khac. Cay chet hang loat khi la van con xanh giong nh b cat ngang goc. c tnh thiet hai cho san xuat cay trong hang nam do vi khuan Ralstonia solanacearum gay ra la 15-95%(Emil Q.Javier, 1994). e khac phuc tnh trang nay, cac bien phap phong tr tac nhan gay benh bang phng phap sinh hoc c nhieu nha khoa hoc quan tam, nghien cu. ay la mot trong nhng phng phap phong chong co hieu qua nhat hien nay kha nang phong tr vi khuan Ralstonia solanacearum, giup cay ca chua sinh trng va phat trien tot hn, lam cho tac nhan gay benh khong khang thuoc, an toan vi moi trng sinh thai cho ket qua nhanh va chnh xac rat phu hp vi xu hng nong nghiep an toan hien nay va ang c rat nhieu nc tren the gii ap dung. Viec nghien cu cac loai vi khuan oi khang nh : Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, e chong lai vi khuan Ralstonia solanacearum la mot van e het sc can thiet hien nay. V vay, nghien cu ve tnh a dang cua cac ca the trong quan the vi khuan gay benh va s lu hanh cua chung SVTH: Ho Thanh Hoang Download Agriviet.com

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cac vung san xuat la rat can thiet, no lam c s khoa hoc cho cac giai phap phong chong va chon tao giong khang benh sau nay. Xuat phat t thc tien tren va e anh gia kha nang phong tr cua cac dong vi khuan Pseudomonas fluorescens. Chung toi thc hien e tai: S dung cac dong vi khuan Pseudomonas fluorescens phong tr vi khuan Ralstonia solanacearum gay benh heo xanh tren cay ca chua.

1.2 Muc ch yeu cau anh gia kha nang phong tr cua cac dong vi khuan Pseudomonas fluorescens vi cac dong vi khuan Ralstonia solanacearum gay benh heo xanh tren cay ca chua.

1.3 Gii han e tai e tai c thc hien trong thang co nhieu ma, at trong b nhiem nhieu loai benh. Cay ca chua la loai cay de man cam vi nhieu loai benh.

1.4 oi tng nghien cu Cac dong vi khuan oi khang thuoc nhom Pseudomonas fluorescens Dong vi khuan gay benh Ralstonia solanacearum Cay trong: ca chua

1.5 Noi dung nghien cu anh gia kha nang oi khang cua cac dong vi khuan Pseudomonas fluorescens vi cac dong vi khuan Ralstonia solanacearum tren a petri trong phong th nghiem.

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anh gia kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum tren cay ca chua trong ieu kien nha li. anh gia kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum tren cay ca chua ngoai ong ruong.

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Chng 2 TONG QUAN TAI LIEU

2.1 Lch s nghien cu benh Vi khuan Ralstonia solanacearum la vi khuan gay benh mach dan gay hai tren 200 loai cay co. Halted a nghien cu benh nay vao nam 1892, nam 1896 E.F.Smith nghien cu, mo ta, nh ten Pseudomonas solanacearum. Nhng nam sau o, benh heo ru c nhieu nha khoa hoc tren the gii i sau, nghien cu mot cach toan dien nh Kelman 1953, Hayward 1964, Cook and Baker 1983, Yabuuchi 1992, 1995(Vu Trieu Man va Le Lng Te, 1998). Benh heo ru do vi khuan Ralstonia solanacearum tren ca chua co ten la: Southern wilt (theo cach goi cua ngi Hoa Ky), Bacterial wilt (theo cach goi cua ngi Anh), Viet Nam goi la benh heo xanh, heo ru.

2.2 Mc o pho bien va gay hai cua Ralstonia solanacearum Ngay nay, Ralstonia solanacearum c ghi nhan la vi khuan gay hai hau het cac chau luc, pho bien la cac nc nh Angola, Trung quoc, Bangladesh, An o, Indonesia, Srilanka, ong An Ethiopia, Lybia, Kenya, Malaysia, Nigieria, Philippin, Nam Phi, ai Loan, Thai Lan, Uganda, Hoa Ky, Viet Nam, Zambia (Subrhmagam, 1994). Viet Nam, benh heo xanh ca chua do vi khuan Ralstonia solanacearum gay ra la mot trong nhng benh gay hai rat pho bien lam chet heo hang loat cay ca chua tren ong ruong, gay ton that ln cac vung trong ca chua trong nc, nhat la vung ong bang song Hong. Benh c ghi nhan tren ca chua, khoai tay, ca tm, thuoc la, au phung, gng, t. Qua mot so ket qua nghien cu cong bo gan ay cho

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thay nc ta vi khuan gay benh heo xanh thuoc division Chau A (Le Lng Te, 2002), race1, biovar 3 va 4 (Nguyen Th Yen, 2002). Hoa Ky, vi khuan Ralstonia solanacearum c phat hien tren khoai tay, thuoc la Florida c xac nh bang phng phap PCR la biovar 1 (Prakash, 2002). Indonesia, benh c phat hien au tien tren au phung vao nam 1905, vung Cirebon. Ve sau benh c ghi nhan tren nhieu loai cay trong khac nh ca chua, khoai tay, thuoc la, t, chuoi. Ket qua nghien cu benh heo xanh tren au phung gia AARD va ACIAR vao nam 1985 cho rang ay la benh chnh tren au phung Indonesia, s thiet hai do benh gay ra co the t 15-90% nang suat (Wright, 1991). ai Loan benh c gay hai tren ca chua, khoai tay, thuoc la, t, ca tm, au phung, dau tay, ta to, thau dau, me, cu cai, rau den va mot so cay trong khac. Tren nhieu loai cay trong s thiet hai do benh gay ra co the t 5-100% nang suat. Cac dong vi khuan Ralstonia solanacearum chu yeu thuoc race1 (Yung-An Lee, 2001). Ai Cap, benh heo xanh vi khuan do Ralstonia solanacearum c ghi nhan la gay hai nang nhat tren ca chua va a ti so trong nhng nam gan ay, s thiet hai gay ra tren a ti so t 5-20% nang suat (Aly, 2000).

2.3 S lc ve vi khuan Ralstonia solanacearum Vi khuan Ralstonia solanacearum co dang hnh gay, hai au hi tron, co mot long roi mot au, kch tht 0,5-1,5, nhuom gram am la loai vi sinh vat ton tai trong at thng xuyen gay ra cac benh chet heo (heo ru, heo xanh) tren nhieu loai cay trong nh lac, khoai tay, ca chua, t, thuoc la pho bien rong hau het khap cac vung, gay tac hai ln cho san xuat. Chung lan truyen theo nc ti, xam nhap vao

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cay qua cac vet thng va di chuyen vao trong bo mach. Benh thng xay ra vao luc cay ang tang trng. Vi khuan phat trien nhiet o tng oi cao, at am, xam nhap qua vet thng, sinh san cac bo mach, k chu va di chuyen cac bo mach t than en la, sinh oc to lam cay heo, chet. Pha bo mach lam tat ngen s van chuyen nc va cac chat dinh dng, benh hai nhieu tren at cat pha, at tht benh nhe hn. Tren nhng ruong trong luan canh vi cay lua nc benh nhe. Vi khuan thch hp nhiet o 30-370C, nhiet o toi thieu la 100C, toi a 410C, nhiet o gay chet la 520C, man cam c moi trng kho.

2.4 S phan chia vi khuan Ralstonia solanacearum theo ac tnh gay hai 2.4.1 Race Da tren pham vi ky chu c chia lam 4 race (Martin va French, 1997) + Race 1: Tan cong tren nhieu vung a ly, loai cay trong khac nhau. ac biet tren cac cay nh khoai tay, ca chua, ca tm, t, thuoc la, au phung. Vi khuan Ralstonia solanacearum thuoc race 1 thng gay hai nhng vung co nhiet o am ap va gay hai nang vung nhiet i. + Race 2: Chu yeu tan cong tren cac cay thuoc ho chuoi nh chuoi tam boi, chuoi la, chuoi si. + Race 3: Chu yeu tan cong tren khoai tay. Khac vi race 1, race 3 thng gay hai nhng vung co nhiet o thap hn va nhng vung co v o cao. + Race 4: c ghi nhan tan cong tren cay dau tam (mulberry) Trung Quoc. S phan chia cac race thng rat phc tap do phu thuoc vao thanh phan cay ky chu va pham vi phan bo cua chung. Theo Martin va French (1997), race 1 lu

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ton nhieu nam trong at, ngc lai race 3 thng co xu hng giam sau vai nam neu khong co khoai tay dai lam ky chu.

2.4.2 Biovar Da vao ac tnh sinh ly, sinh hoa khac nhau cua cac mau phan lap, 5 biovar co the nhan dang da vao kha nang s dung va oxi hoa 3 disaccharides (cellobiose, lactose, maltose) va 3 ru 6 cacbon (dulcitol, mannitol, sorbitol). Biovar 2 thng bao gom race 3, biovar 5 gom race 4 va biovar 1, 3, 5 gom race 1.

2.5 Trieu chng gay hai cua vi khuan Ralstonia solanacearum Trieu chng thng bieu hien ngay sau khi benh xam nhap vao cay. cay b benh ban ngay la mat mau nhan bong, tai xanh, heo cup xuong, giai oan cay con thng bieu hien tren toan cay con giai oan cay trng thanh trieu chng thng bieu hien la ngon trc. 1-2 ngay au cay co the phuc hoi lai c vao luc tri mat hoac ve em, nhng sau 2-3 ngay la heo khong the hoi phuc lai c na va toan cay b heo ru roi chet. Cat ngang oan than cay gan mat at ta thay bo mach b hoa nau, trong ieu kien am o cao than cay b benh dan dan thoi mem, an manh gan mieng cat co the thay dch nhn vi khuan tiet ra mau trang sa. Re co mau nau en va thoi.

2.6 ac iem phat sinh phat trien benh Thi gian vi khuan lu ton trong at phu thuoc nhieu yeu to nh am o, nhiet o, hoa ly at. Ben canh o con phu thuoc vao race gay benh, race 1 thng lu ton lau trong at trai lai race 3 thng giam sau vai nam do kha nang thch ng thap hn. Vi khuan co the lu ton trong at t 5-6 nam, trong c the ky chu thc vat hoac trong hat giong co the song ti 7 thang, con neu bam dnh tren be mat hat ch ton tai 2-7 ngay (Vu Trieu Man va Le Lng Te, 1998). SVTH: Ho Thanh Hoang Download Agriviet.com

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Vi khuan xam nhap vao cay qua re, vet thng c gii, than non, lo h t nhien, do con trung, khi vao ben trong chung sinh san rat nhanh lam bt cac lo mach ong thi tiet ra oc to lam cac bo mach b hoa nau, en va gay ra hien tng heo do cay b thieu nc. Di nhng ieu kien thuan li vi khuan co the di chuyen xuyen qua lp vo va i ra ben ngoai moi trng at, o cung la s tng tac gia at va re, re b nhiem vi khuan t at va ngc lai vi khuan t trong cay i ra moi trng at (Le Lng Te va Vu Trieu Man, 1999). Vi khuan lan truyen chu yeu bang nguon nc, at nh bam dnh vao giay dep, dung cu canh tac (Martin va French, 1997). Benh heo xanh vi khuan phat sinh phat trien phu thuoc vao ieu kien at ai nh tren cac chan at cao benh thng nang hn cac chan at thap, at c luan canh vi lua nc lam giam t le benh ang ke (o Tan Dung, 2001). Thi vu trong cung la yeu to quan trong anh hng en s phat sinh phat trien benh, thi vu trong co ma nhieu, am o cao lam gia tang s phat sinh phat trien benh. Mat o trong cao t le benh thng cao do chung tao mot vung kh hau thuan li cho s phat sinh phat trien benh. V nc la nguon lay lan chu yeu cua benh, do o phng phap ti la mot trong nhng yeu to gia tang t le gay hai cua benh.

2.7 Bien phap phong tr 2.7.1 Bien phap hoa hoc Hien nay, van cha co loai thuoc nao ac hieu e phong tr benh heo xanh tren cay ca chua, tuy nhien bien phap hoa hoc van c xem la can thiet hien nay, nhng xet ve lau dai bien phap hoa hoc ngay cang the hien cac mat trai cua no nh lam cho tac nhan gay benh tr nen khang thuoc, ac biet la gay o nhiem moi

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trng va sc khoe con ngi mot cach nghiem trong. Dung formol, Mocap 10G, Furadan 3H, voi, CuSO4 x ly at. Dung thuoc: Kasuran, Rovral, Dithan. 2.7.2 Bien phap sinh hoc Khi s dung thuoc hoa hoc e bao ve thc vat th thuoc se tao thanh mot lp mong tren be mat la, qua, than cay, mat at, mat nc va mot lp chat lang goi la d lng ban au cua thuoc v vay bien phap sinh hoc trong cong tac bao ve thc vat ngay cang chu y khai thac va v tr cua bien phap nay ngay cang c nang cao. Viec bao ve moi trng song, moi trng san xuat cung nh tien hanh nong nghiep sach th bien phap nay cang co y ngha hn. S dung mot so vi khuan oi khang nh Bacillus sudtilis, Pseudomonas fluorescens e x l hat trc khi gieo, nhung re cay con trc khi trong hoac a vi sinh vat oi khang vao vung re sau trong nham c che, canh tranh va tieu diet vi khuan Ralstonia solanacearum (o Tan Dung, 2001). S dung che pham V58 co cha vi sinh vat oi khang tren ruong trong ca chua Tien Phong - Me Linh - Vnh Phuc, ket qua thu c co cong thc s dung V58 t le chet giam con 25% so vi oi chng 80% (Le Nh Kieu, 2001). Ngoai ra, chung ta can phai lam at vn m sach benh, cay ba ky, bon am va phai, phan chuong phai u hoai, luan canh vi cay lua nc va dung thuoc mot cach hp ly.

2.8 Phong tr sinh hoc benh cay 2.8.1 Khai niem phong tr sinh hoc benh cay Theo Cook va Baker (1983), phong tr sinh hoc benh cay (PTSHBC) la thong qua viec s dung mot hoac nhieu sinh vat (ngoai tr con ngi) e khong che mam benh hay lam giam s sinh trng va phat trien cua mot tac nhan gay hai nao o. en nam 1988, Cook a a ra mot khai niem rong hn ve phong tr sinh SVTH: Ho Thanh Hoang Download Agriviet.com

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hoc. Theo ong PTSHBC la viec s dung sinh vat, gen va cac san pham cua gen e ieu khien tac nhan gay benh. Cac cach ieu khien tac nhan gay benh co the la: (i) duy tr mat so nguon benh mc thap di ngng kinh te, (ii) lam cham hoac loai tr tien trnh xam nhiem cua benh, (iii) kch hoat va tao ieu kien phat huy he thong t ve cua cay.

2.8.2 La chon tac nhan phong tr sinh hoc Chien lc PTSHBC co the chia thanh hai loai (1) chien lc da tren nguyen tac c ban ve sinh thai hoc hay con goi la phong tr sinh hoc co ien (Hokkaness va Lynch, 1995) hay phong tr sinh hoc ch x ly mot lan (Cook, 1993), (2) chien lc s dung vi sinh vat nh la mot loai thuoc sinh hoc va viec x ly co nhng iem gan giong nh x ly thuoc hoa hoc nham muc ch kiem soat benh trong mot khoang thi gian gii han. Chien lc nay con c goi la phong tr sinh hoc tang dan (Hokkaness va Lynch, 1995). S khac nhau ve chien lc phong tr anh hng en s la chon va phng phap sang loc tac nhan phong tr sinh hoc. oi vi chien lc PTSHBC tang dan, ngi ta chu y en nguon goc cua vi sinh vat oi khang. Tuy nhien nhng hieu biet ve kha canh nay la ieu can thiet cho viec anh gia rui ro va la mot trong nhng yeu cau can thiet cho qua trnh ang ky thng mai hoa san pham (Lumsden va Lewis, 1989). Co mot so phng phap sang loc tac nhan phong tr sinh hoc da tren hoat tnh cua men thuy phan cua vi sinh vat oi khang, tng tac gia vi sinh vat gay benh va vi sinh vat oi khang, phng phap co lien quan en cay k chu.

2.8.3 Vi khuan - mot tac nhan phong tr sinh hoc: Nhieu cong trnh nghien cu ve cac tac nhan phong tr sinh hoc a cong bo t dau the ky 20 trong o co vi khuan. ang la nhom vi khuan hoai sinh ma pho SVTH: Ho Thanh Hoang Download Agriviet.com

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bien nhat la Pseudomonas spp., ke en la Bacillus spp. va Streptomyces (Burr va ctv, 1978, Weller va Cook, 1983). Nhng nghien cu ve vi khuan vung re va vi khuan tren than la cay a dan en viec chia chung thanh ba loai: loai co hai cho cay, loai trung tnh va loai co ch cho cay. Anh hng co ch cua nhom vi khuan nay la do chung san sinh ra chat kch thch tang trng cay, cac chat c che hoac lam suy yeu tac nhan gay benh hoac ca hai (Baker, 1988). C che ban au c che tac nhan gay benh la tiet ra cac chat khang sinh (Fravell, 1988). Tuy nhien nhng yeu to khac nh viec tiet ra chat sidrophores, HCN, s canh tranh dinh dng cung co vai tro quan trong trong viec c che tac nhan gay benh (Cook va Baker, 1983). Trong so cac loai Pseudomonas spp., Pseudomonas fluorescens c chu y va tang cng nghien cu hn het, v ngoai kha nang oi khang vi 10 loai nam benh lu ton trong at, loai vi khuan nay co kha nang kch thch s phat trien cay trong. Bang cach x ly hat co hay khong co chat mang hoat chung vao at cung vi gia the lam thc an nen, a lam giam mc tram trong cua benh, lam gia tang s phat trien va tang nang suat cay trong. Vi thuan li la co nhieu c che tac dung nh: nh c, chiem cho, loai tr mam benh ra khoi vung thch hp, tiet chat khang sinh (Pyrrolnitrin, pyoluteorin) ngoai ky sinh, canh tranh dinh dng (hp chat sat), gia tang sc e khang cho cay, Pseudomonas rat co trien vong s dung tren lanh vc thng mai. Bien phap sinh hoc trong phong tr benh cay la ieu khien moi trng, cay trong va sinh vat oi khang mot cach thch hp, e tao nen mot the can bang sinh hoc can thiet, giup giam mat so cua mam benh xuong di ngng gay hai. Nh o, benh cua cay trong ch xuat hien mc o nhe, khong gay anh hng quan trong ve mat kinh te. Bien phap sinh hoc khong co muc ch tieu diet toan bo mam benh, va cung khong co kha nang nay (Pham Van Kim, 1999). SVTH: Ho Thanh Hoang Download Agriviet.com

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2.8.4 Tnh hnh nghien cu trong va ngoai nc: 2.8.4.1Nghien cu trong nc: Le Lng Te, 2002 nghien cu phong chong benh heo xanh vi khuan ca chua co hieu qua kinh te cao bang bien phap s dung rong rai cac giong ca chua khang benh, co nang suat cao CLN-1462A va P.T4719A vung ong bang song Hong. a a vao trong ai tra cac tnh thuoc ong bang song Hong va cac tnh pha Bac nc ta. Le Nh Kieu, Nguyen Ngoc Cng, ao Thu Hang, 2003 nghien cu phat hien biovar 1 va 5 trong quan the vi khuan Ralstonia solanacearum gay benh heo xanh ca chua. ieu nay cho thay quan the gay benh heo xanh ca chua Viet Nam la rat a dang (co 5 biovar). Pham Minh Sang, 2003 nghien cu Hieu lc cua vi khuan oi khang va kch thch sinh trng trong phong tr benh kho van do nam Rhizoctonia solani bc au a khong che c benh kho van cho lua va kch thch cay tang trng manh bang VKK. Nguyen Trung Thanh, 2004 cung a thanh cong trong viec nghien cu Bc au chon loc va anh gia dong VKK, phan lap t at e khong che nam Rhizoctonia solani, Sclerotium rolfsii, va vi khuan Ralsronia solanacearum gay benh tren cay ca chua. Cac dong VKK co kha nang khang vi ca ba tac nhan gay benh trong ieu kien In vitro va trong nha li VKK a khong che c benh do nam Sclerotium rolfsii gay ra. Pham My Lien, 2004 nghien cu ve van e Chon loc va anh gia dong vi khuan Pseudomonas fluorescens oi khang vi nam Sclerotium rolfsii gay benh tren cay ca chua cung a thanh cong trong qua trnh nghien cu.

2.8.4.2 Nghien cu ngoai nc SVTH: Ho Thanh Hoang Download Agriviet.com

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Tai ai Loan, bang phng phap PCR vi cap primer PS-IS ac hieu vi vi khuan Ralstonia solanacearum thuoc race 1, ket qua phan tch cho thay tat ca cac dong vi khuan Ralstonia solanacearum thu thap tren ca chua, khoai tay, t, au phong, ca tm, thuoc la, dau tay, chuoi va nhieu loai cay trong khac eu thuoc race 1 (Yang An Lee, 2001). Phan tch tren 120 dong vi khuan Ralstonia solanacearum phan lap tren ca chua, khoai tay, ca tm, t, chuoi, gng va nhieu loai cay trong khac t Chau A, My, Au, Phi va Chau ai Dng cho thay vi khuan Ralstonia solanacearum thuoc division Chau A thng bao gom biovar 3 va 4, division Chau My thng bao gom biovar 1 va 2. Theo Rindran va Vidhiaekaran (1996) cho rang nhng loai vi khuan Pseudomonas fluorescens c phan lap t vung re, co sac to phat huynh quang vang xanh cung km ham s phat trien cua nam Rhizoctonia solani. Mot trong nhng dong co hieu qua nhat la PFAIR2, phan lap tren than bun c dung e x ly hat, x ly re, rai vao at va phun tren la. Tng nghiem thc rieng le a kiem soat benh co hieu qua. Tuy nhien, s ket hp cua bon cach dan en hieu qua phong tr tot nhat trong nha li. Tren ong ruong, s dung PFAIR2 phong tr benh co hieu qua, gia tang nang suat va co the so sanh vi cac loai thuoc tr nam thong dung nh carbendazim.

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Chng 3

VAT LIEU VA PHNG PHAP NGHIEN CU

3.1 Thi gian va a iem nghien cu 3.1.1 Thi gian e tai c thc hien t ngay 5/6/2005 30/8/2005

3.1.2 a iem - Ni tien hanh nghien cu: Cong ty Hai Mui Ten o thuoc cong ty giong ong Tay, xa Nhuan c, huyen Cu Chi, Thanh Pho Ho Ch Minh. - Ni phan lap va nuoi cay: Phong thc tap Benh cay Bo mon Bao Ve Thc Vat Trng ai hoc Nong Lam Thanh Pho Ho Ch Minh. SVTH: Ho Thanh Hoang Download Agriviet.com

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- Ni phan tch: Trung tam phan tch th nghiem hoa sinh Trng ai hoc Nong Lam Thanh Pho Ho Ch Minh.

3.2 Phng phap nghien cu 3.2.1 Vat lieu nghien cu oi tng nghien cu: cac dong vi khuan Pseudomonas fluorescens oi khang: 18, 73, 85, 123, 124, 187 va cac dong vi khuan Ralstonia solanacearum gay benh: RST004, RST005, RST006, RST01, RSBog3, RSBog4. Hat ca chua vi ty le nay mam 90%. Moi trng phan lap va nuoi cay: - PDA (potato, dextrose, agar) - LB (tryptone, yeast extract, sodium chloride) - WA (water, agar) Vat lieu lay nhiem: hat giong ca chua (1628) 3.2.2 Phng phap anh gia kha nang oi khang cua cac dong vi khuan Pseudomonas fluorescens vi cac dong vi khuan Ralstonia solanacearum tren a petri trong phong th nghiem ieu kien tien hanh: th nghiem c tien hanh trong phong th nghiem tai Phong Thc tap Benh cay Bo mon Bao ve Thuc vat Trng ai hoc Nong Lam Tp. Ho Ch Minh. Vat lieu nghien cu: - Cac dong vi khuan Pseudomonas fluorescens:18, 73, 85, 123, 124. - Cac dong vi khuan Ralstonia solanacearum: RST004, RST005, RST006, RST01, RSBog3, RSBog4. Dung cu th nghiem: Que trang vi khuan, pipet va au tiep a c vo trung.

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Moi trng s dung: PDA (200g Potato, 15g Agar, 20g Dextrose va 1 lt nc cat). Phng phap tien hanh: Lay 100l cac dong vi khuan Ralstonia solanacearum tran eu tren moi trng PDA a chuan b san. Dung pipet hut 20l dch vi khuan va tien hanh cay dch vi khuan thanh 5 iem tng ng vi cac dong vi khuan Pseudomonas fluorescens a chuan b san. Cuoi cung em u nhiet o phong (27-300C), khoang 24 gi sau xem ket qua oi khang cua cac dong vi khuan. Ch tieu theo doi: Kha nang oi khang cua 5 dong vi khuan Pseudomonas fluorescens

3.2.3 Phng phap anh gia kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum tren cay ca chua trong nha li Th nghiem c tien hanh trong ieu kien nha li tai cong ty Hai Mui Ten o. Vat lieu nghien cu: - Nguon vi khuan khang manh nhat va gay oc manh nhat c chon loc trong phong th nghiem. - Hat ca chua vi t le nay mam 90%. - at. - Chau nha 7x12cm (ng knh x chieu cao). - Khay xop (60 lo).

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- Trang thiet b th nghiem tai phong thc tap Benh cay Bo mon Bao Ve Thc Vat va tai trung tam phan tch th nghiem Hoa Sinh trng ai hoc Nong Lam Tp.Ho Ch Minh. Moi trng s dung: LB long (tryptone, yeast extract, sodium chloride). Phng phap tien hanh: Lay nhiem theo phng phap cua Pham ang Minh (2003) co cai tien - Sau khi khao sat tnh oc cua cac dong vi khuan Ralstonia solanacearum trong phong th nghiem, ta chon ra cac dong gay oc cho cay ca chua va chon ra mot dong manh nhat. Tng t, ta cung chon ra mot dong VKK manh nhat. - Nguon vi khuan: t nguon vi khuan tr -800C, nuoi cay tren moi trng LB long khoang 16 gi, ly tam thu vi khuan 3000 vong/phut, hoa vi khuan vao nc cat vo trung. - Hat giong ca chua c ra bang con 70% trong 1 phut, NaOCl 2% trong 5 phut, sau o ra lai nhieu lan vi nc cat vo trung, u hat tren giay tham c lam am bang nc cat vo trung at trong a petri, gi nhiet o 270C. - Lay hat ca chua a nay mam khoang 0,5 2mm ngam trong dch huyen phu a chuan b tren, nong o 109 cfu/ml vi khuan oi khang (khoang 20 phut) a chuan b san, sau o lam kho hat tren giay tham em gieo khay xop a chuan b san at (khay 60 lo). - Khi cay ca chua c 15 ngay tuoi em trong vao chau at a co chung san dung dch vi khuan gay oc, vi mat so vi khuan: 103 cfu/g at, cac cay ca chua c chia thanh 4 nghiem thc, moi nghiem thc 30 cay, vi 3 lan lap lai. Tien hanh theo doi trong vong 20 ngay. Ch tieu theo doi: Chieu cao cay: o t hai la mam len en au ngon la, 5 ngay/lan. So la: em toan bo so la tren cay, 5 ngay/lan. Cac nghiem thc: SVTH: Ho Thanh Hoang Download Agriviet.com

Luan van tot nghiep NT1 (T1): khong x ly vi khuan gay benh va oi khang. NT2 (T2): ch x ly vi khuan gay benh trong ly at.

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NT3 (T3): x ly vi khuan oi khang giai oan hat nt nanh va vi khuan gay benh trong ly at. NT4 (T4): x ly vi khuan oi khang giai oan hat nt nanh, trc khi trong vao ly at a chung san vi khuan gay benh tien hanh nhung cay ca chua vao dch vi khuan oi khang a chuan b san vi mat so 109 cfu/ml, (khoang 20 phut).

3.2.4 Phng phap anh gia tnh oc cua vi khuan Ralstonia solanacearum tren cay ca chua trong ieu kien nha li Cac phng phap anh gia giong nh muc 3.2.3 nhng chung vi khuan gay oc vi mat so vi khuan: 1010 cfu/g at, tien hanh theo doi trong 12 ngay sau trong. Ch tieu theo doi: Ty le chet cua cay ca chua (%) 3.2.5 Phng phap anh gia kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum tren cay ca chua ngoai ong ruong. Th nghiem c tien hanh ngoai ong ruong tai cong ty Hai Mui Ten o. Vat lieu nghien cu: - Nguon VKK manh nhat c chon loc trong phong th nghiem. - Hat ca chua vi t le nay mam 90%. - at c len luong, phu bat va uc lo a chuan b trc. - Khay xop (60 lo). Phng phap tien hanh: Khi cay c 15 ngay tuoi em trong ngoai ong ruong, cac cay ca chua c chia thanh 2 nghiem thc, sau trong 3 ngay ti dch VKK. Tien hanh theo doi trong 20 ngay sau trong. SVTH: Ho Thanh Hoang Download Agriviet.com

Luan van tot nghiep Ch tieu theo doi: Ty le chet cua cay ca chua (%) Cac nghiem thc:

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- NT1 (T1): cac cay ca chua, vi 3 lan lap lai, moi lan lap lai 20 cay. Khong chung vi khuan oi khang va gay benh. - NT2 (T2): cac cay ca chua, vi 3 lan lap lai, moi lan lap lai 80 cay. Co chung vi khuan oi khang.

S o bo tr th nghiem: T1 T2 T2 Ghi chu: T1: oi chng T2 : x ly VKK 3.2.6 Phng phap x ly so lieu Cac so lieu c x ly bang phan mem MSTATC. T1 T2 T 2 T 1

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Chng 4 KET QUA VA THAO LUAN

4.1 Mc oi khang cua cac dong vi khuan Pseudomonas fluorescens vi cac dong vi khuan Ralstonia solanacearum tren a petri Tren a petri co cha moi trng PDA dung que tran vi khuan tran eu 6 dong VKGB, sau o tien hanh cay thanh 5 iem a anh dau trc tng ng vi 5 dong VKK e xac nh mc o oi khang cua 5 dong vi khuan. Ket qua c tnh bay bang 4.1. Bang 4.1 Mc o oi khang cua 5 dong vi khuan tren a petri SVTH: Ho Thanh Hoang Download Agriviet.com

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DVKK 18 73 85 123 124

DVKGB RST004 +++ +++ +++ ++ ++ RST005 +++ ++ +++ + ++ RST006 +++ +++ +++ + + RST01 +++ +++ +++ ++ ++ RSBog3 +++ +++ +++ + + RSBog4 ++ +++ +++ + +

Chu thch: (+++): khang manh (++): khang trung bnh (+): khang yeu DVKGB: dong vi khuan gay benh DVKK: dong vi khuan oi khang Cac dong vi khuan Ralstonia solanacearum: RST004, RST005, RST006, RST01, RSBog3, RSBog4. Cac dong vi khuan Pseudomonas fluorescens: 18, 73, 85, 123, 124. Trong cac dong VKK Pseudomonas fluorescen co 5 dong eu co kha nang oi khang vi cac dong VKGB Ralstonia solanacearum, co 3 dong oi khang manh, 2 dong con lai co mc o khang t trung bnh en yeu. Nh vay, trong 5 dong VKK co 3 dong khang manh, tiep tuc nghien cu 3 dong nay va chon ra mot dong manh nhat.

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Hnh 4.1: Kha nang oi khang cua 5 dong VKK oi vi VKGB RSBog3

Hnh 4.2: Kha nang oi khang cua 5 dong VKK oi vi VKGB RSBog4

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Hnh 4.3: Kha nang oi khang cua 5 dong VKK oi vi VKGB RST01

Hnh 4.4: Kha nang oi khang cua 5 dong VKK oi vi VKGB RST004

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Hnh 4.5: Kha nang oi khang cua 5 dong VKK oi vi VKGB RST005

Hnh 4.6: Kha nang oi khang cua 5 dong VKK oi vi VKGB RST006

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Hnh 4.7: Kha nang oi khang cua dong VKK 85 vi dong VKGB RST01

Hnh 4.8: Kha nang oi khang cua dong VKK 85 vi dong VKGB RST004 SVTH: Ho Thanh Hoang Download Agriviet.com

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4.2 anh gia kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum tren cay ca chua trong ieu kien nha li X ly at bang vi khuan RST01 vi mat so vi khuan 103 cfu/gat. Sau o tien hanh trong ca chua vao chau at e xac nh kha nang oi khang cua dong vi khuan 85. Ket qua c trnh bay bang 4.2, 4.3. Bang 4.2 Sc tang trng ve chieu cao cay Nghiem thc T1 T2 T3 T4 CV% 5 NST 20,2 B 20,3 B 25,3 AB 29,0 A 13,26 Chieu cao trung bnh (cm) 10 NST 30,3 BC 25,2 C 40,3 AB 46,0 A 9,77 15 NST 35,3 B 25,8 B 47,3 A 53,2A 8,99 20 NST 38,0 B 26,8 C 58,5 A 60,8 A 7,83

Ghi chu: Trong cung mot cot cac ch so theo sau cung ky t th s sai biet khong co y ngha ve mat thong ke. T1: oi chng T2: ch x ly VKGB T3, T4 : x ly VKGB va VKK NST: Ngay sau trong CV%: He so bien thien

T ket qua bang 4.2 co mot so nhan xet sau: - Giai oan 5 NST chieu cao gia cac nghiem thc s sai biet co y ngha ve mat thong ke. Cao nhat la T4 (29,0 cm) va khong sai biet ve mat thong ke vi nghiem thc T3. Thap nhat la T1 (20,2 cm) va khong sai biet ve mat thong ke vi nghiem thc T2. SVTH: Ho Thanh Hoang Download Agriviet.com

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- Giai oan 10 NST chieu cao gia cac nghiem thc s khac biet co y ngha ve mat thong ke. Cao nhat la T4 (46,0 cm) va khong sai biet ve mat thong ke vi T3, thap nhat la T2 (25,2 cm) va khong sai biet ve mat thong ke vi T1. - Giai oan 15 NST toc o tang trng chieu cao gia cac nghiem thc s khac biet co y ngha ve mat thong ke. Thap nhat la T2 (25,8 cm) va s khac biet khong co y ngha ve mat thong ke vi T1, cao nhat la T4 (53,2 cm). - Giai oan 20 NST, T4 co chieu cao cao nhat (60,8 cm) va s khac biet khong co y ngha ve mat thong ke vi T3. Thap nhat la T2 (26,8 cm) va s khac biet khong co y ngha ve mat thong ke vi T1.

Bang 4.3 Toc o ra la cua cay So la trung bnh/cay Nghiem thc T1 T2 T3 T4 CV% 19,7 B 16,0 B 21,7 AB 27,0 A 10,46 28,7 B 24,0 B 32 AB 42,7 A 13,64 38,0 BC 25,7 C 45,7 AB 52,7 A 10,74 43,7 BC 31,7 C 60,0 AB 68,0 A 11,70 5 NST 10 NST 15 NST 20 NST

Ghi chu: Trong cung mot cot cac ch so theo sau cung ky t th s sai biet khong co y ngha ve mat thong ke. SVTH: Ho Thanh Hoang Download Agriviet.com

Luan van tot nghiep T1: oi chng T2: ch x ly VKGB T3, T4 : x ly VKGB va VKK NST: Ngay sau trong CV%: He so bien thien

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T ket qua bang 4.3 co mot so nhan xet sau: - giai oan 5 NST so la ca chua gia cac nghiem thc s sai biet co y ngha ve mat thong ke. Nhieu nhat la T4 (27,0 la/cay) va s khac biet khong co y ngha ve mat thong ke vi cac nghiem thc T3. t nhat la T2 (16,0 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T1. - Giai oan 10 NST so la ca chua gia cac nghiem thc s sai biet co y ngha ve mat thong ke. t nhat la T2 (24,0 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T1. Nhieu nhat la T4 (42,7 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T3. - Giai oan 15 NST so la ca chua gia cac nghiem thc s sai biet co y ngha ve mat thong ke. t nhat la T2 (25,7 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T1. Nhieu nhat la T4 (52,7 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T3. - Giai oan 20 NST so la ca chua gia cac nghiem thc s sai biet co y ngha ve mat thong ke. t nhat la T2 (31,7 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T1. Nhieu nhat la T4 (68,0 la/cay) va s khac biet khong co y ngha ve mat thong ke vi T3. Vi mat so 103 cfu/g at, kha nang gay oc cho cay ca chua la rat thap. Cac nghiem thc sinh trng va phat trien bnh thng, rieng nghiem thc T3, T4 phat trien manh, so la va chieu cao cay vt troi hn han cac nghiem thc T1, T2. ieu

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nay chng to VKK co tac dung lam cho benh t hn, re phat trien tot hn, so la nhieu hn va tang chieu cao. Nh vay, dong VKK 85 khong nhng c che s sinh san cua VKGB RST01 ma con giup cho cay sinh trng va phat trien tot hn.

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Hnh 4.9: Tong the cac nghiem thc tren cay ca chua 5 NST

Hnh 4.10: Tng nghiem thc tren cay ca chua 10 NST

4.3 anh gia tnh oc cua VKGB RST01 tren cay ca chua trong ieu kien nha li SVTH: Ho Thanh Hoang Download Agriviet.com

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Khi x ly at bang vi khuan RST01 vi mat so vi khuan 1010cfu/gat. Sau o, tien hanh trong cay ca chua vao chau at e xac nh mc o gay oc cua vi khuan RST01. Ket qua trnh bay bang 3. Sau 3 ngay trong vao chau at, cac nghiem thc T2, T3, T4 co trieu chng bat au heo ru va heo ru hoan toan trong khi o T1 lai sinh trng va phat trien bnh thng ieu nay chng to cac nghiem thc T2, T3, T4 heo ru la do vi khuan RST01 gay ra. T1: Sinh trng va phat trien bnh thng do khong x ly VKK va VKGB. T2: Heo ru nhanh nhat do ch x ly VKGB. T3: Mac du co x ly VKK trc khi gieo hat nhng cay van heo ru kha nhanh ieu nay chng to s sinh san cua VKK la khong kp so vi s sinh san cua VKGB. T4: S heo ru la cham nhat do trc khi trong co x ly VKK, ieu nay chng to VKK co tac ong en VKGB nhng s sinh san cua VKK cung khong kp so vi vi khuan gay benh nen cay bat au heo dan. Bang 4.4 Anh hng cua vi khuan gay benh en ty le chet cua cay Nghiem thc T1 T2 T3 T4 Ghi chu: 3 NST 0 16,7 13,3 0 Ty le chet (%) 6 NST 9 NST 0 0 43,3 86,7 33,3 70 23,33 50 12 NST 0 100 100 86,67

NST: ngay sau trong T1: oi chng T2: ch x ly VKGB T3, T4 : x ly VKGB va VKK

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Trong tng nghiem thc, ty le chet tren cay ca chua la khac nhau, nhng tat ca eu heo ru hang loat ch trong mot thi gian rat ngan. ieu o chng to mat so VKGB trong at la qua day ac lam cho VKK khong sinh san c dan en ty le cay chet 100%. Nh vay, mc o gay oc cua vi khuan RST01 la rat manh. Kha nang phat sinh, phat trien cua chung la rat nhanh, ch trong mot thi gian ngan toan bo cay trong a b heo ru mot cach toan dien. Kha nang oi khang cua vi khuan 85 ch co tac dung trong mot thi gian rat ngan do mat so vi khuan RST01 qua day km ham s sinh san cua dong vi khuan 85.

Hnh 4.11: Tnh oc cua VKGB RST01 tng nghiem thc

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Hnh 4.12: Tnh oc cua VKGB RST01 tong the 4.4 anh gia kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum tren cay ca chua ngoai ong ruong Khi cay ca chua c 15 ngay tuoi, tien hanh trong ra ngoai ong. Sau 3 ngay, ti VKK 85 vao cac hoc a c uc lo (moi hoc 100ml), vi mat so 109 VKK/ml. Ket qua c trnh bay bang 4. Bang 4.5 Ty le chet cua cay Nghiem thc T1 T2 Ghi chu: T1: oi chng T2 : x ly VKK SVTH: Ho Thanh Hoang Download Agriviet.com 5 NST 8,3 6,33 Ty le chet (%) 10 NST 15 NST 41,67 78.33 17,5 52,45 20 NST 100 95

NST: ngay sau trong

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Ch trong mot thi gian ngan, ty le chet cua cay la rat ln, khi nho cay thay re van phat trien. Cay chet co the do quan the sinh vat gay benh trong at qua nhieu, hoac thi tiet co ma dam hoac do benh moc sng gay ra. Nho 10 cay ca chua, cat mot oan than cay nhung phan cat gan sat goc vao nc cat vo trung, quan sat thay cho vet cat co dong trang uc nh sa tiet ra, chng to trong cay ca chua co vi khuan, trong 10 cay, 8 cay co vi khuan tiet ra, chiem ty le 80%. Nh vay, kha nang phong tr cua vi khuan Pseudomonas fluorescens hau nh khong co, cong vi thi tiet co ma to vao moi buoi chieu, ty le benh tren cay ca chua phat trien manh. Cay chet hang loat ch trong thi gian ngan.

Chng 5 KET LUAN VA E NGH

5.1 Ket luan Kha nang oi khang cua cac dong Pseudomonas fluorescens vi cac dong Ralstonia solanacearum tren a petri, trong o co 3 dong khang manh, va 2 dong co mc o khang t trung bnh en yeu. Kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum rat yeu khi x ly vi khuan RST01 vao chau at vi mat so vi khuan: 1010 cfu/g at. Ty le cay chet 12 NST la 100%.

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Kha nang phong tr cua dong vi khuan Pseudomonas fluorescens vi dong vi khuan Ralstonia solanacearum rat manh khi x ly vi khuan RST01 vao chau at vi mat so vi khuan: 103cfu/g at. Ty le cay song 20 NST la 100%. Ty le cay chet ngoai ong ruong la 100%.

5.2 e ngh Tiep tuc nghien cu vi khuan Pseudomonas fluorescens vi so lng mau nhieu hn, ai dien cho nhieu vung va th nghiem oi khang tren cac loai vi khuan khac. Khao sat them cac yeu to anh hng en mat so vi khuan trong ieu kien ng dung thc te e lam tang tnh hieu qua. Khao sat cac yeu to anh hng en viec san xuat cac che pham sinh hoc oi khang.

TAI LIEU THAM KHAO

1. Tieng Viet o Tan Dung, 2001. Benh heo ru hai cay trong can, bien phap phong chong. Nha xuat ban Nong Nghiep, Ha Noi. Nguyen Van Het, 1997. Khao sat benh chet heo cay au phong do vi khuan Ralstonia solanacearum (Smith) huyen Cu Chi Tp.Ho Ch Minh. Luan an thac s Nong Nghiep. ai Hoc Nong Lam Tp. Ho Ch Minh. SVTH: Ho Thanh Hoang Download Agriviet.com

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Vu Trieu Man va Le Lng Te, 1998. Giao trnh benh cay nong nghiep. Nha xuat ban Nong Nghiep, Ha Noi. Le Lng Te va Vu Trieu Man, 1999. Benh vi khuan va virus hai cay trong. Nha xuat ban Giao Duc. Le Lng Te, 2002. Nhan dang Ralstonia solanacearum bang PCR va anh gia tnh khang benh heo xanhcua mot so giong ca chua khang vi cac dong vi khuan Ha Noi Viet Nam. Hoi thao benh cay va sinh hoc phan t Trng ai Hoc Nong Lam Tp. Ho Ch Minh lan th nhat. 21/6/2002. Trng ai Hoc Nong Lam Tp. Ho Ch Minh. Nguyen Th Yen va ctv, 2002. Thanh phan noi, biovar vi khuan gay benh heo xanh cay trong can. Hoi thao benh cay va sinh hoc phan t, Trng ai Hoc Nong Lam Tp. Ho Ch Minh lan th nhat. Trng ai Hoc Nong Lam Tp. Ho Ch Minh. Le Lng Te, 2002. Phong chong benh heo xanh vi khuan ca chua co hieu qua kinh te cao bang bien phap s dung rong rai cac giong ca chua khang benh, co nang suat cao CLN-1462A va P.T 4719A vung ong bang song Hong. Le Nh Kieu, Nguyen Ngoc Cng, ao Thu Hang, 2003. Phat hien biovar 1 va 5 trong quan the vi khuan Ralstonia solanacearum gay benh heo xanh ca chua. Hoi ngh Cong nghe Sinh hoc toan quoc, Ha Noi 2003. Pham ang Minh, 2003. Cau truc quan the vi khuan Ralstonia solanaccearum gay hai tren cay trong mot so tnh pha nam Viet Nam. Luan van tot nghiep ky s nong nghiep. Trng ai hoc Nong Lam Tp. Ho Ch Minh.

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Nguyen Trung Thanh, 2004. Bc au chon loc va anh gia dong vi khuan oi khang phan lap t at e khong che nam Rhizoctnia solani, Sclerotium rolfsii va vi khuan Ralstonia solanaccearum gay benh tren cay ca chua. Luan van tot nghiep ky s nong nghiep. Trng ai hoc Nong Lam Tp. Ho Ch Minh. Pham Minh Sang, 2003. Hieu lc cua che pham vi khuan oi khang va kch thch sinh trng trong phong tr benh kho van (Rhizoctonia solani Kuln) tren lua. Luan van tot nghiep ky s nong nghiep. Trng ai hoc Nong Lam Tp. Ho Ch Minh. 2. Tieng nc ngoai Hayward A.C.1964, Characterics of Pseudomonas solanacearum. J. Appl. Bacteriol. 27: 205 277. Kelman A.1954. The raltionships of pathogencity in Pseudomonas solanacearum to colony appearance on a tetrazolium medium. Phytopathology 44: 693 695. Yung-An L., Shu-Chung F., Ling-Ya C. and Kuo-Chiang H., 2001. Isolate of an insertion sequence from of Ralstonia solanacearum race 1 and its potential use for strain characterization and detection. Applied and Environmental Microbiology 67: 3943 3950.

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PHU LUC

I. Bang 6. Anh hng cua VKK en so la cay ca chua NT LLL 1 T1 2 3 T2 1 So la 5 NST 19 24 16 18 10 NST 33 28 25 25 15 NST 36 39 36 27 20 NST 45 39 47 30

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Luan van tot nghiep 2 3 1 T3 2 3 1 T4 2 3 15 15 21 26 18 27 29 25 24 23 31 38 27 44 51 33 25 25 47 53 37 52 57 49 29 36 60 57 63 62 78 64

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Ghi chu: NT: nghiem thc LLL: lan lap lai NST: ngay sau trong

1. So la 5 ngay sau khi trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------nt 3 189.58 63.194 13.00 0.0049 lll 2 50.17 25.083 5.16 0.0497 Error 6 29.17 4.861 Non-additivity 1 2.03 2.029 0.37 Residual 5 27.14 5.428 -----------------------------------------------------------------------Total 11 268.92 -----------------------------------------------------------------------Grand Mean= 21.083 Grand Sum= 10.46% 253.000 Total Count= 12

Coefficient of Variation=

* Trac nghiem phan hang so la cua ca chua 5 ngay sau trong SVTH: Ho Thanh Hoang Download Agriviet.com

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Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 19.70 16.00 21.70 27.00 B B AB A Mean Mean Mean Mean

Ranked Order 4 3 1 2 = = = = 27.00 21.70 19.70 16.00 A AB B B

2. So la 10 ngay sau khi trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------nt 3 566.33 188.778 10.01 0.0095 lll 2 148.17 74.083 3.93 0.0812 Error 6 113.17 18.861 Non-additivity 1 75.16 75.163 9.89 Residual 5 38.00 7.601 -----------------------------------------------------------------------Total 11 827.67 -----------------------------------------------------------------------Grand Mean= 31.833 Grand Sum= 13.64% 382.000 Total Count= 12

Coefficient of Variation=

* Trac nghiem phan hang so la cua ca chua 10 ngay sau trong

Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 28.70 24.00 32.00 42.70 B B AB A Mean Mean Mean Mean Ranked Order 4 3 1 2 = = = = 42.70 32.00 28.70 24.00 A AB B B

3. So la 15 ngay sau khi trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------nt 3 1203.00 401.000 21.20 0.0014 lll 2 58.50 29.250 1.55 0.2873 Error 6 113.50 18.917 Non-additivity 1 36.99 36.988 2.42 Residual 5 76.51 15.302 -----------------------------------------------------------------------Total 11 1375.00 -----------------------------------------------------------------------Grand Mean= 40.500 Grand Sum= 10.74% 486.000 Total Count= 12

Coefficient of Variation=

* Trac nghiem phan hang so la cua ca chua 15 ngay sau trong SVTH: Ho Thanh Hoang Download Agriviet.com

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41

Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 38.00 25.70 45.70 52.70 BC C AB A Mean Mean Mean Mean

Ranked Order 4 3 1 2 = = = = 52.70 45.70 38.00 25.70 A AB BC C

4. So la 20 ngay sau khi trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------nt 3 2392.33 797.444 22.55 0.0011 lll 2 21.17 10.583 0.30 0.7518 Error 6 212.17 35.361 Non-additivity 1 4.53 4.531 0.11 Residual 5 207.64 41.527 -----------------------------------------------------------------------Total 11 2625.67 -----------------------------------------------------------------------Grand Mean= 50.833 Grand Sum= 11.70% 610.000 Total Count= 12

Coefficient of Variation=

* Trac nghiem phan hang so la cua ca chua 20 ngay sau trong

Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 43.70 31.70 60.00 68.00 BC C AB A Mean Mean Mean Mean Ranked Order 4 3 1 2 = = = = 68.00 60.00 43.70 31.70 A AB BC C

II. Bang 7. Anh hng cua VKK en chieu cao cay ca chua NT LLL 1 T1 2 3 1 T2 2 3 SVTH: Ho Thanh Hoang Download Agriviet.com Chieu cao cay (cm) 5 NST 19,5 24 17 16,5 19,5 25 10 NST 31,5 33 26,5 19,5 23,5 27,5 15 NST 37,5 37 31,5 23,5 25 29 20 NST 40,5 39 34,5 24 26 30,5

Luan van tot nghiep 1 T3 2 3 1 T4 2 3 24,5 27,5 24 27 28,5 31,5 40 42,5 38,5 41 47,5 49,5 48,5 48,5 46 47,5 55 57 57,5 61 57 56 61,5 65

42

Ghi chu: NT: nghiem thc LLL: lan lap lai NST: ngay sau trong

1. Chieu cao 5 ngay sau trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------NT 3 163.73 54.576 5.52 0.0368 LLL 2 20.67 10.333 1.04 0.4080 Error 6 59.33 9.889 Non-additivity 1 0.63 0.632 0.05 Residual 5 58.70 11.740 -----------------------------------------------------------------------Total 11 243.73 -----------------------------------------------------------------------Grand Mean= 23.708 Grand Sum= 13.26% 284.500 Total Count= 12

Coefficient of Variation=

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43

* Trac nghiem phan hang chieu cao cua ca chua 5 ngay sau trong
Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 20.20 20.30 25.30 29.00 B B AB A Mean Mean Mean Mean Ranked Order 4 3 2 1 = = = = 29.00 25.30 20.30 20.20 A AB B B

2. Chieu cao 10 ngay sau trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------NT 3 801.23 267.076 22.27 0.0012 LLL 2 47.54 23.771 1.98 0.2183 Error 6 71.96 11.993 Non-additivity 1 0.68 0.676 0.05 Residual 5 71.28 14.257 -----------------------------------------------------------------------Total 11 920.73 -----------------------------------------------------------------------Grand Mean= 35.458 Grand Sum= 9.77% 425.500 Total Count= 12

Coefficient of Variation=

* Trac nghiem phan hang chieu cao cua ca chua 10 ngay sau trong
Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 30.30 25.20 40.30 46.00 BC C AB A Mean Mean Mean Mean Ranked Order 4 3 1 2 = = = = 46.00 40.30 30.30 25.20 A AB BC C

2. Chieu cao 15 ngay sau trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------NT 3 1346.75 448.917 34.04 0.0004 LLL 2 12.54 6.271 0.48 0.6431 Error 6 79.12 13.187 Non-additivity 1 8.24 8.244 0.58 Residual 5 70.88 14.176 -----------------------------------------------------------------------Total 11 1438.42 -----------------------------------------------------------------------Grand Mean= 40.417 Grand Sum= 485.000 Total Count= 12

SVTH: Ho Thanh Hoang Download Agriviet.com

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Coefficient of Variation=

8.99%

* Trac nghiem phan hang chieu cao cua ca chua 15 ngay sau trong
Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 35.30 25.80 47.30 53.20 B B A A Mean Mean Mean Mean Ranked Order 4 3 1 2 = = = = 53.20 47.30 35.30 25.80 A A B B

2. Chieu cao 20 ngay sau trong

A N A L Y S I S O F V A R I A N C E T A B L E

Degrees of Sum of Source Freedom Squares Mean Square F-value Prob -----------------------------------------------------------------------NT 3 2422.90 807.632 62.09 0.0001 LLL 2 14.29 7.146 0.55 0.6038 Error 6 78.04 13.007 Non-additivity 1 4.81 4.808 0.33 Residual 5 73.23 14.647 -----------------------------------------------------------------------Total 11 2515.23 -----------------------------------------------------------------------Grand Mean= 46.042 Grand Sum= 7.83% 552.500 Total Count= 12

Coefficient of Variation=

* Trac nghiem phan hang chieu cao cua ca chua 20 ngay sau trong
Original Order Mean Mean Mean Mean 1 2 3 4 = = = = 38.00 26.80 58.50 60.80 B C A A Mean Mean Mean Mean Ranked Order 4 3 1 2 = = = = 60.80 58.50 38.00 26.80 A A B C

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