KHOA HOC C6NG NGH£
O U Y TLJ Q T L SALTOL V A O GIOIMG LLJA A 5 g 9 8 BAIXIG PHUOIVG P H A P CHOIM G I O M G IMHCr CHi TH! PHAIXl TIJ
VA LAI HOI GIAO
Uni Thj Ngpc Huy^n^
T6MTAT
Muc ti^u cua nghi^n ciju la g6p phdn chpn tgo gi6ng liia chm man, nSng au^t cao; sCr dyng phirong phap chon gidng nhir chi thi phSn tu k^t h<?p lai h6i giao (MABC-Marker Assisted BackCrossing) nhir 1^ m^t kj' thuat cao trong chon tao gi6ng. T6ng s6 500 clii thj SSR nSm r^ rtc trSn 12 nhi^m sdc M (NST) dupe sii dung de sang loc da hinh cfic gidng h6 m?, trong d6 c6 52 chi thj trong vung gien Saltol. Chi lim dune 63/500 chi thi da hinh, dupe su dung d^ sang Ipc ca th^ ciia cac quAn thI hoi giao BCiF,, BQF, va BC,F|.
Qua ha th^ h^ chon loc, da thu dupe mOt d6ng BC^Fi - P284-112-209 c6 chua viing gien Saitohs, gan 100%
n4n di tniy&n cua giong nhSn gien va bon d6ng BCjF, khac P307-30&- 21, P284-112-195, P284-112-198, P284- 112-213 chi CO mpt locus (noi) di hoii tu trong so 63 chi thi sang loc. Dong BCjF, dS dupe lai tao mang gan 100% hinh thai va nen gien cua cSy n h ^ gien AS996 sSn sang cho phat tnen gidng liia men ASS996-SaItol chiu man ling pho voi bi^n doi khi hau.
Tir kh6a: Giong liia AS996, chi tbj phan tii, chpn giong nha chi thi pban tu k^t hop lai h6i giao (MABC- Marker Assisted BackCrossing), Ilia, Saltol, QTLs.
LB^TVAAIOE
Liia la nguon luong thuc quan trong nh^t cho m6t nua dan so th^ gioi va cung la nguon luong thirc chinh cho 86 trieu dan so Viet Nam. Hon nira, Vi^t Nam con la nuac xuat khau gao dung thii hai the gioi sau Thai Lan. Phat trien giong liia thich ling voi bien doi khi hau (BDKH) va muc nuoc bi^n dang cho dong bang song Hong va dong bang song Cuu Long la rat quan trong doi voi nen kinh te va an ninh luong thuc nuoc nha, dong thoi gop phan vao nen an ninh luong thuc toan cau.
Cac nha khoa hoc ciia IRRI da dua ra mot so giong Ilia cho nang suat cao va chong chiu cdc di^u kifin soc sinh hoc nhu ling ngap hoac han m^n nha su dung ca hai phuotig phap chon gidng tniy^n thong va chon gidng hien dai. Cac giong lua nay c6 th^ giiip giam bat phan nao ton thirong do tac dong cua BDKH [14]. Cong viec chon giong chong chju man tra n^n d^ dang hon sau khi cac vung chiiih tren NST (cac noi tinh trgjig so lirong - quantitative trait loci -QTLs) Hen quan d^n tinh chiu man da dirpc xac djnh. Ket qua nay da dSy nhanh vific ph^t tri^n va su dung he thdng chi thi phan tu trong chpn gidng thong qua cong ngh§ chon gidng nha chi thi tra giiip va lai tra lai (marker assisted backcrossing - MABC)
^ Vi?n Di truyfin Nong nghi?p
[14]. Ca so cua chidn luoc nay la dua (thdng qua lai hiru tinh) mot gien hoac QTL tu dong cho gien vao mot dong nhan gien cung liic voi vi6c sang loc gien dich trfen nen he gien cua dong [12, 13, 14]. Nghien cuu nay ung dung chidn luge MABC d^ chon tao gidng Ilia mdi c6 kha nang chdng chju mSn cho Viet Nam.
L VAT UEU VA PHUONG PHAP NGHIEN CUU i.vatii^u
Giong lua FL478 da diroc sij dung lam vat lieu cho gien chiu man Saltol. Gidng nhan gien la AS996, la mot gidng dang duoc trong phd bien tai cac viing thuoc mien Nam Viet Nam.
2. Phuong phdp nghi&n ciiu
- Trong chien luoc chon gidng MABC, AS996 dupe lai voi FU78 d^ thu hat lai Fi. Th^ he F, dupe lai tra lai voi AS996 de diu mot lupng Ion BCiFi. 573 cay BCiFi da dugc sang Ipc cho gien dich, tai to hop va lira chon nen gien. Nhung cay mang QTL dich va phan lan he gien cua cay nhgn se dupe lira chpn cho th^ h§ hoi giao ke hep. Hon 500 ca th^ BCjFi va 371 ca th^ BC3F1 da duoc sang loc tirong tir nhu da lam doi voi th^ h$ BCiFi. Cac ca th^ BC2F2 va BC3F1 mang gien dich va hau het h6 gien cua gidng nhan gien da dupe xac dinh.
- Xac dinh cac chi thi SSR cho da hinh giua cay cho va nhan gien: khoang 500 chi thi SSR rai rac tren
NONG NGHIEP VA PHAT TRIEN NONG TH$N - KY 2 - THANG 5/2012
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12 nhi^m sac thd cua lua su dijng cho viOc s^ng Ipc ca gien dich, th^ tii td hpp vA n^n gien.
- So li^u phftn tich ki^u gien dim Irfin vi(?c phfln tich ADN cac ca th^ cua cic llic IiO lai inV lai vf'n c.V chi thi SSR bang phirrnig ph<^p sir di^ng ph^n irng PCR tren khay 96 gif'ng. Hi«n tiiih ADN trong 4 phut 94"C. Mdi chu ky c6 1 phiit bi^n Unh ADN 6 94"'C, 1 phiit annealing (tai, u) if SS^C, vi 1 phiit cho tdng hap spi ADN 72''C. 5 phiit d6 tdng hpp ndl d 72'*C sau khi ket tliiic 30 chu ky tdng hpp (Eppendoft Ihemial cyclers). San pham PCR dupe cd djnh trong bromophenol blue gel loading dye (ch£it gien nhudm bromo phenol mau xanh) va din^c phan tich bang di^n di tr&n 4,5% acrylamit gel a 1500 V (Biorad system) ho$c 6% -8% acrylamit gel a 100 v (Dual Triple-Wide Mini-Verlical System, C. B. S.Scientific, CA, USA) va nhuom bac.
- Phan tich sd li^u: Trpng luong phan tu cua cic alen khac nhau dugc xac dmh thong qua ph4n m4n Alpha Ease Fc 5.0. Sd li^u v6 cac chi thi da dupe phan tich su dung phan m^m Graphical Gienotyper (GGT 2.0) ) (Van Berloo, 2008). Dong hpp tu v4 alen cay nhan gien, dong hpp tu v^ alen cSy cho va dj hpp tii dupe ghi nhan lan lupt la 'A', 'B' and 'H'. Phan tram ddng hpp tu alen cay nhan la (96 ^ v^ phan trSm alen cay nhan bao gdm ca alen dong hpp va di hpp la (%
10.
- Danh gia miie dp chiu man theo ti6u chuan cua IRRI
Gieo ma trong khay ndi tr^n m^t dung dich dinh duang Yoshida. Thi nghi^m dupe lap 1^ 3 lan. Cay ma 14 ngay tudi dupe sdc mudi a muc dp EC of 12 dS m ' bang each th6m mudi NaCl trong dung dich dinh duang. Gidng IR29 la ddi chung nhi6m va gidng FL478 lam ddi chimg chju mjn va dong thai la gidng cho gien dich. Dp pH ciia dich nuoi dupe cM6u chinh hang ngay va dung djch nuoi du<yc thay 5 ngay mpt lan. Danh gia dua vao tn^u chung ciia la theo ti^u chuan ctia IRRI (IRRI's Standard Evaluating Score (SES) for rice) khi gidng ddi chiing nhi^m cheft h^t - dat diem 9, voi thang diem danh gia tu 1- khang cao (highly tolerant) ddn 9-nhi^m cao (highly sensitive).
• . KET QUA I K H f N CUU VJi THAO UIAN 1. S^ing Ipc da hinh bd m^
Khoang 500 chi thi phSn hi SSR tr^n 12 NST cua Ilia da duoc silng loc d^ xac dinh cac chi thi cho da hinh giua cac cay bd me cho va nhan gien k h ^ g .
Trong 500 chi thj phan tir SSR rai rdc tr6n 12 NST cua Ilia c6 52 chi thj nam b vung gien Saltol (Saltol region) da dirp^c phfln tich.
Tdng so 63 chi thj cho da hinh giua 2 gidng cho vk nh$n gien AS996/F1478 da dupe ghi nhSn. K^t qu^ nAy cho th^y t ^ sd cho da hinh cua Ccic chi thi SSR da phAn tich vtVi 2 gidng AS996/FU78 U i^t thfip. Tiit cA cac chi Ihj nay da dupe sii dung d^ ph§n (ich sang Ipc cac thd h§ BCiF,, BCzF, va BC3F1.
2. DAnh gi& ki^u gien
.7. K^i quS snng IQC ki^u gien th^hd BC,F, Trong chpn gidng MABC, buoc d^u ti^n ddi voi Inl ca cic th^ lie cAy lai sau khi lai quy tu gien khang vao n6n gien im tii la siing Ipc xac dinh cac ca th^
mang chi Ihj li^n k^t vtVi gien dich (foreground selection). Sau do Id s i n g Ipc vAc dinh cic ca th^ tai td hpp (recombinant selection folowed by background selection). MABC la mot phirong phap dac bi^t hi^u q u i khi dua mot locus gien don di^u khi^n mOt tinh trang quan tarn vao mot n^n gien uu hi [2]. MABC cd 3 uu thd hon han so voi chpn gidng truy^n thdng. Thii nhat la phuong phap nay cho phep chpn loc bang chi thj phan tu li^n k^t mot each hieu qua ddi voi gien dich. Thu hai la kich thuoc cua doan NST mang gien dich dupe dua vao cd th^ han che toi miic ngin nhat ('recombmant selection")- Di^u nay dong nghia voi vi?c han ch^ su anh huong cija vi^c lam thay ddi n^n gien cAy nh^in gien toi miic thSp nhat. Thu ba 1^ khoi phuc l^i n^n gien di truy^
ciia cSy nh&n gien voi toe dp nhanh nhat (thoi gian khoi phyc ngan nh^t) bang chpn Ipc ADN nen gien ('background selection") ma chpn gidng truy^n thdng khfing th^ dgt dupe.
Bang each chpn Ipc trvc ti^p h^ gien cua timg ca th^ trong quAn th^, c6 th^ ki^m tra di truyen ciia dong bd m? %\x di^ng mpt lugiig Ion chi thj phan tii.
Tir dd CO th^ kilm soAt dupe cdc alen quan tam trong cac ck th^ eua qulm th^. D$c bi^t la v i ^ danh gia ti 1^
v^ ham lupng n^n gien gidng nhan gien 6 die cSy t^
td hpp di truyen, k^t hpp lai tra Igi 2 den 3 th^ h^ la cd th^ thu dupe ck th^ mang gien/QTLs din chuy^n vd ti t^ n^n gien cay m? cao nh^t cd th^ qua timg the h^ lai tra lai. D^n thd h^ BC3F, ho$c BC^Fi 4a da c6 th^ cho h; thy, thu dupe cay cd 100% n&i gien uu tii ciia cay nh?n gien wk c6 mang them gien/QTL dich.
Saltol la m6t QTL chinh U6n quan den ham lupng vk ti le Na-K anh huong den tinh chdng chiu
N 6 N G N G H I I P VA PHAT TRIEN NONG T H 6 N • KY 2 - THANG 5/2012
KHOA HOC CdNG N G H £
ciia cay lua trong giai doan ma, da dupe dmh vi trdn NST sd 1 ciia cay liia. QTL nay chiu trach nhiem tdl 43% tinh chdng chiu man cua cay ma. Gidng FL478 OR 66946-3R-178-1-1) voi kha nang chdng chiu m§n cao da dupe phat trien thanh gidng cho gien chdng chiu diing trong chuong trinh chon tao gidng ciia IRRI va la gidng khong nhay cam chu ky quang, ngan ngay hon gidng gdc Pokkali. Hon nua, phan
tich da hinh trong vung SaJtolcho thay rang a FL478 chua mpt doan ADN nhd hon 1 Mb tir Pokkali o' khoang 10,6-11,5 tr^n NST sd 1, rai rac bdi cac alen ciia IR29 [7]. Trong nam 2010, tr^n ea so ket qua nay tir cac nha khoa hpc cua IRRI, thSm mot sd cac chi thj STS da duoc thidt ke cho su dung trong chon gidng MABC, mot ban dd vat ly cho viing Sa/toMuac chi ra duoi day:
11-1 11.2 11.3 114 115 116 I 1 I I 1 I I I M 1 I I I 1 I I I I 1 I M I I I I I I I I I I M I M I I I I I I I I I I I I I
Hinh 1: Ban do vat ly ciia viing gien Saltoltm vi tri 11.1-11.6 Mb tr6n NST sd 1 Tren c a sa ban do viing QTL Saltol, nhung chi
thi trong viing QTL Saltol Ik AP3206 va RM3412, chi thi hiju ich nhat la chi thi nam rai rac 2 phia cua Saltol la RM10694 va RM493, RM10793, trong khi cac chi thi nlm gan dd cd the sii dung cho phep chpn loc am tinh la RM490 nam phia tren Saltol va RM7075 a phia dudi. Cac chi thi microsatellite (ve tinh nhd) khong li^n kdt voi Saltolhao phu tren NST sd 1, cd da hinh giiia 2 gidng cho - nhan gien cung
1 2
dupe sir dung de sang loc cac ca the tai td hop di truyen va chpn loc nen gien th^ nhan. Trong sd 500 chi thi SSR nghien cuu, c6 42 chi thi da dupe phan tich cho sang loc th^ h$ BCiFj. Viec sang loc gien dich da dupe ti^n hanh nha phan tich cac ca di^ voi 3 chi thi AP3206, RM3412 va RM10793. Sau dd, cac chi thi nam 2 phia gan viing Saltol da dupe phan tich trong sang loc xac dinh cac cay tai td hop.
25
Hinh 2: Anh minh hga thi nghi^m sAng Ipc BCiFi (AS996/FL478) sii dung chi thi AP3206 Gidng 1:25bp marker, 2-25 dSn 26-48: cac ca thSBC^Fj. giSng 49:AS996,50: FU78
N 6 N G NGHIEP VA PHAT TRIEN NONG THON - KY 2 • THANG 5/2012
KHOA Hpc C6NG NGHt
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Hinh 3. Anh minh hpa thi nghifm sing Igc BC,F,(AS!»6/FU78) sir dung chi thj RM310 6/.VW /. :«. 51:25 bp marker. 2-25 vi 27-IS:c:k c.-i IM'BC,F,. 4!):AS996. 50: FL478 Trong bang 1 th^ hi^n him l«(mg cSy nhjn di h w tir 14 H% v4 phin tram ham lupng ADN cay glen (R%) cua 12 cay BC,F, tAi iri h<7p, phdn tram " ' * 8'™ I'™ K'*™ "^ ^^ ^l"^" <" ' ' « ' ^ '* » dong hpp tu cua cay nh&n gien la A%; phdn tram cay
Cay BC,F, A H R
65 55,26 34,38 72,45
149 51,43 37,93 70,39
228 60,53 36,36 78,71
238 44,74 34,38 61,92
281 56,25 15,63 64,06
284 78,13 21,88 89,06
305 66,67 33,33 83,33
307 75,76 21,21 86,36
311 63.64 36,36 81,82
401 73,68
0,00 73,68
411 66,67 33,33 83,33
426 63,64 36,36 81,82 Tdng sd 12 cay tai td hpii da dupe siJr dung trong
phan tich sang Ipc nen di truyen cay nhan. Hai ca the P284 va P307 cd li 1^ ham lupng alen cay nhAn cao nhat, toi 89,06 va 86,36%, da dupe xac dmh va sii dung cho viec phat trien quan the BC^Fi- Trong trudng hop su dung chpn gidng MAS thong thuong chi nhan duoc tdi da la 75% alen the nhan, thap hon 16-19% trong nghien cuu nay.
b. Ket qua sang loc kieu gien the he BC-F,
W^HiJ WWi
Eiiil.
I I MHinh 4: Anh minh hpa thi nghi^m sAng Ipc c i c dt th£
BCjFi (AS996xF1478) sir diing chi th) RM10793 b«n trii v4 RM1071I b t a ph4i
Khoang 500 ca the BCgF, cua phep lai (AS996xFL478) da duoc trong va phan tich. Cac chi thi AP3206, RM3412, RM10793, R]^10711 da dupe sii dung trong phan tich s4ng loc gien dich, tuong tu nhu doi voi th^ he BC,F,. CSc chi thi RM10694,RM562, RM7075 nam toa v4 hai b«n virng
gien Ssllol tr6n NST so 1 cung da duoc sii dung trong phan tich sang Ipc cac ca th^ tai to hop di truyen. Trong so 250 ca th^ di hpp tu mang alen boi vi alen la gien tuong ung dich, da tim thay 26 ca th^
tai to hpi).
De phan tich n^n di truyen, cac chi thi cho da hinh ADN nam rai rac tren 12 NST trong phan tich da hinh bo me ban dau da dupe sii dung. Cac ca th^
P307-322, P284-112 va P307-305 da dupe xac dinh la cSy CO ham luong he gien cay nhan gien cao nhat, toi 93,18% trong khi su dung MAS thong thuong chi nhan dupe toi da 87,5% (V th^ he BCjF,.
c. Kpt qua san^' he ki^'u gien theh^ BC^F, Tit lict qua sang Ipc (tii h? BC^F,, 3 quan thi BCjF, da dupe thiet Ijp. Tong s6 371 ca th^ BCjF, da dupe sang Ipc, 94 ca th^ mang chi thi gien dich da dupe phan tich tuong tu nhu vdi th^ h^ BC,F, va BCzF, cho vi^c xac dinh cac ck the t4i to hpp. De sang Ipc nen di truyen cay nhSn, 25 chi thi nam rai ric trSn c i c NST khac nhau ciia liia da duoc phSil tich.
Ca th^ P284-112-209 la ca th^ tot nhsit trong quSn th^ BC3F1 cho thly da mang alen ciia tat ca 63 chi th|
khi phan tich. B6n ca th^ P307-305- 21, P284-112-195, P284-112-198, P284-112-213 chi mang 1 locus dj hpp tir. Ca 5 c4 th^ nay da dupe lua chpn lam dong chpn giong SALT0L!&9m.
NONG NGHllP VA PHAT TRIEN N O N G THON - KY 2 - THANG 5/2012
KHOA HOC C 6 N G N G H £
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a i m ' m':
R.K- BK
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RE 13i ,113) ',112 Mm
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Hinh 5: Sa dd ckc chi thi dung d^ phan tich trfin 12 NST cua cay BC3F1 tot nh& P284-112-209 mang QTL Saltol va toan b6 n^n di truyfe cay nh^n gien phSn tich bang phan m^m GGT2.0
3. Danh gi^ tinh chdng chju man
Hat cua cac ca the da duoc sang loc trong thd he BC2F2 va BC3F1 mang gien Saltol da duoc danh gia muc chdng chiu man. Sdc mudi da dupe tidn hanh ddi vdi cay ma 14 ngay tudi nhir da duoc mo ta trong phan phuong phap nghien cuu. Khi cay ddi chung nhiSm IR29 nhilm di^ni 9, gidng ddi chiing khang ghi diem 3, tat ca cac ca the tir cac cay da chpn loc BC2F2, to ra khang a muc d6 cao tuong duang vdi cay ddi chiing khang. Didu nay cd nghia la gien Saltol a trang thai dong hop tu da hp^t dong tdt a th^
he BC2F2. Mpt sd ca thd BC3F1 cung da dupe danh gik muc khang ghi nhan diem 3-5, dieu nay duoc du doan CO th^ do maah g;*^ Saltol a trang thai di hpp tii.
Nghien ciiu quan the ba^j d^u dupe tao ra giiia IR29 va Pokkali su dung 80 igng RIL, da xac dinh
dupe QTL Saltol khi phan tich ham lupng ti le Na - K cao nh^t hong re vdi LOD - 14,5 va R2 la 64% tren ca SO' chon loc kidu gien [3]. Nghien cuu tidp theo da phan loai cac dong RIL thanh 2 nhdm khang va nhdm nhtim va lap ban dd da dinh vi dupe Saltol nam giua 2 chi thi RM23 va RM140 (10,7 - 12,2 Mb tr^n nhilm sdc the sd 1), va, khang dinh anh huong cua ti 16 Na-K trong rk vdi LOD la 6,6 va R2 la 43% khi su dijfig q u ^ thd RIL [1]. Trong khi dd, chua cd nghien ciiu nao giai thich sir thay ddi ti 16 phan tram cho su th€ hien nhin thay theo each danh gia tifeu chuan SES. Tinh khang hoac su song sot dupe giai thich bdi co chd chia khda dieu khien ben trong dudi^
di^u ki^n stress (lie chd), mudi la nhan td chinh dong gdp vko tinh chdng chiu trong giai doan ma.
Sd U^u cua nghien ciiu gan day khang dinh rang Saltol gop phan vao vi§c dieu chinh ti le ndi can bang NaVK* vdi LOD hkag 7,6 v^ R2 la 27% thong qua 140 N 6 N G NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 5/2012
KHOA HOC C 6 N G N G H C
dong RIL va giam 30% ti le Na-K trong mdm, tir gik tri 1,7-1,2 trong cic ddng lai tra Igi ciia td hpp lR29/Pokkali, trong khi M\ hudng cua S-iltol thd hidn a sd li$u danh giS tidu c h u ^ SES trong cAc quan the QTL va backcross flai ngupe) nhd han r;i(
nhieu. Su that \k Saltol tac ddng d^n li 1^ Na-K nhi^u hon cac tinh trang khac, di6u nky ly giai khS nang gien SKCl van chuy^n mudi duoi su dii^u khi^n cua Saltol QTL [8]. SKCl mk hoa cho nhan id v$n chuy^n mudi giiip cho su di^u chinh nni can bang thong qua vi6c ngan Ctin h^p thu ion Na* trong xylem [9], dieu nay dupe giai thich chiic nang nguySn thuy ciia r^ la lam giam lupng ion Na* van chuydn l^n \k
[5]. Mac dii QTL SKCl da dupe phat hi^n l^n d^u tien o gidng lua Nona Bokra, vi^c nghien cuu them la rat can thiet dd xac d(nh alen ciia SKCl n^u nd cd ciing chuc nang la duy tri ti 10 NaVK' noi can bang trong mam. Dac biet, mpt nghien ciiu g ^ day da dinh vj mpt QTL lien quan ddn ti 1§ Na'/K* nam a vi tri giua 11,1 va 14,6Mb tren NST sd 1 tir gidng liia Japonica Moroberekan [4] va da dua ra y ki^n rang viing Saltol c6 thd cd chiic nang dang kd trong kha nSng chdng chiu mudi 6 ca cac gidng liia Indica va japonica. Trong nghien ciiu nay cac cay dupe sang Ipc P284-112, P307-305 va P307-322 deu c6 di^m khang cao nhu ddi chiing khang. Dieu nay cd nghia la doan gien Saltol ddng hpp tu da boat ddng tdt a nhung cay da sang Ipc d the he BC2F2. Chiic nang cua Saltol a the he BCsF^, BCsF2 se dupe danh gia trong thdi gian tdi.
IV.KETLUAN
- Sang Ipc da hinh gidng cho gien va nhan gien da duoc tien hanh tren 500 chi thi phan tu SSR, trong dd CO 63 chi thi da dupe xac djnh. K^t qua nay cho thay ti le da hinh ADN cua hai gidng AS996/FL478 la rat thap khi su dung cac chi thi phan tu SSR.
- Hai ca thd cua thd h$ BCiFi 1^ P284 va P307 cd ham lupng he gien cay nhan cao nhat tu 89,06 ddn 86,36% tuong ling, cao hon so voi chpn gidng MAS va chon gidng truyen thdng 16-19%. Hai zk thd nay da dupe su dung cho v i ^ thiet lap thd he BCsF,.
- Trong the he BC2F1, ba ca thd cd ham lupTig h^
gien gidng nhan gien la 93,18 ddn 94,03% da dupe x^c dinh bang c^ch sang loc ADN va da dupe sir dung cho viec thidt lap quan th^ BC3F,.
- Ca thd P284-112-209 tdt nhlit cua tiid h^ BC3F1 da duoc sang loc mang 100% ham luong he gien cay
nhan gien khi khSo sat 63 chi thi cho da hinh. Bon cay kh^c la P307-305- 21, P284-112-195, P284-112- 198, P284-112-213 cQng mang x^p xi 100% h^ gien cay nh9n, ciii khac cSy P284-112-209 1^ chiing mang rngt locus dj hpi) tu. Ck 5 cay nay dang dupe tidp tuc lua chpn trdn ddng ru<>ng bang chpn gidng truyen thdng cho vi^c t;io gidng AS996 mang Saltol
TAIUEUTHAMKHAO
1. Bonilla P.. Dvorak J., Mackill D., Deal K., Grcgorio CI, RLI'T and SSLP mapping of salinity tolerance genes in chromosome 1 of rice (Oryza saUva L.) using recombinant inbred lines. Philipp.
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INTROGRESSION OFTHE ,S4i7r?i INTO AS996 USING MARKER ASSISTED BACKCROSSING
Luu Thi Ngoc Huyen Summary
This study focus on developing new salinity tolerance and high yielding rice lines, using markers assisted backrossing (MABC) as a high technology tool for breeding. Total of 500 SSR markers on 12 nee chromosomes were screened for parental polymorphic markers. Of which, 52 primers in the 5a//o/region were checked with the two parents varieties to identify polymorphic primers for screening the 5a/fo/region of the breeding populations. An analysis of 63 SSR markers on approx. 500 plants for each backcross generation of ASS996/FL478 for three steps selection. The two BC,Fi plants P284 and P307 which had the highest recipient alleles up to 89.0696 and 86.3696, were chosen for the next backcrossing. Three BCjFi plants with the recipient alleles up to 94.03 and 93.18% were used to develop BCjFi generation. The best BCjFi plant was P284-112-209 with all the recipient alleles and Sa/to/region. The four plants P307-305- 21, P284-112-195, P284-112-198, P284-112-213 were the second ranking with only one loci heterozygous (applied 63 markei's covered on 12 chromosomes). These five plants were chosen as the breeding lines for result of 5a/fo/AS996 introgression. The breeding line BC4F, having 100% genetic background of donor vanety is ready for develop new salinity tolerant variety ASS996-Saltol to cope with climate change.
Keywords: Variety AS996, marker assisted backcrossing, rice, Saltol, QTLs.
Ngudi phSn bi^n: TS. La T u & Nghia
NONG NGHIEP VA PHAT TRIEN N O N G T H O N - KY2 - THANG 5/2012
