Nguyin VQ Thanh Thanh vd Dtg Tap chi KHOA HOC & CONG NGHE 85(09)/ 148
DAC DIEM CUA GEN GmDREBS PHAN LAP
TIJ MOT SO GIONG DAU TI/OTVG DIA PHUONG VIET NAM
Nguyen Vu Thanh Thanh', Nguyen Thiiy Giang , Hoang Van Manh', Le flii-e Huan', Chu Hoang Mau^
Trudng DH Khoa hgc - DHTN. ' \ 'ten Khoa hoc Sir song - DHTN. 'Dgi hoc Thdi Nguyen
TOM
TATDau tuong {Glycine max (L.) Merrill) la cay tuong ddi man cam vdi digu kien ngoai canh va thudc vao nhdm cay chju han kem. Trong nhifng nam gan day, dign bien khi hau ngay cang phiic tap, lugng mua phan bd khdng dgu giua cac vimg va giira cac thdi ky trong nam nen ban han va nang ndng keo dai. Tinh trang han ban anh hudng Idn dgn nang suat dau tuong, vi vay nghign ciru gen lign quan dgn kha nang chiu ban ciia cay dau tuong lam co sd cho u-ng dung ky thuat chuygn gen nham nang cao kha nang chiu ban ciia cay dau tuong dang dugc quan tam ciia cac nha chon gidng.
Protein GmDREB la nhan td phign ma kich hoat nhdm gen lign quan dgn tinh chiu han ciia dau tuong, khdng phu thudc vao ABA, san pham eiia gen GmDREB dugc tim thay nhigu khi cay gap han, man va lanh. Trong nghien ciru nay, chung tdi trinh bay kgt qua khugch dai gen GmDREB5 tir DNA tdng sd d hai gidng dau tuong Ciic Ldng Phu Binh (CPB) va Vang Ngan Son (VNS) ciia Viet Nam bang phan iing PCR vdi cap mdi duge thiet ke va tdng hgp la DREB5soyF/DREB5soyR. Kgt qua tach ddng va doc trinh tu gen GmDREB5 cho thay trinh tu gen GmDREB5 ciia ca 2 gidng dau tuong CPB, VNS deu dai 924 bp. Ket qua so sanh trinh tu nueleotide ciia hai gidng nay cho thay cd do tuong ddng cao (dat 99.2%), trinh tu amino acid cua protein DREB5 do gen nay ma hda d hai gidng dau tuong nghien ciiu cung cho do tuong ddng cao (dat 98,7%). Khi dem so sanh vdi trinh tu gen GmDREB5 ciia eac gidng dau tuong khac da cdng bd la EF583447, Xanh Tign Dai (XTD) va Ban Gidc (BG) thi do tuong ddng ciia nam gidng nay dao ddng til- 91,9% dgn 99,2%. Trinh tu amino acid cua protein DREB5 ciia nam gidng nay cd do tuong ddng tir 88,4% dgn 99,0%. Can tigp tuc nghign ciiu ve gen GmDREBS lam co sd thiet ke vector mang cSu triic gen GmDREBS, phuc vu chuyen gen tao cay dau tuong bign ddi gen chiu ban.
Til- khoa: Ddu luung dja phuung, ddu luong chuyin gen. chju hgn. gen GmDREBS. Glycine max.
MODAU
Dau tuong {Glycine max (L.) Merrill ) cd vj tri quan trgng trong co' cau cay trdng ndng nghiep ciia the gidi va ciia Viet Nam.Cay dau tuong dugc eoi la cay trdng chien luge ciia niiieu quoc gia tren the gidi. Ham lugng protein trong hat dau tuong rat cao. chiem khoang 32%-56%. bam lugng lipid chiem 12%-25%, ham lugng glucid chiem 10-15%
va chira nhiki loai vitamin...[3], day la ngudn dinh dudng quan trgng eho con ngudi va vat nudi. Trdng cay dau tuong cdn cd tac dung cai tao dat nhd vi khuan cd djnb dam chiia trong cac not san d re. Vdi nhung gia trj to Idn dd, cay dau tuong dugc trdng phd bien d nhieu no-i tir 55" vT B k din 55" vT Nam. tii viing thap hon mue nirdc bien den viing cao tren 2000in so vdi muc nude bien vdi dien tich dat khoang hon 74,7 trieu ba.
' Tel: 0913383289; Email: mauchdhindgmail.com
Nhung nam gan day do bien ddi kbi ban da cd nhimg tac ddng bat Igi den su sinh trudng va phat trien eua cay trdng ndi ehung, trong dd cd cay dau tuo-ng. Dau tuong la cay tuang ddi man cam vdi dieu kien ngoai canh va thudc vao nhdm cay chju ban kem. Han ban da anh hudng Idn den nang suat dau tuo-ng, vi vay nghien ciru gen lien quan den kha nang chju ban ciia cay dau tuo-ng lam co- sd cho img dung ky thuat chuyen gen nham nang cao kha nang chju ban ciia cay dau tuong dang dugc quan tam ciia cac nha chgn gidng. Qua trinh phien ma ciia nhdm gen chju ban chju tac ddng cua cac yeu td dieu khien qua trinh phien ma, trong dd cd nban td DREB. DREB la bg gen tdng hgp protein dugc tim thay nhieu trong te bao sdng khi thuc vat gap cac dieu kien ngoai canh bat lgi nhu ban ban, man va lanh. Nhieu cdng trinh nghien ciru cua cac nha khoa hgc tren the gidi cho rang. DRIT^ hi
Nguyin Vd Thanh Thanh vd Dig Tap chi KHOA HOC & CONG NGHE 85(09)/l: 143- 148 nhan td phien ma tham gia tich cue vao
qua trinh nay bang each kich hoat nhanh su boat ddng ciia cac gen tham gia true tiep chdng lai cac dieu kien bat Igi ciia mdi trudng nhu han han, man va lanh, nhung nhan td phien ma DREB boat ddng khdng can thdng qua sir tac ddng cua ABA (Tran Thj Phuong Lien va dtg) [8].
Nam 2009, Mei Zhang va dtg da nghien cuu nban td phien ma d thue vat va vai trd eiia nd ddi vdi thuc vat chdng lai eac yeu td bat lgi phi sinh hge. Stress phi sinh hge nhu han ban, lanh va do man cao da anh hudng nghiem trgng den sir tang trudng va nang suat eiia eay trdng. Nghien cu'u da de cap den cau true va chiic nang cimg nhu irng dung ciia cac nhan td DREB trong cai thien kha nang chdng chju ciia eay trdng [11]. Li va dtg (2005) phan lap gen GmDLiEB tuong ddng gdm 3 gen GmDREBa, GmDREBb, GmDREBc tir cay dau dau tuong. Ket qua nghien ciiu eho thky ba gen nay ed chuc nang dac biet trong su phan irng vdi stress phi sinh hgc eiia dau tuong [9]. Ming Chen va dtg (2009) eung trinh bay ket qua phan tieh chuc nang eiia gen ma hoa nhan td phien ma DREB3 d dau tuong [13]. Dac diem cau triic ciia gen GmDREB! 6 dau tuong da dugc phan tich bdi Chu Hoang Mau va dtg (2011) [10]. Nghien cuu ciia Stolf-Moreira va dtg (2010) khi xac djnh muc do bieu hien eua yeu to phien ma DREBl cho thay rang, trong dieu kien khd ban GmDICEBl bieu hien gia tang trong la va re [14]. Gen GmDREB2 duoe phan lap tir dau tuong va xep vao nhdm 5-trong phan hg DREB, gia dinh AP2/EREBP. Gen GinDLCEB2 dugc bieu hien bdi han ban va ndng do mudi cao. Phan tich ham lugng proline trong eay thudc la chuygn gen GmDREB2 da ehi ra rang su tich luy proline cao hon so vdi ddi chung trong dieu kien han ban (Minh Chen va dtg (2007) [12]. Nam 2007, Chen va dtg da phan lap gen GmDHEBS tii- mRNA d cay dau tuong, gen cd kich thudc 927 bp [2]. Cao Xin-You va MA You-Zhi (2008) cdng bd kit qua phan lap va nhan dang protein GmGpi tuang tac vdi protein GmDREBS a dau tuong [1], nghien
cuu dac tinh eiia gen GmDREBS phan lap tii- gidng dau tuong dja phuong Viet Nam ciia Chu Hoang Lan va dtg (2010) [7]. Tuy nhien eho den nay nghien ciiu dac diem cua gen GmDREBS va ung dung nd trong chuyen gen van cdn rat it cdng trinh nghien ciru va thao luan ve van de nay.
VAT LIEU VA PHUONG PHAP Vat lieu
Chung tdi su' dung 2 gidng dau tirong dja phuong cd chat lugng tdt do Vien nghien ciru Ngd, Dan Phugng, Ha Ndi cung cap.
Phucng phap nghien ciru
Tach chiet DNA tdng sd theo Gawel va Jarret (1991) [4]. Kiem tra ham lugng va do tinh sach DNA bang phuong phap quang phd hap thu va dien di tren gel agarose 0,8%.
Khuech dai gen GmDREBS bang ky thuat PCR nhd cap mdi DREB5soyF/DREB5soyR dugc thiet ke dua tren trinh tu gen mang ma sd EF583447 tren Ngan hang gen quoc ti (NCBI).
DREBsoy5E:5'-ATGCAATTCCCTCACCAATT-3' DREBsoy5R:5'-TCAATCCTGATCCTTCCACA-3' San pham PCR dugc lam sach bSng bd kit AccuPrep PCR purification (Bioneer - Han Qudc), sau dd dugc gSn vao vector pTZ57R/T va bien nap vao te bao kha bien E.coli chiing DH5a theo phuong phap sdc nhiet. Sir dung enzym eat gidi ban BanMX de kiem tra su ed mat eua gen DREB5 trong plasmid tai to hgp, san pham eat dirge dien di kilm tra tren gel agarose 1,0%. Trinh tii- nucleotide ciia gen DREB5 dugc xac djnh tren thiet bj giai trinh tu nucleotide tu ddng ABI Prism 3130 - USA/Japan.
KET QUA VA THAO LUAN
Ket qua khuech dai va tach dong gen DREB5 cua giong dau tuong Ciic Long
Phu Binh va Vang Ngan Son , La non 7 ngay tudi eiia hai giong dau tirong
Ciie Ldng Phil Binh va Vang Ngan Son duge su' dung di tach DNA tong so. Cac mau DNA cd chat lugng tot dugc khang djnh bang kit qua kilm tra tren may quang phd d budc sdng A,= 260/280mn va dien di tren gel agarose
Nguyin Vit Thanh Thanh vd Dtg Tap chi KHOA HOC & CONG NGHE 85(09)T: 143 - 148 0.8%. Gen GmDREBS ciia gidng dau tuong
CPB va VNS dugc khuech dai bang phan irng PCR vdi cap moi DREBSsoyE/ DREB5soyR, nhiet do gan mdi la 51 C. Sau 30 chu ky phan irng, san pham PCR dugc kiem tra bang dien di tren gel agarose 1,0% (Hinh 1).
San pham PCR tinh sach se dugc gan true tiep vao vector tach ddng pTZ57R/T va sau dd dugc bien nap vao te bao kha bien E.coli chiing DH5a, tien hanh nudi cay tren mdi trudng LB agar cd bd sung ampicillin 50
^g/ml, X-gal 20 mg/ml, IPTG lOOmM, dl qua dem. Chgn lgc cac khuan lac trang nudi trong 3pl LB ldng cd khang sinh ampicillin vdi nong do 50pg/inl, tdc do lac 200 vdng/phiit d 37°C qua dem. Te bao tai td hgp dugc thu lai bang each ly tam va tach chiet plasmid tai td hgp. Kiem tra san pham DNA plasmid bang enzym gidi ban BamVW va dien di tren gel agarose 1,0% (Hinh 2).
Ikb 0,93 kb
Hinh 1. Kgt qua dien di san pham PCR nhan gen GmDREBS ciia 2 ^idnsi dau tuong 1. Marker Ikb; l . C P B ; 2.
li^S^"-*
3kb—•1 1 _ I k b — • r * M i
•"?
VI
- •
^S
L
L _
2,89kb 0.93kb
Hinh 2. Anh dien di san pham cat plasmid bang enzyme gidi han BamHl
M: marker Ikb; Hai ddng I va 2 la DNA plasmid mang gen GmDREBS cd do dai khoang 0.93 kb
Ket qua xac djnh trinh tu gen GmDREBS ciia giong dau tuo'ng CPB va VNS
Plasmid tai td hgp sau kbi tinh sach dugc su- dung trong phan iing xac djnh trinh tu nucleotide. Trinh tu gen GmDREBS ciia hai gidng dau tuong nghien ciru deu cd kich thudc 924 bp. Ket qua so sanh trinh tu nucleotide ciia gen GmDREBS a hai gidng dau tuong CPB va VNS cho thay. hai trinh tu nay chi khac nhau d 7 vj tri la 211, 331. 417, 424,889,890,891 (Bang 1).
Bang 1. Su sai khac trong trinh tu nucleotide eiia sen DREB5 d hai gidng CPB va VNS
\ Vj tri Tgn\ 211 g i d n g \
CPB T VNS G
331 A T
417 C A
424 G A
889 G C
S90 C G
891 A T Do vay, trinh tu gen GmDREBS ciia gidng dau tuong CPB va VNS cd do tuong ddng cao 99,2%. Ket qua so sanh trinh tu amino acid suy dien tir gen GmDREBS ciia hai gidng dau tuong CPB va VNS bieu hien su khac nhau d cac vj tri 71, 142, 297 (Hinh 3). Su tuong ddng ve trinh tu amino acid cua hai giong dau tirong CPB va VNS dat 98,7%.
So sanh trinh ty nucleotide va trinh tu amino acid ciia gen GmDREBS ciia hai giong dau tuong CPB va VNS voi cac giong dau tuong EF583447 (Trung Quoc), Xanh Tien Dai, Ban Gioc (Viet Nam)
Kit qua so sanh trinh tu nucleotide ciia gen GmDREBS giira gidng dau tuong CPB va VNS vdi gen GmDTLEBS ciia gidng dau tuong cd ma so EF583447 trong Ngan hang gen NCBI cd ngudn gdc tir Trung Qudc, gen GmDREBS eiia gidng Xanh Tien Dai va gidng Ban Gidc Viet Nam cho thay, 5 gidng dau tuong cd he sd tuong ddng dao ddng tir 91,9% den 99,2%.
Til- ket qua so sanh trinh tu gen GmDREBS ciia nam gidng dau tirong. chimg tdi thiet lap so dd hinh cay bieu dien mdi quan he di tru>-en eiia nam gidng dau tuong (Hinh 4).
Nguyen VU Thanh Thanh vd Dtg Tap chi KHOA HOC & CONG NGHE 85(09)/1: 143-148
i O 2 0 3-2. 4 0 5<> € 0 . . . . 1 . . . . I . . . . 1 . . . . I . . . . ! . . . . ! . . . . I . . . . I . . . . I . . . . I . . . . I . . . . I
C P B M Q F H < . 2 ! - ' O T S N ' i J - H : i F v ' N Q W H ' J M :, A i K S S S a t i H N N i . A ; ' , ' i; '. A . r U 3 S ; S S : r V H H H H Q a O V T s I S . . . .
—< .Z> 2 0 SJO l O O 1 I . O — 2 0
• . . . 1 - . . . i . . . . I . . - . • - . . - I ; . . . . 1 . . . . f . . . i . . . . I V N . = i ( ' • ) • - • • •
I. S -O 1 q O I S I ' :L 6 O ^"7 0 1 s o
• •-. i - - . . ! . - - - t - - • • ? • - • - i [ . . . . i . . . . \ . . . . { . . . . \ ^
V 7 M S (- ' ) • - •
1.3 0 2 0Q Z-O 2 2 0 2 3 - 3 2 . 3 0 J . . . . I . . . . ! . . . . : . . . . 1 . . . . I . . . . , . . . . ; . . . . 1 . . . . j . . . . i . . . . I . . . . I
V T ^ . S . . . . . ,
2 5 O 2 60 2 ~ 0 2 5 O 2 9 0 3 0 0 C P B : . - W t;.: r j . . . : . o O n - rjvTN C . ' . ' : . N N . . - . , , < , • N E J O O . R ; E ; .
V T - / . S . . . . , . . . . . . . . .
C P B . 0 /
Hinh 3. So sanh trinh tir amino acid ciia gen GmDREBS d gidng dau tuong CPB va VNS Nhdm 1
3.7 Nhom 2
'-9
•;Mb
2 0 Nucleotide Substitutions (xlOO)
Hinh 4. Dd thj bigu thi mdi quan he giua cac gidng dau tuong dua tren phan tich trinh tu nucleotide ciia gen GmDREBS
99,0%. Cac vj tri sai khac duge tbi hien d Ket qua cho thay, 5 gidng dau tuong dugc
chia thanh 2 nhdm: nhdm 1 gom 3 giong CPB, VNS, Xanh Tien Dai; nhdm 2 gom 2 gidng Ban Gidc va EE583447.
Chimg tdi nhan thay, nhdm 1 gdm eac gidng CPB, VNS, Xanh Tien Dai da dugc danh gia kha nang chdng chju, day la eac giong chju ban kem (CPB, VNS) va chju han trung binh (Xanh Tien Dai); nhdm 2 cd gidng Ban Gioc da dugc danh gia la gidng chju ban tdt [5], [6]. Nhu vay, dua tren trinh tu gen cd thi phan chia cac gidng cd su sai khac ve kha nang chju ban. Can tiep tuc md rdng nghien ciru vdi sd lugng gidng nhilu hon d l cd nhiing ket luan ehinh xac vl chi thj phan tu- lien quan den tinh chju han dua tren trinh tu gen DREB5.
Trinh tu amino acid eiia protein DREB5 d gidng CPB va VNS so vdi ba giong EE583447, gidng Ban Gioc va giong Xanh Tien Dai ed do tuang ddng tii- 88,4 din
hinh 5. Hinh 5 cho thay, giira 5 gidng cd tdi 49 vj tri sai khac trong trinh tu amino acid.
Tuy nhien, trinh tu amino acid cua 3 gidng CPB, VNS va Xanh Tien Dai cd sir tuong ddng cao (98,4-98,7%); giong Ban gioc va EF583447 eung ed do tuang dong eao (99,0%) (Bang 2).
Bang 2. He sd tuong ddng vg trinh tu amino acid trong protein DREB5
BG CPB VNS XTD EF583447
BG CPB 100 88,7
100 VNS
87,7 98,7 100
XTD 90,4 98,4 97,4 100
EF583447 99,0 89,4 88,4 91,1 100 Dac biet, khi so sanh giua nhdm chju han tdt va nhdm chju ban kem chimg tdi nhan thay ed su thieu hut ve trinh tu amino acid a cac vj tri 5 8 , 5 9 , 6 0 , 177, 178, 179 (bang 3):
Nguyin VU Thanh Thanh vd Dtg Tap chi KHOA HOC & CONG NGHE 85(09) I: 143-148 Bang 3. Vj tri sai khac trong trinh tu amino acid
cua hai nhdm chiu ban khac nhau 58 59 60 177 178 179 Nhom chju han tot
Nhom chiu han kem
H H Q E 0 0 Nhdm chju ban tdt cd cac amino acid H, H, Q, E, Q, Q lan lugt d cac vj tri 58, 59, 60, 177, 178, 179 edn nhdm chju ban kem khdng cd cac amino acid d vj tri nay. Vi vay. can cd nhirng nghien ciru sau hon de lam rd su khac biet nay.
KET LUAN
- Da khuech dai va xac djnh trinh tu nucleotide ciia gen DREB5 tii- DNA tdng sd ciia hai gidng dau tuo'ng CPB va VNS vdi kich thudc 924 bp.
- Trinh tu nucleotide eiia gen GmDREBS a gidng dau tuong CPB va gidng dau tuong VNS cd do tuong dong eao (99,2%).
- Trinh tu amino acid ciia protein DREB5 o 2 gidng dau tuong CPB, VNS va ba gidng EF583447, Xanh Tien Dai. Ban Gidc cd do tuong dong tir 88,4 din 99.0%. Dac biet. cd su khac biet ve trinh tu gen va trinh tir amino acid giua nhdm chju ban tdt va nhdm chju ban kem.
- Can tiep tuc nghien cii-u ve gen GmDREBS lam ca sd cho viec thiet ke vector mang cau true gen GmDREBS phuc vu chuyen gen tao cay dau tuong bien ddi gen chju ban va xac djnh cbi thj phan tir lien quan den tinb chju ban dua tren trinh tu gen DREB5.
LOI CAM ON: Cdng trinh dugc thuc hien vd hodn thdnh vdi sir hd trg kinh phi cua De tdi DudnTRLG.
EF 56^447 C P B V N S
X a n h t i e n d a i B a n G i o G
Hi-.f-.-'cyvvviAy -•C;.-v- _ . i-.J-TN I i ., HMU r i v .
EF 5 8 3 4 4 7 C P B V N S
X a n h ' t i e i i c l a i B a i i G i o c
R r N ! :'K R' ; R Rl = R
R
EF 08T447 C P B V N S
X a n h t i e i i c l a i B a i i G i o c
R R>TR R K R .KN R N NKx;KKir:r:
EF ••8T447 C P B V N S
X a n h t i e n c i a i B a i i G i o c
. N .
ZK N H N O GD D D - - ; . E", ii . E ! 1 .
2 5 0 EF ':>83447
C P B V N S
X a n h t i e i i c l a i B a n G i o c
DD D 'f'NN C N N ,. E: N LJ K R . E
. . 0 - - . D n . ,., . . . , ,;- . D O . ,. . .
EF 583447 C P B VNS
X a n h t i e n c i a i B a n G i o o
3 1 0 _NT'c; Ci'-' Kn.iy
Hinh 5. So sanh trinh tir amino acid cua CPB. VNS voi EF583447 (Trung Quoc), Xanh Tien Dai, Ban Gioc (Viet Nam)
147
Nguyin Vii Thanh Thanh vd Dtg Tap chi KHOA HOC & C O N G N G H E 8 5 ( 0 9 ) / l : 143 - 148 TAI LIEU T H A M K H A O
[I]. Cao Xin-You. and MA You-Zhi (2008). "Esolation and Identification o f a GmCpi Interacting Protein with GmDREBS Protein in So\bean (Glycine ina.xf. Acta Agronomica Sinica. 34(10); 1688-1695.
[2]. Chen M.. Y. Ma, Z. Xu. L. Li and Q. Wang.
Glycine max deh_\dration-responsive element binding protein 5 (DREB5) mRNA. complete eds. GenBank:
EF583447.I.2007.
13]. Ngo Thi Dan. Trin Dinh Long. Tran Van Lai. Do Thi Dung. Pham Thi Dao (1999). Cay dau luong. Nxb Nong Nghiep.
[4]. Gawel N.J .lart R L (1991). Genomic DNA isolation, hup://\\WW.weihenstephan.de/
[5]. N g u \ i n Hiep Hoa. Nghien ciru gen DREB5 ma hoa protein lien quan den qua trinh phien ma ciia nhom gen chiu han a cay dau tucmg. Luan van thac sL Dai hoc Thai Nguyen. 2010.
[6]. Nguyen Thi Minh Hong. Nghien ciru dac diem cau triic gen DREBl a dau tucmg (Glycine max (L.) Merrill ). Luan \an thac si. Dai hoc Thai Nguyen. 2010.
[7]. Chu Hoang Lan. Ngu>en Tuan Anh. Ngu\en Vu Thanh Thanh. Nguyen Hiep Hoa. Chu Hoang Mau (2010). Characterization of the GinDREBi gene isolated from the soybean cultivar Xanh Tiendai.
Vietnam. Proceedings of 2010 International Conference on Biology. Environment and Chemistty (ICBEC 2010).
28-30. December 2010. Hong Kong. IEEE: 354-358.
[8]. Tran Thi Phucmg Lien (1999). Nghien ciru dgc tinh hoa sinh vd sinh hgc phdn lu cua mgt so giong ddu lirong ed kha ndng chju nong. chju hgn a ('let Nam, Luan an tien si sinh hoc. Vien Cong nghe sinh hoc. Ha Noi. 22-34.
[9]. Li XP. Tian AG. Luo GZ. Gong ZZ. Zhang JS.
Chen SY (2005). Soybean DRE-binding transcription factors that are responsive to abiotic stresses.
Theoretical and Applied Genetics. I I0(8):I355-I362.
[10]. Chu Hoang Mau. Nguyin Vu Thanh Thanh.
Nguyin Thi Minh H6ng. Hoang Van Manh (2011). Dac diem cua gen DREBl phan lap tir giong dau tuong dia phucmg (Glycine max L. Merrill) Xanh la Ba be (Bac Kan)"'. Tgp chi Sinh hgc. 33( I): 74-79.
[ I I ] . Mei Zhang. Wei Liu. Yu-Ping Bi (2009).
Dehvdration-responsive element-binding (DREB) transcription factor in plants and its role during abiotic stresses. Yi chuan. 31(3): 236-244.
[12]. Ming Chen. Qiao-Yan Wang. Xian-Guo Cheng.
Zhao-Shi Xu. Lian-Cheng Li. Xing-Guo Ye. Lan-Qin Xia. You-Zhi Ma (2007). GmDREB2. a soybean DRE- binding transcription factor, conferred drought and high-salt tolerance in transgenic plants. Biochemical and Biophysical Research Communications. 353 (2):
299-305.
[13]. Ming Chen. Zhaoshi Xu. Lanqin Xia. Liancheng Li, Xianguo Cheng, .lianhui Dong. Qiaoyan Wang and Youzhi Ma (2009). Cold-induced modulation and functional analyses of the DRE-binding transcription factor gene. GmDREB3. in soybean (Glycine max L.).
Journal of Experimental Botany. 60 (I): 121-135.
[14]. Stolf-Moreira i L . M E Medri. N Neumaier. N_G Lemos. R L Broi;in. F C Marcelino. M C N de Oliveira.
.1 R B Farias. R V Abdelnoor. A L Nepomuceno (2010).
Cloning and quantitative expression analysis of drought- induced genes in soybean. Genetics and molecular research 9{2):858-67. '
S U M M A R Y
A N A L Y S E O F GmDREBS G E N E S E Q U E N C E S I S O L A T E D F R O M T W O V I E T N A M E S E L O C A L S O Y B E A N C U L T I V A R S
Nguyen Vu Thanh Thanh', Nguyen Thuy Giang', Hoang Van Manh^, Le Due Huan^, Chu Hoang Mau^"
'College of Science - TNU. 'Institute of Life Science - TNU. Thai Nguyen University Soybean (Glycine max (L.) Merrill) is cultivar relatively sensitive to external conditions and is a low drought- tolerant species. Recently, the change climate has been increasingly complex , rainfall has been unevenly distributed among regions and between the periods of the year, drought often prolonged. Drought affected sovbean yield, so the study of genes related to drought tolerance of soybean was very interest ofthe breeders.
DREB proteins are transcription factors activating the gene groups related to drought tolerance of soybean, which is not dependent on ABA. products of GmDREB gene found when cultivars met strees drought, salt and cold. In this study, we presented results of amplification of GmDREBS gene from genome of Vietnamese soybean cultivars (Cue Long Phu Binh and Vang Ngan Son) via PCR reaction using specific primers DREB5so>F/DREB5so> R. The PCR products containing the DREB5 fragments were cloned into pTZ57R/T and sequenced. This gene was 924 bp. The gene sequences of these two varieties had similar high levels reached 99.2%. amino acid sequence ofthe protein encoded by two genes had similar levels reached 98.7%.
When comparing the nucleotide sequencing of GmDREBS gene of two soybean cultivars (Cue long Phu Binh and Vang Ngan Son) to three the soybean cultivars (EF583447. Xanh Tien Dai and Ban Gioc). we found the similarities o f t h e five varieties ranged from 91.9% to 99.2%. Amino acid sequences of DREB5 o f t h e the soybean cultivars ranged from 88.4% to 99.0%. It is necessary for further .studies to designed vector carrying the GmDREBS gene structure for creating transgenic soybeans drought-resistant.
Keywords: Drought tolerance. GmDREBS gene. Glycine max. local soybean, transgenic soybean.
Tel: 0913383289: Email: mauchdhtnQigmail.com
148
