CNG NGH SINH HC THC VT

GV: ThS. BI VN TH VINH

NHM 1 LP 10DSH01: DNG QUC XUN CHUNG NGC MI MI NG TH KIM PHNG O NGUYN THANH VY

CHNG III CNG NGH PHI V TNH HT NHN TO

PHI V TNH

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.

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.

2. MT S KHA CNH GI NH V
S HNH THNH PHI V TNH
S 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).

 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.

Mi t bo phi soma u pht trin qua 3 giai on trc khi trng thnh: Hnh cu Hnh tim Hnh thy li

Hnh tim

Hnh cu

Hnh thy li

3. NGUN GC CA PHI V TNH

Theo 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.

4. C IM CA THC VT TI SINH T CON NG HNH THNH PHI V TNH

Cy 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.

Quy trnh nui cy m cy chui

5. NG DNG CA QU TRNH TO PHI V TNH

S 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.

5.1 S chn lc t bo
S 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 t nh 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

Quy trnh chuyn gene nh vi khun Agrobacterium

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.

5.3 S to cc dng ng hp t v a bi S 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.

5.4 S loi tr virus Phi 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.

5.5 S thnh lp khun cn u 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.

HT NHN TO

Ht nhn to cy a Lan

 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.

CNG NGH HT NHN TO

KHI NIM HT NHN TO

Ht 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 hn To nn nhng c th ng nht v mt di truyn, tnh n nh cng nh cht lng cy trng

KHI NIM HT NHN TO

1. 2. 3. 4.

Cng ngh ht nhn to quan trng vi nhng thc vt: Khng to c ht Ht to thnh vi s lng thp Kh nng sng st rt thp Nhn ging kh khn, cht mm khng th bo qun.

CC NHN T CN THIT TRONG TNG HP HT NHN TO

CC NHN T CN THIT TRONG TNG HP HT NHN TO

Phi 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%.

MU S DNG LM HT NHN TO
Protocorm-like body: Protocorm l mt c quan d tr nh c hnh thnh t phi ang ny mm Protocorm-like body (PLB) l mt cu trc c bit tng ng vi giai on chuyn tip mnh m ca phi thnh cy con

S to PLB t m l

VT LIU LM V BC

C 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 gum S 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

NGUYN TC V IU KIN KHI TO V BC BNG CHT NN SODIUM ALGINATE

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.

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

B SUNG
Do phi v tnh thiu lp ni nh cha cht dinh dng bn ngoi phi, do cn thm: Cht dinh dng Cht iu ha sinh trng Cht khng sinh, cht tr nm, tr su, vi sinh vt. -> Vo cht nn gel