LVTN - Chon Loc Va Nhan Sinh Khoi Nam Trichodema

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1.1 at van e Trong nhng nam gan ay, nen nong nghiep Viet Nam co nhng bc tien vt bac, ngoai viec ap ng nhu cau lng thc - thc pham trong nc, san lng nhieu loai nong san xuat khau cua Viet Nam c xep vao hang au the gii. Tuy nhien, chat lng va hieu qua cua viec san xuat nong san hang hoa nc ta con nhieu han che so vi cac nc trong khu vc. e khac phuc van e nay, chung ta can quan tam en viec xay dng nen nong nghiep theo hng sinh thai ben vng, tang nhanh so lng va nang cao chat lng nong san hang hoa. Theo o, cong tac giong cay trong va bao ve thc vat ong vai tro rat quan trong. Cong tac phong tr sau benh hai cay trong hien nay c ap dung bang nhieu bien phap. Trong o, bien phap hoa hoc van c xem la hu hieu nhat. Mot vai hoa chat tr sau co tnh chon loc cao, t oc hai cho moi trng a c s dung, nhng nhng hoa chat nay thng qua at ch e s dung cho pham vi nong trai nho. Tuy vay bien phap phong tr sau benh hai bang thuoc hoa hoc o at nh hien nay a the hien ro nhng mat trai cua no nh lam cho con trung tr nen khang thuoc, ac biet la gay o nhiem moi trng mot cach nghiem trong. Trc tnh hnh o, bien phap phong tr sau benh hai bang sinh hoc a c nhieu nha khoa hoc quan tam va nghien cu. Nhieu tac nhan ky sinh, ang chu y la mot so loai nam, chung co the oi khang tren mot so benh hai gay ra ton that cho cay trong. ong thi, khong nhng ngan chan c mot so benh hai tren canh ong, nhng che pham nam khang khong anh hng en nhng loai thien ch ban x trong t nhien nh ong vat an tht, ky sinh va con trung co ch. S 1
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bao ton cac loai thien ch t nhien nay la cha khoa vng chac e phong tr sau benh hai tren cay trong mot cach an toan va hieu qua. Cac ket qua a at c cua viec phong tr nam gay benh bang phng phap sinh hoc cho thay tnh hieu qua ln cua no, nam gay benh khong khang thuoc, khong gay o nhiem moi trng. Hien nay, phong tr dch hai cay trong bang bien phap sinh hoc c ay manh nghien cu nhieu nc, c coi nh la mot lnh vc quan trong. Phong tr bang sinh hoc oi vi benh hai chu yeu la khai thac va s dung kha nang oi khang cua mot so loai nam oi vi cac loai nam gay hai cay trong. Nhieu cong trnh nghien cu ve nam Trichodema va san xuat che pham e han che nhng nam gay hai cho cay trong nh nam Rhizoctonia, Sclerotium, Fusarium, Pythium, Verticillium va Botrytis gay benh tren lua, ngo, va mot so cay trong khac a thu c nhng ket qua rat kha quan. Tuy nhien, nam oi khang la mot tac nhan sinh hoc, no co nhng ieu kien song nhat nh va ch phat huy c hieu qua phong tr benh nhng ieu kien nhat nh. Trong khi o, thng do kha nang thch nghi vi moi trng song, cac tac nhan gay benh phat trien manh me, so lng ca the tang nhanh, kha nang chong chu tot, lan lt cac tac nhan oi khang lam cho tnh oi khang mat can bang va ket qua la benh boc phat tren cay trong. e khac phuc ieu nay, viec chon loc, nhan nhanh so lng, tang cng sc song cho tac nhan oi khang va a lai trong moi trng t nhien la het sc can thiet. Xuat phat t nhng van e tren, chung toi d kien thc hien e tai: Chon loc va nhan sinh khoi nam Trichodema oi khang vi nam gay hai cay trong 1.2 Muc tieu va muc ch cua e tai 1.2.1. Muc tieu
- Chon loc va nhan sinh khoi nam Trichodema oi khang cao vi mot so loai nam gay benh cay trong. 1.2.2. Muc ch - S dung cac dong nam oi khang thuoc giong Trichodema sau khi chon loc c nh bien phap sinh hoc e phong tr mot vai tac nhan gay hai tren cay trong nong nghiep. 1.3 Noi dung va oi tng nghien cu 1.3.1 Noi dung - Thu thap va phan lap cac dong nam Trichodema. - anh gia tnh oi khang cua cac dong nam Trichodema vi mot so nam gay hai cay trong trong at tren moi trng chon loc. - Xay dng phng phap nhan sinh khoi Trichodema. - anh gia kha nang phong tr benh cua nam Trichodema c la chon tren mot so loai cay trong trong nha li va ngoai ong ruong. 1.3.2 oi tng - Cac dong nam oi khang thuoc nhom Trichodema. - Cay trong: Ho tieu va sau rieng.
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2.1 Tiem nang s dung Trichodema trong phong tr sinh hoc 2.1.1 Vai tro cua quan the nam Trichodema trong at Trichodema co kha nang tai tao lai quan the, ay la mot hien tng phong tr sinh hoc van con la cau hoi. Theo Bliss (1959), cong bo Trichodema co kha nang thiet lap quan the va tai hoat ong rat nhanh tren at a c x ly kh trung xong hi bang carbon disulfide e diet nam Armillaria mellea tren cay cam, qut, nhng khong cong bo bang chng quan the nam Trichodema phong chong benh. Ohr va cong tac vien (1973), cung cap bang chng thuyet phuc nhat quan the Trichodema trong at co kha nang phong tr nam Armillaria mellea tren at a c x ly xong hi bang methyl bromide. Trichodema khang methyl bromide hn A. mellea, v A. mellea san xuat ra t chat khang (Ohr va cong tac vien, 1975). Them vao nhng bang chng ve vai tro quan the Trichodema trong at trong van e phong tr sinh hoc la them sulfur vao at e duy tr o pH di 3,9 nham phong tr benh thoi re va thoi ngon da Uc. Cach phong tr nay a lam giam tui bao t cua nam Phytophthora va lam tang tnh a acid cua
T.viride.(Cook va Baker, 1983). Kha nang hoat ong phong tr sinh hoc cua Trichodema cac the tiem sinh va si nam c cong bo khong ch trong phong th nghiem (Ayers, 1981 ; Cook va Baker, 1982) ma con trong at (Hubbard va cong tac vien, 1983). Trichodema co kha nang khuyech tan chat oc cua cac nam trong phong th nghiem ke ca cac chat hu c trong at cung nh kha nang keo dai phong tr sinh hoc cua Trichodema. Ngoai ra, kha nang th hai cua nam Trichodema la khang nam. T.hamatum co rat nhieu trong at hu c tai vn m Colombia co kha nang ngan chan nam R.solani (Chet va Baker, 1980 ; 1981) va T.hazianum co nhieu khi phan lap 4
t at tai Mexico co kha nang ngan chan nhieu loai nam at (Lumsden, 1977). Di nhiet o va tia phong xa gamma khong the diet c nam R. solani, ngc lai tren moi trng T. hazianum diet c nam nay (Nelson va cong tac vien, 1983), ay la vai tro chnh cua Trichodema trong viec phong tr sinh hoc. Kha nang ngan can cua at en nhng loai nam benh cay trong at, ac biet la R.solani, Pythium spp., co lien quan en nam Trichodema, a c cong bo rong rai va la van e c nghien cu trong nhieu nam nay. Cac tai lieu Baker (1974 ; 1980) cua Barnett va cong tac vien (1974), cua Cook va Baker (1983) eu cong bo kha nang nay cua Trichodema. 2.1.2 Kha nang lam tang hoac giam tnh khang cua Trichodema trong at ay la van e hap dan c nhieu chuyen gia nghien cu trong nhng nam gan ay. Phong tr sinh hoc bang cach them mot so lng ln bao t T.hazianum cung moi trng nuoi trong vao at c Well va cong tac vien (1962) th nghiem. Cac nha nghien cu nay lan au cong bo s dung mot so lng ln Trichodema nuoi trong tren moi trng ran ra th ngoai ong kiem soat nam Sclerotium rolfsii tren ca chua. Barckman va Rodriguez Kabana (1975) nuoi trong T.hazianum bang phng phap thng mai hoa, la cac hat nho khong hoa tan c gan vi mat ng va rai cac hat nay bang tay doc theo cac hang au phong vi lng 112 140kg/ha sau 70- 100 ngay trong. Vi lng 140 kg/ha T.hazianum co tac dung phong chong S.rolfsii va tang nang suat len trong khoang 3 nam . Trong cac giong Trichodema co T.harzianum nuoi trong tren moi trng ran co tac dung chong c cac benh thoi trang tren hanh (Sclerotium cepivorum) Ai Cap (Abd-El va cong tac vien, 1982) va My (Papavizas va Lewis, 1982) ; benh tren da leo va benh tren cay bong do Verticillium dahliae Lien Xo
(Fedrorinchik va cong tac vien, 1975) ; benh chet rap cay con do Rhizoctonia va benh tan rui (S.rolfsii) tren nhieu cay trong Israel (Chet va cong tac vien, 1982; Elad va cong tac vien, 1982) va benh thoi trai tren da leo (Kommedahi va Windels, 1981). Hieu qua PTSH cua nam nuoi trong tren moi trng ran phu thuoc vao nhiet o, loai moi trng nuoi trong va thi iem cay nam vao at (Elad va cong tac vien, 1980), t le cay Trichodema vao gia the (Elad va cong tac vien, 1980 ; Hadar va cong tac vien, 1979) va mat o cua nam gay benh trong at. Nhng van e phc tap khac lien quan en kha nang chong benh cua Trichodema cung c nghien cu. Kelley (1976), cong bo T.harzianum c cay vao at set vi muc ch phong tr benh chet rap cay con tren da. Ong ta nhan thay rang kha nang khang benh cua nam nay phu thuoc vao d lng dinh dng trong at, ac biet trong at am benh chet rap phat trien rat cao. Khi cay T.harzianum dang vien hu c vao moi trng nhieu dinh dng, lam tang quan the Pythium, hau qua la phat trien benh trong vn da leo (Moody va Gindrat, 1977). Benh chet rap xay ra khi them Trichodema dang vien vao luc trc khi gieo hat da leo. 2.1.3 Kha nang lam tang bao t nam Trichodema trong at Vi cac bao t phan sinh tran, phng phap x ly bang kh trung xong hi hay hi nc tren at co lam tang so lng bao t trong at. Th nghiem phong tr benh Fusarium tren cay hoa cuc a c ghi nhan gan ay khi them vao at dung dch loai T. viride khang thuoc Benomyl. S dung nong o 104 bao t/cm2 dong nam nay tron vao at sau khi a c kh trung bang hi nc (820C trong 2 gi), so bao t cua dong nam nay tang rat nhanh, nam gay benh cay khong xam nhap lai vao at.
Cac nghien cu gan ay (Beagle, 1984 ; Papavizas va Lewis, 1983) ch ra rang san pham cua phng phap len men nh la dang bot, bun than hoac dang vien va cay vao at khong ch tang nhanh so lng kch tnh ma con ngan can benh hieu qua hn la dung bao t tran. Dang vien san pham len men cua T.hamatum, T.harzianum, T.viride, va T. virens co kha nang lam giam kha nang song sot va sinh trng cua R.solani trong at va benh thoi trai tren ca chua (Papavizas va Lewis, 1984). Cach s dung phng phap len men chac chan co anh hng en thi gian song cua bao t, kha nang song sot va nhan sinh khoi trong at va tiem nang PTSH. Du vay phng phap len men se c s dung va cai tien nhieu trong tng lai cho viec s dung nam khang PTSH. 2.1.4 Vai tro cua nam Trichodema trong viec x ly hat giong Kommedahl va Windels (Kommedahi, 1981) co nhieu ghi nhan ve kha nang khang benh phong tr sinh hoc tren cay b benh. Mac du co nhieu loai nam Trichodema co kha nang dung vao PTSH nhng cha co mot san pham thng mai nao c ang ky tai My (Papavizas va Lewis, 1981). Co nhieu ly do, mot trong nhng ly do nay la can mot so lng ln nguyen lieu PTSH tren mot dien tch at th nghiem ln (Hadar va cong tac vien, 1984: Harman va cong tac vien, 1980). S dung Trichodema vao viec x ly hat giong co lien quan en kha nang xam nhap cua Trichodema vao trong at, phng phap nay oi hoi mot so lng bao t ln e ap dung. Tuy nhien, ay la mot phng phap rat co y ngha trong viec phong tr nam gay benh giai oan hat en giai oan cay con. Kha nang PTSH cua nam T. hamatum vi benh chet rap cay con do nam R.solani va Pythium spp. co hieu qua tren hat giong au Hoa Lan va cu cai ng (Harman va cong tac vien, 1980). Dong nam T.harzianum x ly qua tia t ngoai (Papavizas va Lewis 1982) va T. viride co hieu qua trong PTSH (Papavizas va Lewis, 1981). Hat au nanh c x ly T. pseudokoningii va hat giong bap c x ly
T.hazianum co hieu qua lam ngan chan nguon benh va lam tang nang suat trong viec phong tr nam Rhizoctonia tren canh ong nhiem nam nay va co hieu qua khi dung nam T. harzianum x ly hat bong phong tr nam R.solani tai Israel. Hieu qua thanh cong trong viec dung Trichodema x ly hat giong b anh hng t nhieu yeu to phan lap (Papavizas va Lewis, 1981): tuoi cua hat giong gieo trong (Kommedahi va Windels, 1981), nhiet o cua at va tai hoat ong cua at (Harman va cong tac vien, 1981), loai at va vi sinh vat hien dien trong at (Hadar va cong tac vien, 1984), dinh dng trong qua trnh cay nam (Harman va cong tac vien, 1981), mat o nam khi cay vao at (Nelson, 1975), tiem nang benh gay hai cay trong at, va tuoi cua cay trong (Kommedahi va Windels, 1981). Trichodema co hieu qua nhat trong viec phong tr benh chet rap cay con, kha nang tao sinh khoi trong at va he re ngan can benh gay hai cay bang cach canh tranh, ky sinh tren nam hoac khang sinh hoc. Ngoai ra chung con gay anh hng manh en vi khuan (Hadar va cong tac vien, 1984) va cac loai nam khac trong at. 2.2 ac iem cua nam Trichodema 2.2.1 Hnh thai, s sinh trng va s hnh thanh bao t cua Trichodema - ac iem hnh thai: Trichodema la mot loai nam at, phat trien tot tren cac loai at giau dinh dng hoac tren tan d thc vat. ac iem hnh thai cua nam nay la canh bao t khong mau, si nam khong mau, co vach ngan, co kha nang phan nhanh nhieu va cho lng bao t rat ln. Bao t thng co mau xanh, n bao hnh trng, tron, elip hoac hnh oval tuy theo tng loai. Bao t nh nh cua canh. - S sinh trng cua Trichodema: la mot loai nam hoai sinh trong at nen Trichodema co kha nang s dung nguon hon hp carbon va nitrogen. Nguon cacbon va nang lng Trichodema s dung c la Monosaceharides va
Disaccharides, cung vi hon hp Polysaccgarides, puriness, pyrinidines, acide amin, tanmins va caechins co ong; Aldehydes va acide hu c. ac biet la acide beo (E.B.Nelson, G.E. Harman), methanol methylamine, formate va NH3 la nguon am bat buoc phai co trong moi trng nuoi trong Trichodema. Nhng nguon nitrogen nao cung ho tr cho moi trng co nhieu dinh dng. Muoi, cac nguon sulfur va cac hon hp nh vitamin cung co anh hng ln en kha nang sinh trng cua Trichodema. Nhng muoi sodium chloride se lam giam s sinh trng va phat trien cua mot so loai Trichodema. Do o trong moi trng nuoi trong khong c co mat cua muoi nay. Nong o CO2 trong moi trng nuoi trong cung anh hng en s sinh trng cua nam oi khang trong at. Tuy nhien anh hng cua CO2 en kha nang sinh trng va san xuat cua Trichodema phu thuoc vao nong o pH cua moi trng at. CO2 nong o 10% khong anh hng en toc o sinh trng cua Trichodema. Toc o moc nhanh cua Trichodema nong o CO2 cao trong moi trng kiem, co the giai thch tai sao Trichodema thng song trong moi trng at phen, am t, t hien dien tren at kiem. V the CO2 co anh hng en sinh trng cua Trichodema tai o pH co gia tr cao. - S hnh thanh bao t tren moi trng: Phan ln cac loai Trichodema co cam quang, de nay mam nhieu ieu kien moi trng t nhien va nhan tao di ieu kien toi sang lan lon, hay bao t co the xuat hien trong ieu kien sang. Moi trng agar trong khoang 20-30 giay anh sang 85 Lux lam tang hieu qua nay mam. The bao t phialoconidio cam ng vi anh sang nhat se xuat hien nhieu di anh sang ban ngay ch trong khoang 3 phut hoac gan tia cc tm (bc song 366nm) trong khoang 10 30 giay. Cac tac gia a cong bo Trichodema khong hnh thanh bao t bc song di 254nm hoac tren 1.100nm va hnh thanh bao t nhieu nhat bc song 380nm en 440nm. Cac bao t cam quang han che phat trien di anh hng cua cac hoa chat. Cac hon hp nh azaguanine, 5fluorouracil, actiomycin D, Cycloheximide, phenethyl alcohol va ethidium 9
bromide ngan can s hnh thanh cac hau mo bao t, ay la 1 cau truc ac biet cua c the rat quan trong trong hnh thai hoc, lam tang tiem nang trong phong tr sinh hoc. T. hamatum, T.hazianum, T.viride va T. virens trong ca moi trng long va ran co acide thch hp cho bao t nay mam hn la moi trng trung tnh. 2.2.2 Sinh thai hoc cua Trichodema Sinh thai hoc cho biet s phan bo cua Trichodema trong ieu kien c hoc cua bao t nam Trichodema trong at. - at khang nam: at t nhien co kha nang khang nam va kha nang nay se mat dan. ieu nay co lien quan en s xuat hien va mat o phan bo c hoc cua Trichodema. Bao t phan sinh cua Trichodema co kha nang khang nam cao va lien quan en hien tng giam kha nang khang nam trong at. o nhay cua at khang nam c cong bo tren at trung tnh, at kiem chua va at acide. Cac bao t phan sinh khang nam nhieu hn hau mo bao t, si nam t khang nam hn bao t phan sinh. - Thiet lap quan the va hien tng nay mam trong at: Vi sinh vat trong at khong va hoat ong phu thuoc vao nhieu loai chat nen trong at, co nhieu phng phap xac nh khac nhau. Trong nhieu trng hp cho thay van e nay khong thch hp vi Trichodema va tang len nhieu trong nhieu loai at khac nhau. Khi cay si nam non (cha co bao t) vao at eu lien quan mat thiet vi tnh trang thanh phan moi trng at. Bao t sinh soi nay n (mat o 100) va thiet lap quan the can bang trong at (mat o duy tr can bang trong at t 9 36 tuan sau khi cay nam vao at). ieu nay phu thuoc vao tuoi nam va nh vay co lien quan en thanh phan thc an, va viec hnh thanh quan the si nam Trichodema t thanh phan nuoi trong khong lien quan en loai at. Vien Alginate cha bao t phan sinh c san xuat bang phng phap len men tao quan the bao t t hn so vi phng phap nhan sinh khoi bang at (chu yeu la hau mo bao t). Them vao
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o, viec len men at c them vao at chat Pyrax kho giup lam tang quan the t 5.103 len 6-7.106 bao t / gram at. - Thiet lap quan the tai vung re cay: Trichodema a c phan lap t re cay va co kha nang dung vao viec phong tr sinh hoc tai vung re cay b benh. Hieu qua cua Trichodema khong ch x ly hat ma con tiep tuc thiet lap quan the di vung re cay sau khi x ly hat. Trichodema x ly hat phat trien nhanh xung quanh he re tao cac bao t ngan can benh xam nhiem cay trong. Neu Trichodema c cay vao at vi tac dung chong benh cho cay bat buoc phai cay doc theo be mat re nhng cach xa la mam. Trichodema co kha nang diet tr benh thoi re, hat va benh chet cay con. Nhng nghien cu gan ay cho thay T.hazzianum khong thiet lap quan the xung quanh he re cay ho au va cay au Hoa Lan con. Quan sat bao t tren vung re cay gom re, vo, hat b thoi va la mam, so lng bao t tren moi gram at xung quanh he re luon luon t. W.L.Chao, G.E. Harma va E.B.Nelson tren so lieu khong cong bo, cho rang bao t cua Trichodema t thiet lap quan the hay t di chuyen vao vung re cay. Vi
T. hazzianum vai bao t c tm thay cach xung quanh he re cay 10cm, tren cay c x ly hat. Ngc lai, so lng bao t tm thay nhieu tren la mam au Hoa Lan b thoi va vo hat giong ke ca mau benh xung quanh re. Co nhieu giai thch ve viec so lng bao t Trichodema tang hoac giam trong at va Trichodema khong co kha nang thiet lap quan the vung re cay, do co nhieu ly do bao gom: thieu dinh dng, hien dien chat oc trong re cay hay hien dien cua chat khang hoac s hien dien cua vi sinh vat oi lap vi Trichodema (W.L. Chao, G.E. Hazman, E.B. Nelson tren so lieu khong cong bo) tai vung re hay mc o re cua cay. V du: Pseudomonas oi lap vi tac nhan phong tr sinh hoc nhng khi co hien dien cua chat sat trong vung re cua cay hay Pseudomonas san xuat chat oc chuyen oi gay anh hng en bao t cua Trichodema. 11
2.2.3 Kha nang oi khang cua nam Trichodema Nam oi khang la nhng thanh vien pho bien cua he vi sinh vat at (Pomsch va cong tac vien 1920). Chung thng tiet ra cac men, khang sinh gay oc cho nam gay benh hoac nam khang canh tranh ieu kien song vi nam gay benh. S phan biet cua chung phu thuoc vao vung a ly, loai at, ieu kien kh hau, va tham thc vat tng khu vc. Nam oi khang co the km ham s sinh trng, phat trien cua nam gay benh, giup cay hoi phuc, sinh trng va phat trien, mot so loai nam oi khang a c tm thay: Penicillium axalicum. P. frequetans. P. Vermiculata, P. nigricans, P. chregsogetum la oi khang cua nam pythiumspp, phizoctotia solani, Sclerotium Cepivorum, Verticillium alboatrum (Martin va cong tac vien 1995). Nam Aspergillus niger oi khang vi nam Fusaium sokeni, Rhizoctonia solani, Aeteraria alternata. Nam Aureobasidium, pollulans va Kikuchii la oi khang cua nam Diaporthe phaseolorum var. sojage (Egurazdova et at, 1979). oi vi nam Trichodema, cung la mot trong nhng loai nam co kha nang c che mot so nam gay benh khac nh: Sclerotium rolfsii, phytopthora, Fusarium Pythium, Rhizoctonia gay benh tren nhieu loai cay trong: Cay ho au, cay an trai, hoa thao, cay cong nghiep va cay hoa kieng. 2.2.4 C che oi khang cua nam Trichodema S oi khang cua nam Trichodema thong qua nhieu c che. Vao nam 1932 Weinding a mo ta hien tng nam Trichodema ky sinh nam gay benh va at ten cho hien tng o la Giao thoa si nam (Cnyder, 1976). Hien tng giao thoa gom ba giai oan nh sau: (1) Si nam Trichodema vay quanh si nam gay benh .(2) Sau s vay quanh, si nam Trichodema that chat lay cac si nam gay benh cay. (3) Cuoi cung la si nam Trichodema am xuyen lam thung lp te bao cua nam gay benh, dan en s gay benh lam cho chat nguyen sinh trong nam gay
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benh b phan huy va dan en nam benh chet. Sau nay quan sat di knh hien vi, hien tng ky sinh cua nam Trichodema c mo ta nh sau: Tai nhng iem nam Trichodema tiep xuc vi nam gay benh a lam cho nam gay benh teo lai va chet (Dubey, 1995; Rousscu va cong tac vien, 1996). Ngc lai nhng iem khong co s tiep xuc cua nam Trichodema vi nam gay benh van chet th cac nha nghien cu cho la tac ong cua chat khang sinh t nam Trichodema sinh ra gay oc cho nam gay benh (Agrowcal va cong tac vien, 1979; Michrina va cong tac vien 1996). 2.2.5 Cac san pham trao oi cua nam Trichodema Weidling la tac gia au tien cong bo san pham c trao oi cua Trichodema. Weidling va Emerson a phan lap c chat ket tinh t chat trao oi hu c rat oc khi pha loang nhieu lan khi dung. Trichodema chong R.solani. Chat nay ten thong thng: Gliotoxin chat oc th 2 do Brian va Mc Growan cong bo la viridin san xuat t T. viride. Webster va Lomas ghi nhan la hai chat khang sinh nay eu hien dien tren moi trng c loc t T. viride sau o Weidling co lap ra san pham gliotoxin, Brian va Mc Growan co lap viridin. Dennis va Webstre ghi nhan Trichodema spp. co san pham khang, khac vi gliotoxin va viridin, san pham o la chat khang sinh, co the hoa tan c trichlorofore ten thong thng la T.viride va T. polysporum va 1 chat khang peptied t T. hazianum. Ngoai chat oc la chat trao oi va khang sinh ra, Trichodema con co the tiet ra nhieu enzym khac nh exo va endoglucanases, cellobiase va chitinase co kha nang phan huy thanh te bao cua nam gay benh. Nam Trichodema cung nh mot so nam moc khac nh Gliocladium, Calvatia cho lng enzym chitinase cao, chitinase co nhieu chc nang, mot trong nhng chc nang chnh la kha nang phan huy chitinase la thanh phan chat dnh
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cau tao vach te bao nam, yeu to rat quan trong trong hoat ong ky sinh nham oi khang lai cac loai nam gay benh thc vat, bang 2.1 cho thay cac thanh phan cau tao cua thanh te bao nam. Chitine co cau tao va chc nang giong Cellulose, trong thien nhien Chitine la thanh phan hu c chiem th hai sau Cellulose ve so lng, Chitine co the thay the mot phan hay toan bo Cellulose trong thanh te bao cua mot so loai thc vat. Chitine la chat ran vo nh hnh, no khong tan trong nc va hau het cac acide cung nh kiem, alcohol va cac dung moi hu c khac. Chitine co the b phan huy bi acide vo c manh (HCL am ac, H2SO4 am ac) hoac bang enzym sinh vat. Qua trnh thuy phan se cat cac lien ket - 14glycoside cho ra olisaccharide. Trong qua trnh ky sinh tren nam benh, Trichodema co the tiet ra cac loai enzym e phan huy thanh te bao cua nam gay benh. Bang 2.1 Cau tao cua thanh te bao cac nhom nam chu yeu Nhom phan loai Ten khoa hoc Myxomyces Plasmodiophora Oomycetes Hyphochtridiomycetes Chytridiomycetes Zygomycetes Ascomycetes Basidiomycetes Deuteromycetes Saccharomycetes 14 Glucan Mannan Chitin Glucan Cau tao chnh cua thanh te bao Cellulose Chitin Cellulose - Glucan Cellulose - Glucan Chitin - Glucan Chitin Chitosan
Nam nhay Nam noan Nam co Nam tiep hp Nam nang Nam am Nam bat toan Ngoai le
Rhodotorulaceae
Chitin Mannan
Ngoai ra, 3 loai enzym ngoai bao khac sinh san ra t nam Trichodema a c tach chiet, tinh sach cung co hoat tnh phan huy Chitine. Cac enzym o la: N-acetylglucosamidase, Chitosesidase va Endochitinase. 2.3. Tnh hnh benh hai tren tieu va sau rieng 2.3.1 Benh hai tren tieu Thanh phan benh hai tren tieu rat a dang va phong phu, chung lam cho cay suy yeu, heo vang hoac lam cay chet rat nhanh, pho bien tat ca cac vung tieu nguyen lieu tren the gii nh Malaysia, Indonesia, An o, Brazil, Srilauka, Thailand va Viet Nam. Trong cac benh hai tren cay tieu: Than th (Collectotrichum gleoeposrioides); en la (Lasiondiplodia theo bromce); om la (Rosellina sp); kho van (Rhizoctonia solani); Benh chet nhanh day tieu (Phytophthora parasitica); Benh tieu ien (MLO) va benh do tuyen trung gay ra. Trong o benh chet nhanh day tieu that s la mot tai hai cho nha vn. Benh xuat hien va lay lan rat nhanh, thng lam tieu chet hang loat gay mat trang hoac giam nang suat tram trong. Theo ket qua nghien cu cua bo mon Bao Ve Thc Vat, Vien Ky Thuat Khoa Hoc Nong Lam Nghiep Tay Nguyen nam 1999 th benh nay xuat hien vi ty le rat thap (0,11% Gia Lai), cha c coi la benh nguy hiem cho cay tieu vung Tay Nguyen nhng en nam 2000 2001 benh a gay thanh dch mot so vung EaHLeo, Easup, C Mgar thuoc tnh aklak. tnh Gia Lai, vung b hai nang la Mang Yang (Tap ch thong tin khoa hoc ky thuat Nong Lam Nghiep, So 1/2001). T khi cay tieu ru la vang va rung hang loat ch trong vong 5 7 ngay. Ca vn tieu co the b hai trong vong vai tuan hay vai thang. Khi thay trieu chng heo day th bo re a b nam tan cong trc t 1 2 thang, do o benh 15
nay rat kho phong tr. Trong vn co khoang 5 7% cay chet th phan ln cac cay khac a b nam tan cong. Thc tai, cac vn tieu chuyen canh Bnh Phc, Ba Ra Vung Tau, ong Nai hien ang b benh nay tan pha d oi, co vn hau nh b chet hoan toan, gay mat trang. Benh nay hai hau het cac bo phan cua cay tieu: than, la, re, co re va trai. Bo re va phan than ngam b nam tan cong thoi en, vo bong ra khoi re, phan day tren mat at b heo, la chuyen qua mau vang va rung hang loat trong vong 7 14 ngay, e lai canh tr trui, sau o toan day b heo en va chet. Vao mua ma benh xuat hien nhng la di. Nhng vong nau en, tap trung au la, cac om ln dan, co mau nau sam va rat de rung. Khi benh tan cong vao day, than, la b benh va rung, cung luc o long cung rung. Bien phap phong tr benh chet nhanh day tieu, hien nay rat kho tr nen cha co bien phap nao ngan can c. oi vi benh nay, cong tac phong benh la chnh . 2.3.2 Benh tren cay sau rieng Cay sau rieng la loai cay an trai co gia tr kinh te cao, ngay cang c m rong dien tch nhieu ni. Do o thanh phan sau benh hai cung phat trien khong kem. Malaysia, co trng hp 50% so cay con trong vn b chet do benh nt chay nha than. Thai Lan (1994) 20% so cay b chet thi ky ang cho trai do benh nc than chay mu (Nguyen Minh Chau 2003). Benh pho bien va nguy hiem nhat tren cay sau rieng o la benh nt than chay mu do nam Phytophthora sp. gay ra ong thi nam nay con hai them bo phan trai lam trai sau rieng b thoi. oi vi benh nt than chay mu, phan b hai chu yeu phan than 1m t goc len. au tien, tren vo than co cac vet am mau hi t, sau co mau nau o, cho
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vo benh nt ra va chay mu mau vang. Lau ngay vet benh lan khap vung than va an sau vao phan go, lam cay ky chu heo va rung cac la va mot so canh pha ngon b kho chet, tiep theo la cac canh pha di va cuoi cung la ca cay b chet. Neu nam tan cong vao phan re se lam thoi re va sau o cay canh chet nhanh hn (Tap ch Khuyen Nong Tay Ninh so 4/2002). phan trai, vet benh thoi thng co cac si nam mau trang tren vet benh, benh lam thoi mot phan hoac thoi ca trai va de lay sang nhng trai khac. Cac loai thuoc hoa hoc e ap dung phong tr gom : Ridomil MZ 72 WP, Mancozed 80WP, Alliet 80WP vi nong o 2% va dung dch KmNO4 1% (thuoc tm) (Vo Th Thu Oanh, 1999). Theo tap ch Khuyen Nong Tay Ninh, so 4/2000 cho biet, co the s dung thuoc Alliete 80WP va Ridomyl MZ 72WP. Ngoai ra, can phai ket hp vi bien phap canh tac va ve sinh ong ruong. 2.3.3 Nam Phytophthora La loai nam a ky chu, ngoai gay hai tieu, sau rieng con gay hai tren rau, hoa kieng thuoc ho Pythiaceae, bo Pernoporales lp Omycetes, si nam khong mau, khong vach ngan, n bao kch thc khong eu, bao t mang hnh trng va hnh qua chanh, tren au co nuom hoac khong co nuom, khong mau trong suot. Bao t hnh cau hoac hnh than co hai long roi, di chuyen rat nhanh trong nc, nhiet o thch hp e nam sinh trng va phat trien 25-300C, pH: 6-7. Tren cay tieu, dong nam thng gay hai c xac nh nam co ten la Phytophthora sp., gay hai chu yeu trong mua ma, nhat la vao cuoi ma, khi co kh hau am va am. Nam Phytophthora sp. co the tan cong rieng le nhng a so la co s ket hp vi cac nam khac nh: Fusarium, Pythium, Rhizoctiona cung tan cong lam cay sau rieng chet nhanh chong. Tren cay sau rieng, nam Phytophthora sp. gay hai nang nhng vn trong day, am o cao, nhat la am o quanh goc cao. Benh phat hien manh tren
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at xau, at thoat nc kem. Nam benh xam nhap vao cay qua vet thng do con trung pha hay xay xat trong qua trnh cham soc. 2.4 Cac phng phap len men tao che pham sinh hoc Hien nay tren the gii co nhieu phng phap len men tao che pham sinh hoc e tr nam benh, sau hai cay trong trong nong nghiep. Ngi ta a xay dng nhng quy trnh e thu nhan san pham len men kha hoan chnh va c ap dung vao thc te san xuat ln quy mo cong nghiep. Tuy nhien, quy trnh len men van ang con nam trong giai oan tm kiem mot phng phap thch hp, chon la ieu kien va moi trng nuoi cay toi u e at so lng bao t gom chat kho cao, gia thanh san pham re ong thi san pham tao ra phai de bao quan, gi c hoat tnh lau ben nhiet o bnh thng. Mot so phng phap len men che tao che pham sinh hoc a c nghien cu va ng dung nh sau: 2.4.1 Phng phap len men chm Nhieu tac gia a ap dung phng phap len men chm e nuoi cay cac nam diet sau nh nam Beanveria bassiana (Samsinakova, 1961), Metarrhizium anisopliae (Adamek, 1965) va cac tac gia khac (Weiser, 1966). Nam nuoi cay chm phat trien cho dang Chlamydospores, con khi nuoi be mat hoac nam phat trien cho dang conidia. Khi nuoi cay chm nam thng phat trien qua 6 giai oan: Giai oai 1: Bao t phong len, sau o tao thanh mot hay nhieu ong mam Giai oan 2: Cac si nam phan nhanh Giai oan 3: Tao thanh Chlamydospores lan th nhat Giai oan 4: Cac Chlamydospores phat trien lai tao thanh si nam Giai oan 5: Si nam phat trien bat au tao Chlamydospores lan th hai Giai oan 6: Tao thanh Chlamydospores lan 2.
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Neu tiep tuc nuoi cay se xay ra s phan li hoan toan cua si nam. Lng Chlamydospores lan th hai on nh mot thi gian, sau o cung giam i. Nhc iem cua phng phap len men chm la ch thu c che pham dang bao t choi Chlamydospores, khong ben vng, de b mat hoat tnh v co thi gian song ngan, co cau truc khong ben vng. 2.4.2 Phng phap len men be mat khong vo trung tao che pham nam Phng phap len men be mat khong vo trung tao che pham nam a c thc hien bi A.A Evlacchova (1968), a ap dung phng phap len men nay e tao che pham Boverin, chat diet sau vi sinh tren c s nam Beauveria bassiana. Moi trng dinh dng c nau soi 1000C va khi nguoi cho them chat khang sinh Streptomycine (0,01%) e ngan can s phat trien cua vi khuan trong qua trnh len men. Tuy nhien, e thc hien phng phap nay co hieu qua can thc hien 5 nguyen tac chu yeu sau: (1) Bao t nam c cay phu kn khap be mat moi trng dch (a un soi 1000C/20 phut). Cac bao t nam sau khi cay se t ieu hoa khuyech tan thanh mot mang mong khap be mat moi trng, ngan nga cac vi sinh vat la phat trien. (2) Lng bao t tren mot n v be mat moi trng c cay vi mot lng ln u ap ao c phat trien ban au cua cac vi sinh vat la (1-2 ty bao t/cm2) (3) Ngay sau khi nay mam, bao t cua cac nam diet sau se tiet ra cac chat trao oi chat, giong khang sinh e c che s phat trien cua vi khuan va nam la (Calvish, 1972 ; Scherf denberg, 1965; Evlachova, 1966 ;1974; Evlachova va Tarasov, 1968). (4) Tao moi trng pH thap 5 5,5 thuan li hn cho s phat trien cua nam, c che s phat trien cua vi khuan.
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(5) S dung dung cu, thiet b va phong nuoi cay sach se han che toi thieu s nhiem cua cac nam nuoi. 2.4.3 Phng phap len men hai giai oan tao che pham nam phng phap nay, cac bc thc hien c tien hanh nh sau (s o 3): Nh vay, phng phap len men 2 giai oan thc chat la phng phap len men chm (giai oan 1) va sau o chuyen sang len men be mat (giai oan 2). Nh a neu trong phng phap cay chm, s phat trien cua nam xay ra gom 6 pha phat trien, thuan li nhat cho s chuyen sang giai oan 2 nuoi be mat la luc ket thuc pha 3 va bat au pha 4, ngha la luc trong dch nuoi cay tao nhieu Chlamydospores lan th nhat, bat au phat trien thanh si nam. Muc ch cuoi cung khi san xuat nam la thu c mot lng ln dang conidia (bao t co cau truc ben vng, co thi gian song lau, gi hoat tnh ben lau hn) tren mot the tch moi trng dinh dng. V vay, can thiet phai nghien cu ieu kien tao conidia khi nuoi be mat giai oan 2. Khi nhiet o nuoi trong khoang 26-320C co hieu suat tao conidia cao nhat. Khi nhiet o thap hn 240C va cao hn 350C hieu suat tao conidia giam. Am o khong kh cao Rh = 90 100% thuc ay nhanh s tao thanh mang nam tren be mat dch nuoi va tao nhieu conidia. Am o khong kh cao Rh = 70 80% se can tr s tao thanh mang nam va tao t conidia. 2.4.4 Phng phap len men xop tao che pham nam La phng phap c ap dung rong rai nhat v ay la qua trnh len men n gian, de thanh cong hn cac quy trnh khac. Trong quy trnh nay, cac loai c chat dung e lam moi trng cho nam khang phat trien la cam trau, bot gao, bot bap cung vi dch dinh dng e nuoi cay nam.
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Len men xop se thu nhan c che pham sinh hoc dang nh bao t (conidiospore) cua cac nam khang, on nh va ben vng hn dang Chlamydospores (bao t choi), v vay phng phap nay a c nhieu nha khoa hoc tren the gii quan tam t lau. Khi nuoi nam khang tren moi trng xop (hat) theo Solivey F.F (1984) a at hieu suat bao t so vi cac phng phap len men khac e chong sau benh. Tuy nhien, theo ong kha nang song cua bao t trong che pham phu thuoc khong ch vao ieu kien bao quan ma con phu thuoc vao s say kho va c chat dinh dng. Ket qua cho thay cac che pham sinh hoc nam diet sau Metarrhizium anisopliae va Beauveria bassiana neu bao quan nhiet o 5 100C co the gi c hoat tnh trong 6-8 thang. Ngc lai bao quan nhiet o phong th ch gi c 6-8 tuan. Nh vay cac che pham sinh hoc t nam c san xuat ra can phai c bao quan ieu kien lanh, ni kho rao va se co kha nang gi c hoat tnh cua che pham trong khoang thi gian 6-8 thang. 2.4.5 Phng phap len men xop tao che pham nam Trichodema Ngi ta cung thng ap dung qui trnh len men xop e tao sinh khoi, a ap dung rong rai trong san xuat e diet cac loai nam gay hai cay trong. Bi v, moi trng len men xop cho lng bao t / gram che pham cao, quy trnh n gian de thc hien, gia thanh san pham tao ra thap, ap ng c nhu cau cua ngi nong dan. e co c san pham tao ra co nhieu bao t, cac ieu kien moi trng nh o am tng oi khong kh Rh = 95%; nhiet o at: 30 320C va thi gian nuoi cay la 5-8 ngay. Thanh phan cac loai moi trng dung e len men xop, cung la tao che pham nam Trichodema nh sau: (1) Bot ngo + ba au phung
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(2) Cam gao + bot ngo manh (3) Cam gao + bot ngo + bot au nanh (4) Cam gao + bot ngo + ba au kho (5) Lua nau chn (6) Gao nau chn - Ngoai ra, cac nghien cu cua Nguyen Ngoc Tu (1997) con th nghiem tren 3 loai moi trng khac e nhan sinh khoi Trichodema o la: (1) moi trng gom cac thanh phan la cam, trau ; (2) moi trng than, bun ; (3) moi trng bot thach cao tam mat r 10%. Th nghiem c tien hanh trong cung ieu kien ve am o, nhiet o va thi gian nuoi cay va hai dong Trichodema c chon la Trichodema 1 va Trichodema 2 . Ket qua cua th nghiem nh sau: Moi trng cam trau cho san pham Trichodema co so lng bao t/g che pham kho at 109 bao t / gram, so lng bao t cao nhat va bao quan nhiet o phong. Moi trng than bun va bot thach cao tam mat r cho so lng bao t / gram che pham kho thap hn, khoang 107, 108 bao t / gram. Tiep tuc th nghiem ve thi gian bao quan cua che pham Trichodema -1 va Trichodema -2 a len men t moi trng cam trau sau 3 thang bao quan nhiet o phong (29-320C) so lng bao t tnh c / gram chat kho nh sau: Trichodema 2 con 2,72.109 bao t / gram va Trichodema 1 con 0,65.109 bao t / gram so vi ban au la 5,88.109 bao t / gram Trichodema 2 va 2,24.109 bao t / gram Trichodema - 1. Tuy nhien, che pham Trichodema tot nhat la khong nen e qua 9 thang. Vi che pham nhan cha bao quan so lng bao t / gram la cao nhat nhng sau qua trnh bao quan, lng bao t giam dan, nhat la sau 12 thang th lng bao t trong che pham giam ang ke. Nh vay, thi gian bao quan cang lau th cang anh hng ti chat lng cua che pham. 22
2.5 Mot so che pham t Trichodema a c san xuat va ng dung Nen nong nghiep hien ai vi phng thc ch oc canh mot hoac vai loai cay trong tren mot dien tch rong ln, xem thuoc hoa hoc nh la mot bien phap tot nhat e phong tr dch hai tren cay trong a dan en mot loat cac hau qua ma con ngi va thien nhien phai ganh chu o la: o nhiem moi trng; o nhiem nguon nc gay chet cho cac ong vat thuy san, gia suc, gia cam ; lam mat can bang sinh thai trong t nhien ; tao noi mi khang thuoc con trung lam tang them mc o tan pha tren cay nhieu hn; ton d lng thuoc hoa hoc qua mc cho phep tren san pham nong nghiep, gay hai ti sc khoe con ngi ; hn na s dung thuoc hoa hoc chi ph au t cao nhng ngay cang khong co hieu qua. Do o, can phai nhanh chong giam bt lng thuoc s dung hoac chuyen sang che pham vi sinh nham khac phuc cac hau qua tren. T nhng thc tai o, tren the gii hien nay a co nhieu quoc gia s dung che pham vi sinh e tr sau benh hai cay trong. Theo Dunin (1979) Lien Xo (cu) a s dung che pham Trichodema (t nam Trichodema lignorum) tren cay bong vai lam giam 15-20% benh heo do nam Verticillium va lam tang nang suat len 3-9 ta bong / hecta. S dung che pham Trichodema cung lam giam 2,5 3 lan benh thoi re cay con thuoc la va rau mau. Trong nhng nam gia thap ky 80, che pham Trichodema Lien Xo cu a s dung tren dien tch 3.000 hecta (Filippa, 1987), s dung 30-40g/m2 che pham (S.V. Badai), 1986). Che pham nam Trichodema Lien Xo (cu) co ten thng mai Trichodermin vi 4 dong che pham, Trichodermin-1: nam nhan nuoi tren moi trng dinh dng giau chat am va chat bot; Trichodermin-2: nam nhan nuoi tren moi trng cac phe lieu thc vat; Trichodermin-3: nam nhan tren than bun say kho va Trichodermin-4 la nam c nhan theo phng phap cay sau cac nguon nam Trichodema lignorum (Tran Th Thuan, 1999).
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Ngoai ra, con mot so che pham khac t nam Trichodema ang c cac quoc gia tren the gii s dung. nc ta, trong nhng nam gan ay, a co mot so cong trnh nghien cu san xuat che pham Trichodema. Mot so san pham cua Vien Sinh Hoc Nhiet i a vao th nghiem tren cay rau Cu Chi - Thanh pho Ho Ch Minh ; Che pham cua Bo mon Bao Ve Thc Vat Trng ai Hoc Can Th th nghiem tren cay xa lach xoong Vnh Long, eu cho nhng ket qua rat kha quan. Tuy nhien, cho ti nay, theo tai lieu c cong bo mi nhat cua Bo Nong Nghiep va Phat Trien Nong Thon so 49/2004/Q BNN ngay 13/10/2004, mi co duy nhat mot san pham vi ten thng mai TriB1, ham lng Trichodema 3.2 x 109 bao t /gram, c ang ky tr benh heo do nam Rhizoctonia, Sclerotium, Fusarium hai ca chua, khoai tay, au o, thuoc la va ho tieu Viet Nam.
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Chng 3 NOI DUNG VA PHNG PHAP NGHIEN CU

3.1 Noi dung 1. Thu thap va phan lap cac dong nam Trichodema cac vung sinh thai khac nhau. 2. anh gia tnh oi khang cua cac dong nam Trichodema c phan lap vi mot so nam gay hai cay trong trong at tren moi trng chon loc. 3. Nhan sinh khoi dong nam Trichodema co tnh khang cao bang phng phap len men long va len men xop e tm ra quy trnh len men thch hp nhat, tao ra san pham cho hieu qua phong tr cao, gia thanh thap. 4. S dung che pham t qui trnh len men e anh gia kha nang khang nam cua Trichodema oi vi nam Phytophthora gay benh cho cay tieu trong ieu kien nha li va gay benh x mu tren cay sau rieng ngoai ong ruong. 3.2 Thi gian va a iem thc hien Thi gian: T thang 2 nam 2003 en thang 8 nam 2004 25
a iem: - Th nghiem thu thap, phan lap va anh gia kha nang oi khang cua cac dong Trichodema thc hien Bo mon Bao Ve Thc Vat, Khoa Nong hoc, ai hoc Nong Lam Tp. HCM va Trung tam Khoa hoc Nong Nghiep va Chuyen giao cong nghe trng ai hoc Nong Lam Tp HCM. - Th nghiem nhan sinh khoi Trichodema thc hien Trung tam Khoa hoc Nong Nghiep va Chuyen giao cong nghe trng ai hoc Nong Lam Tp HCM. - Th nghiem anh gia kha nang phong tr benh cua che pham Trichodema tren cay tieu thc hien Nha li ai hoc Nong Lam Tp. HCM. - Th nghiem anh gia kha nang phong tr benh cua che pham Trichodema tren cay sau rieng giai oan vn m thc hien xa Long nh - Chau Thanh, Tien Giang. - Th nghiem anh gia kha nang phong tr benh cua che pham Trichodema tren cay sau rieng ngoai ong ruong thc hien xa Tam Bnh - Cai Lay Tien Giang. 3.3 Phng phap nghien cu 3.3.1 Vat lieu nghien cu - Cac dong nam Trichodema va cac nam gay benh c phan lap va lam thuan t cac mau at va mau cay benh lay t cac vung va cac loai cay trong khac nhau (xem bang 3.2 ; bang 3.3). - Cay tieu giong Kalimanda 4 thang tuoi mua Trai giong Chau c, Tnh Ba Ra Vung Tau, c trong trong chau nha en kch thc 20cm x 20cm. - Cay sau rieng cm vang hat lep Ri 6 chuan b xuat vn Vien Cay an qua Mien Nam. Vn sau rieng giong kho qua xanh, 5 nam tuoi xa Tam Bnh Cai Lay Tien Giang.
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- S dung may lac hieu IKA WERKE AS 1.9 cho phng phap len men long, cac hop xop kch thc 15cm x 20cm dung cho phng phap len men xop. - Ngoai ra con co tac dung khac can thiet c s dung viec phan lap, nuoi cay va quan sat vi sinh vat trong phong th nghiem. 3.3.2. Thu thap va phan lap nam + Thu thap mau nam Trichodema: Tuy theo loai cay trong, a ban lay mau, trong mot ruong hay mot vn cay chon nhng ni at mun, nhieu xac cay chet dung dao xen lay 50g at. Tren Bang 3.2a Nguon goc cua cac dong nam Trichodema c phan lap STT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Ma T.14 T.15 T.16 T.17 T.29 T.30 T.31 T.32 T.33 T.34 T.35 T.36 T.37 T.41 a iem SR a Oai Trng H NL Tp.HCM Trng H NL Tp.HCM Trng H NL Tp.HCM Lam ong Sun Agro - An o Can Th Bnh Phc Kien Giang Ha Noi Bnh Phc Cu Chi ong Nai Trng H NL Tp.HCM Ni lay mau Vn sau rieng at trai khoa Nong hoc at trai khoa Nong hoc at trai TT N-Lam Ng Vn che San pham th nghiem Vn cam, quyt Vn tieu Ruong da Ruong au phong Ruong da Ruong ca chua Vn sau rieng at trai khoa Nong hoc
Bang 3.2b Nguon goc nam gay benh dung trong cac th nghiem 27
STT 1 2 3 4
a iem a Lat Tay Ninh Ba Ra Vung Tau Tien Giang
Ten nam Fusarium Sclerotium Phytophthora Rhizoctonia
Cay trong a Lan Ca chua Tieu Ca chua
cay, chon nhng ni vet benh mi va lanh, cao lay khoang 5g. Tat ca mau c lay cho vao tui nylon. Ghi ly lch mau va lu tr 4oC cho en khi phan tch. S dung cac ch cai T va so th t ky hieu vung lay mau. Ngoai ra con chon them chon them mot mau san pham t nc ngoai ay la san pham cua Cong ty Sun Agro An o gi mau gii thieu Viet Nam. + Phan lap nam Trichodema: Mau at, mau tai vet benh tai moi iem thu c tron eu. Can 10 gram at (hay 1 gram mau benh) cho vao bnh tam giac dung tch 250ml vi lng nc cat theo ty le 1:10, lac manh tren may lac hieu ORBITAL INCUBATOR (SI 50) 150-180 vong/phut, nhiet o 28oC. Dung micro pipet hut 1ml dung dch trch cho vao ong nghiem co cha san 9ml nc cat, lac eu va tiep tuc pha loang ty le 10-5. moi nong o khac nhau dung micro-pipet hut 0,1ml dung dch roi cho vao a petri, ng knh 10cm cha moi trng PGA. Nuoi trong tu nh on nhiet o 250C. Sau 48 gi bao t nay mam va phat trien tren be mat moi trng, chung c nhan dang roi cay chuyen sang moi trng PGA tren a petri. Cay chuyen en khi khong con tap nhiem cac loai nam khac. Kiem tra di knh hien vi va phan loai chung. Lu gi nguon nam trong moi trng bot bap, 2% ng Dextrose trong ong nghiem.
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+ Cac dong nam at s dung trong th nghiem c thu thap tren cac loai cay trong, cac a iem neu tren va phan lap tng t nh phan lap nam Trichodema. 3.3.3 anh gia kha nang oi khang cua Trichodema vi 4 loai nam gay hai cay trong: Fusarium, Sclerotium, Phytophthora va Rhizoctonia - Dung 14 dong nam Trichodema a phan lap c e th nghiem tnh oi khang vi 4 loai nam gay hai trong at: Phizoctonia solanni, Sclerotium rolfsii, Phytophthora va Fusariuum.
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Hnh 3.1: Phng phap anh gia tnh khang cua Trichoderma tren a pertri vi cac nam gay benh, A: nam Trichoderma ; B: cac loai nam gay benh. a. Trichoderma khang manh vi cac loai nam gay benh (+++) b. Trichoderma khang trung bnh vi cac loai nam gay benh (++) c. Trichoderma khang yeu vi cac loai nam gay benh (+) d. Trichoderma khong khang vi cac loai nam gay benh (-) - Th nghiem oi khang: Nam Trichodema va nam gay benh c cay oi xng nhau qua ng knh a, theo s o cua hnh 3.1 lap lai 4 lan, moi lan lap lai la 1 a Petri trong moi trng bot bap. Th nghiem c quan sat 2 ngay mot lan, sau 7 ngay anh gia ket qua khang hoac khong khang. * C s anh gia tnh khang cua nam Trichodema: Sau 7 ngay nuoi cay: - Phan nam Trichodema phat trien bao phu qua phan nam gay hai. - Phan nam gay hai b bao mon dan mep khuan lac. - Phan nam Trichodema phat trien va khong che lam cho phan nam gay hai khong phat trien c.
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- Chung toi phan loai tnh khang cac mc khang manh: Nam Trichodema tan cong va phan huy hoan toan nam gay hai, ky hieu +++ (xem hnh 3.1a); khang trung bnh: Nam Trichodema tan cong va phan huy mot phan nam gay hai, ky hieu ++ (xem hnh 3.1b) ; khang yeu: Nam Trichodema ngan chan s phat trien cua nam nam gay hai, ky hieu + (xem hnh 3.1c) va khong khang: nam gay hai gan nh phat trien bnh thng, xam nhap vao vung phat trien cua nam Trichodema, ky hieu - (xem hnh 3.1d). 3.3.4 Phng phap nhan sinh khoi nam Trichodema 3.3.4.1 Tren moi trng long Chon 2 dong Trichodema co tnh khang tot sau th nghiem anh gia kha nang khang cua cac dong Trichodema e len men. Th nghiem c th nghiem tren thanh phan moi trng chu yeu la ng va gia au xanh. Tien hanh vi hai cong thc moi trng co lng ng glucose va lng gia khac nhau. Cong thc moi trng (1): Cac nghiem thc khac nhau co lng ng khac nhau, lng gia song khong oi. Th nghiem co 5 nghiem thc, 4 lan lap lai va cong thc c trnh bay qua bang 3.3. Bang 3.3 Thanh phan moi trng cac nghiem thc trong th nghiem khao sat anh hng cua ham lng ng en viec nhan sinh khoi Trichodema dang long Nghiem thc Thanh phan moi trng Gia (g) ng (g) Ure (g) KH2PO4 (g) MgSO4 (g) Nc (ml)
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P1 P2 P3 P4 P5
100 100 100 100 100
0 10 20 30 40
1 1 1 1 1
1 1 1 1 1
0,5 0,5 0,5 0,5 0,5
1000 1000 1000 1000 1000
Bang 3.4 Thanh phan moi trng cac nghiem thc trong th nghiem khao sat anh hng cua ham lng gia au xanh en viec nhan sinh khoi Trichodema dang long Nghiem thc G1 G2 G3 G4 G5 Thanh phan moi trng Gia (g) 0 50 100 150 200 ng (g) 20 20 20 20 20 Ure (g) 1 1 1 1 1 KH2PO4 (g) 1 1 1 1 1 MgSO4 (g) 0,5 0,5 0,5 0,5 0,5 Nc (ml) 1000 1000 1000 1000 1000
Cong thc moi trng (2): Cac nghiem thc khac nhau co lng gia song khac nhau, lng ng khong oi. Thanh phan moi trng cua cac nghiem thc cong thc moi trng (2) c trnh bay qua bang 3.4. Th nghiem co 5 nghiem thc, 4 lan lap lai, bo tr theo kieu hoan toan ngau nhien. - Phng phap thc hien: Cho 100ml dung dch moi trng vao bnh tam giac 250ml, moi bnh tam giac la 1 lan lap lai. Dung autoclave hap kh trung nhiet o 1200C, ap suat 1atm trong thi gian 30 phut. Cat 5 khoanh thach nam Trichodema, ng knh khoanh 5mm, nghien nat, cay vao bnh tam giac, lac 150 32
vong/1phut trong thi gian 72 gi (xem hnh 4.5). Sau khi ngng lac, loc lay si nam, tham kho bang giay tham, va can tnh trong lng si nam. - Ch tieu theo doi: Can xac nh khoi lng si nam. Theo doi thi gian phat trien cua nam Trichodema bang cach quan sat si nam phat trien bao t xanh tren mat moi trng. 3.3.4.2 Len men tren moi trng xop Chon 2 dong Trichodema co tnh khang cao sau th nghiem anh gia tnh oi khang. Th nghiem vi thanh phan c chat lam moi trng gom: Cam, trau, vo ca phe va cm. Th nghiem c tien hanh vi 7 nghiem thc va 4 lan lap lai, bo tr theo kieu hoan toan ngau nhien. Cong thc th nghiem xem bang 3.5. - Phng phap thc hien: Kh trung cac loai c chat nhiet o 1200C, ap suat 1atm, trong thi gian 30 phut. S dung cac hop xop kch thc 15cm x 20cm e len men, moi hop xop la mot lan lap lai. Can cac thanh phan c chat theo ty le cua tng nghiem thc. Cho them vao 15ml nc cat. Cat 5 khoanh thach nam Trichodema c chon nghien nat cho vao moi trng va tron eu. U cac hop xop trong phong kn cho en khi bao t nam moc xanh ay mat hop em say kho nhiet o 350C en khi o am con khoang 12% cho vao coi xay nhuyen va e vao vao hop nha gi nhiet o phong (250C) (xem hnh 4.6a, 4.6b va 4.6c). Moi thang kiem tra kha nang song sot cua che pham 1 lan tren moi trng PGA.
Bang 3.5 Thanh phan moi trng cac nghiem thc trong th nghiem khao sat anh hng cua cac loai c chat khac nhau en viec nhan sinh khoi Trichodema dang ran Nghiem thc N1 N2 Thanh phan moi trng 15g Cam + 15g Trau 10g Cam + 20g Trau 33 Ty le 1:1 1:2
N3 N4 N5 N6 N7
20g Cam + 10g Trau 15g Cam + 15g Vo cafe 10g Cam + 20gVo cafe 20g Cam + 10g Vo cafe 30g Cm
2:1 1:1 1:2 2:1
- Ch tieu theo doi : * Tnh lng bao t / gram che pham sau khi hoan thanh qua trnh len men theo cong thc: Trong o: D = (4000 x a x 103 x 10 n)/b
a : So lng bao t em trong 16 o ln. b : So o con trong 16 o ln ( 256 o con) n : Nong o pha loang cua dung dch bao t.
* Theo doi thi gian song cua che pham nam Trichodema: 1 thang 1 lan, lay che pham lu gi, pha loang 10-5 lan, dung 1ml dung dch cay vao a petri co cha lan moi trng PGA, xem xet s nay mam cua bao t. 3.3.5 anh gia kha nang phong tr benh cua che pham nam Trichodema tren mot so loai cay trong trong nha li, ngoai ong ruong 3.3.5.1 Trong nha li - anh gia kha nang khang cua che pham nam Trichodema vi nam Phytophthora sp., gay benh chet cay tieu trong nha li. So sanh hieu qua cac che pham cua tng dong Trichodema va hieu qua gia cac cach x ly. - Bo tr th nghiem: Th nghiem c bo tr theo kieu hoan toan ngau nhien, gom 8 nghiem thc va 4 lan lap lai. Moi lan lap lai co 5 chau, kch thc chau 20cm x 25cm. Trong 2 day tren 1 chau. Trc khi x ly, moi chau khong che ch con 3 nhanh moi nhanh co 7 la. Tren moi cay trung bnh co 15 ot (long). Cong thc th nghiem theo bang 3.6.
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Bang 3.6 Cong thc bo tr th nghiem anh gia kha nang phong tr benh cua che pham Trichodema tren cay tieu trong nha li Ky hieu T1 T2 T3 T4 T5 T6 T7 T8 Nghiem Thc A1B1 A1B2 A1B3 A2B1 A2B2 A2B3 B01 B02 Cach x ly Dung che pham T.32 (A1) e bon (B1) Dung che pham T.32 (A1) e phun (B1) Dung che pham T.32 (A1) bon va phun (B3) Dung che pham T.41 (A2) e bon (B1) Dung che pham T.41 (A2) e phun (B1) Dung che pham T.41 (A2) bon va phun (B3) C co chung Phytophthora khong co Trichodema C khong chung Phytophthora khong co Trichodema
- Phng phap thc hien: Dung che pham Trichodema thu c phng phap len men e s dung cho th nghiem. Dang bot dung e bon, dang long dung e phun. Ruong th nghiem c lay benh nhan tao bang cach chung nam Phytophthora. Sau khi bon che pham dang ran 2 ngay cac nghiem thc bon va bon + phun tien hanh chung nam gay benh Phytophthora len cay tieu cac nghiem thc (tr nghiem thc oi chng khong chung Phytophthora). Sau chung 5 ngay dung che pham dang long phun cac nghiem thc phun va bon + phun, 7 ngay phun mot lan. Lieu lng bon 5g che pham / chau, lieu lng phun dung che pham dang long co ham lng si nam 100g/lt phun t eu len cay th nghiem. + Cac ch tieu theo doi: - Ty le la benh: em so la benh va so la ieu tra t khi chung Phytophthora en khi benh ngng khong tien trien hoac la rung. Tnh ty le % la benh cac ngay 2, 4, 6, 8 va 41 ngay sau chung Phytophthora. 35
- Theo doi s gia tang ng knh vet benh tren la: Sau khi chung Phytophthora, moi o th nghiem chon 3 la benh, o kch thc vet benh en khi la rung hoac vet benh ngng phat trien. - Ty le day chet: em so day benh va so day ieu tra t khi chung Phytophthora en khi benh ngng khong phat trien hoac day chet hoan toan. Tnh ty le % day benh cac ngay 8, 15, 22, 29 va 41 ngay sau chung Phytophthora - So choi non mi moc: em so choi non mi moc cac ngay 8, 15, 22, 29 va 41 ngay sau chung Phytophthora. - Ty le choi non chet: em so choi non chet va so choi non luc ieu tra. Ghi nhan ket qua cac ngay 8, 15, 22, 29 va 41 ngay sau chung Phytophthora. - Anh hng en re cay: Can trong lng ti toan bo - bo re cua tat ca cac nghiem thc khi ket thuc th nghiem. 3.3.5.2 anh gia kha nang phong tr benh cua nam Trichodema tren cay sau rieng trong giai oan vn m - anh gia kha nang oi khang cua che pham Trichodema dang ran vi nam Phytophthora gay hai tren sau rieng trong giai oan vn m vi cac lieu lng khac nhau. - Bo tr th nghiem: Th nghiem c bo tr theo kieu khoi hoan toan ngau nhien 3 lan lap lai, moi nghiem thc x ly 10 cay. ay la nhng cay chuan b xuat khoi vn m e trong ngoai ong. ng knh canh goc ghep 2,5cm va ng knh canh mat ghep 1,2cm. Moi trng dinh dng trong moi bau cay giong trc khi x ly hon hp che pham gom co: cat xay dng, tro trau, trau muc, s da va phan chuong hoai vi ty le 1:3:3:2:1. ay la cong thc moi trng m cay cua Vien cay an qua Mien Nam. Kch thc bau 15 x 30 cm. Cong thc th nghiem gom co 6 nghiem thc (xem bang 3.7). Tat ca cac nghiem thc eu co chung Phytophthora, trong o co 4 nghiem thc co x ly che
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pham Trichodema, 1 nghiem thc oi chng co phan chuong, khong x ly che pham Trichodema va 1 nghiem thc oi chng khong co phan chuong, khong x ly che pham Trichodema. Phan chuong dung trong th nghiem c mua t Vien cay an qua Mien Nam, c u san dung e m cay. Bang 3.7 Cong thc bo tr th nghiem anh gia kha nang phong tr benh cua che pham Trichodema tren cay sau rieng vn m Nghiem thc T1 T2 T3 T4 C1 C2 Lieu lng / Bau 10g che pham Trichodema + 800g phan chuong hoai muc 20g che pham Trichodema + 800g phan chuong hoai muc 40g che pham Trichodema + 800g phan chuong hoai muc 25g che pham Trichodema + 0g phan chuong hoai muc 0g che pham Trichodema + 800g phan chuong hoai muc 0g che pham Trichodema + 0g phan chuong hoai muc Ch s dung che pham Trichodema Ch s dung phan chuong hoai. Cach x ly Hon hp phan chuong va che pham cho vao gia the cay con giong. Chung nam Phytophthora ngay khi x ly che pham
- Phng phap chung nam: Nam Phytophthora palmivora c thu thap t vn sau rieng b benh, nuoi cay va lam thuan tren moi trng CMA. Nhan nhanh nam tren moi trng CMA, sau 10 ngay nuoi cay tren moi trng CMA, moi bau cay giong chung 4 mau thach co cha nam (1cm2) vao 4 hng phan di ay bau, roi cho vao moi bau cay giong 0,kg hon hp phan va che pham Trichodema nh cong thc th nghiem bang 7. Ti nc cho cay 1 lan/ngay sau
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khi chung nam. Ghi nhan bieu hien cua cay 15, 30, 45 va 60 ngay sau khi chung nam Phytophthora palmivora va x ly che pham Trichodema. Ch tieu theo doi - Ty le chet canh, heo ot: theo doi sau khi x ly en luc canh va ngon heo co dau hieu hoi phuc. - Ty le cay hoi phuc: theo doi t sau khi x ly hon hp en luc canh heo b chet han hoac c hoi phuc hoan toan. - Ty le cay chet: theo doi t sau chung en luc cay chet hoac ngng nhiem benh. 3.3.5.3 anh gia kha nang phong tr benh cua nam Trichodema tren cay sau rieng ngoai ong ruong - anh gia kha nang oi khang cua che pham Trichodema dang ran vi nam Phytophthora gay hai tren cay sau rieng ngoai ong cac lieu lng va cach x ly khac nhau. - Th nghiem thc hien vn Sau rieng kho qua xanh 5 nam tuoi Tam Bnh, Cai Lay, Tien Giang. Tnh trang cay trc ay a c trong tren nen at thap, cay sinh trng kem, la vang, khao sat re thay t re t, re b thoi t. Cac mau at c thu thap e khao sat nam benh eu ghi nhan co s hien dien cua nam Phytophthora bang phng phap bay bao t cua Linderman va Zeitoun (1977) - Bo tr th nghiem: Th nghiem c bo tr theo kieu khoi hoan toan ngau nhien 3 lan lap lai, moi nghiem thc mot cay. Dung che pham nam Trichodema dang bot va phan chuong hoai theo cac cong thc th nghiem (xem bang 3.8). Th nghiem co 6 nghiem thc, trong o co 4 nghiem thc co x ly che pham Trichodema, 1 nghiem thc oi chng ch co phan chuong (C1) va 1 nghiem thc oi chng c x ly theo cach truyen thong cua dan a phng (C2).
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Nguon goc phan chuong hoai mua Vien cay an qua Mien Nam, loai phan dung bon lot. Cach bon theo ranh c ao quanh goc. Bon xong lap at lai. Bang 3.8 Cong thc bo tr th nghiem anh gia kha nang phong tr benh cua che pham Trichodema tren cay sau rieng ngoai ong ruong Stt 1 2 3 4 5 6 Nghiem thc T1 T2 T3 T4 C1 C2 Lieu lng 125g che pham Trichodema + 10kg phan chuong hoai 250g che pham Trichodema + 10kg phan chuong hoai 500g che pham Trichodema + 10kg phan chuong hoai 200g che pham Trichodema + 0kg phan chuong hoai 0g che pham Trichodema + 10kg phan chuong hoai 5kg s da, 1kg NPK 16168, 0,5kg Ure
Trc khi x ly, tien hanh don co, xi nhe vung at di tan, tao mot g at nho rong bang tan cay chung quanh vung re e tranh ra troi khi x ly hon hp che pham. Thu mau at e phan tch nam benh. Moi cay lay 4 mau at (0,5 kg/mau) 4 hng trong vung re non, di lp at mat 5 1cm. Nam c phan lap theo phng phap bay bao t cua Linderman va Zeitoun (1977) Ch tieu theo doi: - Quan sat sc sinh trng cua cay cac thi iem 15, 30, 45 va 60 ngay sau khi x ly e anh gia kha nang phuc hoi cua cay sau khi c x ly hon hp tren. - Ham lng diep luc to trong la: c the hien qua ch so SPAD (Soil Plant Analysis Development). S dung may o ham lng diep luc to tren la: SPAD 502 cua hang Minolta Nhat Ban. Phng phap o ham lng diep luc: Chon cac la banh te (co o tuoi ong eu nhau), c phat trien ra sau khi x ly hon hp
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che pham e o. Moi cay o 10 la, tnh ch so trung bnh cua 10 la cho mot lan lap lai. - Kch thc la: c the hien qua ch so dien tch la (LAI). Tien hanh o chieu dai la va chieu rong la khi la at c kch thc toi a, moi cay o 10 la, tnh tr so trung bnh cua 10 la. - Tan suat xuat hien cua nam Phytophthora: Sau khi c x ly hon hp che pham, moi cay lay 4 mau at 4 hng cua cay. Tnh ty le % mau at thu c co xuat hien nam Phytophthora. 3.3.6 Phng phap anh gia ket qua So lieu thu thap c t cac th nghiem eu c chuyen oi thanh so lieu thong ke tng ng va bieu th bang X SE. anh gia cac ket qua bang phan tch ANOVA, ANOCOVA, trac nghiem LSD, trac nghiem Duncan (Gomez, 1984). S dung phan mem thong ke Statgraphic 7.0 e x ly.
Chng 4 KET QUA VA THAO LUAN

4.1 Ket qua 40
4.1.1 Thu thap va phan lap nam Trichodema Viec tien hanh lay mau c thc hien tren cac loai cay trong khac nhau nhieu vung sinh thai khac nhau. Chung toi chon iem lay mau vung co lch s nhiem benh, ruong nhiem benh va cay trong co benh va phuc hoi. Tren c s cac mau benh lay t at va vet benh ngng phat trien, a kho. ong thi, e khao sat them kha nang oi khang cua dong nam Trichodema nhng vung sinh thai khac biet vi ieu kien moi trng Viet Nam, chung toi phan lap them mot dong Trichodema t che pham th nghiem t An o. ay la che pham cua Cong ty Sun Agro e ngh chung toi th hieu lc Viet Nam. Tong cong thu c 14 dong a vao th nghiem khao sat tnh oi khang. Quan sat di knh hien vi, da vao mot so ac iem ve hnh thai mau sac, mui v tren moi trng chon loc, hnh dang canh bao t, bao t, va si nam e nh danh chung. Bc au nh danh da tren c s phan loai cua Giao s Gary J.Samuels (2003). Trong cac mau Trichodema phan lap c chung toi xac nh c 13 dong. Dong Trichodema thu thap c tren cay ca chua Cu Chi chung toi cha nh danh c. Ngoai ra, trong cac dong thu thap c co 1 dong tren sau rieng ong Nai chung toi xac nh ay la dong T. sinensis, cha c khuyen cao s dung trong phong tr sinh hoc. V vay dong nay co the khao sat tnh khang trong phong th nghiem, khong nen e phat tan ra ngoai ong ruong. Ket qua nh danh cua cac dong phan lap c trnh bay trong bang 4.9. Bang 4.9 Ket qua nh danh cua cac dong Trichodema phan lap c STT 1 2 3 4 Ma T.14 T.15 T.16 T.17 a iem Trng H NL Tp.HCM Trng H NL Tp.HCM Trng H NL Tp.HCM Trng H NL Tp.HCM 41 nh danh T.harzianum T. asperellum T. asperellum T. sinensis
5 6 7 8 9 10 11 12 13 14
T.29 T.30 T.31 T.32 T.33 T.34 T.35 T.36 T.37 T.41
Lam ong An o Can Th Bnh Phc Kien Giang Tay Ninh Bnh Phc Cu Chi ong Nai Trng H NL Tp.HCM
T. asperellum T.harzianum T. asperellum T. viride T. asperellum T. asperellum T. asperellum Cha nh danh c T. sinensis T.harzianum
4.1.2 anh gia kha nang oi khang cua Trichodema vi 4 loai nam Fusarium, Sclerotium, Phytophthora va Rhizoctonia Chung toi dung 14 dong nam Trichodema a phan lap c e th nghiem tnh oi khang vi 4 loai nam trong at gay hai trong at: Phizoctonia Solanni, Sclerotium rolfsii, Phytophthora va Fusarium. Th nghiem c thc hien ong loat tren a petri vi moi trng bot bap co 2% ng Dextrose va ieu kien th nghiem ong nhat. Quan sat s oi khang cua Trichodema vi tng loai nam tren sau 7 ngay nuoi cay. Bang 4.10 Kha nang oi khang cua Trichodema vi mot so loai nam gay hai cay trong trong at STT 1 2 3 Ma T.14 T.15 T.16 Kha nang oi khang cua Trichodema vi cac nam Phytophthora Fusarium +++ +++ +++ 42 Rhizoctonia +++ +++ + Sclerotium -
4 5 6 7 8 9 10 11 12 13 14
T.17 T.29 T.30 T.31 T.32 T.33 T.34 T.35 T.36 T.37 T.41
+++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++
+ + + +
+++ +++ +++ + +++ +++ ++ +++ + +++ +++
+++
* Ky hieu: khang manh (+++), khang trung bnh (++), khang yeu (+) va khong khang (-). Qua ket qua thu c nh bang 4.10, cung nh quan sat trc tiep tren a petri chung toi nhan thay tnh khang nam co s khac biet gia cac dong Trichodema ang c th nghiem (xem hnh 4.2 ; 4.3 ; 4.4 va 4.5). ac biet, s khac biet the hien ro hn khi so sanh tnh khang oi vi tng loai nam gay hai khac nhau.
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a c
d Hnh 4.2: Kha nang oi khang cua mot so dong nam Trichodema vi cac loai nam Fusarium, Rhizoctonia, Sclerotium va Phytophthora ( t trai sang phai) a. Dong Trichodema so 29 (T29) b. Dong Trichodema so 30 (T30) c. Dong Trichodema so 32 (T32) d. Dong Trichodema so 41 (T41)
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c Hnh 4.3: Kha nang oi khang cua mot so dong nam Trichodema vi nam gay benh Phytophthora. a. Dong Trichodema so 16 (T16): Khang trung bnh (++) b. Dong Trichodema so 33 (T33): Khang trung bnh (++) c. Dong Trichodema so 36 (T36): Khang manh (+++) d. Dong Trichodema so 47 (T41): Khang manh (+++)
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d Hnh 4.4: Kha nang oi khang cua mot so dong nam Trichodema vi nam gay benh Sclerotium a. Dong Trichodema so 36 (T36): Khong khang (-) b. Dong Trichodema so 30 (T33): Khong khang (-) c. Dong Trichodema so 29 (T36): Khang yeu (-) d. Dong Trichodema so 41 (T41): Khang manh (+++)
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So sanh gia cac dong nam Trichodema khac nhau, chung toi thay rang ch co dong T.41 la co kha nang oi khang vi ca 4 loai nam gay benh trong th nghiem (xem hnh 4.2d). Mot so khang yeu vi Fusarium. Hau het eu khang vi Rhizoctonia, tuy nhien, mc o khang khac nhau gia cac dong. ay la ieu kien e la chon nhng dong cu the s dung tiep tuc cho viec nghien cu. ieu nay con noi len tnh chuyen biet cua nam Trichodema trong viec oi khang vi tng loai nam gay hai cu the. Mot dong nam co kha nang oi khang cao mot loai benh, mot cay trong hay mot vung sinh thai nay cha han a co hieu qua khi s dung cho ieu kien khac. ay la mot han che cua viec s dung san pham sinh hoc so vi cac san pham thuoc hoa hoc khac, pho rong hn. V vay, muon nang cao hieu qua cua che pham sinh hoc, ma cu the la che pham Trichodema can khac phuc nhc iem nay. Tc la can phai tuyen chon nhng nguon nam tren tng ieu kien cu the e ap dung ngay chnh trong ieu kien o. oi vi nam Fusarium, ch co 4 trong 14 dong nam Trichodema co kha nang khang yeu con lai hau nh khong khang. Tuy nhien, theo Godwin (2002) cho rang kha nang oi khang cua nam Trichodema vi nam Fusarium thay oi tuy theo nguon dinh dng co trong moi trng. oi vi nam Sclerotium hau nh ch co nam Trichodema dong 41 co kha nang khang manh con cac dong khac eu khang yeu va khong khang (xem hnh 4.4d) oi vi nam Phytophthora, hau het cac dong nam Trichodema eu co kha nang khang cao vi loai nam nay (xem hnh 4.3). ieu nay cho thay kha nang oi khang cua nam Trichodema vi nam gay benh do Phytophthora rat ln. V vay, trong ieu kien viec s dung che pham nam Trichodema con cha pho bien va cha thuyet phuc ngi nong dan thay oi tap tuc s dung thuoc. Nen a che pham vi khuyen cao tr benh do nam Phytophthora gay hai tren mot so cay trong chnh, co gia tr kinh te cao va ang b benh gay hai rat nghiem trong, gay 47
hoang mang cho nha vn ma ngi nong dan goi vi nhng ngon t nh la tieu ien hay sau rieng t t. 4.1.3 Ket qua nhan sinh khoi nam Trichodema 4.1.3.1 Ket qua nhan sinh khoi Trichodema dang long Qua th nghiem khao sat tnh oi khang cua nam Trichodema, chung toi nhan thay dong T.41 la u viec hn ca. No co kha nang khang vi ca 4 loai nam a ra khao sat. V vay chung toi quyet nh chon dong nay a vao th nghiem nhan sinh khoi. Tuy nhien, theo Godwin (2002), kha nang oi khang cua nam Trichodema con phu thuoc vao ieu kien moi trng no ton tai. V vay chung toi chon them 1 dong trong so nhng dong nam Trichodema co kha nang khang cao e khao sat them viec nhan sinh khoi. Do d kien cua chung toi se khao sat kha nang oi cua nam Trichodema tren cay tieu va sau rieng. V vay, chung toi chon them dong thu phan lap c vn tieu cua tnh Bnh Phc, co ky hieu T.32 e a vao th nghiem nhan sinh khoi. Th nghiem nhan sinh khoi dang long, chung toi s dung moi trng chu yeu la gia the ng glucose va gia au xanh. Trong moi trng chung toi cho them Ure, KH2PO4 va MgSO4. ay la nhng loai khoang thiet yeu cho viec phat trien cua te bao nam cung nh giup cho viec am bao o pH thch hp cho nam phat trien. Nhng loai khoang nay ch can mot lng nho, nen anh hng cua s thay oi lieu lng cac chat nay len khoi lng khuan ty thu c la khong ln. V vay, trong moi trng d kien khao sat ch con 2 yeu to hu c la ng va gia can phai xac nh c lieu lng thch hp. Chung toi lan lt co nh mot yeu to e khao sat s thay oi cua yeu to kia. Th nghiem, cua chung toi tien hanh vi 2 dong nam Trichodema. V vay, chung toi bo tr th nghiem dang ngau nhien a yeu to vi 2 dong, 5 nghiem thc va 4 lan lap lai. Tong cong co 40
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bnh tam giac cho mot th nghiem. Th nghiem c thc hien trong ieu kien hoan toan giong nhau. 4.1.3.1.1 Khao sat yeu to ng thay oi, co nh lng gia Trc tien chung toi so sanh gia cac nghiem thc khac nhau xem co s khac biet nh the nao? Phan tch ket qua bang ANOVA va qua so lieu phan tch, so sanh, nhan thay rang lng ng trong thanh phan moi trng tang len cang tang lng khuan ty thu c, sau o giam, nghiem thc co 30g ng trong 1 lt moi trng (P.4) co gia tr cao nhat. Tuy nhien, viec tang khoi lng khuan ty nhng nghiem thc co lng ng ln khong ang ke va khong co y ngha thong ke. V vay, co the s dung lng ng nghiem thc co 20g ng trong 1 lt moi trng (P.3) e s dung trong cac tr nghiem tiep theo va san xuat che pham dang long. Bang 4.11 So sanh khoi lng khuan ty thu c cac nghiem thc moi trng co lng ng khac nhau trong th nghiem nhan sinh khoi Trichodema dang long Nghiem thc 0g ng + 100g gia 10g ng + 100g gia 40g ng + 100g gia 20g ng + 100g gia 30g ng + 100g gia Khoi lng khuan ty TB (g) So sanh 0,4 0,1 1,7 0,1 2,3 0,1 2,3 0,1 2,5 0,1
x x x x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan.
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4.1.3.1.2 Khao sat yeu to gia thay oi, co nh lng ng -Tren c s th nghiem khao sat lng ng thu c. Chung toi thay oi lng gia trong moi trng th nghiem. Tien hanh nh th nghiem khao sat ham lng ng. Qua ket qua th nghiem bang 4.12 cho thay s khac biet cua cac nghiem thc vi lng gia khac nhau rat co y ngha ve phng dien thong ke. Lng gia cang ln thu c khoi lng khuan ty cang ln. Tuy nhien, xet ve yeu to hieu qua kinh te, chung ta co the chon nghiem thc G.3, 100g gia song cho 1 lt moi trng hn la chon G.5, 200g gia song cho 1 lt moi trng. Bang 4.12 So sanh khoi lng khuan ty thu c cac nghiem thc moi trng co lng gia khac nhau trong th nghiem nhan sinh khoi Trichodema dang long Nghiem thc 0g gia + 20g ng 50g gia + 20g ng 150g gia + 20g ng 100g gia + 20g ng 200g gia + 20g ng Khoi lng khuan ty TB (g) So sanh 0.41 0,05 1.23 0,07 1.41 0,04 1.43 0,03 1.59 0,05
x x x x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. + So sanh 2 dong nam Trichodema dung trong th nghiem nhan sinh khoi long. Tren c s khoi lng khuan ty thu c 2 th nghiem nhan sinh khoi long tren, lay nghiem thc chnh la 2 dong nam Trichodema T.41 va T.32 trong ieu kien thay oi lng ng, lng gia va lap lai. Phan tch ANOCOVA, ket qua thu c theo bang 4.13.
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Qua ket qua phan tch so lieu bang 4.13, ta thay rang th nghiem khao sat lng ng s khac biet ve khoi lng khuan ty gia 2 dong nam Trichodema thu c la rat khac nhau va co y ngha ln ve phan tch thong ke. Dong T.32 cho ket qua cao hn dong T.41 rat nhieu. Tuy nhien, ieu o lai khong lap lai th nghiem khao sat lng gia. Chng to moi trng khac nhau va dong nam khac nhau s phat trien cua chung cung khac nhau. V vay, nen tiep tuc s dung ca 2 dong nam nay trong cac th nghiem tiep theo. Bang 4.13 So sanh khoi lng khuan ty thu c cua 2 dong T.41 va T.32 trong cac th nghiem nhan sinh khoi Trichodema dang long Nghiem thc T.41 T.32 Th nghiem ng KL TB (g) 1,65 0,11 2,04 0,11 So sanh
x x
Th nghiem gia KL TB (g) 1,21 0,06 1.21 0,06 So sanh
* Tren cung mot cot theo phng thang dng th s khac biet mc = 0,05 theo trac nghiem Duncan. 4.1.3.1.3 Trac nghiem che pham dang long Qua ket qua thu c t 2 th nghiem khao sat lng ng va lng gia, chung toi chon thanh phan moi trng la 20g ng gluco, 100g gia song, 1g Ure, 1g KH2PO4 va 0.5g MgSO4 e khao sat them kha nang ton tai cua che pham sau khi len men cua 2 dong nam Trichodema. Th nghiem c bo tr 2 nghiem thc (2 dong) vi 10 lan lap lai trong 10 bnh tam giac. Sau khi len men xong, chung toi quan sat hang ngay va nhan thay sau thi gian 1 tuan che pham bat au chuyen mau. Phan di dung dch co mau vang sam. Tren mat ong vang, xuat hien mau xanh, mau ac trng cua nam Trichodema phat trien. Sau o tren mat chuyen mau sam dan. Sau 2 thang chuyen mau nau en, ay la hien tng Trichodema phan huy (xem hnh 4.5b ; 4.5c). 51
b Hnh 4.5: Nhan sinh khoi, san xuat che pham Trichodema dang long a. S dung may lac len men chm san xuat che pham Trichodema b. Che pham dang long cua dong Trichodema so 32 (T.32) ngay sau khi len men
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c. Che pham dang long cua dong Trichodema so 32 (T.41) 60 ngay sau khi len men
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4.1.3.2 Ket qua nhan sinh khoi Trichodema dang ran Th nghiem c tien hanh ong thi viec len men dang long vi 2 dong Trichodema T.41 va T.32 c chon. Dung cac gia the ran, xop nh cam, trau, vo ca phe va cm nguoi. Vi ty le gia the khac nhau to hp thanh 7 nghiem thc. Trong ieu kien th nghiem nh nhau. Th nghiem c bo tr theo khoi ngau nhien a yeu to, vi 4 lan lap lai. Sau khi len men xong, em lng bao t bang buong em hong cau Thomas vi n v tnh la bao t /ml che pham. Lu lai tat ca cac mau len men e theo doi kha nang nay mam cua bao t trong thi gian lu tr (xem hnh 4.7). Ket qua thu c sau khi len men, phan tch, c ket qua qua bang 4.14. Bang 4.14 So sanh so lng bao t thu c cac nghiem thc co c chat khac nhau trong th nghiem nhan sinh khoi Trichodema dang ran Nghiem thc 30g cm 10g cam + 20g vo ca phe 15g cam + 15g vo ca phe 20g cam + 10g vo ca phe 20g cam + 10g trau 15g cam + 15g trau 10g cam + 20g trau So lng bao t / ml x 108 1,12 0,03 2,6 0,4 4,9 0,6 6,9 0,5 18,5 0,7 29,0 5,0 33,5 5,8 So sanh
x x x x x x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Qua bang 4.14, chung ta thay rang cac loai gia the dung e len men khac nhau anh hng rat ln en so lng bao t tren mot n v khoi lng. Nghiem thc 30g cm (N.7) co gia tr nho nhat. ay la nghiem thc ma gia the len men la cm, be mat thong thoang t, dan en s phat trien cua nam Trichodema kem, 53
kha nang tao bao t thap. Cac nghiem thc gia the len men la hon hp cam va vo ca phe (N.4, N.5 va N.6), cho lng bao t mc o trung bnh va gan nh nhau cac ty le, khong co s khac biet ve phng dien thong ke. Cac nghiem thc cam va trau cho ket qua cao nhat (N.1 , N.2 va N.3). ong thi co s khac biet ang ke gia cac ty le. Nghiem thc 10g cam + 20g trau (N.2) co gia tr cao nhat nhng khong khac biet lam so vi nghiem thc 15g cam + 15g trau (N.1). Hai nghiem thc nay khac biet co y ngha vi nghiem thc 20g cam + 10g trau (N.3). Chng to cung trong thanh phan cam trau nhng trau hi nhch hn mot chut cho ket qua tot hn. Bang 4.15 So sanh so lng bao t thu c gia cac dong T.32 va T.41 trong th nghiem nhan sinh khoi Trichodema dang ran Nghiem thc T.41 T.32 So lng bao t /ml x 108 10,0 1,6 17,6 1,6 So sanh
x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Qua ket qua bang 4.15, so lieu thong ke cho thay co s khac biet rat y ngha ve so lng bao t thu c sau khi nhan sinh khoi gia 2 dong Trichodema T.32 va T.41. ay la c s cho viec la chon dong Trichodema nhan sinh khoi s dung trong cac th nghiem tiep theo va ngoai ong ruong. Ben canh so sanh ve so lng bao t thu c tren mot n v khoi lng sau khi len men, viec xem xet thi gian ton tr che pham rat quan trong. Lng bao t ln cha han a phat huy tot kha nang sinh trng cua nam ma cai chnh la lieu bao t o co kha nang nay mam hay khong sau mot thi gian lu tr. Va thi gian lu gi che pham len men c bao lau?
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a c
Hnh 4.6: Nhan sinh khoi, san xuat che pham Trichoderma dang ran a. Che pham Trichoderma dang ran sau 7 ngay len men tren moi trng cam va trau. b. Che pham Trichoderma dang ran sau 7 ngay len men tren moi trng cm. c. Che pham Trichoderma dang ran c lu tr 9 thang sau khi len men cac nghiem thc th nghiem. 55
Chung toi tien hanh viec trac nghiem kha nang nay mam cua tat ca cac mau che pham trong th nghiem tren bang phng phap pha loang nhieu lan roi cho vao a petri e xac nh chung con co kha nang nay mam hay khong. Trac nghiem thc hien 1 lan / thang. Ky hieu con nay mam (+) khong nay mam (-). Ket qua c the hien qua bang 4.16. Bang 4.16 Trac nghiem s nay mam cua cac che pham nam Trichodema theo phng phap nhan sinh khoi dang ran Nghiem thc 15g cam + 15g trau 10g cam + 20g trau 20g cam + 10g trau 15g cam + 15g vo ca phe 10g cam + 20g vo ca phe 20g cam + 10g vo ca phe 30g cm Thi gian sau len men (Thang) 1 + + + + + + + 2 + + + + + + + 3 + + + + + + + 4 + + + + + + + 5 + + + + + + + 6 + + + + + + 7 + + + + + 8 + 9 -
Qua cac th nghiem va trac nghiem len men tao che pham dang long va dang xop, ve s khac biet gia cac nghiem thc, chung toi chon c cac phng phap len men phu hp, lam vat lieu cho cac th nghiem tiep theo. Tuy nhien, so lng bao t hoac khoi lng khuan ty ln cha u ma can phai co cach x ly hp ly. ong thi, s khac biet gi 2 dong Trichodema kha ro th nghiem nhan sinh khoi dang ran nhng khong ro lam th nghiem dang long. V vay, chung toi tiep tuc khao sat tiep 2 dong nay nhng th nghiem tiep theo trong nha li cung nh ngoai ong ruong.
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4.1.4 Khao sat hieu lc cua che pham Trichodema tr benh do nam Phytophthora gay hai tren cay tieu trong ieu kien nha li Trong th nghiem nay, ngoai viec khao sat hieu lc cua 2 dong che pham Trichodema thu c t viec nhan sinh khoi dang long va dang ran, chung toi khao sat them hieu qua cua cac cach x ly khac nhau. Th nghiem c tien hanh trong ieu kien nha li vi nhng nghiem thc x ly Trichodema c chung nam Phytophthora gay benh (xem hnh 4.7b va 4.7c), oi chng la nghiem thc co chung Phytophthora khong x ly Trichodema va oi chng khong chung Phytophthora khong x ly Trichodema (xem hnh 4.8). Nh vay, to hp cac yeu to th nghiem ta co 2 kieu bo tr th nghiem. So sanh gia cac nghiem thc vi oi chng la th nghiem n yeu to, anh gia ket qua bang phan tch ANOVA. So sanh gia cac cach x ly bon, phun va bon ket hp phun cung nh so sanh hieu qua che pham cua 2 dong Trichodema la th nghiem a yeu to, anh gia ket qua bang phan tch ANOCOVA. Sau khi tien hanh th nghiem, nhn tong the toan bo th nghiem, chung toi nhan thay co s khac biet rat ang ke gia cac lo th nghiem (xem hnh 4.9) va tren tng cay tieu cua tng nghiem thc (xem hnh 4.10.1 va 4.10.2). Ket qua cua tng ch tieu theo doi nh sau: + Ty le la benh: Sau khi chung Phytophthora, chung toi theo doi, em so la benh va so la ieu tra. Quan sat ruong th nghiem, chung toi nhan thay 1 ngay sau khi chung a co trieu chng la b nhiem benh. Benh tren la tien trien rat nhanh trong vong 1 tuan, sau o gan nh dng lai. Quan sat vet benh tren la chung toi thay trieu chng gay hai do Phytophthora tren cay la tieu rat a dang (xem hnh 4.11). Co vet benh au ngon la, lam la rung hoac con mot phan tren cay. Co vet benh cuong la, lam la rung khi con xanh. Co vet benh gay hai mep la, la hoac toan bo phien la. ieu tra 2, 4, 6, 8 va 41 ngay sau khi chung Phytophthora, ket qua nh sau:
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Hnh 4.7: Hnh dang bao t nam Phytophthora c dung trong th nghiem a. Sporangia cua nam Phytophthora palmivora tren moi trng PGA sau 7 ngay nuoi cay (40X) dung nhiem benh tren sau rieng. b. Sporangia cua nam Phytophthora sp. tren moi trng PGA sau 7 ngay nuoi cay (40X) dung nhiem benh tren cay tieu c. Sporangia cua nam Phytophthora sp. tren moi trng PGA sau 7 ngay nuoi cay (40X) dung nhiem benh tren cay tieu phong ln 58
b Hnh 4.8: Vn tieu chuan b th nghiem. a. Toan canh vn tieu luc mi sang chau. b. Phan o bo tr theo tng nghiem thc.
c. Trc khi chung Phytophthora va x ly Trichoderma.
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e g h
Hnh 4.9: Cac nghiem thc tieu 60 ngay sau x ly theo tng o th nghiem a. C khong x ly b. C co Phytophthora c. T.32 bon d. T.41 bon e. T.32 phun f. T.41 phun g. T.32 bon + phun h. T.41 bon + phun
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Hnh 4.10.1: Hnh dang cay tieu 60 ngay sau x ly theo tng cong thc cua th nghiem phong tr benh tren cay tieu a. Tong the cac cay cua 8 nghiem thc bo tr th nghiem b. Cay tieu cua nghiem thc oi chng khong chung Phytophthora c. Cay tieu cua nghiem thc oi chng co chung Phytophthora
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Hnh 4.10.2: Hnh dang cay tieu 60 ngay sau x ly theo tng cong thc cua th nghiem phong tr benh tren cay tieu a. Cay tieu cua nghiem thc phun che pham T.41 b. Cay tieu cua nghiem thc bon che pham T.41 c. Cay tieu cua nghiem thc bon + phun che pham T.41 d. Cay tieu cua nghiem thc phun che pham T.32 e. Cay tieu cua nghiem thc bon che pham T.32 f. Cay tieu cua nghiem thc bon + phun che pham T.32
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Hnh 4.11: Trieu chng benh do nam Phytophthora gay hai tren la tieu a. Cac dang la b benh do nam Phytophthora va la khong benh b. Benh gia la e. Benh au la c. Benh phien la f. benh 2 mep la d. Benh cuong la g. la khong b benh
63
- So sanh gia cac nghiem thc vi nhau va vi oi chng: Ket qua phan tch thong ke cho thay 2, 4 ngay sau chung s khac biet khong ro rang. Bat au t ngay th 6 sau chung, co s khac biet ro gia cac nghiem thc co x ly Trichodema va cac nghiem thc oi chng co chung Phytophthora. S khac biet gia cac nghiem thc nay duy tr cho en khi chung toi ieu tra lan cuoi vao thi gian 41 ngay sau chung Phytophthora (xem bang 4.17). Bang 4.17 So sanh ty le la tieu b benh gia cac nghiem thc qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu 2 NSC Nghiem thc Ty le la benh T.41 bon + phun T.41 bon T.32 bon + phun T.32 bon T.32 phun T.41 phun C co chung P. So sanh 6 NSC Ty le la benh So sanh 41 NSC Ty le la benh 18,3 2,6 x 21,0 2,8 x x 30,5 3,5 x x x 32,6 2,5 33,3 6,0 37,2 6,3
x 89,2 5,3 x x x x x x
So sanh
12,8 1,3 x 15,7 3,0 x x 17,1 3,1 x x 21,2 1,6 24,0 1,4 21,0 2,5 24,0 1,7
x x x x x x
17,9 2,3 x 18,8 2,6 x 23,1 3,5 x x 28,3 3,4 x x 30,5 4,4 30,5 5,8 46,4 3,1
x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Trng hp nghiem thc BO2, khong chung Phytophthora va khong x ly Trichodema (T8), chung toi khong a vao bang phan tch do o la nghiem thc oi chng khong chung Phytophthora, trong ieu kien nha li luc bo tr th nghiem khong thay xuat hien benh. - So sanh gia cac cach x ly: tat ca cac giai oan ieu tra eu thay co s khac biet. Ty le benh thap nhat nghiem thc va bon va phun, cao nhat 64
nghiem thc phun. Tuy nhien, theo ket qua x ly thong ke, vi 3 cach x ly va 8 lan lap lai, s khac biet khong co y ngha (xem bang 4.18). Bang 4.18 So sanh ty le la tieu b benh gia cac cach x ly Trichodema qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu 2 NSC Nghiem thc Bon + phun Bon Phun Ty le la benh So Sanh 4 NSC Ty le la benh So sanh 41 NSC Ty le la benh So sanh
15,0 1,7 x 18,5 1,7 x x 22,5 1,7

x
16,8 2,4 x 21,1 2,4 x x 27,3 2,4

x
24,4 2,9 x 26,8 2,9 x x 35,2 2,9

x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. - So sanh gia cac dong che pham Trichodema: Theo doi tat ca cac giai oan ieu tra eu thay co s khac biet. Ty le benh nhng nghiem thc x ly che pham t dong nam T.41 co ty le benh thap hn nhng nghiem thc x ly che pham t dong nam T.32. Tuy nhien, theo ket qua x ly thong ke, vi s khac biet ch co y ngha lan ieu tra cuoi cung, tc 41 ngay sau khi lay nhiem benh (xem bang 4.19). Bang 4.19 So sanh ty le la tieu b benh gia 2 dong T.32 va T.41 qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu 2 NSC Nghiem thc T.41 T.32 Ty le la benh 17,5 1,6 19,8 1,6 8 NSC Ty le la benh 22,38 27,30 41 NSC Ty le La benh 33,4 2,7 24,2 2,7 x
x
So sanh
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. 65
+ S gia tang vet benh tren la: Sau khi chung Phytophthora, moi o th nghiem chung toi anh dau 3 la benh, quan sat trieu chng va o kch thc vet benh. cac ngay 2, 4, 6 vet benh tat ca cac nghiem thc eu gia tang sau o co trieu chng sap rung. en ngay th 8 sau khi chung Phytophthora co en 9/21 la quan sat b rung. Kch thc vet benh cac la cha rung cung rat ln, t 3-4 cm, tc hn dien tch la. Mac du vet benh tang rat nhanh trong nhng ngay au sau khi chung Phytophthora, nhng qua phan tch thong ke, s khac biet gia cac nghiem thc la khong ln (xem bang 4.20). Cung nh s khac biet khong co y ngha thong ke khi so sanh gia 2 dong Trichodema dung th nghiem va cach x ly che pham nh the nao. Bang 4.20 So sanh s gia tang vet benh tren la tieu gia cac nghiem thc qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu (mm) Nghiem thc T.32-bon + phun T.41-bon T.32 bon C co chung P. T.41-bon + phun T.41-phun T.32-phun 2 NSC 6,7 2,0 8,0 0,6 10,7 2,3 11,7 0,9 14,3 2,8 14,7 6,8 15,3 4,0 4 NSC 19,0 1,7 21,0 1,0 22,7 2,0 24,3 2,0 27,3 2,4 27,7 6,3 26,7 3,0 6 NSC 27,3 2,0 30,0 0,6 29,7 1,5 33,7 1,8 34,3 2,5 36,0 6,1 36,0 3,2
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. + Ty le day chet: quan sat ruong th nghiem, chung toi nhan thay 8 ngay sau khi chung Phytophthora hau het cac nghiem thc co xuat hien day chet. Ch co nghiem thc T.41-bon + phun la cha co hien tng chet. nghiem thc oi 66
chng co chung Phytophthora khong x ly Trichodema (T7) co gan 1/3 day chet. Qua 15 ngay, tat ca cac nghiem thc eu xuat hien day chet, ty le day chet tang len so vi luc 8 ngay. Ca biet, C co chung Phytophthora co hn 50% day chet. T 15 en 34 ngay sau chung Phytophthora, hau het cac nghiem thc co x ly Trichodema day khong b chet them va sau o co bien ong nhe. Rieng T7, ty le day chet tang dan va at hn 80% 41 ngay sau x ly. So sanh gia cac nghiem thc, chung toi thay khong co s khac lam gia nhng nghiem thc co x ly Trichodema ma eu khac biet rat ro vi nghiem thc ch chung Phytophthora. ieu o c the hien rat ro qua ket qua phan tch thong ke. Ket qua phan tch mot so thi iem co bien ong ln c the hien qua bang 4.21. Bang 4.21 So sanh ty le day tieu chet gia cac nghiem thc qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu 8 NSC Nghiem thc T.41-bon + phun T.32-bon T.41-bon T.41-phun T.32-phun T.32-bon + phun C co chung P. Ty le day chet So sanh 15 NSC Ty le day chet So sanh 41 NSC Ty le day chet So sanh
0,0 0,0 x 3,3 2,0 x 3,3 2,0 x 3,3 2,0 x 5,0 1,7 x 8,3 5,0 x 35,0 9,2
X
13,3 4,7 x 11,7 3,2 x 5,0 1,7 x 13,3 4,7 x 10,0 3,3 x 10,0 4,3 x 51,7 10,3
x
15,8 4,2 x 18,3 1,7 x 16,7 5,8 x 13,3 4,7 x 31,7 12,0 x 15,0 7,4 x 83,2 7,0
x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. So sanh gia cac cach x ly, chung toi nhan thay t thi gian au sau khi chung Phytophthora nghiem thc bon co ty le day chet thap nhat. Sau 29 ngay
67
chung Phytophthora nghiem thc va bon va phun co ty le day chet thap hn. Tuy nhien qua phan tch thong ke, s khac biet nay khong co y ngha. So sanh gia cac dong Trichodema, qua ket qua phan tch thong ke, s khac biet ve ty le day chet gia 2 dong khong co y ngha. Ve trieu chng day chet ca t ngon, goc va gia than (hnh 4.12, 4.13) + So choi non mi moc: Quan sat trong suot qua trnh theo doi, chung toi thay cac nghiem thc T.32-bon, T.32-phun, T.32-bon + phun co s phat trien choi non nhieu hn so vi cac nghiem thc con lai. thi gian 8 ngay sau khi chung Phytophthora, cac nghiem thc T.41-bon, T.41-phun, T.41-bon + phun cha co s khac biet vi oi chng. Nhng ngay sau quan sat bat au thay bat au co khac biet nhng khong nhieu lam (xem bang 4.22). Bang 4.22 So sanh so choi mi moc gia cac nghiem thc qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu Nghiem thc C co chung P. T.41-bon T.41-phun T.41-bon + phun T.32-bon T.32-bon + phun T.32-phun 8 NSC So choi SS 13,8 1,5 x 17,3 1,3 x 17,8 0,8 x 18,3 2,4 x 30,3 5,2 32,0 6,5 32,3 1,7 15 NSC So choi 17,5 1,3 x 20,5 0,9 x 25,0 1,4 x x 24,0 2,4 x x
X 33,8 4,4 X 36,0 7,3 X 42,3 2,9
41 NSC SS So choi 23,5 1,6 x 28,0 2,5 x x 29,5 1,3 x x 33,8 5,0 x x x
x x x x x x
SS
x x 39,5 5,2 x 43,3 5,8 x 50,3 4,3
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. So sanh so choi non mi moc gia cac cach x ly khac nhau, phun, bon va va phun va bon, trong suot qua trnh theo doi chung toi nhan thay khong co 68
a.
Hnh 4.12: Trieu chng benh do nam Phytophthora gay hai tren ngon tieu a. Cac dang trieu chng chet ngon sau khi chung nam Phytophthora va ngon cay tieu khoe b. Ngon cay tieu trc khi chung Phytophthora c. Ngon cay tieu b benh 4 ngay sau khi chung Phytophthora d. Ngon cay tieu b benh 30 ngay sau khi chung Phytophthora 69
Hnh 4.13: Trieu chng benh do nam Phytophthora gay hai tren day tieu a. Day tieu chet phan goc khi chung nam Phytophthora b. Day tieu chet phan gia khi chung nam Phytophthora c. Day tieu chet phan ngon khi chung nam Phytophthora d. Day tieu khoe khong b nhiem benh 70
khac biet ln. Qua phan tch thong ke, s khac biet hoan toan khong co y ngha gia cac cach x ly che pham Trichodema. So sanh gia cac dong che pham Trichodema, qua bang 4.23, chung toi nhan thay co s khac biet kha ln gia cac nghiem thc. Bang 4.23 So sanh so choi mi moc gia 2 dong T.32 va T.41 qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu Nghiem thc T.41 T.32 8 NSC So choi 17,8 2,0 31,5 2,0
x x
15 NSC SS So choi 23,16 2,1 37,33 2,1

x x
22 NSC SS So choi SS 29,1 2,3 41,8 2,3

x x
41 NSC So choi SS 30,4 2,5 44,3 2,5

x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. + Ty le choi non chet: Quan sat s sinh trng cua choi non cung vi s lay nhiem benh cua chung, chung toi thay a so cac choi non nghiem thc oi chng co chung Phytophthora eu b nhiem va chet gan het vao cuoi t ieu tra (xem hnh 4.14). Trong khi o cac nghiem thc co x ly Trichodema ch dng lai mc khoang 15% so choi non b nhiem benh. Theo ket qua phan tch thong ke, s khac biet gia cac nghiem thc co va khong x ly Trichodema ch thc s co y ngha 29 ngay sau chung Phytophthora. Ty le choi chet mot so giai oan ieu tra the hien qua bang 4.24. So sanh gi cac cach x ly Trichodema, chung toi thay co khac biet trong thi gian au, sau o s khac biet nay t bieu hien hn. Qua x ly thong ke, bang ket qua cho thay hau het s khac biet nay eu khong co y ngha. Theo doi gia cac nghiem thc x ly nhng dongTrichodema khac nhau, ong thi vi viec gia tang choi mi khac nhau, ty le cac choi chet cung khac nhau rat ro trong gan ca qua trnh ieu tra. Ch lan ieu tra cuoi cung s khac 71
Hnh 4.14: Trieu chng benh do nam Phytophthora gay hai choi tieu mi a. Choi tieu non b nam Phytophthora gay hai tren la b. Choi tieu mi hnh thanh b nam Phytophthora hai chet c. Choi tieu non b nam Phytophthora gay hai tren ngon d. Choi tieu non khoe khong b nhiem benh 72
biet nay khong c ro ve phng dien thong ke. Ket qua phan tch so lieu thong ke the hien qua bang 4.25. Bang 4.24 So sanh ty le choi tieu mi moc b chet gia cac nghiem thc qua mot so ky ieu tra cua th nghiem phong tr benh tren cay tieu 15 NSC Nghiem thc Ty le choi chet T.32-bon + phun T.32-bon T.41-bon T.41-bon + phun T.32-phun T.41-phun C co chung P. SS 22 NSC Ty le choi chet SS 34 NSC Ty le choi chet SS
2,3 2,3 x 0,0 0,0 x 1,4 1,4 x 6,0 4,8 x 3,8 1,6 x 16,0 3,1 x x 29,0 15,2
4,3 3,3 x 9,3 2,4 x 9,3 2,6 x 12,7 4,4 x 12,0 3,2 x
7,2 3,7 x 11,9 2,6 x 11,9 3,7 x 13,0 4,5 x 14,1 3,0 x
20,7 3,4 x x 14,4 3,9 x

x 90,4 4,8 x
x 40,7 16,2
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Bang 4.25 So sanh ty le choi tieu mi moc b chet gia 2 dong T.32 va T.41 cua th nghiem phong tr benh tren cay tieu Nghiem thc T.32 T.41 8 NSC Ty le choi chet 2,0 1,9 9,6 1,9
x x
15 NSC SS Ty le choi chet 8,5 2,1 16,6 2,1

x x
22 NSC SS Ty le choi chet 9,3 3,4 17,0 3,4 SS

x x
41 NSC Ty le choi chet 11,0 4,9 20,7 4,9 SS

x x
* Tren cung mot cot theo phng ngang th s khac biet mc = 0,05 theo trac nghiem Duncan.
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+ Anh hng en re cay: Sau khi ket thuc qua trnh ieu tra theo doi cac ch tieu la, day, choi, chung toi tien hanh nho cay e khao sat them anh hng cua Trichodema va Phytophthora en viec sinh trng cua re cay (xem hnh 4.15). Ket qua thu c the hien qua bang 4.26. Bang 4.26 So sanh khoi lng bo re sau th nghiem phong tr benh tren cay tieu Nghiem thc C co chung P. T.41-bon T.41-phun T.32-bon T.32-phun T.32-bon + phun T.41-bon + phun C khong chung P. Khoi lng re (g/chau) So sanh
21.8 1,1 x 31.0 0,8 32.2 0,7 32.7 0,8 33.7 0,8 34.3 2,0 37.9 1,8 46.7 3,2
x x x x x x x x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Qua bang 4.26, x ly thong ke ve so sanh, co s khac biet rat co y ngha gia nghiem thc oi chng co chung Phytophthora (B01) vi tat ca cac nghiem thc khac. Trong khi s khac biet gia nhng nghiem thc co x ly che pham Trichodema vi nghiem thc oi chng khong chung Phytophthora (B02) khong khac biet lam. Trong o nhng nghiem thc va bon ket hp phun gan tng ng vi nghiem thc oi chng khong chung Phytophthora. ieu nay cung the hien ro khi phan tch thong ke so sanh khoi lng re gia cac cach x ly khac nhau qua bang 4.27, nghiem thc va bon ket hp phun che pham Trichodema bo re tieu c bao ve tot hn. 74
Hnh 4.15: Hnh dang cay va bo re tieu cac nghiem thc sau th nghiem a,b. C khong chung Phytophthora c,d. C co chung Phytophthora 75 e,f. X ly Trichodema T.32 g,h. X ly Trichodema T.41
Bang 4.27 So sanh khoi lng bo re cac nghiem thc co cach x ly Trichodema khac nhau sau th nghiem phong tr benh tren cay tieu Nghiem thc Bon Phun Bon + phun Khoi lng re (g/chau) So sanh
31.8 0,9 x 32.9 0,9 x 36.1 0,9

x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Tuy nhien, khi so sanh khoi lng re gia cac nghiem thc x ly cac dong Trichodema khac nhau (T.41 va T.32) hau nh s khac biet ve khoi lng re khong co y ngha (xem bang 4.28) Bang 4.28 So sanh khoi lng bo re cac nghiem thc x ly cac dong Trichodema khac nhau sau th nghiem phong tr benh tren cay tieu Nghiem thc T.32 T.41 Trong lng re (g/chau) 33.6 0,8 33.7 0,8 So sanh
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Qua th nghiem x ly 2 dong che pham Trichodema khac nhau T.41 va T.32, vi 3 cach x ly khac nhau la dung dang ran e bon, dung dang long e phun va dung ket hp va bon va phun tren ruong th nghiem a lay nhiem nhan tao nam gay benh Phytophthora so vi oi chng co va khong lay nhiem Phytophthora. Vi mot so ch tieu a theo doi, anh gia sau phan tch chung toi thay rang:
76
Hau het eu co s khac biet rat co y ngha gia cac nghiem thc co x ly Trichodema so vi cac nghiem thc oi chng co lay nhiem Phytophthora, ch co ch tieu s gia tang vet benh la khong co y ngha, ieu nay cho thay Trichodema co tac dung cham hn so vi s tan cong cua Phytophthora. ong thi mot so la vet benh gay hai khong ln nhng a rung sau khi theo doi, ay co the la mot c che t bao ve cua cay trong trc s tan cong cua cac tac nhan gay hai. So sanh gia cac cach x ly Trichodema, bon, phun va bon ket hp phun, hau het cac ch tieu theo doi, s khac biet khong ln. Qua o, khi a ra s dung ai tra tren ong ruong ta nen chon phng phap nao va tien dung va co hieu qua kinh te cho tng loai cay trong cu the nh ti che pham cho cac loai rau, bon dang ran cho cac cay lau nam. So sanh gia hai dong Trichodema dung th nghiem, a so s khac biet khong ln. Trong mot so ch tieu dong T.32 co nhnh hn co the s dung dong nay cho cac th nghiem tiep theo e khang nh lai hieu lc cua no khi a ra ong ruong. 4.1.5 Khao sat hieu lc cua che pham Trichodema tr benh do nam Phytophthora sp. gay hai tren cay sau rieng. Qua viec chon loc, th oi khang cac dong Trichodema vi cac loai nam gay hai. Qua th nghiem nhan sinh khoi Trichodema dang long va dang ran. ac biet, qua th nghiem khao sat hieu lc cua che pham Trichodema trong nha li, chung toi quyet nh chon che pham dang ran cua dong T.32 tiep tuc th nghiem ngoai ong ruong. Vi muc ch kiem tra hieu lc cua che pham, xac nh lieu lng thch hp va cach x ly hp ly, chung toi bo tr co nhieu nghiem thc co x ly che pham Trichodema vi lieu lng khac nhau va co hay khong ket hp vi phan chuong so vi 2 nghiem thc oi chng la co bon phan chuong va khong bon phan chuong.
77
Cay trong dung th nghiem la sau rieng, khac trong nha li (cay tieu), e kiem tra them pho tac ong cua che pham. Chung toi tien hanh ong thi tren cay sau rieng vn m va ong ruong e xem xet them viec ap dung che pham Trichodema trong thc tien phong tr dch hai sau nay. 4.1.5.1 Khao sat trong giai oan cay sau rieng chuan b xuat vn - Ty le cay chet canh, heo ot: sau khi chung nam 1 tuan hau het cac cay sau rieng con eu co bieu hien b vang, rung la chan, kho va chet cac canh ben di, mot so kho va chet ot (xem hnh 4.16a va 4.16b). Ket qua ieu tra cho thay, 15 ngay sau khi chung nhiem c x ly hon hp phan chuong va che pham Trichodema, ty le cay b kho canh ben di va chet ot nghiem thc 2, nghiem thc khong x ly che pham Trichodema cao nhat (93,33%). Nghiem thc ch x ly 25 gram che pham Trichodema / cay (T4) co ty le cay b kho canh va chet ot thap nhat va khac biet co y ngha so vi cac nghiem thc khac. Cac nghiem thc x ly che pham Trichodema va phan chuong co bieu hien gan giong nhau va ty le cay b kho canh va chet ot tng ng nhau (bang 4.29). - Ty le cay hoi phuc: Cay bat au the hien s hoi phuc 30 ngay sau khi x ly, chung ra nhieu ot non mi, nhng cho en 45 ngay sau khi x ly cac cay the hien s hoi phuc kha ro ret, ac biet oi vi cac nghiem thc co x ly che pham Trichodema. Nghiem thc ch x ly phan chuong (T5) cung hoi phuc tuy nhien mc hoi phuc thap hn so vi cac nghiem thc co x ly che pham Trichodema. 60 ngay sau khi x ly cay gan nh hoi phuc hoan toan oi vi cac nghiem thc co x ly che pham Trichodema. Qua x ly thong ke, khong co s khac biet co y ngha gia nhng nghiem thc nay (xem hnh 4.17, va 4.18).
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Hnh 4.16: Trieu chng benh do nam Phytophthora gay hai tren sau rieng a,b. Trieu chng la sau rieng b benh 15 ngay sau khi chung nam Phytophthora palmovora c. Trieu chng la sau rieng b benh ngoai ong ruong d. Re sau rieng vn m nam Phytophthora palmovora gay hai 79
Hnh 4.17: Sau rieng giai oan cay vn m 2 thang sau chung nhiem Phytophthora palmovora va x ly che pham Trichodema a. Toan canh ruong th nghiem b. Nghiem thc x ly 10g Trichodema + 800g phan chuong / bau c. Nghiem thc oi chng khong x ly Trichodema va phan chuong 80
Hnh 4.18: Cay sau rieng giai oan cay vn m 2 thang sau chung nhiem Phytophthora palmovora va x ly che pham Trichodema cac nghiem thc a. 10g CP Tr. + 800g PCh. c. 40g CP Tr. + 800g PCh. e. 0g CP Tr. + 800g PCh. 81 b. 20g CP Tr. + 800g PCh. d. 25g CP Tr. + 0g PCh. f. 0g CP Tr. + 0g PCh.
Bang 4.29 So sanh ty le cay b chet canh, heo ot gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m Nghiem thc 20g Tr. 10g Tr. + P.Ch 40g Tr. + P.Ch 20g Tr. + P.Ch C co P.Ch C khong P.Ch Ty le benh 15 ngay SXL So sanh
33,3 3,3 x 53,3 3,3 60,0 5,8 73,3 3,3 93,3 3,3 93,3 3,3
x x x x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Bang 4.30 So sanh ty le cay hoi phuc gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m Nghiem thc C khong P.Ch C co P.Ch 20g Tr. 10g Tr. + P.Ch 20g Tr. + P.Ch 40g Tr. + P.Ch 45 NSXL Ty le hoi phuc 13,3 3,3 x 33,3 3,3 46,7 3,3 50,0 5,8 53,3 3,3 56,7 6,7
x x x x x x
60 NSXL SS Ty le hoi phuc 13,3 3,3 x 33,3 6,7 83,3 3,3 76,7 3,3 76,7 3,3 86,7 3,3
x x x x x
SS
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. - Ty le cay chet han: Sau 45 ngay chung nhiem nam benh va x ly hon hp phan chuong va che pham Trichodema, hau het cac nghiem thc eu co 82
cay b chet, ty le cay chet cac nghiem thc co x ly Trichodema eu rat thap so vi nghiem thc ch bon phan chuong va nghiem thc khong phan chuong. Quan sat re cay chet co trieu chng thoi (xem hnh 4.16d) 60 ngay sau chung nhiem nam x ly hon hp phan chuong va che pham Trichodema ty le cay b chet cac nghiem thc co x ly Trichodema va ke ca nghiem thc ch co phan chuong eu khong gia tang. Nghiem thc oi chng khong phan chuong ty le chet la 96,66%. Bang 4.31 So sanh ty le cay chet han gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m Nghiem thc 10g Tr. + P.Ch 20g Tr. + P.Ch 40g Tr. + P.Ch 20g Tr. C co P.Ch C khong P.Ch 45 NSXL Ty le cay chet 10,0 0,0 x 10,0 0,0 x 10,0 0,0 x 10,0 0,0 x 20,0 0,0 46,7 3,3
x x
60 NSXL SS Ty le cay chet 10,0 0,0 x 10,0 0,0 x 10,0 0,0 x 10,0 0,0 x 20,0 0,0 73,3 26,7
x x
SS
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Qua phan tch thong ke, s khac biet gia cac nghiem thc co x ly che pham Trichodema vi nghiem thc khong x ly che pham Trichodema va gia nghiem thc co bon phan chuong vi nghiem thc khong bon phan chuong la rat y ngha. Nhng s khac biet gia cac nghiem thc co bon phan chuong ket hp che pham Trichodema cac lieu lng khac nhau khong ln, khong co y ngha ve phng dien thong ke. 83
- Dien tch la: ve kch thc la cac nghiem thc cung co s khac biet. 30 ngay sau x ly nghiem thc x ly 25 gram che pham Trichodema / cay (T4) kch thc la to nhat. Tuy nhien en 60 ngay sau khi x ly th nghiem thc 40g Trichodema + 800g phan chuong co dien tch la to, cay xanh tot va ep nhat (xem bang 4.32). Ket qua phan tch thong ke 60 ngay sau x ly cho thay co s khac biet co y ngha gia cac nghiem thc co bon phan ket hp che pham Trichodema vi cac nghiem thc khong bon phan chuong hoac khong x ly Trichodema. Nhng s khac biet khac thc s khong co y ngha. Bang 4.32 So sanh dien tch la gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m (cm2) Nghiem thc C khong P.Ch 20g Tr. C co P.Ch 10g Tr. + P.Ch 20g Tr. + P.Ch 40g Tr. + P.Ch 30 NSXL Dien tch la So sanh 45,7 3,3 x 77,2 2,7 58,0 6,2 x x 63,3 2,4 63,4 4,2 66,6 7,1
x x x x x x x
60 NSXL Dien tch la 0,3 0,3 x 67,7 5,9 68,3 4,1 81,7 2,8 85,3 2,8 94,0 0,6
x x x x x x
So sanh
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. 4.1.5.2 Khao sat tren cay sau rieng ngoai ong ruong 15 ngay sau khi x ly, cay bat au ra ot non hau het cac nghiem thc, khao sat vung re trong at cung thay co s ra re mi hau het tat ca cac cay sau khi c x ly. o ch so SPAD, cho en 1 thang sau khi x ly, gia cac 84
nghiem thc cha the hien s khac biet ve ham lng diep luc to trong la nhng thang th 2 sau khi x ly th a co s khac biet ro ret gia cac nghiem thc co x ly che pham Trichodema va cac nghiem thc khong x ly che pham Trichodema (xem bang 4.33). Ruong th nghiem va cach x ly che pham xem hnh 4.19 va 4.20. Bang 4.33 So sanh ch so diep luc to tren la gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng ngoai ruong Nghiem thc 125g Tr. + P.Ch 250g Tr. + P.Ch 500g Tr. + P.Ch 200g Tr. C co P.Ch C nh Nd 30 NSXL Ch so SPAD 23,9 1,1 25,7 1,2 24,9 1,1 23,7 1,0 23,4 1,3 21,9 1,1 60 NSXL Ch so SPAD 26,8 1,3 32,1 1,5 32,4 1,6 31,3 1,5 21,5 1,0 x 20,3 1,0 x So sanh
x x x x x x x
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Khao sat kch thc la qua 2 thang x ly, chung toi cung ghi nhan co s khac biet ve kch thc la gia cac nghiem thc. Nghiem thc co kch thc la cao nhat van la cac nghiem thc co x ly phan chuong ket hp vi Trichodema lieu lng cao. Tuy nhien, qua phan tch thong ke, s khac biet nay khong thc s co y ngha (xem bang 4.34). - Tan suat xuat hien cua nam Phytophthora palmivora sau khi x ly: Ket qua cho thay sau khi x ly hon hp che pham phan chuong va Trichodema, tan suat xuat hien cua nam qua cac lan phan lap eu giam, ac biet la nghiem thc x ly phan chuong va Trichodema lieu lng 500g, co tan suat xuat hien cua nam la thap 85
nhat va khac biet co y ngha so vi tat ca cac nghiem thc con lai. So sanh gia cac nghiem thc co x ly ket hp che pham Trichodema va phan chuong eu khac biet co y ngha vi cac nghiem thc khac. ac biet co s khac biet ln gia co bon phan chuong vi cach x ly nh nong dan ang thc hien (xem bang 4.35; bang 4.36). Bang 4.34 So sanh dien tch la gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng ngoai ruong (cm2) Nghiem thc C nh Nd C co P.Ch 250g Tr. + P.Ch 125g Tr. + P.Ch 200g Tr. 500g Tr. + P.Ch 30 NSXL Dien tch la So sanh 32,7 2,5 34,8 2,6 37,7 2,5 38,0 2,5 40,7 2,5 44,1 2,5
x x x x x x x x x x x x
60 NSXL Dien tch la 60,6 5,4 x 65,9 5,6 x x 82,7 5,4 x x 72,2 5,3 x x 66,4 5,4 80,0 5,4
x x
So sanh
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Qua th nghiem tren cay sau rieng giai oan vn m, hau het cac ch tieu lien quan en hieu lc cua che pham Trichodema, eu co s khac biet rat co y ngha gia nhng nghiem thc co x ly va khong x ly che pham nay. nhng s khac biet gia cac nghiem thc co lieu lng s dung khac nhau va cach x ly khac nhau khong co y ngha lam. ong ngha vi viec la chon nhng nghiem thc lieu lng s dung thap e tang hieu qua kinh te, hay tiep tuc th nghiem nhng lieu lng thap hn na e xac nh lieu lng toi u.
86
Hnh 4.19: Vn sau rieng 5 nam tuoi trong th nghiem anh gia hieu qua phong tr benh cua che pham Trichoderma ngoai ong ruong a. Vn cay sau rieng trc th nghiem b. Vn cay sau rieng ang lam ve sinh chuan b bo tr th nghiem c. Vn cay sau rieng ang x ly Trichoderma va phan chuong d. Vn cay sau rieng 15 ngay sau x ly Trichoderma va phan chuong 87
Hnh 4.20: Cach x ly che pham Trichoderma va phan chuong trong th nghiem phong tr benh tren cay sau rieng ngoai ong ruong a. anh ranh quanh goc cay sau rieng b. Bon hon hp che pham Trichoderma va phan chuong c. Rac che pham Trichoderma nghiem thc khong co phan chuong d. lap a lai sau khi x ly che pham Trichoderma va phan chuong 88
Bang 4.35 So sanh tan suat xuat hien Phytophthora gia cac nghiem thc 15 va 30 ngay sau x ly cua th nghiem phong tr benh tren cay sau rieng ngoai ruong (%) Nghiem thc 500g Tr. + P.Ch 250g Tr. + P.Ch 125g Tr. + P.Ch C co P.Ch 200g Tr. C nh Nd 15 NSXL Tan suat 45,5 4,7 x 58,4 4,6 x x 58,4 4,6 x x 64,4 4,6 62,4 4,5 83,5 4,6
x x
30 NSXL SS Tan suat 17,1 3,6 x 27,6 3,5 x x 29,7 3,5 38,7 3,5 43,6 3,5
x 84,6 3,6 x x x x x x
SS
* Tren cung mot cot theo phng thang ng th s khac biet mc = 0,05 theo trac nghiem Duncan. Bang 4.36 So sanh tan suat xuat hien Phytophthora gia cac nghiem thc 45 va 60 ngay sau x ly cua th nghiem phong tr benh tren cay sau rieng ngoai ruong (%) Nghiem thc 500g Tr. + P.Ch 250g Tr. + P.Ch 125g Tr. + P.Ch C co P.Ch 200g Tr. C nh Nd 45 NSXL % 14,7 2,7 x 21,5 2,6 x x 23,6 2,6 36,4 2,6 39,3 2,6 87,0 2,6
x x x
60 NSXL SS % 13,6 1,7 x 21,1 1,7 25,4 1,7 41,1 1,7 41,5 1,7
x 86,1 1,7 x x x x x
SS
* Tren cung mot cot theo phng thang dng th s khac biet mc = 0,05 theo trac nghiem Duncan. 89
ca th nghiem cay vn m va th nghiem cay ngoai ong ruong, cac nghiem thc co bon ket hp phan chuong va che pham Trichodema khac biet rat co y ngha vi nhng nghiem thc khong ket hp trong nhng ch tieu theo doi en sinh trng cua cay trong va s ton lu cua mam benh, nhng s khac biet nay lai khong co y ngha oi vi viec tang trng cua dien tch la. ieu nay cung kha de hieu bi ay la yeu to quyet nh do yeu to giong hn la tac ong bi yeu to ben ngoai. 4.2 Thao luan Qua ket qua cac th nghiem chung toi a thc hien, t viec anh gia kha nang oi khang cua nam Trichodema vi bon loai nam gay hai cay trong pho bien trong at, en cac th nghiem anh gia kha nang phong tr benh cua che pham nam Trichodema tren cay trong trong nha li, trong vn m va ngoai ong ruong, Trichodema co kha nang khang mot so loai nam gay benh tren cay trong th a ro. ieu o cung a c chng minh bang nhieu cong trnh nghien cu cua nhieu tac gia tren the gii (Bliss, 1959 ; Baker, 1974 ; Barnett, 1974 ; Ohr, 1975 ; Lumsden, 1977 va Cook, 1980), trong nc (Lu Hong Man, 1977 ; Nguyen Ngoc Tu, 1977 ; Tran Th Thuan, 1995 ; Tran Th Ngoc Mai, 1999). Tuy nhien, th nghiem anh gia tnh oi khang cua chung toi ch dng lai 14 dong nam Trichodema vi 4 loai nam gay hai cay trong Phytophthora, Rhizoctonia, Fusarium va Sclerotium, tat yeu se con nhieu han che trong viec tm c mot dong nam Trichodema co hieu qua oi khang tot nhat e a vao san xuat tao che pham s dung ai tra cho cac loai cay trong. Qua ket qua nhan sinh khoi, vi phng phap len men chm va len men xop, chung toi a tao c nhng che pham cac dang khac nhau (dang long va dang ran), vi nhng dong Trichodema khac nhau (T.32 va T.41), khang nh lai viec san xuat ra che pham che pham Trichodema khong phai la kho. Che pham
90
Trichodema cung a c ng dung t lau tren the gii (Dunin 1979 ; Filippa 1987 ; Badai 1986). Nhng viec s dung chung nh mot bien phap sinh hoc trong phong tr dch hai hien nay nc ta con rat han che. Phai chang con nhng bat cap trong qua trnh san xuat, ton tr, hieu qua kinh te cung nh cong tac khuyen nong e thay oi c tap quan s dung thuoc hoa hoc cua ngi dan. Trong qua trnh th nghiem, chung toi nhan thay viec len men dang long s dung may lac, lng che pham mot lan san xuat ra con thap. Chung toi ang tiep tuc th nghiem mot phng phap len men mi, n gian hn t nhng vat lieu rat de kiem tren th trng. o la phng phap len men bang cach sut kh. Vi thiet b c dung la may sut kh cho nhng be nuoi ca va vo bnh nc tinh khiet loai ln sau khi s dung (xem hnh 4.21). Viec len men xop, tao che pham Trichodema dang ran, bang nhng hop xop, phng phap nay kha n gian, de lam. Nhng ngc lai, khong the san xuat ai tra bang phng phap o c, lng che pham thu c thap. Tuy nhien, ay la c s e chon c loai moi trng dung cho viec nhan sinh khoi dang ran. Trau va cam la nhng phu pham trong che bien lng thc rat pho bien nc ta. Vi 2 loai moi trng nay, cung vi phng phap len men kha n gian, co the san xuat che pham Trichodema dang ran rat de dang cac nha may nong dc, tren nhng day chuyen san xuat thuoc tr sau benh dang hat. Kh trung moi trng bang nhng noi hap bang hi nc co cong suat ln. Cu the phng phap nay co the san xuat ai tra tren day chuyen cong nghe cua Cong Ty Lien Doanh San Xuat Nong Dc Vi Sinh Viguato Tp. Ho Ch Minh. Th nghiem anh gia kha nang phong tr benh do nam Phytophthora gay hai tren cay tieu cua che pham nam Trichodema, chung toi thc hien c t khau chung nam Phytophthora gay benh en viec bo tr th nghiem n va a yeu to. So sanh c nhieu yeu to nh cac dong Trichodema, cac cach x ly
91
Hnh 4.21: Th nghiem sut kh trong phng phap len men chm tao che pham Trichodema dang long a,b,c la cac bnh len men nhn cac goc o khac nhau
92
Th nghiem anh gia kha nang phong tr benh do nam Phytophthora gay hai tren cay sau rieng cua che pham nam Trichodema, chung toi cung thc hien c t khau chung nam Phytophthora gay benh en viec bo tr th nghiem so sanh c nhieu yeu to nh lieu lng che pham Trichodema, cach x ly co hay khong ket hp vi phan chuong, theo doi nhieu ch tieu phong tr benh va anh hng en sinh trng cay trong ca trong vn m va ngoai ong ruong. Tuy nhien, th nghiem tren cay sau rieng ngoai ong ruong chung toi cung ch thc hien c trong thi gian ngan, nen ch anh gia c mot so ch tieu lien quan en sinh trng cay trong va han che mam mong nam gay hai ton lu trong at khi s dung che pham nam Trichodema. Xuyen suot lai cac th nghiem trong toan bo e tai, chung toi muon a ra mot cach nhn n gian hn ve s dung Trichodema trong phong tr sinh hoc. o la: Phai am bao san xuat va s dung chung mot cach n gian, gon nhe, hieu qua kinh te cao va thuan li trong viec ng dung vao thc tien. V vay, nhng th nghiem cua chung toi tap trung vao viec ng dung vao thc tien. Phan lap, chon dong nam khang t nhng tam iem vung benh, nhan sinh khoi, a che pham ve ung cay trong va moi trng song cua no. Th nghiem anh gia tnh khang cua Trichodema vi mot so loai nam tren da petri ch dng ket qua anh gia cam quan. Phng phap san xuat che pham Trichodema cung het sc tho s, co the ap dung no trong nhng ieu kien c s vat chat cac a phng con thieu thon. Th nghiem khao sat hieu lc cua che pham, mac du trong mot thi gian ngan, chung toi khao sat c hieu qua cua no oi vi dch benh tren 2 loai cay trong pho bien hien nay, xem xet lieu lng s dung, cach x ly va anh hng en sinh trng cay trong, ton lu mam benh trong at. Vi hy vong rut ngan hn na mot qui trnh khep kn san xuat che pham Trichodema chuyen biet cho tng loai cay trong, tng vung sinh thai.
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Tom tat cac bc c ban trong chon loc, nhan sinh khoi va s dung che pham Trichodema 1. 2. Phan lap nam Trichodema t nhng vung ang b dch benh. Khao sat tnh oi khang tren da petri, chon dong nam Trichodema oi khang cao vi nam ang gay dch. 3. Nhan sinh khoi bang phng phap len men long hay len men ran tuy ieu kien s dung cho tng loai cay trong va tap quan cua a phng. 4. anh gia hieu lc, chon lieu lng s dung va cach s dung bang th nghiem tien hanh tren chnh cay trong va vung phan lap nam.
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Chng 5 KET LUAN VA E NGH

5.1 Ket luan e tai chon c dong Trichodema co kha nang oi khang cao, nhan c cac dang sinh khoi co ham lng bao t hoac si nam trong che pham ln, a c vao nghien cu va thc tien san xuat che pham. anh gia hieu lc che pham, cach s dung, ap dung trong phong tr benh cay trong bang phng phap phong tr sinh hoc. Cu the nh sau: 1. Thu thap va phan lap c 14 dong Trichodema cac a iem khac nhau. 2. Chon ra c hai dong Trichodema (T.31 va T.41) co kha nang khang vi nam benh cao nhat trong 14 dong Trichodema noi tren trong a petri. 3. Khoi lng khuan ty cua nhom Trichodema thu c trong phng phap nhan sinh khoi long, sau 72 gi nuoi cay, nghiem thc 30g ng, 100g gia au xanh, 1g ure, 1g KH2Po4, 0,5g MgSO4 va 1000ml nc cat (P4) cao nhat at 2,48g khuan ty. Tuy nhien, nghiem thc s dung 20g ng (P3) ma khoi lng khuan ty at 2,32g. Khoi lng khuan ty cua nam Trichodema thu c trong th nghiem khao sat lng gia nghiem thc s dung 200g gia au xanh (G5) at 1,55g va trong nghiem thc ch s dung 100g gia song (G3) lng khuan ty thu c 1,43g sau 72 gi nuoi cay. Qua o, chung toi chon cong thc moi trng gom 20g ng, 100g gia au xanh, 1g ure, 1g KH2Po4, 0,5g MgSO4 va 1000ml nc cat la moi trng thch hp nhat e nhan sinh khoi Trichodema dang long. 4. th nghiem so sanh khoi lng khuan ty cua 2 dong nam Trichodema T.41 va T.32 th lng khuan ty cua dong T.32 cao hn T.41 (2,04g so vi 1,65g) sau 72 gi nuoi cay. 95
5. Nhan sinh khoi dang ran 7 nghiem thc khac nhau va at mot so bao t / ml cao nhat nghiem thc 15g cam + 15g trau (N.1) at 33,46 x 108 bao t / ml, thap nhat nghiem thc 30g cm (N.7) c 1,12 x 108 bao t / ml sau 6 ngay nuoi cay. So lng bao t thu c dong T.32 cao hn T.41 la 7,9 x 108 bao t / ml. Nghiem thc 15g cam + 15g trau c chon cho viec nhan sinh khoi Trichodema dang ran. 6. Th nghiem khao sat hieu lc che pham 2 dong nam Trichodema T.41 va T.32 trong viec phong tr benh tren cay tieu cho thay hau het cac ch tieu theo doi eu co s khac biet rat co y ngha gia cac nghiem thc co x ly Trichodema so vi cac nghiem thc oi chng co lay nhiem Phytophthora. Gia hai dong Trichodema dung th nghiem (T.41 va T.32), co khac biet nhng khong ln. Trong mot so ch tieu dong T.32 co nhnh hn. Gia cac cach x ly Trichodema: Bon, phun va bon ket hp phun, cac ch tieu theo doi, s khac biet khong ln. 7. S dung che pham Trichodema dang ran dong T.32 trong viec phong tr benh do nam Phytophthora gay ra tren cay sau rieng vn m, cac ch tieu: Ty le cay sau rieng chet canh, heo ot, ty le cay hoi phuc sau khi x ly va ty le cay chet han sau khi x ly cac nghiem thc co x ly che pham Trichodema eu cho ket qua tot hn han so vi cac nghiem thc oi chng khong x ly che pham Trichodema 8. S dung che pham Trichodema dang ran dong T.32 trong viec phong tr benh do nam Phytophthora gay ra tren cay sau rieng ngoai ong ruong, cac nghiem thc co bon ket hp phan chuong va che pham Trichodema khac biet rat co y ngha vi nhng nghiem thc khong ket hp trong nhng ch tieu theo doi en sinh trng cua cay trong va s ton lu cua mam benh Phytophthora. 5.2 e ngh
96
Tiep tuc nghien cu ve kha nang phong tr benh cua cac dong nam Trichodema oi vi cac loai nam gay nhng loai benh kho tr tren nhng cay trong khac. Nghien cu tiep nhng phng phap tao che pham Trichodema n gian hn, co the ap dung san xuat tng a phng, tng trang trai cho tng loai cay trong cu the. Tren cay dai ngay, can co nhng nghien cu vi thi gian dai hn e co c nhng ket qua toan dien hn, bi nhng bien phap phong tr sinh hoc thng co tac dung cham nhng keo dai va ben vng hn.
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Chng 6 TAI LIEU THAM KHAO

1. Nguyen Th Ngoc An, 1999. Ky thuat trong va cham soc vn cay va cac van e lien quan. Nha xuat ban Nong Nghiep, Thanh pho Ho Ch Minh, trang 111-153. 2. Nguyen Van Cam va Pham Van Lam (Trung tam au tranh sinh hoc Vien Bao ve Thc vat), 1996. Tuyen tap cong trnh nghien cu bien phap sinh hoc phong tr dch hai cay trong, tap 1. Nha xuat ban Nong Nghiep. 3. Ta Kim Chinh, 1996. Nghien cu tuyen chon mot so chung vi nam diet con trung gay hai Viet Nam va kha nang ng dung. Luan an PTS, Vien cong nghe sinh hoc - TT. Khoa hoc T nhien va Cong nghe Quoc gia, Ha Noi. 4. ng Hong Dat va ctv, 1979. Giao trnh vi sinh vat trong trot. Nha xuat ban Nong Nghiep, Ha Noi, Viet Nam. 5. Nguyen Lan Dung, 1981. S dung vi sinh vat e phong tr sau hai cay trong. Nha xuat ban Khoa hoc va Ky thuat, Ha Noi. 6. o Tan Dung va cvt, 2001. ac tnh sinh hoc va kha nang phong chong mot so benh nam hai re cay trong can cua nam oi khang Trichodema viride. Tap ch Bao ve Thc vat 4: 12-16. 7. Bui Xuan ong, 1977. Mot so van e ve nam hoc. Nha xuat ban Khoa hoc va Ky thuat, Ha Noi. 8. Bui Xuan ong, 1984, 1986. Nhom nam Hyphomycetes Viet Nam, tap 1 va 2. Nha xuat ban Khoa hoc va Ky thuat, Ha Noi. 9. Bui Xuan ong va Nguyen Huy Van, 2000. Vi nam dung trong cong nghe sinh hoc. Nha xuat ban Khoa hoc va Ky thuat, Ha Noi. 10. Vu Cong Hau, 2000. Trong cay an qua Viet Nam. In lan th ba. Nha xuat ban Nong Nghiep, Thanh pho Ho Ch Minh.
98
11. Tran Van Hoa va ctv, 2000. Sau benh hai cay trong, cach phong tr. 101 cau hoi thng gap trong san xuat Nong Nghiep, tap 4. Nha xuat ban Tre. 12. Pham Van Lam, 1995. Bien phap sinh hoc phong chong dch hai nong nghiep. Nha xuat ban Nong Nghiep, Ha Noi. 13. Tran Van Mao, 1998. S dung sau nam co ch. ai hoc Lam Nghiep. Nha xuat ban Nong nghiep, Ha Noi. 14. Le Van Nhng va Nguyen Lan Dung, 1992. Cong nghe sinh hoc - Mot c hoi cho tat ca. Nha xuat ban Nong nghiep, Ha Noi. 15. Vo Th Thu Oanh, 1999. Benh cay chuyen khoa. 16. Le Thanh Phong, 1994. Cay sau rieng. Nha xuat ban Nong Nghiep, 19 trang. 17. Tran Th Thanh, 2000. Cong nghe vi sinh. Nha xuat ban Giao duc. 18. Le Duy Thang, 1999. Ky thuat trong nam. Nuoi trong mot so nam an thong dung Viet Nam, tap 1. Nha xuat ban Nong Nghiep, Thanh pho Ho Ch Minh. 19. Tran Th Thuan, 1998. Anh hng cua ieu kien chieu sang en s sinh trng va phat trien cua nam. Tap ch Bao ve Thc vat 3: 7-10. 20. Tran Th Thuan, 1998. Hieu qua oi khang cua nam Trichodema oi vi nam gay benh hai cay trong. Tap ch Bao ve Thc vat 4: 35-38. 21. Tran Th Thuan, 1998. Chat trao oi do nam Trichodema sinh ra va sinh trng phat trien cua mot so cay trong. Tap ch Bao ve Thc vat 5: 39-40. 22. Tran Th Thuan, 1999. Phng phap san xuat va s dung nam Trichodema e phong tr benh hai cay trong. Tap ch Bao ve Thc vat 4: 33-34. 23. Nguyen Ngoc Tu va Nguyen Th Hng Giang, 1997. Bao ve cay trong bang cac che pham t vi nam. Nha xuat ban Nong Nghiep, Thanh pho Ho Ch Minh, trang 111-153. 24. Nguyen Van Tuat va Le Van Thuyet, 2000. San xuat, che bien va s dung thuoc bao ve thc vat thao moc va sinh hoc. Vien bao ve thc vat. Nha xuat ban Nong nghiep, Ha Noi. 99
25. Nhieu tac gia, 2004. Tap ch khoa hoc ai hoc Can Th. Nha xuat ban Nong Nghiep. 26. Amy Y. Rossman, John M. McKemy, Rebecca A. Pardo-Schultheiss and HansJosef Schroers, 2000. Molecular studies of the Bionectriaceae using large subunit rDNA sequences. Mycologia, 93 (1), 2001. The Mycological Society of America, pp. 100-110. 27. C. R. Howell and R. D. Stipanovic, 1995. Mechanisms in the Biocontrol of Rhizoctonia solani-Induced Cotton Seedling Disease by Gliocladium virens: Antibiosis. Phytopathology 85 (4), 1995. Texas, USA, pp. 469-472. 28. Crop Protection Handbook, 2004. <URL: http:/www.meisterpro.com> 29. Developing Expeertise in the Management of cocoa. Difeases in Viet Nam. Workshop No.1. 11-14 November 2003. Nong Lam University. 30. D. J. Rae, V. C. Nguyen, M. D. Huang, S. Leong, P. Nanta and G. A. C. Beattie, 2000. Integrated Control of Citrus Pests in China and Southeast Asia. University of Western Sydney, Hawkesbury. 31. Elad.Y.Hadar.Y, Hadar. EChet, I and Henis., 1981. Biological control of Rhizoctonia Solani by T.hazzianuni in Carnation. 32. Evans, H. C., 1982. Entomogenous fungi in tropical forest ecosystems : an appraisal. Ecol. Entomol 7: 47-60. 33. Ferron, P., 1978. Biological control of insect pests by entomogenous fungi. Annu. Rev. Entomol 23: 409-422. 34. Fuxa, J. R., 1987. Ecological considerations for the use of entomogenous in IBM. Annu. Rev. Entomol 32: 225-251. 35. Gary J. Samuels, 1998. The Hypocrea schweinitzii complex and Trichodema sect. Longibrachiatum. United States Dept. of Agriculture, Agricultural Research Service, Systematic Botany and Mycology Lab., Rm. 304, B011A, BARC-West, Beltsville, MD 20705 U.S.A. 36. Gary J. Samuels, 2004. Trichodema a guide to identification and biology. United States Dept. of Agriculture, Agricultural Research Service, Systematic Botany and Mycology Lab., Beltsville, MD 20705 U.S.A. 100
37. Gary E. Harman and Christian P. Kubicek, 1798-1998. Trichodema and Gliocladium. Basic biology, taxonomy and genetics, 1. Taylor & Francis. 38. Gary E. Harman and Christian P. Kubicek, 1798-1998. Trichodema and Gliocladium. Enzymes, biological control and commercial application, 2. Taylor & Francis. 39. Gary J. Samules and Amy. Rossman; Clark T. Rogerson and Rosalind Lowen, 1999. General of Bionectriaceae, Hypocreaceae and Nectriaceae (Hypocreales, Ascomycetes). 40. George N.Agrios, 2000. Plant Pathology, Third edition. Academic press. 41. Hans Josef Schroers, 2001. A monograph of Bionectria (Ascomycota, Hypocreales, Bionectriaceae) and its Clonostachys anamorphs. Centraalbureau voor Schimmelcultures, Fungal Biodiversity Centre, Utrecht, The Netherlands. 42. Hans-Josef Schroers, Gary J. Samules, Keith A. Seifert and Walter Gams, 1998. Classification of the mycoparasite Gliocladium roseum in Clonostachys as C. rosea, its relationship to Bionectria ochroleuca, and notes on other Gliocladium-like fungi. Mycologia 91 (2), 1999. The Mycological Society of America, pp. 365-385. 43. Humber, R. A., 1981. An alternative view of certain texonomic criteria used in the Entomophthorales (Zygomycetes). Mycotaxon 13: 191-240. 44. Ignoffo, C. M., D. L. Hostetter, K. D. Biever, C. Garcia, G. D. Thomas, W. A. Dickerson and R. Pinnell, 1978. Evaluation of an entomopathogenic bacterium, fungus, and virus for control for Heliothis zea on soybeans. J. Econ. Entomol. 71: 165-168. 45. John Bissett, George Szakacs, Carol Ann Nolan, Irina Dzuhinina, Cornelia Gradinger, and Christian P. Kubicek, 2003. New species of Trichodema from Asia. Can. J. Bot. 81:570-586. 46. King, D. S., 1979. Systematics of fungi causing entomophthora - mycosis. Mycologia 71: 731-745.
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47. Kuruvilla, S. and A. Jacob, 1980b. Pathogenicity of the entomogenous fungus Paecilomyces farinosus (Dickson ex Fries) to several insect pests. Entomon 5: 175-176. 48. Maniania, N. K. and J. Fargues, 1984. Specificite des Hyphomycetes entomopathogenes pour les larves des lepidopteres Noctuidae. Entomophaga 29: 451-464. 49. Marques, E. J., A. M. Vilas Boas and C. E. F. Pereira, 1981. Para producao do fungo entomogeno Metarhizium anisopliae (Metsch.) em laboratories setoriasis. Bol. Tec. Planalsucar (Piracicaba) 3 (2): 1-23. 50. McCabe, D. and R. S. Soper, 1985. Preparation of an entomopathogenic insect control agent. United States Patent # 4,530,834, 23 July, 1985. pp. 4. 51. Petch, T., 1931. Notes on entomogenous fungi. Trans. Br. Mycol. Soc. 16: 5575. 52. Petch, T., 1935. Notes on entomogenous fungi. Trans. Br. Mycol. Soc. 19: 161194. 53. Pickford, R. and P. W. Riegert, 1964. The fungus disease caused by entomophthora grylli and its effect on grasshopper populations in Saskatchewan in 1963. Can. Entomol. 96: 1158-1166. 54. Riba, G., S. Marcandis, G. Richard and I. Larget, 1983. Sensibilite de la pyrale du mais (Ostrinia nubilalis Hubner) (Lep. Pyralidae) aux Hyphomycetes entomopathogenes. Entomophaga 28: 55-64. 55. Roberts, D. W. and R. A. Humber, 1981. Entomogenous fungi. In Cole, G. T. and B.Kendrick(eds.), Biology of Conidial Fungi 2, Academic Press, New York. 56. Roffey, J., 1968. The occurrence of the fungus entomophthora grylli F. on locusts and grasshoppers in Thailand. J. Invertebr. Pathol. 11: 237-241. 57. Rombach, M. C., R. M. Aguda and D. W. Roberts, 1986b. Biological control of the brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae) with dry mycelium applications of Metarhizium anisopliae (Deuteromycotina : Hyphomycetes ). Philipp. Entomol. 8: 613-627. 102
58. Rombach, M. C., R. M. Aguda, B. M. Shepard, and D. W. Roberts, 1986b. Infection of the rice brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae) by field application of entomopathogenic hyphomycetes (Deuteromycotina). Environ. Entomol. 15: 1070-1073. 59.Yoshimichi Doi, 1972. Revision of the Hypocreales with Cultural Observations. IV. The Genus Hypocrea and its Allies in Japan, 15(4). Bulletin of the national Science museum, Tokyo, Japan. 60.Trichodema Treated Black Pepper Vines <URL:www.iisr.org/photogallary/biologicalcontrol/ pages/Trichodema%20Treated%20Black%20Pepper%20Vines.htm> 61.Management of nursery wilt of black pepper ( Piper nigrum L.) with ... <URL:www.ias.ac.in/currsci/sep102002/561.pdf > 62.Mycoflora and mycotoxins in Brazilian black pepper, white pepper ... <URL:www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve& db=PubMed&list_uids=11229375&dopt=Abstract> 63.The Indian Institute of Spices Research <URL: www.ncipm.org.in/Trichodermo%20harzianum.htm> 64.Durian root and stem rot (Phytophthora palmivora) <URL:www.ipmthailand.org/en/Pests/Root_and_stem_rot.htm - > 65.Durian Germplasm Evaluation <URL:www.rirdc.gov.au/reports/NPP/02-091.pdf >
Chng 7 PHU LUC
103
Phu luc 7.1 Khoi lng khuan ty thu c cac nghiem thc moi trng co lng ng khac nhau trong th nghiem nhan sinh khoi Trichoderma dang long (g) Nghiem thc Lap lai 0 g ng + 100 g gia 10 g ng + 100 g gia 40 g ng + 100 g gia 20 g ng + 100 g gia 30 g ng + 100 g gia 1
0,2 1,9 2,5 2,6 2,8
Dong T.32 2
0,9 1,9 2,5 2,6 2,8
Dong T.41 4
0,2 1,6 2,0 3,1 2,7
3
0,3 2,1 2,8 2,8 2,4
1
0,5 1,3 2,3 2,2 2,0
2
0,5 1,6 2,3 2,1 2,0
3
0,5 1,9 1,8 2,1 1,5
4
0,4 1,4 2,3 2,1 2,0
Analysis of Variance for LNG KHUAN TY THU C TRONG TH NGHIEM LEN MEN LONG CO LNG NG THAY OI ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES DUONG.DG 1.5484225 1 1.5484225 20.851 .0001 MAIN EFFECTS A:DUONG.NT 22.274265 4 5.5685663 74.986 .0000 RESIDUAL 2.5248900 34 .0742615 ----------------------------------------------------------------------TOTAL (CORRECTED) 26.347578 39 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.2 Khoi lng khuan ty thu c cac nghiem thc moi trng co lng gia khac nhau trong th nghiem nhan sinh khoi Trichoderma dang long (g) Nghiem thc Lap lai 0 g gia + 20 g ng 1
0,2
Dong T.32 2
0,4
Dong T.41 4
0,4
3
0,6
1
0,2
2
0,4
3
0,6
4
0,4
104
50 g gia + 20 g ng 150 g gia + 20 g ng 100 g gia + 20 g ng 200 g gia + 20 g ng
1,5 1,5 1,6 1,4
1,2 1,3 1,3 1,7
1,1 1,5 1,4 1,6
1,1 1,5 1,4 1,7
1,5 1,5 1,6 1,4
1,2 1,3 1,3 1,7
1,1 1,5 1,4 1,6
1,1 1,5 1,4 1,7
Analysis of Variance for LNG KHUAN TY THU C TRONG TH NGHIEM LEN MEN LONG CO LNG GIA THAY OI ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES GIA.DG .00000E0000 1 .00000E0000 .000 1.0000 MAIN EFFECTS A:GIA.NT 6.9352400 4 1.7338100 81.829 .0000 RESIDUAL .7204000 34 .0211882 ------------------------------------------------------------------------TOTAL (CORRECTED) 7.6556400 39 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error. Analysis of Variance for LNG KHUAN TY CUA 2 DONG TRICHODERMA T.32 VA T.41TRONG TH NGHIEM LEN MEN LONG ----------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ----------------------------------------------------------------------COVARIATES DUONG.NT 15.797531 1 15.797531 64.934 .0000 MAIN EFFECTS A:DUONG.DG 1.5484225 1 1.5484225 6.365 .0161 RESIDUAL 9.0016238 37 .2432871 -----------------------------------------------------------------------TOTAL (CORRECTED) 26.347578 39 -----------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.3 So lng bao t thu c cac nghiem thc co c chat khac nhau trong th nghiem nhan sinh khoi Trichoderma dang ran (108 bao t /ml) Nghiem thc Lap lai 30 g cm 10 g cam + 20 g vo ca phe 1
1,2 1,5
Dong T.32 2
1,2 1,5
Dong T.41 4
1,2 2,9
3
1,3 4,3
1
1,1 3,2
2
1,0 3,3
3
1,0 2,1
4
1,1 1,7
105
15 g cam + 15 g vo ca phe 20 g cam + 10 g vo ca phe 20 g cam + 10 g trau 15 g cam + 15 g trau 10 g cam + 20 g trau
58,0 57,0 21,0 50,0 21,0
61,0 56,0 19,0 43,0 52,0
62,0 47,0 25,0 51,0 43,0
73,0 67,0 26,0 47,0 38,0
2,7 6,3 17,0 25,0 27,0
3,6 8,2 15,0 14,0 15,0
3,8 8,9 17,0 17,0 16,0
4,2 8,2 17,0 17,0 16,0
Analysis of Variance for LNG BAO T THU C CUA CAC NGHIEM THC TRONG TH NGHIEM LEN MEN DANG RAN ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES NSKR.DG 809.21010 1 809.21010 15.475 .0003 MAIN EFFECTS A:NSKR.NT 8411.1077 6 1401.8513 26.809 .0000 RESIDUAL 2509.9209 48 52.290020 ------------------------------------------------------------------------TOTAL (CORRECTED) 11730.239 55 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error. Analysis of Variance for LNG BAO T THU C CUA 2 DONG TRICHODERMA T.32 VA T.41 TRONG TH NGHIEM LEN MEN DANG RAN ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES NSKR.NT 7052.2120 1 7052.2120 96.610 .0000 MAIN EFFECTS A:NSKR.DG 809.21010 1 809.21010 11.086 .0016 RESIDUAL 3868.8166 53 72.996541 ------------------------------------------------------------------------TOTAL (CORRECTED) 11730.239 55 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.4 Ty le la tieu b benh cua cac nghiem thc 2, 4 va 6 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu (%) Nghiem thc Lap lai T.32 bon
I
20,0
2 NSC
4 NSC
6 NSC
II
23,8
III
23,8
IV
17,1
I
20,0
II
24,8
III
36,2
IV
21,0
I
24,8
II
27,6
III
38,1
IV
22,9
106
T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P. C khong P.
20,0 23,8 19,1 24,8 11,4 19,1 0
25,7 20,0 13,3 20,0 16,2 24,8 0
14,3 9,5 8,6 26,7 13,3 26,7 0
23,8 15,2 21,9 24,8 10,4 25,7 0
23,8 23,8 20,0 24,8 12,3 25,2 0
32,4 21,0 13,3 21,0 21,0 34,3 0
15,2 11,4 10,5 26,7 15,2 28,6 0
34,3 17,1 22,9 40,0 12,4 34,3 0
26,7 29,5 20,0 28,6 15,2 46,7 0
36,2 28,6 14,3 21,9 23,8 53,3 0
16,2 16,2 15,2 28,6 19,1 38,1 0
42,9 18,1 25,7 42,9 13,3 47,6 0
Phu luc 7.5 Ty le la tieu b benh cua cac nghiem thc 8 va 41 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu (%) Nghiem thc Lap lai T.32 bon T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P, C khong P,
I 29,5 38,1 34,3 25,7 24,8 15,2 74,3 0 II 29,5 44,8 37,1 17,1 22,9 24,8 59,1 0
8 NSC 41 NSC
III 40,0 19,1 29,5 15,2 37,1 20,0 53,3 0
IV 31,4 46,7 21,0 25,7 48,6 13,3 76,2 0
I 29,5 38,1 34,3 25,7 24,8 15,2 100, 0
II 29,5 44,8 37,1 17,1 22,9 24,8 95,2 0
III 40,0 19,1 29,5 15,2 37,1 20,0 85,7 0
IV 31,4 46,7 21,0 25,7 48,6 13,3 76,0 0
One-Way Analysis of Variance for TY LE LA TIEU B BENH CUA CAC NGHIEM THC 41 NGAY SAU KHI CHUNG NAM GAY BENH ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d,f, Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 13634.433 6 2272.4054 29.038 .0000 Within groups 1643.395 21 78.2569 ------------------------------------------------------------------------Total (corrected) 15277,828 27
107
0 missing value(s) have been excluded,
Analysis of Variance for TY LE LA TIEU B BENH CUA CAC CACH X LY 41 NGAY SAU KHI CHUNG NAM GAY BENH ------------------------------------------------------------------------Source of variation Sum of Squares d,f, Mean square F-ratio Sig, level ------------------------------------------------------------------------COVARIATES TB5B,DG 508,94460 1 508,94460 7,314 ,0136 MAIN EFFECTS A:TB5B.CXL 518.83691 2 259.41845 3.728 .0421 RESIDUAL 1391.6058 20 69.580291 ------------------------------------------------------------------------TOTAL (CORRECTED) 2419.3873 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Analysis of Variance for TY LE LA TIEU B BENH NT X LY 2 DONG TRICHODERMA T.32 VA T.41 SAU KHI CHUNG NAM GAY BENH 41 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB5B.DG 508.94460 1 508.94460 7.314 .0136 MAIN EFFECTS A:TB5B.CXL 518.83691 2 259.41845 3.728 .0421 RESIDUAL 1391.6058 20 69.580291 ------------------------------------------------------------------------TOTAL (CORRECTED) 2419.3873 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.6 S gia tang vet benh tren la tieu cua cac nghiem thc qua cac ky ieu tra th nghiem phong tr benh tren cay tieu (mm) Nghiem thc Lap lai T.32 bon
I 10
2 NSC 4 NSC 6 NSC 8 NSC
II 7
III 15
I 23
II 19
III 26
I 30
II 27
III 32
I R
II R
III R
108
T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P. C khong P.
10 3 7 6 12 13
23 10 8 28 20 12
13 7 9 10 11 10
22 16 20 19 26 28
32 22 20 40 32 24
26 19 23 24 24 21
29 24 29 28 34 37
40 31 30 48 41 33
34 27 31 32 33 31
34 30 36 34 R R
37 37 36 R R R
R 33 38 39 40 39
One-Way Analysis of Variance for S GIA TANG VET BENH TREN LA TIEU CUA CAC NGHIEM THC 8 NGAY SAU KHI CHUNG NAM GAY BENH ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 48.583333 5 9.7166667 1.315 .3696 Within groups 44.333333 6 7.3888889 ------------------------------------------------------------------------Total (corrected) 92.916667 11 9 missing value(s) have been excluded.
Analysis of Variance for S GIA TA7NG VET BENH TREN LA TIEU CAC CACH X LY TRICHODERMA SAU KHI CHUNG NAM GAY BENH 8 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB3B.CXL .6627451 1 .6627451 .090 .7750 MAIN EFFECTS A:TB3B.DG 19.766254 1 19.766254 2.682 .1402 RESIDUAL 58.970588 8 7.3713235 ------------------------------------------------------------------------TOTAL (CORRECTED) 83.636364 10 ------------------------------------------------------------------------7 missing values have been excluded. All F-ratios are based on the residual mean square error.
Analysis of Variance for S GIA TANG VET BENH TREN LA TIEU NT X LY 2 DONGTRICHODERMA T.32 VA T.41 SAU KHI CHUNG NAM GAY BENH 8 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level -------------------------------------------------------------------------
109
COVARIATES TB3B.CXL MAIN EFFECTS A:TB3B.DG
.6627451 19.766254
1 1
.6627451 19.766254
.090 2.682
.7750 .1402
RESIDUAL 58.970588 8 7.3713235 ------------------------------------------------------------------------TOTAL (CORRECTED) 83.636364 10 ------------------------------------------------------------------------7 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.7 Ty le day tieu chet cua cac nghiem thc 8, 15 va 22 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu (%) Nghiem thc Lap lai T.32 bon T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P, C khong P,
I
6,7 46,7 0
8 NSC
15 NSC
22 NSC
II
6,7 20,0 6,7 13,3 0
III
6,7 6,7 13,3 6,7 26,7 0
IV
6,7 6,7 6,7 53,3 0
I
13,3 6,7 6,7 6,7 6,7 80,0 0
II
20,0 13,3 20,0 6,7 26,7 6,7 53,3 0
III
6,7 13,3 13,3 6,7 6,7 13,3 40,0 0
IV
6,7 13,3 13,3 26,7 33,3 0
I
13,3 6,7 6,7 6,7 6,7 80,0 0
II
20,0 13,3 20,0 6,7 26,7 6,7 53,3 0
III
6,7 13,3 13,3 6,7 6,7 13,3 40,0 0
IV
6,7 13,3 13,3 26,7 33,3 0
Phu luc 7.8 Ty le day tieu chet cua cac nghiem thc 29, 34 va 41 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu (%) Nghiem thc Lap lai T.32 bon
I
13,3
29 NSC
34 NSC
41 NSC
II
20,0
III
6,7
IV
6,7
I
20,0
II
20,0
III
13,3
IV
20,0
I
20,0
II
20,0
III
13,3
IV
20,0
110
T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P, C khong P,
6,7 6,7 6,7 6,7 80,0 0
13,3 20,0 6,7 26,7 6,7 53,3 0
13,3 13,3 6,7 6,7 13,3 40,0 0
13,3 13,3 26,7 33,3 0
6,7 6,7 6,7 16,7 86,7 0
26,7 33,3 13,3 26,7 6,7 60,0 0
53,3 20,0 13,3 6,7 13,3 53,3 0
46,7 33,3 13,3 26,7 33,3 0
6,7 6,7 6,7 16,7 100, 0
26,7 33,3 13,3 26,7 6,7 86,0 0
53,3 20,0 13,3 6,7 13,3 80,0 0
46,7 33,3 13,3 26,7 66,6 0
One-Way Analysis of Variance for TY LE DAY TIEU B CHET CUA CAC NGHIEM THC 41 NGAY SAU KHI CHUNG NAM GAY BENH ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 15235.433 6 2539.2389 13.808 .0000 Within groups 3861.776 21 183.8941 ------------------------------------------------------------------------Total (corrected) 19097.209 27 0 missing value(s) have been excluded.
Variance for TY LE DAY TIEU B CHET NT X LY 2 DONG TRICHODERMA T.32 VA T.41 SAU KHI CHUNG NAM GAY BENH 34 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB4B.CXL 17.388900 1 17.388900 .093 .7663 MAIN EFFECTS A:TB4B.DG 245.12042 1 245.12042 1.315 .2644 Analysis of RESIDUAL 3914.6538 21 186.41209 --------------------------------------------------------------------TOTAL (CORRECTED) 4177.1631 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Analysis
for TY LE DAY TIEU B CHET CAC CACH X LY TRICHODERMA SAU KHI CHUNG NAM GAY BENH 34 NGAY ------------------------------------------------------------------------of Variance
111
Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB4B.DG 245.12042 1 245.12042 1.318 .2645 MAIN EFFECTS A:TB4B.CXL 211.95743 2 105.97872 .570 .5746 RESIDUAL 3720.0853 20 186.00426 ------------------------------------------------------------------------TOTAL (CORRECTED) 4177.1631 23 ------------------------------------------------------------------------0 missing values have been excluded.
Phu luc 7.9 So choi mi moc cua cac nghiem thc thc 8, 15 va 22 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu Nghiem thc Lap lai T.32 bon T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P.
I 22 34 20 16 16 25 17
8 NSC 15 NSC 22 NSC
II 21 29 49 21 19 17 10
III 42 30 35 15 19 14 15
IV 36 36 24 17 17 17 13
I 25 50 20 21 21 26 19
II 29 39 55 22 27 30 14
III 45 37 38 18 25 20 17
IV 36 43 31 21 27 20 20
I 25 59 29 25 27 37 21
II 36 49 50 31 27 42 21
III 47 40 43 21 28 29 21
IV 22 34 20 16 16 25 17
Phu luc 7.10 So choi mi moc cua cac nghiem thc thc 29 va 41 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu Nghiem thc Lap lai T.32 bon
I 42
29 NSC 41 NSC
II 26
III 36
IV 50
I 42
II 27
III 36
IV 51
112
T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P.
48 34 27 33 22 28
61 31 28 30 39 21
49 59 32 27 43 22
40 44 21 28 29 21
48 35 28 33 22 28
62 34 30 30 40 21
49 59 33 27 44 23
41 45 21 28 29 22
One-Way Analysis of Variance for SO CHOI TIEU NON MI MOC CUA CAC NGHIEM THC 41 NGAY SAU KHI CHUNG NAM GAY BENH ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 2131.4286 6 355.23810 5.385 .0017 Within groups 1385.2500 21 65.96429 ------------------------------------------------------------------------Total (corrected) 3516.6786 27 0 missing value(s) have been excluded. Analysis of Variance for SO CHOI TIEU NON MI MOC CAC CACH X LY TRICHODERMA SAU KHI CHUNG NAM GAY BENH 41 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB1B.DG 1162.0417 1 1162.0417 15.490 .0008 MAIN EFFECTS A:TB1B.CXL 165.25000 2 82.625000 1.101 .3517 RESIDUAL 1500.3333 20 75.016667 ------------------------------------------------------------------------TOTAL (CORRECTED) 2827.6250 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error. Analysis of Variance for SO CHOI TIEU NON MI MOC NT X LY 2 DONGTRICHODERMA T.32 VA T.41 SAU KHI CHUNG NAM GAY BENH 41 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB1B.CXL 90.250000 1 90.250000 1.203 .2851 MAIN EFFECTS A:TB1B.DG 1162.0417 1 1162.0417 15.491 .0008 RESIDUAL 1575.3333 21 75.015873
113
------------------------------------------------------------------------TOTAL (CORRECTED) 2827.6250 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.11 Ty le choi tieu mi moc b chet cua cac nghiem thc 8, 15 va 22 ngay sau khi chung Phytophthora th nghiem phong tr benh tren cay tieu Nghiem thc Lap lai T.32 bon T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P,
I
35,3
8 NSC
15 NSC
22 NSC
II
10,2 21,1 -
III
6,7 -
IV
2,8 5,9 23,5 15,4
I
19,0 3,8 73,7
II
7,7 9,1 14,8 21,4
III
2,7 5,6 8,0 5,9
IV
4,7 22,2 20,0 15,0
I
12,0 5,1 3,4 8,0 14,8 18,9 85,7
II
11,1 20,4 13,8 16,1 14,8 28,6
III
2,1 10,0 9,5 27,1 13,8 9,2
IV
11,9 12,5 3,7 26,1 18,2 39,3
One-Way Analysis of Variance SO CHOI TIEU NON MI MOC B CHET CUA CAC NGHIEM THC 34 NGAY SAU KHI CHUNG NAM GAY BENH ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 21153.991 6 3525.6651 60.810 .0000 Within groups 1217.555 21 57.9788 ------------------------------------------------------------------------Total (corrected) 22371.546 27
Phu luc 7.12 Ty le choi tieu mi moc b chet cua cac nghiem thc 29 va 34 ngay sau khi chung Phytophthora nghiem phong tr benh tren cay tieu Nghiem thc Lap lai
I
29 NSC 34 NSC
II
III
IV
II
III
IV
114
T.32 bon T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P,
15,3 6,6 3,2 10,7 13,1 17,9 100,0
11,1 20,4 15,3 15,6 14,8 63,6
2,0 10,0 9,5 5,3 13,8 47,6
11,9 12,5 2,9 3,6 18,2 18,2 66,7
18,5 9,7 8,8 13,3 13,1 20,0 100,0
11,1 22,4 16,9 21,2 14,8 91,3
5,9 10,0 9,5 5,3 13,8 77,3
12,0 14,3 2,9 3,6 24,2 18,2 92,9
Analysis of Variance for SO CHOI TIEU NON MI MOC B CHET CAC CACH X LY TRICHODERMA SAU KHI CHUNG NAM GAY BENH 34 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB2B.DG 559.50727 1 559.50727 1.894 .1840 MAIN EFFECTS A:TB2B.CXL 400.73726 2 200.36863 .678 .5189 RESIDUAL 5909.5294 20 295.47647 ------------------------------------------------------------------------TOTAL (CORRECTED) 6869.7739 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error. Analysis of Variance for SO CHOI TIEU NON MI MOC B CHET NT X LY 2 DONG TRICHODERMA T.32 VA T.41 SAU KHI CHUNG NAM GAY BENH 34 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES TB2B.CXL 368.54401 1 368.54401 1.303 .2666 MAIN EFFECTS A:TB2B.DG 559.50727 1 559.50727 1.977 .1743 RESIDUAL 5941.7227 21 282.93917 ------------------------------------------------------------------------TOTAL (CORRECTED) 6869.7739 23 ------------------------------------------------------------------------0 missing values have been excluded.
Phu luc 7.13 Khoi lng bo re sau th nghiem phong tr benh tren cay tieu Nghiem thc Lap lai T.32 bon
I 34,3
Khoi lng re trung bnh (g) / chau

II 32,7 III 30,7 IV 33,3
115
T.32 phun T.32 bon + phun T.41 bon T.41 phun T.41 bon + phun C co chung P, C khong chung P,
36,0 29,3 30,0 32,7 38,7 19,5 42,4
32,7 39,3 32,7 32,7 41,3 24,8 52,0
33,7 34,7 29,3 30,3 39,0 22,5 52,4
32,3 34,0 32,0 33,3 32,7 20,8 40,0
One-Way Analysis of Variance KHOI LNG RE CAC NGHIEM THC SAU KHI KET THUC TH NGHIEM (60 NSC CHUNG NAM GAY BENH ) ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 1349.0967 7 192.72810 17.845 .0000 Within groups 259.2083 24 10.80034 ------------------------------------------------------------------------Total (corrected) 1608.3050 31 0 missing value(s) have been excluded. Analysis of Variance for KHOI LNG RE CAC CACH X LY TRICHODERMA SAU KHI KET THUC TH NGHIEM (60 NSC CHUNG NAM GAY BENH ) ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES KLRB.DG .1044560 1 .1044560 .013 .9101 MAIN EFFECTS A:KLRB.CXL 78.153356 2 39.076678 5.025 .0171 RESIDUAL 155.51968 20 7.7759838 ------------------------------------------------------------------------TOTAL (CORRECTED) 233.77749 23 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error. Analysis of Variance for KHOI LNG RE NT X LY 2 DONG TRICHODERMA T.32 VA T.41 SAU KHI KET THUC TH NGHIEM (60 NSC CHUNG NAM GAY BENH ) ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES KLRB.CXL 72.250000 1 72.250000 9.399 .0059 MAIN EFFECTS A:KLRB.DG .1044560 1 .1044560 .014 .9095
116
Phu luc 7.14 Ty le cay b chet canh, heo ot cua cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m (%) Nghiem thc Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I 50 70 50 30 90 90
Ty le benh 15 ngay SXL

II 60 80 60 30 90 90 III 50 70 70 40 100 100
One-Way Analysis of Variance TY LE CAY CHET CUA CAC NGHIEM THC TRONG TH NGHIEM SAU RIENG VN M SAU CHUNG NAM BENH 60 NGAY ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 9643.4223 5 1928.6845 5.424 .0077 Within groups 4266.6670 12 355.5556 ------------------------------------------------------------------------Total (corrected) 13910.089 17 0 missing value(s) have been excluded.
Phu luc 7.15 Ty le cay hoi phuc cua cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m (%) Nghiem thc Lap lai Ty le cay hoi phuc 45 ngay SXL
50 40 60
117
10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
50 50 50 30 10 50
50 50 50 30 20 40
60 70 40 40 10 60
Phu luc 7.16 Ty le cay chet han gia cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m (%) Nghiem thc Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I
10 10 10,01 10,01 20,01 50
45 ngay SXL
II
10,.01 10 10 10 20 40
60 ngay SXL
III
10 10,01 10 10 20 50
I
10,01 10,01 10 10 20 90
II
10 10 10,01 10,01 20,01 100
III
10 10 10 10 20 100
One-Way Analysis of Variance DIEN TCH LA CUA CAC NGHIEM THC TRONG TH NGHIEM SAU RIENG VN M SAU CHUNG NAM BENH 30 NGAY ------------------------------------------------------------------------Means plot: LSD Confidence level: 95 Range test: LSD Analysis of variance ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------Between groups 1613.2360 5 322.64720 4.964 .0108 Within groups 779.9587 12 64.99656 ------------------------------------------------------------------------Total (corrected) 2393.1946 17 0 missing value(s) have been excluded.
118
Phu luc 4.17 Dien tch la cua cac nghiem thc th nghiem phong tr benh tren cay sau rieng vn m (cm2) Nghiem thc Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I 66 72 79 82 68 48
45 ngay SXL
II 58 58 66 76 47 39 III 66 61 55 74 59 50 I 85 83 95 75 69 0
60 ngay SXL
II 76 82 94 56 75 1 III 84 91 93 72 61 0
Phu luc 4.18 Ch so diep luc to tren la cua cac nghiem thc th nghiem phong tr benh tren cay sau rieng ngoai ruong Nghiem thc Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I 23,7 28,8 23,4 24,6 21,1 21,7
45 ngay SXL
II 22,4 22,9 24,6 23,8 26,3 23,0 III 25,5 25,5 26,6 22,6 22,7 21,1 I
60 ngay SXL
II 31,1 32,8 33,4 32,5 23,3 20,7 III 25,2 32,5 35,0 34,4 21,1 18,7
24,2 30,9 29,0 26,9 20,0 21,4
Phu luc 7.19 Dien tch la cua cac nghiem thc th nghiem phong tr benh tren cay sau rieng ngoai ruong (cm2) Nghiem thc Trc XL 45 ngay SXL 60 ngay SXL
119
Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I 24 28 36 32 30 30
I 28 28 30 30 21 32
II 30 30 21 24 24 24
III 36 35 38,5 42 36,8 36
II 36 36 42 43,5 32 32
III 42 42 51,6 36,6 36 30
I 70 90 79,5 85,2 64,5 50,4
II 75 70 83,2 56 65 64,5
III 71 88,8 78,4 58,8 65,3 67,5
Analysis of Variance for DIEN TCH LA CUA CAC NGHIEM THC TRONG TH NGHIEM SAU RIENG NGOAI ONG SAU X LY CHE PHAM 30 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES DTLSRR.TRUOCXL .8840045 1 .8840045 .048 .8327 MAIN EFFECTS A:DTLSRR.T 243.61928 5 48.723856 2.653 .0824 RESIDUAL 202.02266 11 18.365697 ------------------------------------------------------------------------TOTAL (CORRECTED) 447.86000 17 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error. Analysis of Variance for DIEN TCH LA CUA CAC NGHIEM THC TRONG TH NGHIEM SAU RIENG NGOAI ONG SAU X LY CHE PHAM 60 NGAY ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES SB260N.TRXL 25.237338 1 25.237338 .296 .6032 MAIN EFFECTS A:SB260N.T 1110.8501 5 222.17002 2.602 .0865 RESIDUAL 939.24933 11 85.386303 ------------------------------------------------------------------------TOTAL (CORRECTED) 2135.3250 17 ------------------------------------------------------------------------0 missing values have been excluded. All F-ratios are based on the residual mean square error.
Phu luc 7.20 Tan suat xuat hien Phytophthora cua cac nghiem thc 15 va 30 ngay sau x ly cua th nghiem phong tr benh tren cay sau rieng ngoai ruong (%) 120
Nghiem thc Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I
Trc XL
I 75 75 100 81,2 81,2 75 II 75 75 81,2 87,5 81,2 68,7
15 ngay SXL
III 62,5 50 37,5 62,5 62,5 87,5 II 62,5 62,5 50 62,4 62,5 68,7 III 50 62,5 50 62,5 68,7 93,8
30 ngay SXL
I 31,3 31,3 12,5 50 50 87,5 II 25 25 25 37,5 31,3 75 III 31,3 25 18,7 43,7 37,5 87,5
87,5 87,5 75 75 87,5 87,5
Phu luc 7.21 Tan suat xuat hien Phytophthora cua cac nghiem thc 45 va 60 ngay sau x ly cua th nghiem phong tr benh tren cay sau rieng ngoai ruong (%) Nghiem thc Lap lai 10 g Tr. + P.Ch 20 g Tr. + P.Ch 40 g Tr. + P.Ch 20 g Tr. C co P.Ch C khong P.Ch
I 24 28 36 32 30 30
Trc XL
I 28 28 30 30 21 32 II 30 30 21 24 24 24
45 ngay SXL
III 25 25 12,5 43,7 43,7 87,5 II 25 18,7 25 31,2 37,5 87,5 III 18,7 18,7 12,5 43,7 31,2 81,2
60 ngay SXL
I 25 25 12,5 43,7 43,7 87,5 II 25 18,7 18,7 37,5 43,7 81,3 III 25,4 18,7 12,5 43,7 37,5 87,5
Analysis of Variance for TAN SUAT XUAT HIEN NAM PHYTOPHTHORA CAC NGHIEM THC TRONG TH NGHIEM SAU RIENG NGOAI ONG 60 NSXL ------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level ------------------------------------------------------------------------COVARIATES SB3.TRXL 33.557251 1 33.557251 3.865 .0751
121
MAIN EFFECTS A:SB3.T
9628.5217
1925.7043
221.772
.0000
122
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LVTN - Chon Loc Va Nhan Sinh Khoi Nam Trichodema

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Chng 2 TONG QUAN

Chng 3 NOI DUNG VA PHNG PHAP NGHIEN CU

a iem a Lat Tay Ninh Ba Ra Vung Tau Tien Giang

Ten nam Fusarium Sclerotium Phytophthora Rhizoctonia

Cay trong a Lan Ca chua Tieu Ca chua

100 100 100 100 100

0,5 0,5 0,5 0,5 0,5

1000 1000 1000 1000 1000

2:1 1:1 1:2 2:1

Chng 4 KET QUA VA THAO LUAN

+++ +++ +++ + +++ +++ ++ +++ + +++ +++

Th nghiem gia KL TB (g) 1,21 0,06 1.21 0,06 So sanh

c. Trc khi chung Phytophthora va x ly Trichoderma.

15,0 1,7 x 18,5 1,7 x x 22,5 1,7

16,8 2,4 x 21,1 2,4 x x 27,3 2,4

24,4 2,9 x 26,8 2,9 x x 35,2 2,9

x x 39,5 5,2 x 43,3 5,8 x 50,3 4,3

15 NSC SS So choi 23,16 2,1 37,33 2,1

22 NSC SS So choi SS 29,1 2,3 41,8 2,3

41 NSC So choi SS 30,4 2,5 44,3 2,5

20,7 3,4 x x 14,4 3,9 x

15 NSC SS Ty le choi chet 8,5 2,1 16,6 2,1

22 NSC SS Ty le choi chet 9,3 3,4 17,0 3,4 SS

41 NSC Ty le choi chet 11,0 4,9 20,7 4,9 SS

31.8 0,9 x 32.9 0,9 x 36.1 0,9

Chng 5 KET LUAN VA E NGH

Chng 6 TAI LIEU THAM KHAO

Chng 7 PHU LUC

50 g gia + 20 g ng 150 g gia + 20 g ng 100 g gia + 20 g ng 200 g gia + 20 g ng

1,5 1,5 1,6 1,4

1,2 1,3 1,3 1,7

1,1 1,5 1,4 1,6

1,1 1,5 1,4 1,7

1,5 1,5 1,6 1,4

1,2 1,3 1,3 1,7

1,1 1,5 1,4 1,6

1,1 1,5 1,4 1,7

58,0 57,0 21,0 50,0 21,0

61,0 56,0 19,0 43,0 52,0

62,0 47,0 25,0 51,0 43,0

73,0 67,0 26,0 47,0 38,0

2,7 6,3 17,0 25,0 27,0

3,6 8,2 15,0 14,0 15,0

3,8 8,9 17,0 17,0 16,0

4,2 8,2 17,0 17,0 16,0

20,0 23,8 19,1 24,8 11,4 19,1 0

25,7 20,0 13,3 20,0 16,2 24,8 0

14,3 9,5 8,6 26,7 13,3 26,7 0

23,8 15,2 21,9 24,8 10,4 25,7 0

23,8 23,8 20,0 24,8 12,3 25,2 0

32,4 21,0 13,3 21,0 21,0 34,3 0

15,2 11,4 10,5 26,7 15,2 28,6 0

34,3 17,1 22,9 40,0 12,4 34,3 0

26,7 29,5 20,0 28,6 15,2 46,7 0

36,2 28,6 14,3 21,9 23,8 53,3 0

16,2 16,2 15,2 28,6 19,1 38,1 0

42,9 18,1 25,7 42,9 13,3 47,6 0

III 40,0 19,1 29,5 15,2 37,1 20,0 53,3 0

IV 31,4 46,7 21,0 25,7 48,6 13,3 76,2 0

I 29,5 38,1 34,3 25,7 24,8 15,2 100, 0

II 29,5 44,8 37,1 17,1 22,9 24,8 95,2 0

III 40,0 19,1 29,5 15,2 37,1 20,0 85,7 0