HOl NGH! KHOA HOC CONG NGHE SINH HQC TOAN QUOC 2013
THltT Kfe VECTOR CHUVeN GEN MANG GEN MA HOANHAN t O PHIEN MA
NAC2 Dl£u K H | £ N T I N H C H 6 N G CHI.U HAN CUA CAY LACNguyen Thj Thu Nga^', Le Van So^', Le Trin Binh' ' Tnmng 091 hoc Thai Nguyen
' Vien Cong nghe sinh hoc. Vi6n H^ Iam Khoe hoc va Cdng nghe V7#( Nam TOM TAT
H<? protein NAC (NAM, ATAF va CVC) la m9i trong nhihig hp Ion nhSt co cltira cac nhan tfi phien mii dac timg clio thtrc vat, Chiing (Jong vai (ro quan trong trong cac giai doan phat triln ciia cay tr6ng Da co nhigu cong trinh nghien ciru cua cac nha klioa I19C ve chirc nang cua nhom nhan to phien ma nay trong ana trinh didu Ichicn tinh ch6ng chiu eiia thtrc vat. Nhieu niian to pliicn ma trong ho da dupe tim hieu chirc nang va chuyin vilo mot so dfii nrpng khae nhau nhu lua. d|u nrong, lac, Arabidopsis..., buoe dau da thu dirge nhimg kit qua nhat dmh. Trong nghien evni nay, nhan 16 phien ma NAC2 diroc tach dong tir gifing lac chiu han tfit L12 (da duoc chimg tfil cfing bo tren tap chi Sinh hoc so thing 6 (2013)). Vfii viec sir dung ngufin gen NAC2 tir gifing lac chiu h?n lot L12. chung toi di thilt t l thanh cong vecior mang ciu mic gen phii hop bilu hien 6 thiie vat {pBII2IHAC2). Be dinh gii kha nang hoat dgng cua ciu triic nay trong thuc vat chiing toi da chuyin thir nghiem vao cay mfi hinh. Gifing thuoc la C9I duoc lua chon de chuyen gen.
Kit qua da thu dui?e eay IhuSe la chuyin gen mang ciu Iriic NAC2. day la ngufin nguyen lieu dau tien cho viec phin lich kha nang bilu hien ciia gen chuyin va la co so buoc diu dc dinh gia tinh chfing chju hgn ciia th\rc vaL Vol nhimg ket qua thu dirge tren cay mo htnh, chiing tfii se chuyin ciu triic vao mfit so gifing lac chju hen kem vol hy vgng nang cao kha nang chfing chiu cua cac giong nay nhim muc dich tao ra cac gifing lac co kha nang chiu han tfil phvc vy cho eong tac chon giong.
Tir khfia: Cay lac, chong chiu han, ehuyen gen, NAC2. nhan tfi phien ma NAC
Cac lac dong bat loi cua moi tru'O'ng, die biel l i tinh tr?ng khan hiem nuoc. co the lam giam khoang 70% san luong nong nghiep tren toan the gioi. Cic host dong nong nghifp ma ton li/ong n u * ; nhieu nhat trong so cac hoal dong cua con nguoi. va a cac vung khd han Iran the gioi, djc biet la 6 cac nuoc dang phat trien, luong nuoc dung Irong nong nghiep c6 the vuot 90% luong nude dung trong slnh hoat cua ngudi dan (Kazuo etal., 2012)
Cav tr6ng chuyen gen tren the gioi xult hien iJ My nam 1990 de lir dd phat bieii nhu vu bao, toan diacau c6 tren IbU trilu ha da O'ng dung. Cd r4t nhiiu linh trang duoc Iso ra nhd cdng ngh« c*uy§n gen nhu khang thuoc tru co, khang sSu due than, khang sSu benh, ch6ng chiu (James, 2011) Viec t?o ta cay trong chiu han van 14 dieu dang duoc nghi&n ciru v4 thi> nghiem Tang kha nang chiu han vdi cSy tring thuc su la van de kho khan va phuc tap boi day 14 tinh trang phy thuOc vao hoat ddng cua nhieu gen khae nhau (Kazuo et al, 2003).
Trong qua Irinh tao giSng nhd cong nghe ChuySn gen cin xac d|nh cac gen chu chdt quydinh ' ' * ' ^ " 9 f j " 5 ' " ' ' ° " i cac loai cay, rdi dua nhOng gen nay vao trong cay Ir6ng vdl mong muon lam tang kha nang_ch|u han^ Han han gSy ra hang loat L phan irng smh ly, sinh hda d cay tring, anh huong din ho,l dgng cua rat nhjeu Sen: cac nghien cuu v6 biiu hien gen da xac dinh duoc hang Iram gen hoat ddng trong khi gap ban (Kazuo e( al.. 2003), (Kazuo el ai.. 2007).
Hg protein NAC (NAM, ATAF va CUC) la mdt tn=ng nhdng hg Idn nhat cd chda M C nhan « Pt-!*" ™ = ; ' ^ f ! " " 9 * ° Ihuc vSL Mat s6 hrong Idn cac protein NAC da dugc xac dinh tnjng Svc vat (117 thanh vien o Arabidopsis. 15 thanh S o l i i a . ?52 thanh vien trong he gen d|u luong...) (S»ati « a/.. 2012) Da cd nhieu nghien cuu v | J? P^ten NAC Chung minh cho su lien quan cOa nhdm nay din kha nang ching chiu h|n '^<^'^\''"^.,V^^;^f-f^^^"''"^'"^' elal, 2009). (Kazuo e( a/.. 2012). (Liu el a/.. 2011). (Mao e(a/., 2012), (Swab etal., 2012), (Zheng etai.. 2009).
Trong bai bao nay, chung tdi trinh bay k i l qua thiit k i thanh cdng vector chuytn gen mang gen ma hda nhan t i phien
ZN^2 nhim a i u khiin Unh ching chiu han cua cJy 1,0 va '''^^''?^^^''^1^^"^^^Z£*it"^^a inic nay vao cay thuic la. Nhdng kit qua nay se la Bin d i cho vieo chuyen cau true chiu han vao cay lac nham tao ra
cac giong lac cd kha nang chiu ban tit phuc vg cho cdng tac chon giong.
N G U Y £ N LIEU V A PHU'ONG PHAP NguySn lieu thi,pc vat
^ f c ? , E 1 ^ X / ^ S " S a r c ^ " * ^ ^ 6 n g n g h f i t i bao thucvat-Vidn Cdng nghd sinh hoccung cip.
Chung vi khuan \k cac nguyen iieu DNA
Vector DTRA7/3 pBll21 do ph6ng Cong ngh§ tk bao thi/c v|l - Vi?n Cong nghe sinh hoc cung cip.
clcShOngvi k h ' i n E coli (DHSa) va chOng Agrobacterium tumefaciens CV58C1, do phong Cong nghf l^ bko Ih^c v$l - Vi^n C6ng nghf sinh hpc cung cSp.
Hoa chSt . • • u w», Cac toai hoa chit tinh khiit va thdng dung cd nguin gic tir Anh, Bdc, Tmng Qudc, Han Qudc: Tag-polymerase buSer PCR piling dac hieu cua hang Invitrogen, EDTA SDS, Tris... kit thdi gel va tach plasmid. cac loa. enzyme han di4 cua B ^ o ' S n ^ Y S S l x b a c l , NaCI, Agarose. Tryplon. Tris HCI. EDTA Glycerol. Elhanol 70%. nude deion. Cac ioa, khSng S h kanamycin. rifamicine. cefotaxime, cariiecillin va ckc hoa chit thdng dung khae ... cua cac hang Femienlas.
Invitrogen. Merok. Sigma....
HQI NGH! KHOA HOC C 6 N G N G H E SINH HOC T O A N Q U 6 C 2013 Ptiu-omg phap nghiin CLFU
Thiet fte vector chuySn gen l/iirc vat mang cau true chiu han pBI121:NAC2
Gen NAC2 du-ac khu§ch dai b^ng ky thuat PCR vi/i cap m6i dkc hieu NAC2Ncol/NAC2Notl theo chu trinh nhiet nhu- sau: 94''C/3 phiil, 94"C/30 giay, 57 "C/SO giay, 72<'C/1 phut 30 giSy; 72°C/7 phill. 4°C/30 phut. 35 chu ki.
Thanh ph^n phan ung c l l NAC2 vk vector pRTRA7/3 bing NotI va Wco/: Buffer O (2 pi), NAC2 (pRTRA7/3) (3 (10) pi).
Enzyme NotI (0,2 pi). Enzyme Ncol (0,4 pi), HzO khCr ion (14,4 (7.4) pi). U 37 C trong 2h, di^n di kiem tra, cat lay bang quan tam va thoi gel.
Thanh phan phan i>ng ghep n6i pRTRA7/3: NAC2- NAC2 (7,5pl). pTRA7/3 (Spl), Buffer T4 ligase (2pl), T4 ligase (Ipl), H2O khu ion (4,5pl). Hon hgp phan li-ng dugc u a nhiel do 22''C trong 2h. D9I phan Crng tren da v^ thuc hi&n qua trinh bien nap vao te b^o kha biin B.coll DH5a.
Thanh phan phan i>ng clt WAC2 va vectorpB/J2t bing XbaL Sad: Buffer Tango (2 |jl), NAC2 (pBI121) (10 pi). Enzyme Xbai (1 pi), Enzyme Sad (1 pi). H3O khi> ion (6 pi). 0 37°C trong 2h, dien di kiem tra, cat lay b3ng quan tam v^ thoi gel.
Thanh phan phin i>ng ghep n6i NAC2:pBI121: NAC2 (8pl), pB1121 (4pl), T4 buffer (2pl), T4 ligase (1pl),^H20 khu- ion (Spl). H6n hi7p phan Crng ducrc u d nhigt do 22''C trong 2h, flat phan iJng tren dk va thuc hi#n qua trinh bien nap vao tk bao kha bien E coli 0H5a.
DNA plasmid du-pc bien n?p vao tk bao kha biln E. coli (DH5a) theo phuwng phap soc nhi^t cua Cohen va dong tac gia (1972) va bien ngp vao Agrobacterium bang phucng phap xung dien. DNA plasmid duoc tach chiit va ikm sgch theo phuong phkp ciia Sambrook vk Russell (2002) DNA tai to hg-p du'oc kiim tra bing phu-ang phap PCR vd-i cap m6i dac hi^u NAC2XbaI/NAC2Sacl
Bien n^p gen vao cay ttiudc 14
Giong thudc la C91 du-p'c cung d p tir phdng Cong nghe l l bao thyc vat vai cay con da phat triin duwc 3-4 ik that duwc su dyng de tien hanh c^c thi nghiem phyc vy cho chuyen gen.
T^i sinh da ch6i vk chuyen gen; Cac manh ik tir cky con du-o-c clt vai kich thu-ac khoang Icm^ va dat len mfii tnj-o-ng cam Crng GM (MS + sucrose 30g/i + Agar 8g/l + BAP 1mg/l). Sau hai ngay tr§n moi tru-ong cam ung, manh ik se du-o-c ngam vdi djch huyin phu vi khuan cd chiJa vector chuyin gen mang gen quan tkm. Sau 10 phut cac manh la se du^c dong nudi cly vd'i vi khuin Iren moi tnro'ng GM khong co khang sinh (BAP 1 mg/l) trong 2 ng^y. Sau do. cac manh la tilp tyc du-p-c chuyen I6n mfii tnj-ang GM chu-a kh^ng sinh cefotaxim 500mg/l vk kanamycine 50mg/l (BAP 1 mg/l) d l tai sinh. Sau khoing 4 tuin cac cum ch6i hinh thanh du-oc chuyin sang moi truo-ng GM mai (khdng co BAP)
Ra r|- Sau 3-4 tuin cac choi phat triin cao khoang 2-3 cm du-o-c c l t va chuyen sang m6i tru-6-ng ra r l RM (MS + sucrose 30g/l + Aga 8g/l + IBA 0,1 mg/l) cdchi>a khdng smh chpn Ipc va khang sJnh diet khuln.
Ra blu va nha lu-ai. Sau 3 - 4 tuin c3y con ra r l va phat tnen hoan chfnh vd-i 3-4 la that s i n sang cho ra b l u trlu : cat (1.1). CSy phat tnln vd'i 4-5 la th§t thi chuyin ra trong trong blu 40% dat, 30% triu hun ho|c mCin muc vli 30%i phan chuong muc du-pc trOn deu va xO ly sgch binh trong dilu kien ty nhien.
Cac dong cay thuic 1^ chuyin gen du-pc kiim tra bing phuang ph^p colony-PCR vai cgp moi dac hi§u NAC2Xbal/NAC2SacL
KtJ QUA VA T H A O L U A N
Thilt k l vector tai t l hpp chi>a c l u true gen NAC2
Dya trfin trinh ty gen AhNAC2 da cdng b i tren ngan hang gen (No. EU755023). chung toi tiln hanh thiit k l m i i nhan gen NAC2 d gilng Igc chju han l i t L12
Trinh ty hai c3p moi nhu- sau;
Trinh ty mii
^ NAC2NC01F CCCCATGGATGGGAATTCAAGAGAAAGA NAC2NotlR TTGCGGCCGCTTGCCTG/VACCCGAACCCAACC 2 NAC2XbaiF GCTCTAGAATGGG/WTTCAAGAGAAAGAC
NAC2SaciR CGAGCTCTCAGAGTTCGTCTTTGGATCCA
Gen NAC2 dupe nhkn len bing c|ip mdi dac hieu NAC2Ncol/NAC2Notl, san phim PCR dien di co kich thu-ac khoang lOSObp.
H O l N G H ! K H O A H O C C O N G N G H E S I N H H Q C T O A N Q U O C 2 0 1 3
Hlnh 1 . Ket qua nhan gen NAC2 tir gidng I9C L12. M: Thang DNA chuan 1 hb.
f ^ " i ' l ' l D ^ ^P^ .^'•^^'•^ " " ^ ^^^^ ^^ ^^ 'y ^^ enzyme c i j n g v 6 i vecior pRTRA7/3 b i n g 2 enzyme Ncol/NotI V e c t o r o d i i p i i d i i i r\^n Q u p c [inn s g c n va x u ly cat enzyme c u n g v o i
p R T R A 7 / 3 c d chu-a cac g e n m a hda peptide chi>c nSng n h i r s a u :
mo hirth v e c t o r pRTRA7;3 - — S/J::; HIT,; I '..-L
Mo h b i h v e c t o r p R T R A 7 / 3 ; N A C 2
g-, ;;?^H'-'=H-";-R'l
Hinh 2. Mo hlnh mo ta qua trinh t h i i t ke vector
Vector tai t l h p p pRTRA7/3:NAC2 sau phSn u-ng ghep n l i d u p e kiem tra b i n g phan i>ng colony-PCR v a i d p m i l flgc h i | u NAC2Ncol/NAC2Notl v a b i n g phdn i r n g cat bd-i e n z y m e Ncol/Noll.
Hinh 3. D l f n d i san pham phan i>ng cat vector pRTRA7/3:NAC2 b i n g
NoO/Ncol
Hinh 4. Dien di san ph^m colony-PCR v 6 i Cfip moi dac hieu NAC2Ncol/NAC2Notl
NACZ-Notl/ 'NAC2-ticol tu- vector
H i n h knh dikn di s i n p h i m c l t tren H i n h 3 c h o I h l y k i t q u a thu du-pc hai phSn d o a n g e n c d kich t h u w c khoang 1,1kb va 3,5kb. t i r o n g i>ng v 6 i kich t h u o c cua g e n NAC2 ( n i i t h ^ m d o g n c-myc va KDEL) v ^ vector pRTRA7/3 C h i i n g toi d o n g
&\di t h y c hien p h i n i>ng colony-PCR vd'i c g p moi d # c hi#u NAC2Ncol/NAC2NotL k i t qua trfin H i n h 4 cho t h i y , san p h i m thu du-pc p h i n d o g n g e n cd kfch thu-oc tu-ang u-ng vd'i t i n h t o i n theo ly t h u y i t (1,1kp).
Vector pBI121 d u v c si> d u n g d e c h u y ^ v a o t l b a o t h y c v§t t h d n g qua Agrobacterium tumefaciens. M o hinh d u p e m o la nhu- sau:
Hinh 5. M6 hlnh vsctor 1^1121
V e c t o r p S / 1 2 t l i vector m a n g promoter 35S d i l u k h i l n d o g n peptide ma hda cho p-glucunanida vd-i hai d a u bj c l l bd'i hai e n z y m e g i d i h g n Xbal/SacL D i t h y c hi$n q u a Irinh ghep n l i d o g n g e n NAC2/c-myc/KDEL tir vector pRTRA7/3 v a o v e d o r pBI121 c h i i n g tfli si> dgng c | p m i i /VAC2XbaMVAC2Sac/ d e nhan dogn gen NAC2/c-myc/KDEL tir vector pRTRA7/3. K l l q u i d i r p c the hi$n trong H i n h 6.
HOl NGH! KHOA HOC C O N G NGHE SINH HOC TOAN Q U 6 C 2013
xOOObp
Hinh 6. Di#n di san pham colony-PCR vdi c^p moi die hiSu NAC2XbailMAC2Sa<A Xir vector pRTTiA7/3:NAC2 Phin ung clt dogn gen NAC2/c-myc/KDEL vk vector pBl121 duac tiin hanh v6i enzyme Xba//Sac1. San phim c l l duoc thoi gel, tmh sach d i tiin hanh phan Crng ghep nil tgo vecior tai to hap pBI121:NAC2
Kit qua kilm tra vector tai to hpp bing phan img colony-PCR vai cgp m i l dac hieu NAC2Xbal/NAC2Sacl va clt enzyme XballSaci cho thiy sin phim PCR vd-i cgp mii die hi§u cho bing cd kfch thy6c khoang 1,1kb San phim clt vector bing XballSaci thu di/pc 2 phan dogn gen khoang 1.1 va l l k b , kich thu-ac nay phij hpp vd'i tinh toan theo W thuylt cua gen NAC2 vk vector pBI121. Nhir vgy clu true gen NAC2 6a du-pc thilt ke thanh cdng vio vector pBI121 su dyng trong chuyin gen thyc vkt.
Hinh 7. Dlfn di san pham colony-PCR v6i c^p m6i
d^c hieu NAC2XbailNaC2Saci td vector pBl121:NAC2 Hinh S. Dien dl san phim cat vector pBI121:NAC2 bang Xbal/Sacll
' K n H i ^''^- h>j:'i^j!
Hinh 9. Md hinh vector pBI121:NAC2
Su- dyng plasmid ti> dong khuln duvc kiim tra va lya chpn d l tiln hanh bien ngp vio Agrobacterium va biin nap vao
ciy thuoc l i . "
Hinh 10. San pham colony-PCR v « cap moi dac hi?u NAC2-Xbal/NAC2-Sacl 8 d6ng plasmid tich tir Agrobacterium Bien ngp cau true gen v i o ciy thuic la
?E,*Dn°D ^i':^^'^^" 1^"^"^?^ "^^n loc tren moi Inffirng chua khang sinh kanamycine v i da du^c kilm tra gen chuyin bang PCR vai cip moi die hieu/V/lC2Xbal//VAC2Sacl. • a y
M 1 3 3 4 5 6 7 9 lu 11 n 13 U 1 5 16 17 IS BC Hinh 11. Hinh knh difn di sin phim PCR cac dong
thu&c la chuyen gen.
M: marker Ikb; 1-18: cac dong thu6c la chuyin gen- E)C: dOi chijng khfing chuyin gen
Kit qui di^n di san pham PCR cho thiy, bang dgc higu khong xuat hiSri 6 mlu cly chuyin gen so 3 v i 5, cie dong cay Chuyen gen con Igi diu xull hi?n bing c6 kieh thude phii hpp voi tinh loin 1^ Ihuyit khoing l.lkb. Cic ddng cay thuic
^cnuyen gen cho kgt qui duwng tinh dang du-pc tring va tilp luc theo d6i d l phin tfch khi nang biiu hign cua gen
HOi NGH! KHOA HOC CONG NGHE SINH HOC TOAN Q U 6 C 2013
Lfng dung ky thuat chuyin gen trong vigc nghien CLPU cac nhan t l phien ma NAC d l chuyin vao cay tring da du-pc (M'^" ^'^ ^'^ ^^ ^^'^ ^^'^ ^''^" '^^ ^^' '""^"^ "^'^^ "*^ ^^^^ "^^^ (Arabidopsis, diu tuang, Igc. lua...) (Lu el al., 2011), tMao-eLaL-2012) (7hanq ft gi_ 2009) Bud-c diu da cd nhirng thinh cdng nhlt dmh trong viic giai thich CCT chl liin quan cua nhdm nhin 16 phien ml nay den kha ning chlng chju han cua cay trongr- ^ Sau khi phin lap, nhi>ng gen thuOc ho NAC duvc thiit k i voi cac promoter manh. dgc hieu (CaMV35S promoter, nd29A promoter, ..) va ^chuyin thanh cdng vao mdl s i loai thue v i t nha ky thuat chuyin gen M61 s5 nghien cu-u cdng b i da thu dLFpc cay trong chuyen gen ma hda NAG lang cirong kha nang chong chiu khi gap tinh trang thilu nud^: cua moi tru-d-ng nhu-/i\ra6/c/ops/s, thuic la, Iiia. lua m^... [Uuetal.. 2011), (Mao e( a/, 2012), (Swali efa/., 2012), {Zheng efa/., 2009). O lua, ONAC045, SNAC1 dugc cam ung bd'i han han. ning dp muoi cao, nhigt dp thip. ABA trong la va re (Zheng et al, 2009). OsNACd tang cu-d'ng kha nang ching chiu han han va muii trong lua. NAC TF 6- lua mi, TaNAC2, TNAC4, Taf^ACS da dupe tim Ihly trong phin ung vai han. Ignh. muli, bi Ihupng tin va ABA (Mao ef a/., 2012).
T3NAC2. mpt thanh vien trong hp NAC TF duve tach dong thinh cdng d' liia mi dap u-ng vai sy thilu nud-c, ning dp muoi eao, nhiet dp thap. Tht nghigm chuyen gen da chi ra ring, TaNAC2 lim tang kha nang chdng chju vai hgn, muoi va Ignh 6- cay Arabidopsis (Mao ef al., 2012). 6 cky lac. gen AhNAC2, AhNACS duac tach ddng thanh cong va bleu hign cua chiing cho thiy du-oc cam u-ng bai tinh trang thilu nirirc, ndng dg muoi cao v i nhiet dd thip Gen AhNAC2 da duvc chuyin thanh cdng vao Arabidopsis. Ting cird-ng kha nang ching diju voi hgn va muoi da du-pc quan sat 6- ciy
•Arabidopsis chuyen gen qua phan tich Northern blot v i RT-PCR (Liu ef al.. 2011).
KST LUAW
Vdl vi^c si> dung nguin gen NAC2 tir giong Igc chiu hgn tit L12, chiing tdi da Uiiet k l thanh cdng vector mang c l u triic gen phu hpp bilu hiSn d- thyc vgt.
Da thu dupe cay thuoe la chuyin gen mang clu tnic NAC2, day la nguon nguygn ligu eho viee phan lich khi ning bilu hien iOa gen chuyin va la ca so- bud-e diu d i dinh gia tinh ching chiu hgn ciia thyc vit.
TAI L I E U THAM K H A O
n factors Plant
G U i l h e i n e r L . P . , n a m l m a S M M a v i m i l l p n i m k J S f l f n i l k t J p n n O J i"nillji/^fu.,|i.pf,f,-,,y,-.f ^.^j.hr^nnflfHI-' f f ' ; n , J , m n ^ o r f ^ n c o n , a l . r , • ^ and expression analysis uncover their distinct roles in stress response. Gene. 444.10-23.
James C. (2011) Tinh tr^ng cay chuyin gen/ eay tring CNSH difgc dira vao canh tkc dgi trk Iren tokn thi giiri frong nSm 2005. Bao cao tdm t i l s6 43 cua ISAAA: Ithaca, NY.
Kazuo N , Hlronori T , Junya M., Kazuo S , Kazuko Y.S. (2012). NAC transcnption factors in plant abiotic stress responses Biochimica el Biophysica Acta (BBA)- Gene Reguialoiy Mechanisms, 1819(2)'97-103.
Kazuo S , Kazuko Y.S. (2007). Gene networks involved in drought stress response and tolerance. Experimental Botany, 58(2):221- 227.
Kazuo S.. Kazuko Y.S., Moloaki S (2003). Regulatory network of gene express/on in the drought and cold stress responses Current opinion in Plant Biology, (6):410417,
Uu X , Hong L., Li X.Y., Yao Y , Hu B , Li L. (2011). Improved drought and salt tolerance in transgenic Arabidopsis overexpressing a NAC transcriptional factor from Arachis hypogaea. Biosci. Biotechnol Biochem., 75{3):443-450.
lirtao X., Zhang H., Qian X , Li A., Zhao G , Jing R (2012) TaNAC2, a NAC-type wheal TF conferring enhanced multiple abiotic stress tolerance in Arabidopsis Joumal of Expenmenlal Botany, 63(6): 2933-2946.
Swati P.. Pranav P.S., Prem S S , Manoj P. (2012). NAC proteins, regulation and role in stress tolerance. Trends in plant science, 7(6):369-381
Zheng X , Chen 6., Lu G., Han B. (2009). Overexpressmn of a NAC TF enhances rice drought and salt tolerance. Biochemical and Biopfiysical research communications, 379:985-989.
VECTOR CONSTRUCTION CONTAINS GENES ENCODING TRANSCRIPTION FACTORS NAC2 CONTROL DROUGHT TOLERANCE IN PEANUT
Nguyen Thi Thu Nga'*, Le Van Son^, Le Tran Binh^
'Thai Nguyen University
%stitute of Biotechnology SUMMARY
Tlie NAC (NAM, ATAF, and CUC) protein is one of the biggest femilies contain transcription factors specific to plants They play an important role in the development stage of the crop. There have been many research works of scientists on the functions of tliis group oftranscriptionRictors in the control tolerance of plants Many of these transcription factors has been to leara and function into a number of different objects such as nee. soybean, peanut, Arabidopsis, initially obtained certain results. In this study, transcription factore NAC2 line is separated from die drought-resistant varieties well LI2 (were we announced in a previous article). With the use of genetic resources NAC2 from varieties of drought tolerant L12, we have successfully designed appropriate vecior carrying ilie genetic expression m plants (pBI121: NAC2) To evaluate the performance of this structure in plant test we have moved into the model. C91 tobacco varieties selected for gene transfer. Results obtamed transgenic tobacco plants canying NAC2 structure, this is the first resource for the analysis of transgene eiqiression and ability is the basis of the first step to assess the drought tolerance of plants. With tlic results obtained on the model, we will transfer the structure into a number of drought-resistant varieties less in the hope of improving tolerance of these vaneties aims to create vaneties resistant good term for breeding work.
Keywords: Arachis hypogaea, drought tolerance, gen transfer, NAC2, NAC transcnption factors.
" Author for corresspondence [email protected]