B GIO DC V O TO TRNG I HC NNG LM THNH PH H CH MINH

BI BO CO MN CNG NGH SN XUT THUC BVTV TI: NM BEAUVERIA BASSIANA V NM ENTOMOPATHOGENIC (EPF) KHC TRONG QUN L CN TRNG GY HI GVHD: PGS.TS.Phan Phc Hin Nhm bo co: 4 1. V Vn Nhn 2. Nguyn Hoi Nam 3. Phm Thanh Vn 4. L Mu Hong

TPHCM 11/2012

Mc Lc
Mc Lc....................................................................................................................... 2 CHNG 1: LI M U.............................................................................................3 5. Cng ngh sn xut thuc bo v thc vt t nm...............................................10 5.1 Phng php ln men chm..............................................................................10 5.3 Cc giai on trong quy trnh sn xut nm....................................................11 5.3.1 Chun b nguyn liu ( hay mi trng ln men).......................................11 5.3.2 Phn lp ging nm thun.........................................................................12 5.3.3 Cc mi trng nhn ging........................................................................13 5.4 Cc thng s cn ch trong qu trnh sn xut.............................................13 5.5 Thnh phm.....................................................................................................14 6. Mt s ch phm thuc tr su cha beauveria bassiana.....................................14 6.1 Thuc tr su Muskardin..................................................................................14 6.2 Ch phm Biovip..............................................................................................15 6.3 Thuc tr su sinh hc AT vi sinh.....................................................................15 Kt lun.....................................................................................................................16

CHNG 1: LI M U
1. t vn
-

Nh c gii thiu bi bo co trc th thuc tr su sinh hc c sn xut t bn ngun chnh l: vi khun, nm, tuyn trng v virus. c s phn cng v hng dn ca thy Phan Phc Hin nn hm nay nhm xin c trnh by v ti: Nm Beauveria bassiana v nm Entomopathogenic (EPF) khc trong qun l cn trng gy hi.

2. Mc tiu
-

Gii thiu s lc v nm Beauveria bassiana v nm Entomopathogenic (EPF). Gii thiu quy trnh sn xut 2 loi nm trn Mt s ch phm ph bin Tho lun nhm Tm hiu ti liu trn mng. Tm c cc ti liu lin quan trn th vin, ti liu ca GVHD.

3. Phng php

CHNG 2: NI DUNG
1. Mt s loi nm c kh nng gy bnh cn trng hi cy trng - C khong 200 loi c th k sinh gy bnh ln cn trng tp trung ch yu ngnh nm tht Mycobionta, ngnh ph lp nm bt ton Deuteromycetes (Fungi imperfecti). - Da theo c im hnh thi, lp nm bt ton c chia lm 4 b: + B nm ti gi +B nm a gi +B nm bt th +B nm bng - H nm bng Moniliaceae c rt nhiu loi k sinh v gy bnh cho cn trng tp trung ch yu mt s chi: + Chi Beauveria + Chi Nomuraea + Chi Metarhizium + Chi Paecilomyces + Chi Hirsutella + Chi Verticilium 2. Mt s c im c bn ca chi nm Beauveria v Metarhizium A. Metarhizium (nm lc cng) Gm c hai loi chnh: + M.anisopliae + M.flavoviride c im hnh thi: C si nm v bo t, lc u mu trng sau chuyn sang mu xanh. Cun sinh bo t ngn, bo t trn, dng hnh oval, hnh trng, c chai hoc hnh tr.
-

Kch thc bo t khong 3.5-4 m c t:

+ Ngoi c t c tn l Destruxin A: C29H47O7N5, B: C30H51O7N5, C hoc D. + Cc ngoi c t l sn phm th cp, vng peptit. C ch gy bnh ca nm M.anisopliae ln cn trng: Khi bo t nm lc cng bm trn b mt cn trng trong khong 24h th bo t s ny mm to thnh ng mm xuyn qua v cn trng, sau tip tc phn nhnh to nn mt mng si nm chng cht bn trong c th cn trng, cng ging nh nm Beauveria Bassiana. Nm Metarhizium anisopliae (Ma) tit ra cc c t Destruxin A,B v chnh cc c t trn gy cht cn trng. B. Chi Beauveria (nm bch cng) Gm 3 loi chnh c kh nng tiu dit cn trng: + Beauveria bassiana + Beauveria tenella + Beauveria brongniartii 3. c im ca nm Beauveria bassiana (Bb)
3.1

c im hnh thi

- Nm Bb sinh ra nhng bo t trn n bo, khng mu, hnh cu hoc hnh trng, ng knh t 1-4 m, si nm c ng nm ngang t 3-5 m pht trin mnh trn mi trng nhn to hoc trn c th cn trng. - Chng mang nhiu gi sinh bo t, phng to di vi kch thc 3-5 x 3-6 m. - Cc gi bo t trn thng to thnh cc nhnh phn ngn hoc trc tip to thnh cc nhnh ca gi, phn ngn ca bo t c dng cung hp hnh zch zc khng u. 3.2 c t ca nm Beauveria - L Boverixin, vng Depxipeptit c im si 93-940 C - Cng thc nguyn l C45H57O9N3. Nu nui cy trong 1 lt mi trng cc nh khoa hc tch c 1,5-3,8 gam c t Boverixin. 3.3 C ch tc ng ln c th cn trng

Trong t nhin, khi bo t nm Bb ri vo c th cn trng, gp iu kin

thi tit thch hp, ch sau 12-24h th bo t nm ny mm, chng hnh thnh si

m xuyn qua lp v kitin sau pht trin bn trong c th ca cn trng, cn trng phi huy ng cc t bo bch huyt chng , nhng nm Beauveria tit ra c t Boverixin c cha proteaza v mt s cht khc ph hy ngay c t bo bch huyt lm cho su cht, si nm mc rt nhiu trong c th su v sau chui ra ngoi, to 1 lp bo t ph trn c th su. Nm Metarhizium anisopliae gy hi cho b ry, b xt v nhiu loi su hi. Khi bo t nm ri trn c th cn trng s ny mm v mc vo trong c th cn trng. Metarhizium c th gii phng cc bo t (conidia) ngay c trong iu kin m thp (50%). Ngoi ra, Metarhizium anisopliae c th c c dinh dng t lipid trn lp da ngoi ny ca cn trng. Cc loi nm cn sn xut ra cc cht chuyn ha th cp, nh destruxins l ngoi c t. Nm pht trin bn trong c th cn trng v n cht b ca c th cn trng. Sau khi thm nhp vo, chng pht trin nhanh chng bn trong lm cn trng cht. Cc cn trng nhim nm tip xc vi cn trng khc s lm cho nhng cn trng ny b nhim Metarhizium anisopliae. Khi cn trng cht, nm xut hin. Lc u hnh thnh mt lp trng nhng ch ni gia cc t trn c th cn trng. Nu l nm M.anisoplae, chng s chuyn thnh mu xanh lc m. Bo t xut hin t k ch cht li ly lan sang k ch mi nh gi hoc nc. Khng ch ph hy chc nng sinh l ca cn trng, c t ca nm cn lm nh ch s dinh dng ca cn trng, gy hin tng t lit, ph hy qu trnh h hp, chn p c hc s tun hon mu do cc th si nm pht trin v sinh sn trong xoang thn cn trng. Theo s liu ca Sekhrina T.A (1960), khi cho nhng hay con ci b cnh cng nhim nm th thy mn ca cn trng gim ng k. Cc con ci b Chelonia khi x l Boverin (ch phm sinh khi nm Beauveria bassiana), s lng trng gim hn i chng 41,3% trong iu kin phng th nghim. Theo Gamper N.M.etal (1986), qua nghin cu, nhiu tc gi cc nc u thy rng trng giai on non ca s pht trin bo thai s mn cm cao hn nm. Nm cng nh hng ln giai on u trng ca cn trng. Thng bo ca Prasertphon S. (1997) cho thy c t c to thnh mt s nm Entomophthora:

E.apculata, E.cornata khi tim vo xoang c th nhng Galleria mellonella lm chng b c ch qu trnh bin thi v cht. Nm cng nh hng n s hp th oxy ca cn trng. Bnh l ca cn trng b nhim cng ging nh cc ng vt c xng sng, mt trong nhng du hiu ch yu ca bnh l s bin i h hp. Cng trnh nghin cu ca Sussman A. (1992) cho bit nhng b nhim nm ch t lu sau s hp th oxy ca chng tng ln 7 ln. c gi thit cho rng cn trng b cht do cc kh qun v l th b ph hy trc tip hay bng cch ph hy h thn kinh.

V bnh l m hc, s tn cng ca nm gy bnh vo thn ch c th l c thc hin bng s xm nhp trc tip qua v, qua thnh ng tiu ha, qua l th hay ch b tn thng v cuticun. Cc nm dit su xm nhp vo c th cn trng qua ton b v cuticun ngoi (exocuticun). S xm nhp ny c thc hin bng lc c hc, khi tin su vo lp cuticun trong (endocuticun) l nh vai tr ch yu ca hot ng enzym do nm sn sinh ra. Ngoi vic tit enzym, cc nm cn tit ra c t trc tip vo huyt tng trong qu trnh sinh sn ca n.

Trong qu tnh pht trin nm Bch cng tit ra mt c t, chnh c t ny lm cho cn trng b cht. Khi bo t nm gp phi c th cn trng, chng s ny mm mc thnh si nm xuyn qua v kitin v pht trin trong c th lm tiu hao cc t bo bch huyt v cui cng lm cn trng b cht, trn c th ph kn lp phn trng. Khi cht c th cn trng cng li, cc bo t tip tc pht tn trong khng kh. Nm Lc cng, sau khi ri trn b mt c th cn trng trong 24 gi nm s mc si m xuyn qua v cn trng, pht trin thnh cc nhnh chng cht trong c th, trong qu trnh pht trin chng tit ra cc c t lm cho cn trng cht. Sau khi cht cn trng c lp mu xanh trn b mt c th

4. ng dng mt s nm entomopathogenic (EPF) trong qun l cn trng gy hi - Cc chng nm ca EPF: Beauveria bassiana, Metarhizium anisopliae, Nomuraea rileyi, Paecilomyces spp., Lecanicillium, - EPF thuc v ba ngnh: Ascomycota (cc lp :Laboulbeniales v Pyrenomycetes) Deuteromycota (class Hyphomycetes), v Zygomycota (lp nm tin hp), chia thnh 12 lp. 4.1 Tng quan EPF - Hai ging, Aschersonia v Tetranacrium, ca lp Coelomycetes l tc nhn gy bnh quan trng trn rui trng v cn trng khc. - Lp Hyphomycetes bao gm hn 40 ging entomopathogenic to ra trn th gii. N bao gm Verticillium lecanii, loi c th hnh thnh bo t. - Sn xut Viviparous tr c th phn lp trn rp b nhim bnh. - Culicinomyces clavisporus v Tolyplocadium cylindrosporum ly nhim u trng mui os.

4.2 ng dng
-

Nm entomopathogenic c th pht trin bng bo t hoc si nm. Vic s dng nm entomopathogens thay th cho thuc tr su hoc ng dng kt hp thuc tr su vi entomopathogens nm c th s rt hu ch cho vic qun l kh nng khng thuc tr su.

Chng sinh enzym thy phn ngoi bo cht, cht nhy kt dnh, thm nhp vo dch hi, ly lan ca si nm v sinh cht c hi. nghin cu tng hp phn lp 36 Bb, vi s hin din ca 9 enzyme ngoi bo tham gia vo vic ph hy lp chitin, phn t protein v polysaccharides.

5. Cng ngh sn xut thuc bo v thc vt t nm 5.1 Phng php ln men chm.

S cng ngh
Nguyn liu Kh trng Lm ngui Cy ging cp 1 Lc trn my lc Ging cp 2 Thu sinh khi Cy vo mi trng ln men xp (Ly tm, sy..) Thnh phm (dng bt hoc lng) Hnh 1: Quy trnh sn xut nm bng phng php ln men chm. (Phm Th Thy, 1994) Nguyn liu Kh trng Lm ngui

5.2 Phng php ln men xp.

S cng ngh:
ng ging thun Mi trng nhn ging cp 1 Mi trng nhn ging cp 2 Ri ra nia hnh thnh bo t trn ( 2 ngy) Thu sinh khi, sy 40 450C trong 6 8h Hn hp ph gia to ch phm nm. Kim tra cht lng bo t nm Th hot lc trn su ng gi, bo qun v s dng

Hnh 2: Quy trnh sn xut nm Ma theo phng php ln men b mt. - Mi trng nhn ging cp 1 kh trng 0.8 at trong 30 pht. - Mi trng cp 2 v mi trng ln men xp phi kh trng 1 at trong 30 pht. 5.3 Cc giai on trong quy trnh sn xut nm. 5.3.1 Chun b nguyn liu ( hay mi trng ln men) - Mi trng bao gm: bt cm go bt ng bt u tng tru ( hoc b ma..) theo t l 5:3:1:1. - Nguyn liu trc khi vo ln men cng vi nm thun chng phi qua s ch, loi b tp cht, qua thanh trng nhm hn ch s pht trin ca cc vi sinh vt khng c li trong cng ngh.

- Thanh trng l qu trnh gia nhit n 60-90C, nhm tiu dit nhng vi sinh vt gy hi nh vi khun, virut, nm men, nm mc. n v PU th hin hiu qu ca qu trnh thanh trng trong mt khong thi gian xc nh. - Qu trnh thanh trng gip lm gim lng vi sinh vt trong cc sn phm. Mt trong nhng phng php thanh trng ph bin l s dng hm thanh trng c trang b cc ng phun. V cn thit phi xc nh c thi gian v nhit ti thiu qu trnh thanh trng va c hiu qu tiu dit vi sinh vt, va khng to ra cc mi v khng mong mun cho sn phm, ng thi tit kim c nng lng s dng. - n v thanh trng (PU) l n v c dng nh gi hiu qu ca mt qu trnh thanh trng mt sn phm gi iu kin nhit 60C trong thi gian 1 pht. Khi nhit t n 60C trong 1 pht th gi tr PU l 1 n v. 5.3.2 Phn lp ging nm thun. - Sn xut nm Metarhizium flavoviride phi chn ging chng l Metarhizium flavoviride c phn lp trn co co, chu chu. - Sn xut nm Metarhizium anisopliae phi chn ging chng l Metarhizium anisopliae c phn lp trn ry nu hoc l trn b hi cy da. - Hoc c th phn lp Beauveria bassina phn lp t su rm thng. - Cc loi nm ny phn lp trn cn trng sau khi chng cht. Mi trng phn lp.
-

Mu xc cn trng b cht ha nc theo phng php pha long, gieo cy trn mi trng thch petri, nui 5 7 ngy nhit t 28 - 300C. Mun hn ch vi khun khng c li ta b sung vo mi trng phn lp mt s cht nh sau: + NaCNS: 0.24 0.4M/l + H tetracyclin: 2 -5mg/l + Neonicin, polymycin, bacytracin: 50mg/l

5.3.3 Cc mi trng nhn ging. 5.3.3.1 Mi trng nhn ging cp 1 Mi trng Sabouraud - Agar - Pepton 20g 10g

- Glucoza 40g - MgSO4.7H2O 0.5g - KH2PO4 - H2O - pH 6 5.3.3.2 Mi trng nhn ging cp 2. Mi trng Sabouraud Dextroza Agar Cao nm men (i vi ln men chm) - Cao nm men - Pepton - Sacaroza - Agar - H2O - pH 10g 15g 10g 20g 1000ml 6.5 5.4 Cc thng s cn ch trong qu trnh sn xut. - Nhit : nhit thch hp trong khong 25 - 30 0C, nu nhit qu cao hay qu thp th bo t s b cht hoc khng sinh bo t. - m : trong phm vi 80 90% - Mi trng nui cy: phi b sung thm cht kitin v glucose hnh thnh s lng bo t ln. - Kitin trong mi trng gip cho chun nm sinh kitinaza c tt hn, gip cho nm xuyn qua lp biu b su, gip cho nm pht trin cng nh hnh thnh bo t nh. 1g 1000ml

- Ngoi yu t ngun C v N cn ch ti cc nguyn t khong nh Co, Zn, cng vi vitamin. - pH: pH ti thch nm trong khong 5,5 n 6.5. - Thng kh: do qu trnh l ln men hiu kh nn cn phi to ra thng thong cn thit, mi trng c xp, thng kh tt ngi ta thng b sung tru vo. 5.5 Thnh phm. - Sn phm sau qu trnh ln men c hon thnh dng lng, dng bt mn hoc mnh, ht. Sn phm dng bo t l tt nht, d s dng. - S lng bo t t t 5 t bo t/1g ch phm. - Hm m trong sn phm phi m bo t 7 10%. - pH : 6 - Thi gian bo qun: 12 thng - Hiu qu dit su: 70 90% sau 15 ngy th nghim. - Cn bo qun ni thong mt, trnh tip xc vi nh nng mt tri. - khi s dng ch phm dng bt phi tin hnh pha long mt t bo vo khong 107-108/ml ri phun vo t hoc trc tip ln cy. 6. Mt s ch phm thuc tr su cha beauveria bassiana 6.1 Thuc tr su Muskardin - Muskardin ch phm mi ca CPC. Sn phm Muskardin l thuc tr su sinh hc, thnh phn chnh l nm Beauveria Bassiana (nm trng), c ch bin di dng bt thm nc. - Muskardin c ph tc ng rt rng trn nhiu loi cn trng nh : Su t, Su xanh trn rau mu; su c thn, su cun l, ry nu trn la; su c thn hi bp; B xt mui, B cnh t, ry xanh trn cy Ch.

6.2 Ch phm Biovip - Vin La BSCL nghin cu thnh cng 2 ch phm Ometar v Biovip, 2 ch phm ny c hiu qu cao trong phng tr mt s su hi cy trng nh: ry nu, ry xanh, b xt hi la, ry xanh v ry mm hi cy bng vi. Ometar cn c hiu qu rt cao khi dng tr b cnh cng hi da. - Ch phm sinh hc Ometar v Biovip c sn xut t 2 chng: nm xanh, Metarhizium anisopliae c phn lp t con b xt hi hi la v nm trng, Beauveria bassiana c phn lp t con ry nu hi la. 6.3 Thuc tr su sinh hc AT vi sinh - Ch phm AT tr su sinh hc dng lng c thnh phn Metarhizium anisipoliae, Beauveria bassiana v Isaria sp - Hiu lc phng tr i vi cc loi su c qu 15ong, su sng ng, su c qu, b nhy trn l, su c qu u, nhng ha bm, su bm gi, su c ci, su khoang, su ct l, rp vng, b tr, su cun l, su c l, b xt trn l, b chch ht, ry phn trng

Kt lun Thuc tr su vi sinh t nm hin nay v ang c p dng rng ri dit tr cc loi su bnh c hi cho cy trng. Trong nm bch cng v nm lc cng l 2 loi nm ph bin ang c cc nh nghin cu cng nh cc cng ty cho ra nhiu sn phm mi c th dit tr su bnh bo v cy trng. Vic s dng thuc tr su vi sinh t nm gp phn gim thiu nhim mi trng, h chi ph, khng gy hi cho ngi s dng. Ti liu tham kho
1. Gio trnh cng ngh sinh hc trong bo v thc vt PGS.TS Phm Th Thy.

2. Lng c Phm, Sn xut v s dng ch phm sinh hc trong nng nghip, Nh xut bn gio dc Vit Nam.
3. Beauveria bassiana and other

entomopathogenic fungi in the

management of insect pests -G.G.Khachatourians, E.P.Valencia1 and G.S.Miranpuri2

4. http://baigiang.violet.vn/present/show/entry_id/4195966 5. http://www.tstcantho.com.vn/?mod=article&id=23&section_id=21

6. http://www.khoahoc.com.vn/doisong/ung-dung/26027_Su-dung-Ometar-phattrien-nong-nghiep-ben-vung.aspx