Trng H Nng Lm TP.

HCM
NG DNG CNG NGH VI SINH TRONG SN XUT THUC TR SU VI SINH (Bacillus thuringiensis var)
GVHD : Th.S Vng Th Vit Hoa H v Tn Trn Cao Hiu MSSV 10139061

Nguyn Vn Giu
Nguyn Vn Phc Trn Nguyn Thanh Tm L Th Hng Thy

10139048
10139179 10139199 10139235 TP.HCM thng 10/2013

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NI DUNG
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Thuc tr su sinh hc Bt Vi khun Bacillus thuringiensis

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C ch gy c ca Bt
Cng ngh sn xut thuc tr su sinh hc Bt (Bacillus thuringiensis)
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HNH NH

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THUC TR SU Bt
Lch s pht trin: Nm 1901, vi khun Bacillus thuringiensis (Bt) c pht hin bi Ishwatari. Nm 1911, Ernst Berlinner nghin cu ra tc nhn gy bnh trn loi su xm tnh Thuringia, c ca loi Bacillus thuringiensis Nm 1938, Bacillus thuringiensis c s dng lm thuc tr su ti Php v n nhng thp nin nm 1950, Bt c s dng rng ri ti M.
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THUC TR SU Bt
Bt c nghin cu v ng dng rng ri trn nhiu quc gia nh : Hoa K, Lin X C, Php, c, Trung Quc, Nht Bn nc ta, cui thp k 80 mt s c quan nghin cu khoa hc bt u sn xut cc ch phm dit su t Bt. Hiu lc ca cc ch phm Bt sn xut trong nc i vi su t, su xanh, su cun l loi nh tng ng t : 60% 100% , 12% 32%, 28% 100% .
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THUC TR SU Bt
Cc lai sn phm thng mi c trn th trng nh Vi-BT 32000WP, 16000WP; BT Xentary 35WDG, Firibiotox P dng bt; Firibiotox C dng dch c c ...

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VI KHUN Bt
Trc khun Gram (+), kch thc 1-1,2 x 3-5 m Ph tim mao, di chuyn c Sinh bo t, kch thc 1,6 2 m Hiu kh hoc hiu kh khng bt buc

T bo ng ring hoc xp chui

To ra tinh th protein trong giai on to bo t, kch thc 0,6 2 m Ph bin trong t nhin, c tr trong t, trn b mt cy v trn xc su
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VI KHUN Bt
Nhu cu dinh dng: Ngun cacbon: tinh bt, maltose, glucose. Ngun nit: cao tht b, pepton, bt men, bt bnh lc. Mui v c: K2HPO4, MgSO4, CaCO3

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VI KHUN Bt
Vi khun Bt sinh trng nhit 12-400C. Nhit thch hp 27-320C. 35-400C chng sinh trng nhanh nhng chng lo ha. Nhit thp chng sinh trng chm. pH thch hp l 7,5. 8,5 vn c th hnh thnh bo t. pH=5 th khng hnh thnh bo t.

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VI KHUN Bt
Lm ngng kt sa Trong ng gluco glyxerol s to axit Phn ng dng vi metyl

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VI KHUN Bt
Chu trnh sinh trng v pht trin Th sinh dng Nang bo t Bo t v tinh th

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VI KHUN Bt
Th sinh dng: Dng que, hai u t, kch thc 1,2-1,8 m x3-5 m, bt mu gram dng. Lng mc xung quanh, hi ng hoc khng ng, thng tn ti 1 hoc 2 c th lin nhau. Sinh sn theo kiu phn chia ngang. Trong thi k sinh sn c 2,4,8 th dinh dng lin nhau thnh chui.

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VI KHUN Bt
Nang bo t: khi cc th vi khun gi, mt u no trong c th hnh thnh bo t hnh bu dc, cn u kia hnh thnh tinh th hnh thoi. Nang bo t hnh trng di, to hn th sinh dng.

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VI KHUN Bt
Bo t v tinh th: kch thc bo t t 0,8-0,9 m x 2 m. Tinh th c kch thc t 0,6-2um. Hnh thoi, cng c loi hnh trn, bu dc. Tinh th l mt protein l cht dit su hiu qu ch yu.
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C T CA Bt
Ngoi c t (alpha- exotoxin) Ngoi c t (beta- exotoxin) Ngoi c t y(gamar-exotoxin)

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Ngoi c t (phospholipase C)
1953, ln u tin Toumanoff pht hin thy BT var. elesti sn sinh enzyme lethinase. Phn hy mang tnh cm ng ca Phospholipit trong m ca cn trng, lm cn trng b cht. u tin enzyme ny lin kt vi t bo rut ca cn trng, sau tch ra v c hot ha bi mt cht khng bn nhit, cht ny c trng lng phn t thp, c th l lipit.

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Ngoi c t
c tm ra vo nm1959 khi Halt v Arkawwa nui u trng rui nh bng thc n c cha BT. Thnh phn gm adenin, riboza v phospho (1:1:1). Ha tan trong nc, bn vng nhit cao,c th chu c nhit 120-1210C trong 10-15 pht. V th gi l ngoi c t chu nhit.

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Ngoi c t (tt)
Xut hin trong giai on vi khun pht trin mnh, trc khi hnh thnh bo t. L mt nucleotit c trng lng phn t thp, c adenin, riboza, phospho vi t l bng nhau.

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Ngoi c t (tt)
Tc ng km hm nucleotidaza, ARNpolymeraza. Cn cng hng vi ni c t gy thay i sinh l dn n ci cht nhanh cho u trng. Hiu qu trong vic chng su non (gy tr tr trong vic chuyn ha lt xc).

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TINH TH C
Ni c t (delta- endotoxin) Tinh th c Cry c to ra vi mt lng ln hn nhiu so vi cht c Cyt v l tc nhn c hiu qu chnh trong vic gy c cho cn trng.

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Tinh th c
Tinh th c khng ha tan trong nc hoc cc cht hu c nhng c th ha tan trong dung dch kim. C hn 50 gene m ha cc protein tinh th c, c th chia protein tinh th c thnh 15 nhm da trn s ging nhau trong trnh t gene

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Ni c t
c hn trong 50 gen m ha cho protein tinh th c c gii m cho php phn loi cc cht c ny thnh 15 nhm Khi lng Hnh dng
Gen tinh th protein tinh th (kDa) i tng dit

Cry I (nhiu nhm ph: Hnh thoi A(a), A(b), A(c), B, C, D, E, F, G) Cry II (nhm ph: A, B, Hnh khi C) Cry III (nhm ph: A, B, C) Cry IV (nhm ph: A, B, C) Cry V IX
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Cc u trng b cnh vy (Lepidoptera) Cn trng b cnh vy (Lepidoptera), b hai cnh (Diptera) Cn trng b cnh cng(Coleoptera) Cn trng hai cnh (Diptera) Nhiu loi

69 71 73 74

Hnh phng hoc v nh hnh Hnh thoi a dng

73 134 35 129

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Ni c t (tt)
Cc dng tinh th ni c t: Dng nh thp Dng hnh cu Dng hnh vung Dng khng n nh Dnh hnh lm

( theo Rn. Gaixin, Feng Xichang v Feng Weixiong, 1983)

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Vng chc nng ca Cry

Vng I: l mt b gm 7 chui xon , mt vi chui hoc tt c cc chui c th ci vo mng t bo rut, to ra cc l cc ion qua li t do Vng II: cha 3 di khng song song tng t nh vng gn khng nguyn ca globulin min dch, vng ny c chc nng gn vi th th trn b mt t bo biu m rut.

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Vng chc nng ca cry

Vng III: bo v c t c hot ha khi b phn hy bi protease rut. Vi cu trc v hot tnh nh vy, tinh th c

lin kt mt cch c hiu vi mng t bo biu m rut ca su

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C CH GY C

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C CH GY C
Khi su n phi l c vi khun, tinh th c cng vi bo t xm nhp vo c th. Trong iu kin bnh thng, tinh th khng ha tan. Khi i vo rut gia ca su ni c pH>9,5 lm cho tinh th c tan ra. Dng ha tan ny cha phi l dng hot ng. Dng tin c t ny c protease ha trong rut gia ca su hot ha thnh c t

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C CH GY C
c t ny gn vo t bo biu m thnh rut, lm cho rut su b t lit dn n ngng n. ng thi c t ny m qua mng to thnh l xuyn mng dn n mt cn bng ion lm vi nhung mao trng phnh ln. y l nguyn nhn lm su cht. pH trong rut gim xung bng pH ni mi trong huyt tng. pH thp ny cho php cc bo t ny mm, xm chim vt ch cui cng gy cht.

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C CH GY C
Thuc tr su BT Nguy him cht su !

Su ngng n,mng nc ln v cht

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1 s loi su m BT c th dit tr

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Su Lymantria dispa

Helicoverpa armigera

B cnh cng Coleoptera

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B 2 cnh Diptera. Click to edit Master subtitle style

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CNG NGH SN XUT Bt

Sn xut ch yu theo 2 cch: ln men thng (ln men b mt) v ln men chm c sc kh. Cng ngh ln men xp t hiu qu khng cao nn ch yu dng phng php ln men chm.

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CNG NGH SN XUT Bt

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CNG NGH SN XUT Bt

Ging: thng s dng chng Bacillus thurgiensis. Ging c nhn ln sau mi cp gp khong 1015 ln ging c. Chun b mi trng: Mi trng thng s dng nui cy Bt bao gm cc thnh phn sau: Ngun cacbon l tinh bt, maltose, glucose. Nu l tinh bt thng dng l go, ng, sn.

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CNG NGH SN XUT Bt

Tinh bt qua giai on thy phn to ng n v ng i s dng amylase hoc H2SO4 trong iu kin nhit cao thy phn. Ngun N: thng s dng l bt c, bt ng, bt banh lc, bt bnh u, cao tht b, peptone, bt men,. Mui v c thng b sung vo l cc mui sau: K2HPO4, MgSO4,,CaCO3

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CNG NGH SN XUT Bt

Kh trng: Sau khi chun b mi trng xong nn i kh trng vi cc ch kh trng sau: Kh trng bng phng php gin on: tin hnh p sut d 0.05-0.1Mpa vi nhit t 110-1200C trong vng 1 1,5 gi. Hoc kh trng lin tc: tin hnh nhit cao hn 140-145 v thi gian ngn hn 5- 15 pht

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CNG NGH SN XUT Bt

Ln men: Tin hnh trong ni ln men c dung tich 500l, 1000l, 2000l , iu kin nhit t 27320C iu chnh pH =7,5 trong khong thi gian t 48-72h. Trong qu trnh ln men tin hnh thi kh. Ch thi kh: ngng thi kh ln nht cho qu trnh ln men l 0,5-0,6m3 mi trng/m3 khng kh.

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CNG NGH SN XUT Bt

Cc yu t nh hng n qu trnh ln men: Nhit : nh hng n s hnh thnh bo t. Nu nhit qu cao hoc qu thp s ko di hoc rt ngn qu trnh ln men. Nhit thch hp l 300C. Cc ch lun chuyn ging: nh hng n s hnh thnh bo t v tinh th c. Nu s dng lin tc s xy ra hin tng nhim thc khun th. V vy ln men khong 10-15 ln ging c th cn phi thay ging mi.
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CNG NGH SN XUT Bt

nh hng ca pH: vi khun Bt thch hp trong mi trng kim pH=7.5, iu kin 8,5 vn c th hnh thnh bo t, nu pH =5 khng hnh thnh bo t. nh hng ca oxi: vi khun loi hiu kh, phi c oxi mi sinh trng tt, nht l khi hnh thnh bo t. Nu thiu oxi khng hnh thnh bo t hoc hnh thnh chm. Lng oxi thng cung cp cho qu trnh ln men l 0,5-0,6m3 mi trng/m3 khng kh l tt nht.
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U NHC CA THUC TR SU Bt
u im
Khng c hi cho ngi,gia sc,mi trng

Nhc im
B nh hng bi thi tit

Khng nh hng n cht lng nng sn

Khng tn hi n nhng sinh vt c ch Hiu qu ko di qua nhiu th h su hi Hiu qu kinh t cao

Ph tc ng hp Hiu qu ban u khng cao

Tc ng chm (1-3 ngy)

Sn xut phc tp,gi thnh cao

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KT LUN
Bacillus thuringiensis c ng dng ht sc to ln trong vic sn xut cc ch phm tr su sinh hc. Trn th gii hin nay, cc loi thuc tr su ha hc ang c xu hng c thay th dn bi cc loi thuc tr su sinh hc. Nhu cu bc thit l phi pht trin cc loi thuc bo v thc vt mi, c tc dng dit su cao hiu lc nhanh chng, khng lm nh hng n mi trng, khng to nn tnh khng thuc ca su hi. V Bacillus thuringiensis l mt trong nhng i tng nghin cu ht sc thun li. Nhng thnh tu t c ca thuc tr su Bt khng ch mang li nhng li ch r rng m cn ha hn tng lai vi nhiu ng dng mi nhm pht trin nhng bin php c hiu qu hn na trong vic khng ch dch hi.
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TP.HCM thng 10/2011