5. Ung Dung Ky Thuat Hat Nhan Trong Chon Tao Ginog Lua Chat Luong Cao - Tran Duy Quy - Vn

8/3/2019 5. Ung Dung Ky Thuat Hat Nhan Trong Chon Tao Ginog Lua Chat Luong Cao - Tran Duy Quy - Vn

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Trn Duy Qu, Bi Huy Thy, Nguyn Vn Bch , oTh Thanh Bng v cs.

NG DNG K THUT HT NHNTRONG CHN TO GING LA

CHT LNG CAO


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NI DUNG CHNH BO CO

1. Lch s ngnh di truyn v chn ging t bin.

2. Cc tc nhn gy t bin v c ch tc ng ca chngn vt liu di truyn.

3. Mc tiu, nh hng ca ngnh di truyn v chn gingt bin.

4. Nhng kt qa ni bt ca ngnh di truyn v chn gingla t bin Vit Nam

5. Chin lc pht trin lnh vc di truyn v chn ging la Vit Nam


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Lch s ngnh di truyn

v chn ging t bin


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Tnm 1900 - 1905, nh bc hcngi o Huygodveri phthin ra cy tbin tnhin cy Oenothera lamarkiana. ng

nghin cu quy lut pht sinh cc tbintnhin v hnhthnh nn thuyttbin.

Cc nghin ca cc nh sinh hc t thk 19, 20 nhLamark,Darwin, Menden.., khngnh cc tbin trong tnhin lngunbin di phong ph cho qu trnh chnlctnhin v nhnto hnh thnh nn cc loi mi v gingmi, thch ng ttviiukintnhin v pngtt nhu cuca con ngi.

Tuy nhin tbintnhin xy ra vitnsrtthpt1/vnn 1/triu, v th lm th no tm ra cc tc nhn nng cao

tbintnhin nn.

Phng php gy t bin nhn to c bit n t nm1925 khi Natxon v Philippp pht hinrng tia Roentgen c khnng gy ra bind di truynNmHng.

A. Trn th gii


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Nhng phi sau, cc cng trnh ca Muller, Xapegin,Stadle.. trn rui dm v ng (1926-1935), Xakharop,

Rapoport, Gustafson v cs trn la m, la mch, mchba gc (1938-1940) thu crtnhiutbin c gi trtrong nghin cu di truyn v chn ging. Chnh nhngcng trnh ny l cscho sra i ngnh di truynhcphng x lm nn tng cho sra i ngnh chnbintging phng x.

Ngay tnhngnm 1970, Cquan nng lng nguyn tquct (IAEA) v Tchc nng lngthgii (FAO) titrmrnghng nghin cu gy tbincitonhngging cy nng nghip v cy cng nghipnhiunc trnthgiinhmto ra hng lotgingminh: la, la m,la mch, to, chanh, ma, chui v nhng loi cy trngkhc.

A. Trn th gii


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Cho ti thng 7/2010 (FAO/IAEA Mutant Varieties Database),trn 2970 ging cy trngcto ra bng gy tbinthcnghim trn phm vi 62 nc. Vicngdngkthutht nhncitin cy trng mang lihiuqucck to lnv kinh tnng nghip. c tnh hng trm t la v hng trm triuhecta gieo trngbngnhngging cy trngc to ra ttbin.

Nhng thnh tu to ln m gy tbinthcnghimemli trnth gii l mt s ging cy trng nh: c Bermuda, l Nhtbn,ging la na ln Remei Nhtbn, khoai lang, khoai ty, hoacc, hoa hngvi mu sc khc nhau v hnh dng cnh hoa adng cam khng ht ca ran,Vi khng ht A4 ca TrungQuc. cbitTrung qucging la tbin Zhefu 802 c

trng vi din tch ln nht th gii (trn 10,5 triu hecta) vcsdngrng ri trong snxut trong thi gian kh di trn10 nm.

Mts thnh tuchngingtbin trn thgii qua cc nmc trnh by biudiy:

A. Trn th gii


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Slngging

Nm

Hn 2970 ging cy trng

FAO/IAEA Mutant Varieties Database

Slngging cy trngcto ra nh gytbin trn thgii (Tnh n 7/2010)


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Cc nghin cu di truyn v chntogingvit nam ctin hnh chmhnnhiu so vithgii.

Nm 1966, cc nghin cu v nh hng ca ria Gamma,ngun coban 60, DES, DMS n cc bin d di truyn la,du tm tib mn Di truyn khoa Sinh iHcTngHp

H ni (Trnh B Hu, Phan Phi, L Duy Thnh). T nm1968 trn u H Lan ca Trn Minh Nam, trnNigenladamastica ca Phan Phi (1969-1972), trn la ChnChu ln v Trung Quc 2 ca L Duy Thnh, Trn Duy Qu(1969-1970), tip theo l cc nghin cu trn cy c chua, to,

la Vin cy lngthc v thcphm(V Tuyn Hong vcs.. 1975-1980).

Cc nghin cunhhngca tia gama ca Thi Cng Tng,NguynVnMng (1971-1974) trn mts cy trng.

B. Vit Nam


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B. Vit Nam Sau gii phng Min Nam cc nghin cuchntogingtbin pht trinmnh

nhiuvin nghin cu v cc trngihc: ihcTngHp H Ni,ihc

sphm H ni I,II,Nng nghip I, Vin Di truyn Nng Nghip,Vin la B sngCu Long, Vin Khoa Hc k thut Nng Nghip Min Nam, Vin Cy Lngthc v cy thcphm,Vin Nghin cuht nhn Lt

cbit trong nhngnmgny cc Vini tin phong v c nhng thnh tusutsc trong nghin cu di truyn v chngingtbincc cy trng khcnhau nhla, ng, utng, rau hoa: Vin di truyn Nng Nghip (Phan Phi,

Trn Duy Qu, NguynHung, Mai Quang Vinh v cc cs., 1984- 2009), VinKhoa hc Nng nghip Min Nam ( Khc Thnh v cc cs., 1995-2009), Vinnghin cu la B sng Cu Long (onVn Ro, Bi ChBu v cc cs., 1990-2009), Vin Cy Lngthc v thcphm(V Tuyn Hong v cs., 1990-2009),Trng i hoc s pham H ni I (Nguyn Minh Cng v cs 1990-2009),VinNghin cuht nhn Lt (L Xun Thm v cs, 1998-2009).

Cho n nay cc nh khoa hcVit nam chntochn 57 cc gingtbintrn cc itng la, ng, utng, c chua, du tm, to, hoa cc, hoa hng.tr thnh nc th4 vchngingtbinca chu sau Trung Quc,NhtBn, n v ng th8 th gii sau Trung Quc,n ,Nht,Lin X c,HLan, c,M.


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Cc tc nhn gy tbin v cchtc ng ca chng n vt liu di

truyn


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Cc phng php gy to t bin

Tc nhn vt l: tia gamma, tia X, cc chmneutron nhanh, neutron chm

Tc nhn ho hc: EMS, EI, DES, DMS, NMU,NEU

Kt hp x l tc nhn vt l v tc nhn ho hc-Sdngphngphp no tuthuc: itng,

kiu sinh sn ca cy trng, tnh c hiu ca loitnh trngmong ito ra


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TCNHN VT L

Chiu x tia Cc tm (UV)

Chiu x tia in t trng (electromagnetic radiation)+ Tia X t my gia tc tia X.+ Tia Gamma t ngun Co-60, ngun Cs-137.

Chiu ht in t+ Chiu tia nhit hoc chiu chm tia ntron chm tl phn ng ht nhn+ Chiu chm tia ntron nhanh t l phn ng ht

nhn.+ Cc tiu phn Beta: Phosphorus 32 hoc Sulfur- 35.

Chm tia ion.


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TC NHN HA HC

Dimethyl Sulfate (DMS) Diethyl Sulfate (DES) Ethyl Methane Sulfonate (EMS) Ethylenimine (EI)

N- Nitroso Methyl Urea (NMU) N-Nitroso Ethyl Urea (NEU) Sodium Azide (NaM3) Colchichine


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C CH TC NG Nhiu th h cc nh khoa hc trn th gii v Vit nam khi

nghin cucch tc ngca cc tc nhn vt l v ha hcnphn tADN, NST, t bo, cth, chng gy nn cc phnngha hcvi phn tADN to nn cc dimertimin to nn sthay th cc gcbaznitca nhm purin bng pyrimidin, hocto nn cc ton, sai hnh cc NST Ni tm li l to nnnhng thay ivcu trc ca gene, nhngcu trc ny c ditruyn cho cc th h sau thng qua cc qu trnh phn bo,nguyn nhim v gimnhim. Cui cng to nn cc c thmimang nhngcim di truyn khc vibm.

Chng ta tin hnh chnlc cc tbinc tao ra theo nhngc tnh mong munnhnngsut cao, khnngchngchu su

bnh,hn, ngto thnh cc gingmi.


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CC PHNG PHP CI TIN GINGCY TRNG

Chn lc v bo tn nhng vt liu cy trng sn c trongt nhin c ci tin mt cch t pht p ng nhngtiu ch ca nh chn ging.

Lai to kt hp chn lc.

Gy to t bin kt hp chn lc.

K thut sinh hc: chuyn gen to ra cc c th c tnhtrng mi GMO (Genetics Modified Organism).


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Mc tiu, nh hng ca ngnhdi truyn v chn ging t bin


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Mc tiu ci tin ging cy trng

To ra ngun gene khi u a dng phc v ccnghin cu lthuytv di truynhc v chntogingcy trng:

- Nghin cubnchtca gene quy nh cc tnh trng

- Lp bn cc gene quy nh cc tnh trng: nngsut, cht lng, tnh chng chu su bnh, kh nngchungivi strees ca mi trng(hn, ng, chuamn, nng, lnh gen btdcc TGMS..)

Toc

ccging

mi

theo yucu

ca

ngi

tiudng.


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u im ca chn ging t bin

- To ra s da dngvmt di truyncavtliukhiu nhanh v hiuqu.

- Tocgingmi trong mt thi gian ngn,cbitiviloi cy sinh sn v tnh.

- Thay imthocmt vi tnh trng m khng lmnhhngtinhng tnh trng cn lica cy trng.

- Nng cao hiuqu cng tc chn toging so viccphng php chngingcin v hini.


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Nhng kt qa ni bt ca ngnh di

truyn v chn ging la t bin


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Trong bo co ny, chng ti tp chung trnh by

nhngktquchyu thu c khi x l cc gingla a phng, nhp ni hay cc dng mi lai toc nh cc ging: Tm Thm Thi Bnh, TmXun i,DHng, Chim bu,Cm,Np ci Hoavng, NpHiHu,McTuyn, IR8, C4-63, TB1, cc

dng mi lai to nh: ST3, ST5, ST13, ST16, ST19,ST20bng cc tc nhn vt l v ha hc nh tiaGama, ngun Coban60 vi liu lng 15, 20 v 25Krad. Cc cht ha hc nh NMU, NEU, EI, DES,DMS vi cc nng 0,01; 0,015; 0,025; 0,030% trongthi gian t18 -24h ht kh hochtnymm.


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T nm 1980 n nay, chng ti thu c rt nhiucc tbin mang nhngcim nng sinh hc qu

nh kiu cy cy ci tin, cc yu t cu thnh nngsut cao nhcu trc bng, khilng 1000 httngcao, chtlnggo cao, dothm. cbitnhiutbin chn smcyc hai v/nm, khng cnhiu

su bnhnho n, bc l, kh vn,ry nu..

Nhiu trong cc stbin nu trn ctrctipchn lc qua mt s th h tr thnh ging hoc

c lai tch lyvibmhoc lai gia cc dng tbinvi nhau v thng qua chn lc tr thnh gingphcvsnxut.


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Tnm 1985 n nay, chng ti a ra c hng chcging latbincchn lcbngphng php thcnghimnhDT10;DT11; DT13; DT14; DT17; DT33, DT21; DT22; DT37; DT38; A20;

DT122; D271; CM1, Tm thmtbin, ST3; MT4; MT2 Nhiugingn nay vn tn ti trong snxut. Ngoi cc tbin c licho cng tc chn to ging cn c cc t bin phc v cho ccnghin culpbn di truynv gen nhgen btdcc TGMS,gen khng o n, bc l, ry nu, gen mi thm, gen khng mncmvi quang chu k..

Mt v d in hnh l ging la t bin DT10 ca Vin Ditruyn Nng Nghipnhnc tvicchiuxht kh cagingC4-63 liu 20Kr. Ging ny c cng nhn tiu chunging

quc gia nm 1990. Khc vigingbm,ging DT10 c khnngchulnh,chu ng, chngchubnhtthnbm v cho nngsutkh cao v nnh(nngsutt 5,5 - 7,0 tn /ha). Ging la nyctrngnhiu vng minbcVit Nam vidin tch khong 1triu hecta, chim 33% din tch trng la thpk 90. Mts dngtbin c trinvngcsdngnhngunvtliu ban u cho

cc php lai nhtrnghpcaging la A20, DT22. 22


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Phnln cc gingtbincto ra bngchnlctrctiptcc dng tbintrinvngtcc thhM3 tri, chn ra nhng dng c nhng tnh trngtt hn ging gc v mt s c tnh nng hc nh

nngsut,chtlng (Hnh 2, Hnh 3, 4).

Mo AABBCC


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Mo AABBCC..

M1 T bo khi sinh th 1 AABBCcT bo khi sinh th 2 AABBCCT bo khi sinh th 3 AaBbCc

M2 (Cc th h con M1)

Cc bin i di truyntrong loci

Plants from:

1st sector1/3AABBC.3/4AABBcc.1/4

2nd sector1/3

AABBCC(Khng phn ly)

3rd sector1/3A.B.C. 9/16A.bbC 3/16

aaB.CC 3/16aabbC 1/16aa- Thn ln; bb- mm bch tng; cc: lp phn trn l

Chn lc cc cy thn ln t bin

Tn s ca cc cy thn ln M2

Kiu hnh: aaB.CC 1/3 x 3/16=3/48 (1/16)Kiu gen: aaBBCC 1/3 x 1/16=1/48

Tt c nhng c th c kiu hnh aaB.CC c trng ring bit M3 tip tcchn lc

Hnh 2: S k thut chn t bin truyn thng cy trng sinh sn hu tnh(la, ng, u tng)


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Hnh 3: B NST la O.sativa. L


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Hnh 4: Cc kiu t bin NST hu k nguyn phn


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Nhng tnh trngccitin thng qua tbin

thng l nngsut,gimchiu cao, chngchu,su, bnh (i vi la: DT10, DT11,...); ph vtnh cm quang (tm thmtbin, DT21); Chtlng( la CM6); Chu stress ca mi trng(la CM1), c t bin thay i mu sc ca ht

go lm tng tnh thng phm trn th trngnhtrnghpging la Chim buaphnghtgo,nngsutthp,gingtbin CM1v CM6 htgochuyn sang mu trng lm tnggi trthngphm (Bng 1; 2,3); Hnh 5,6).


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Hnh 5: Cc kiu t bin tnh trng cy la


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Hnh 6: Cc kiu t bin tnh trng cy la


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Bng 1. Cc ging la mi c to ra bng phng php gy tbin nhn to

PD2 Htt +

Cc gingt bin

Phng php x l t binChiu x (1) Ha cht(2) Kt hp (1 )

v 2Kt hp 1 v

lai to

DT10 20 Kr iviht kh M2: 0,025%NEU1

+

DT11 20 Kr iviht kh M2: 0,025%NEU

+

A20 20 Kr iviht kh 0,015% NMU2

CM1 20 Kr iviht kh

CM6 20 Kr iviht khDT33 20 Kr iviht kh

DT36 Chiux callusTm thm tbin

10-20 Kr vihtt

DT21 NEU 0,025% +

DT22 NEU 0,025% +Khang Dn tbin

10-20 Kr vihtt

DT38 25 Kr iviht kh

CL9 Ht kh +

B 2 Ph h h l


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Bng 2. Phngphpchnlcccginglatbin

Teen ging Phng php chn lcTrc tip Gin tip

DT10 +DT11 +A20 +CM1 +CM6 +DT33 +DT17 +DT36 +DT21 +Tm Thmtbin +

DT22 +Khang Dn tbin +DT38 +CL9

ST3

+PD2 +

+

h i i i l bi


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GingTnh trng c citin nh t bin

Nm c cng nhntin b khoa hc kthutSTT Ngun gc t bin

1 C4-63 DT10Nng sut cao, chu lnh,chu su bnh 1990 (Ging quc gia)

2 C4-63 DT11Cht lng tt, ngoncm hn DT10

1995 (Ging quc gia)

3 DT10/CR203 DT13cht lng cm kh hnDT10, nng sut cao

1998 (Ging quc gia)

4 A8 A20 Chu hn, chu phnmn, cht lng tt

1993 (Ging quc gia)

5 Chim bu CM1Chu phn mn, i muht t sang trng, chnsm

1999 (Ging quc gia)

6 Chim bu CM6Chu phn mn, mu ht,

cht lng, mi thm 2000 (Ging tm thi)

7 CR203 DT33

Nng sut cao hn ginggc 20%, chu c bnhbc l

1994 (Ging quc gia)

8VN-01/ b BG-90-2 (F6)

DT36Nng sut cao, khng subnh

2002 (Ging tin bKHKT)

Bng 3: Cc tnh trng c ci tin ging la t bin

B 3 C h i i i l bi


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9 DT10/ OM80 DT17 Nng sut cao, cht lng tt 1999 (Ging tm thi)

10 DV2/ np 415 DT21Ph v tnh cm quang, nngsut 2000 (Ging quc gia)

11 Tm thm Hi huTm thmt bin

Ph v tnh cm quang, ngnngy 2000 (ging quc gia)

12 DT21Dai loan DT22 Nng sut cao, cht lng tt 2002 (Ging tm thi)

13 Khang dn DT38Nng sut cao, cng cy,chng 2007 (Ging tm thi)

14 Khang dn Khang dnt bin

Nng sut cao 2007 (Ging Quc gia)

15 Bc thmBc thmt bin

Ph v tnh cm quang, chtlng tt hn 2006 (Kho nghim quc gia)

16Gy t bin con laiIR64/Khang dn 18

CL9Ci thin cht lng go

2006 (Ging quc gia)

17Np t bin415/KT90

PD2Ph v tnh cm quang, chtlng tt hn 2006 (Ging quc gia)

18 Sc Trng (ST) ST3 B Nng sut cao, cht lng tt 2006 (gingKN quc gia)

19 IR64 VN 95-20Nng sut cao, cht lng tt c cng nhn ging quc

gia nm 1999

Bng 3: Cc tnh trng c ci tin ging la t bin


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Ngoi ra, kthp nghin cutbinvi nghin cu sinh

hc phn tcng l mthngicnc pht huy.

Vickthpgiakthut sinh hc phn tvi nghin cugy totbincthchinmt cch chtch, c thcung cpnhngphng php nghin cu chnh xc, hiu

qu, nhanh v kinh thn trong cng tc citin cy trngtheo hngchngingtbin vdnhnghin culpbn di truynv gen nhgen btdcc TGMS, genkhng o n, bc l, ry nu, gen mi thm, gen khngmncmvi quang chu k (Hnh 7,8).


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TILLING (Targeting Induced Local Lesion in Genome) phng php sng lc nhanh v chnh xc cc dng t bin mc phn t (Till

et al. 2003).

S dng cc ch th phn t v PCR xc nh t bin mc phn t


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The TGMS-VN1 gene ina Vietnamese rice variety

was mapped.

Hnh 7: Bn gene bt dc c nhy cm vi nhit (TGMS-VN1) cy la


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Marker assisted selection against

rice blast

Hnh 8: Bn gen khng o n cy la


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Chn lc cc ging la t bin nng sut cao,cht lng tt, c sn: Tm Thm t bin


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Np DT22


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Chin lc pht trin lnh vc di

truyn v chngingVit Nam


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Cc ktqu trnh by trn chng tphng php chn togingtbinnhkthutht nhn l rthiuqa:

- Tocnhiutbin gi tr trong nghin cu l thuyt ditruyn v lpbn gene cy trng:

- Tocnhiuvtliukhiu cho cng tc chntoging:

+ Chnlctrctip thnh gingmi

+ Lai togia cc tbingia cc ging ban u hay gia cctbinvi nhau to nn gingmi.

Nng cao hiuqa v rt ngn thi gian togingmi so vicc phng php chntotruynthng khc hay phng phpchuyn gene.

D d i h h h i h h h


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D s dng, gi thnh thp so vi cc phng php chn toginghini khc.

V vychinlc xy dng v pht trinlnhvc nghin cu ditruyn v chngingtbinnhsdngk thutht nhn nc ta theo chng ti phinhsau:

- Nh nccn xy dng v thchinchng trnh di hntnay n 2015 v tm nhn 2030 v ng dng k thut ht nhntrong nng nghip trong c lnhvc gy tbinthcnghimcy trng. Hng nm c nhng ti trngimcp nh nctp chung vo cc cy trng ch lc nh la, ng, u tng,caf, ch, rau hoa..

- Nh nccn nhanh chng utmththngcsvtchtgm nh xng, my mc nhmy chiux,hoc xy dngmt

trng Gamma l ttnht v c thchiuxnhiuloi cy trngcng mt lc, nhiu giai on khc nhau ca chu trnh phttrin c thhtphn,hpt,tin phi, ht chn sinh l, cy angsinh trng v pht trin Do sto ra cnhiuvt liukhiu trn nhiuvtliu khc nhau cng mt lc phcvcng tc nghin cu di truyn v chntoging cy trngnc

ta.


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Cn tp chung o tongun lc cn bkthutvx lchiux,vn hnh cc my mc thitbtrng Gamma field, cn b nghincuv di truynchntoging. scahng nghin cngdnght nhn trong nng nghip ngang tmvi cc nc tintinkhu vc v thgii

Nh nccn c nhng chnh sch thu ht ngunlc trong v ngoinc, cc doanh nghip trc tip vo mt s khu trong qu trnhnghin cu chn to ging nh k thut ht nhn nh chnh sch

lngbngvi cc nh nghin cu, chnh sch taithuvi ccdoanh ngipu tvo vic trin khai cc ktqa nghin cucalnhvcngdngkthutht nhn trong nng nghip.

Cnkthpchtchgiuphng php gy tbin truyn thngvi cc kthutca CNSH nhnui cy m t bo, kthut marker

phn t,kthut tch v chuyn gene nng cao hnnahiuqacakthutht nhn trong nghin cu di truyn v chnging cytrngnc ta tin ln mtbcmi,pngc cc mc tiukinh t,mbo an ninh lngthc v nng cao chtlngcamtsmt hng nng snnhla, go, ch, caf.


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5. Ung Dung Ky Thuat Hat Nhan Trong Chon Tao Ginog Lua Chat Luong Cao - Tran Duy Quy - Vn