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CNG NGH SINH HC THC VTGV: ThS. BI VN TH VINHNHM 1 LP 10DSH01:DNG QUC XUNCHUNG NGC MI MING TH KIM PHNGO NGUYN THANH VY
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CHNG IIICNG NGH PHI V TNH HT NHN TO
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PHI V TNH
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1. KHI NIM V PHI V TNH
Phi v tnh, phi soma, phi sinh dng hay phi th h u l khi nim m t mt cu trc lng cc bt nh bao gm cc chi v cc r, m di nhng iu kin thch hp th c th pht trin thnh mt c th c chc nng hon chnh.
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Cc t bo c kh nng pht sinh phi c cc c tnh c bn sau:
T bo c kch thc nh, ng knh.T bo cht m c, hnh nhn ln.Hot ng bin dng mnh m.Hm lng protein v RNA cao.C mt s lng ln ribosome, ty th v ht tinh bt.
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2. MT S KHA CNH GI NH V S HNH THNH PHI V TNHS cm ng hnh thnh phi v tnh phi lm kt thc s biu hin ca mt gene trong m thc vt ti thi im v thay th bng chng trnh biu hin ca gene sinh phi. Theo Sharp v cc ng nghip (1980) s to phi soma c th theo 2 con ng:Phi pht sinh trc tip khng qua giai on to m so (con ng PEDC).S to phi soma cn c giai on to m so ban u (con ng IEDC).
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Hai danh t t bo tin phi PEDC (Preembryogenic determined cell) v t bo pht sinh phi IEDC (induced embryogenic determined cell) dng phn loi m, nhng thc cht l 1 qu trnh tip ni nhau, kt thc s pht trin l s h thng nhng t bo phi (EC- Embryogenic cell).Nu m cy c cc PEDC th ch cn mt s kch thch phn chia t bo l hnh thnh phi => hnh thnh phi trc tip.Nu m cy l nhng t bo phn ha khng cn kh nng sinh phi th chng phi tri qua nhiu ln phn chia t bo lin tip di s cm ng ca auxin to m so => hnh thnh phi gin tip.
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Mi t bo phi soma u pht trin qua 3 giai on trc khi trng thnh: Hnh cuHnh tim Hnh thy li
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Hnh cuHnh timHnh thy li
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3. NGUN GC CA PHI V TNHTheo Kohlenbach (1978) cc th phi soma xut hin trong nui cy invitro t 3 ngun t bo nh bi:T bo sinh dng ca cy trng thnh.Cc t bo sinh sn khng phi hp t.Trc h dip v t dip ca phi hp t v cy con khng c thng qua s pht trin m so no.
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4. C IM CA THC VT TI SINH T CON NG HNH THNH PHI V TNHCy con c to thnh vn duy tr c tnh ca cy m v ng nht v mt di truyn.Phi v tnh pht trin thnh c th hon chnh nhanh, cy ra hoa kt tri nhanh hn l bin i c th.Cy ti sinh t phi v tnh c sc sng cao, d sng khi di chuyn ra mi trng t v sinh trng.
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Quy trnh nui cy m cy chui
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5. NG DNG CA QU TRNH TO PHI V TNHS lng ln phi v tnh l ngun nguyn liu ng k phc v cho ng dng thc tin quan trng nh : sn xut ht nhn to, ci thin cht lng cy trng, bo qun cht mm, loi tr virus, sn xut cc cht bin dng in vitro v thnh lp khun cn in vitro.
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5.1 S chn lc t boS dng nhng h thng ti sinh thc vt c kh nng ti to nn cy mi t t bo n nhm trnh s xut hin bin d gene. ng thi chn lc nhng c tnh m ta mong mun.V d: ng bin i gene khng su c r, nng sut cao, hiu qu s dng phn N tt. Phong Lan khng thuc dit c nh vi khun Agrobacterium.Nhn ging lan H ip bng nui cy m
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Quy trnh chuyn gene nh vi khun Agrobacterium
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5.2 S ti sinh cc thc vt chuyn gene:Bin np vo mu cy v sau cho ti sinh thng qua to phi v tnh trc tip hoc gin tip.Bin np vo t bo c kh nng sinh phi v ti sinh bng con ng sinh phi gin tip.Bin np vo phi v tnh v ti sinh thng qua con ng sinh phi th cp hoc sinh phi gin tip.Bin np vo phi v tnh, tng sinh thng qua con ng hnh thnh phi th cp v ti sinh qua con ng hnh thnh c quan.
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5.3 S to cc dng ng hp t v a biS dng mu cy n bi nh ht phn, bo t to phi n bi v k thut nhn i b NST to phi ng hp t.S dng mu cy ni nh to phi tam bi.X l colchicine hoc amiprophos-methyl to phi a bi.
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5.4 S loi tr virusPhi v tnh c 1 h mao mch rt pht trin, h thng ny khng ni lin vi h mch dn ca m cy nn to c cc dng cy sch bnh virus.
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5.5 S thnh lp khun cnu tin c a ra do Redenbaugh v cng s vo nm 1987. Nghin cu cho thy vic nm khun cn ngoi vo mi trng thc y pht trin cy con ngun gc phi v tnh.
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HT NHN TOHt nhn to cy a Lan
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CNG NGH HT NHN TO
c nghin cu nhm to v bao ci thin cht lng phi. c gii thiu u tin nm 1977, nhng n nm 1986 Redenbaugh v cng s mi tm li tt c cc nhn t cn thit to nn v bc hiu qu nht.
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KHI NIM HT NHN TOHt tng hp hay ht nhn to l thut ng ch phi v tnh hay nhng th nhn ging khc c bc bi 1 lp gel, gip cho vic bo qun trong thi gian di v vn chuyn d dng hnTo nn nhng c th ng nht v mt di truyn, tnh n nh cng nh cht lng cy trng
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KHI NIM HT NHN TOCng ngh ht nhn to quan trng vi nhng thc vt:Khng to c htHt to thnh vi s lng thpKh nng sng st rt thpNhn ging kh khn, cht mm khng th bo qun.
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CC NHN T CN THIT TRONG TNG HP HT NHN TO
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CC NHN T CN THIT TRONG TNG HP HT NHN TOPhi v tnh i hi: s lng ln, cht lng cao, sc sng tt, pht trin ng b.S dng phi v tnh giai on hnh tim mun v hnh thy li lm ht nhn to cho t l ny mm cao n 100%.
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MU S DNG LM HT NHN TOProtocorm-like body: Protocorm l mt c quan d tr nh c hnh thnh t phi ang ny mmProtocorm-like body (PLB) l mt cu trc c bit tng ng vi giai on chuyn tip mnh m ca phi thnh cy con
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S to PLB t m l
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VT LIU LM V BCC nhiu tc nhn to gel c s dng lm v bc cho phi: agar, alginate, polyco2133, carboxyl methyl cellulose, carrageenan, gelrite, guargum, sodium pectate, tragacanth gum, dextran, xanthan gumS dng alginate nhiu v: dnh va phi, khng gy c cho phi, c tnh tng hp sinh hc, to gel nhanh, r tin, lu c, cng gel va phi
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NGUYN TC V IU KIN KHI TO V BC BNG CHT NN SODIUM ALGINATE
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Nguyn tc trong qu trnh to v bc alginate l sodium alginate cha phi s to thnh tng ht nh, trn, cng khi c nh vo dung dch CaCl2 nh tro i in Na+ v Ca2+. V bc cng nhiu hay t ph thuc vo s lng ion Na+ v Ca2+ trao i.
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Nhc im ca sodium alginate: cht dinh dng c th mt i khi v bao, s trao i kh km -> s dng than hot tnh tng kh nng sng st ca phi v tnh
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B SUNGDo phi v tnh thiu lp ni nh cha cht dinh dng bn ngoi phi, do cn thm:Cht dinh dngCht iu ha sinh trngCht khng sinh, cht tr nm, tr su, vi sinh vt.-> Vo cht nn gel
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