On tap Sinh hoc

KHI NIM V GING

Ging vt nui, cy trng, vi sinh vt l nhng nhom sinh vt do con ngi to ra, c cc c im di truyn nht nh, cht lng tt, nng sut cao v n nh, c cc phn ng cng kiu i vi iu kin ngoi cnh, thch hp vi cc iu kin kh hu, sinh thi, dinh dng v k thut sn xut nht nh.

CAC PHNG PHAP CHN GING

1. K thut di truyn - Khi nim: K thut di truyn l k thut thao tc trn vt liu di truyn da vo nhng hiu bit v cu trc ho hc ca cc axit nuclic v di truyn vi sinh vt. - Phng php c s dng ph bin hin nay l k thut cy gen, tc l chuyn mt on ADN t t bo cho sang t bo nhn bng cch dng plasmit lm th truyn. K thut cy gen c 3 khu ch yu: + Tch ADN nhim sc th ca t bo cho v tch plasmit ra khi t bo. + Ct v ni ADN ca t bo cho vo ADN plasmit nhng im xc nh, to nn ADN ti t hp. Thao tc ct tch on ADN c thc hin nh enzim ct (restrictaza). Cc phn t enzim ny nhn ra v ct t ADN nhng nucltit xc nh nh ngi ta c th tch cc gen m ho nhng prtin nht nh. Vic ct t ADN vng ca plasmit cng c thc hin do enzim ct cn vic ghp on ADN ca t bo cho vo ADN plasmit th do enzim ni (ligaza) m nhim. + Chuyn ADN ti t hp vo t bo nhn, to iu kin cho gen ghp c biu hin. Plasmit mang ADN ti t hp c chuyn vo t bo nhn bng nhiu phng php khc nhau. Vo t bo nhn, n t nhn i, c truyn qua cc th h t bo sau qua c ch phn bo v tng hp loi prtin m ho trong on ADN c ghp. T bo nhn c dng ph bin l vi khun ng rut E.Coli. T bo E.Coli sau 30 pht li t nhn i. Sau 12 gi, 1 t bo ban u s sinh ra 16 triu t bo, qua cc plasmit trong chng cng c nhn ln rt nhanh v sn xut ra mt lng ln cc cht tng ng vi cc gen ghp vo plasmit. Trong k thut cy gen ngi ta cn dng th thc khun lm th truyn. N gn on ADN ca t bo cho vo ADN ca n v trong khi xm nhp vo t bo nhn n s em theo c on ADN ny vo . 2. ng dng k thut di truyn K thut di truyn cho php to ra cc ging, chng vi khun c kh nng sn xut trn quy m ln to ra nhiu loi sn phm sinh hc c gi tr nh axit amin, prtin, vitamin, enzim, hoocmn, khng sinh...lm gim gi thnh chi ph sn xut ti hng vn ln. c nhng thnh tu ni bt nh vic chuyn gen m ha hoocmn Insulin ngi, hoocmn sinh trng b, chuyn gen khng thuc dit c t loi thuc l cnh Petunia vo cy bng v cy u tng (1989), cy gen quy nh kh nng chng c mt s chng virut vo mt ging khoai ty (1990). 3. Phng php gy t bin nhn to a) Gy t bin nhn to bng cc tc nhn vt l: Cc tc nhn gy t bin c s dng ph bin hin nay l cc loi tia phng x, tia t ngoi, sc nhit gy nn cc t bin gen, t bin NST to ra ngun nguyn liu cho to ging cy trng, vi sinh vt. Ty thuc vo tnh bn vng ca vt cht di truyn mi ging m s dng cng sut liu lng phng x khc nhau. b) Gy t bin nhn to bng cc tc nhn ho hc: 1

On tap Sinh hoc S dng cc tc nhn ha hc nh 5 - brmuraxin (5 BU), EMS (tylmtal sunfonat), consixin, cc ha cht siu t bin. NMU (nitrz mtyl ur), NEU, EI... tc ng vo ADN, NST khi chng ang trn con ng nhn i hnh thnh s to nn cc t bin gen, t bin NST. Thng to nn nhiu t bin phi tc ng vo cc thi k phn bo mnh nht , vo ht ny mm, giai on hp t, tin phi... Cc tc nhn gy t bin nhn to c ng dng c hu qu trong chn ging vi sinh vt, chn ging cy trng to c hng trm ging c gi tr v nng sut, phm cht v kh nng thch nghi. 4. Cc phng php lai a) Lai gn ng vt (t th phn thc vt): - Lai gn l phng php lai gia cc c th c quan h rt gn gi v mt di truyn (lai gia cc c th sinh ra trong cng mt la, lai gia con ci vi b m, thc vt l php t th phn). - Lai gn lin tc nhiu ln lm cho d hp t gim, ng hp t tng, th h con chu c sc sng, kh nng thch nghi km dn, nng sut gim, qui thai nhiu. - Trong chn ging lai gn cng c vai tr nht nh nh cng c cc tnh trng qu him, nh gi hu qu ca mi dng to ra, lm nguyn liu khi u cho to u th lai v lai to ging mi. b) To u th lai: - u th lai l hin tng c th lai F1 c sc sng hn hn b m v cc ch tiu sinh trng nhanh, pht trin mnh, chng chu tt, nng sut cao vi iu kin bt li ca mi trng. Tuy nhin u th lai biu hin cao nht F1, sau gim dn qua cc th h, v th d hp t gim, ng hp t tng. - C s di truyn ca hin tng u th lai, y l vn phc tp, c 3 cch gii thch nh sau: + Gi thuyt v trng thi d hp: Tp giao gia cc dng thun chng, F1 d hp v cc gen mong mun, mu thun ni b gia cc cp gen cao, trao i cht tng cng, kh c tc dng gy hi ca cc gen ln t bin. AABBCC x aabbcc AaBbCc + Gi thuyt v tc dng cng gp ca cc gen tri c li: Cc tnh trng a gen c chi phi bi nhiu gen tri c li khi lai tp trung c cc gen tri c li, tng cng hiu qu cng gp. AAbbCC x aaBBcc AaBbCc + Gi thuyt siu tri: l kt qu ca s tng tc gia 2 alen khc nhau v chc phn ca cng mt lcut dn n hiu qu b tr, m rng phm vi biu hin kiu hnh. AA < Aa > aa - Phng php to u th lai: Lai khc dng n, lai khc dng kp, lai thun v lai nghch gia cc dng t th phn mt cch cng phu d tm ra t hp lai c gi tr kinh t nht (ng lai F1, la lai F1). c) Lai kinh t: c s dng trong chn nui to u th lai. l php lai gia cc dng b, m thuc 2 ging thun khc nhau to ra F1, ri dng con lai F1 lm sn phm, khng dng n nhn ging tip cc i sau. Ph bin nc ta hin nay l dng con ci thuc ging trong nc cho giao phi vi con c cao sn thuc ging thun nhp ni. Con lai c kh nng thch nghi vi iu kin kh hu v chn nui ca ging m, c sc tng sn ca ging b (ln lai kinh t F1, b lai sinh, c chp lai...). d) Lai ci tin ging: S dng mt ging cao sn ci tin mt ging nng sut thp. nc ta thng dng nhng con c tt nht ca ging ngoi cho phi vi nhng con ci tt nht ca ging a phng. Con c ging cao sn c s dng lin tip qua nhiu i lai. V mt di truyn hc, phng php lai ci tin ging ban u lm tng t l th d hp, sau tng dn t l th ng hp v cc gen c li. e) Lai khc th v vic to ging mi: s dng u th lai, ng thi to ra cc ging mi ngi ta dng phng php lai khc th (lai gia 2 th hoc lai tng hp nhiu th c ngun gen khc nhau). Sau phi chn lc rt cng phu to ra ging mi, v trong cc th h lai c s phn tnh. f) Lai xa: l cc hnh thc lai gia cc dng b m thuc 2 loi khc nhau hoc thuc cc chi, cc h khc nhau nhm to ra cc bin d t hp mi c gi tr. - Nhng kh khn trong lai xa: + Thc vt khc loi thng kh giao phn: ht phn khc loi khng ny mm trn vi nhy hoc ny mm c nhng chiu di ng phn khng ph hp vi chiu di vi nhy nn khng th tinh c. ng vt khc 2

On tap Sinh hoc loi thng kh giao phi, do chu k sinh sn khc nhau, h thng phn x sinh dc khc nhau, b my sinh dc khng ph hp, tinh trng khc loi b cht trong ng sinh dc ci. + Kh khn ch yu v mt di truyn l c th lai xa thng khng c kh nng sinh sn (bt th). Nguyn nhn ca hin tng ny l b NST ca 2 loi b, m khc nhau v s lng, hnh dng NST, kch thc, cch sp xp cc gen trn NST, s khng ph hp gia nhn v t bo cht ca hp t. S khng tng hp gia b NST ca 2 loi nh hng ti s lin kt cc cp NST tng ng trong k u ca gim phn I, do qu trnh pht sinh giao t b tr ngi, c th lai xa khng pht sinh c giao t, hay giao t to c nhng khng tham gia c vo qu trnh th tinh... - Cch khc phc hin tng bt th c th lai xa: S dng phng php gy a bi th bng tc nhn consixin (gi l phng php song nh bi) lm tng i b NST ca loi b v loi m, to iu kin xp thnh cp tng ng, th qu trnh gim phn s din ra bnh thng, c th lai tr nn hu th (th nghim thnh cng u tin ca G.D.Cacpsenk (1927) khi lai ci bp (2n = 18) vi ci c (2n = 18)). Cy lai F1 (2n = 18) c b NST t hp 2 b NST n bi khng tng ng ca 2 loi nn khng c kh nng sinh sn. Tc gi to ra dng 4n = 36 lm cho cy lai sinh sn c. - ng dng phng php lai xa: Phng php lai xa km theo a bi ho to c nhng ging la m, khoai ty a bi c sn lng cao, chng bnh gii. Hin nay ngi ta rt ch lai gia cc loi cy di chng chu tt, khng su bnh vi cc loi cy trng nng sut cao, phm cht tt v cc php lai gia cc loi ng vt to c nhiu dng lai c gi tr. g) Lai t bo sinh dng: - Lai t bo sinh dng l phng php dung hp 2 t bo trn khc loi to ra t bo lai cha b NST ca 2 t bo gc. - Cc bc c bn ca lai t bo sinh dng: + Tch t bo trn thuc 2 loi khc nhau d nh a lai. + Trn ln 2 dng t bo trn thuc 2 loi trong mi trng dinh dng nhn to c b sung thm cc virut Xene lm gim hot tnh, tc ng nh mt cht kt dnh hoc dng keo hu c polietylen glycol hay xung in cao p. + Dng cc mi trng chn lc to c nhng dng t bo lai pht trin bnh thng. Dng cc hoocmn ph hp, ngi ta kch thch t bo lai pht trin thnh cy lai. - Thnh tu: Theo hng ny c nhng thnh cng bc u trn thc vt trong nhng nm 70 nh to c cy lai t 2 loi thuc l khc nhau, cy lai gia khoai ty v c chua. Cng to c nhng t bo lai khc loi ng vt nhng cc t bo ny thng khng c kh nng sng v sinh sn. Bng k thut lai t bo trn, trong tng lai, c th to ra nhng c th lai c ngun gen rt khc xa nhau m bng lai hu tnh khng th thc hin c, c th to ra nhng c th khm mang c tnh ca nhng loi rt khc nhau, thm ch gia thc vt vi ng vt. 5. Cc phng php chn lc a) Chn lc hng lot: - Cch tin hnh: Trong 1 qun th vt nui hay cy trng, da vo kiu hnh ngi ta chn ra mt nhm c th ph hp nht vi mc tiu chn lc lm ging. Tu theo vt liu khi u, yu cu v hiu qu chn lc, c th tin hnh chn lc hng lot 1 ln hay phi lp li nhiu ln. - Phm vi ng dng: i vi nhng cy t th phn, c khi ch chn lc 1 ln mang li hiu qu. i vi nhng cy giao phn v qun th c kiu gen khng ng nht, cc th h sau c s phn tnh, nn thng phi chn lc hng lot nhiu ln. Chn lc hng lot l phng php hu hiu duy tr cht lng v nng sut ca ging khi a vo sn xut i tr qua nhiu v, phc trng nhng ging khu vc ho va` cung cp ging cho sn xut. - u im: 3

On tap Sinh hoc Phng php chn lc hng lot n gin, d lm, t tn km thi gian, cng sc, khng i hi trnh khoa hc k thut cao nhng a li hiu qu tt, nn c th p dng rng ri. Phn ln cc ging tt cc a phng l do nhn dn sng to ra trong thc tin sn xut nng nghip bng phng php . - Nhc im: Khi chn lc ch cn c trn kiu hnh, khng kim tra c kiu gen ca c th nn vic cng c, tch lu cc bin d tt, chm a n kt qu. Phng php chn lc hng lot thng ch d c hiu qu i vi tnh trng c h s di truyn kh cao. b) Chn lc c th - Cch tin hnh: Trong qun th khi u ngi ta cng chn ly mt s t c th tt nht nhng iu sai khc cn bn so vi chn lc hng lot l chn lc c th con chu ca nhng c th ny c nhn ln mt cch ring r theo tng dng, do kiu gen ca mi c th ban u s c kim tra qua nhiu th h. S so snh gia cc dng v so snh vi ging khi u s cho php chn c nhng dng tt nht, loi b nhng dng khng p ng mc tiu chn ging. Phng php chn lc c th c th c tin hnh 1 ln hay nhiu ln. - Phm vi ng dng: Khi mc tiu chn lc l loi tnh trng c h s di truyn thp th phi p dng phng php chn lc c th. Chn lc c th mt ln c p dng cho cc cy nhn ging v tnh v cc cy t th phn. Dng t th phn c kiu gen kh ng nht v n nh nn c khi ch chn lc c th 1 ln la` c kt qu. i vi cy giao phn, nu mun p dng chn lc c th th phi tin hnh nhiu ln. Trong qun th giao phn rt kh xc nh cy b, v con chu ca 1 cy ban u thng khng ng nht v kiu gen v kiu hnh, do chn lc c th 1 ln khng nh gi. i vi vt nui, ngi ta kim tra c ging qua i sau. Con c khng th cho sa, trng, nhng nh hng n 1 s lng ln con chu, trong c c c v ci, thun li cho vic nh gi. Ngy nay phng php kim tra qua i con c b sung bng nhng phn tch ho sinh, t bo trn con c ging. Trong chn nui gia cm, ngi ta cn p dng phng php kim tra qua i sau i vi con mi. - u im: Chn lc c th kt hp c vic nh gi da trn kiu hnh vi vic kim tra kiu gen, do nhanh chng t hiu qu, nht l khi mc tiu chn lc l nhng tnh trng ch c li cho ngi m t c li cho bn thn sinh vt nh hm lng du trong ht hng dng, t l b trong sa b, ging to ra c tnh n nh v di truyn cao. - Nhc im: Tuy nhin chn lc c th i hi cng phu, mt nhiu thi gian theo di cht ch, kh p dng rng ri.