Cong nghe sinh hoc & Gidng cav tronp
CHUYEN GEN CODA MA HOA CHOLINE OXIDASE VAO CAY XOAN TA
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(Meiia azedarach L . ) T A N G CLTONG K H A N A N G C H I U H A NBui Van Tfadng^, Le Van S o u ' v a Chu Hodng H a '
ThS. Viin Cong nghe sinh hoc Ldm nghiip. Trudng Dai hoc Ldm nghiep TS. Vien Cong nghe Sinh hoc, Vien Hdn Ldm Khoa hoc vd Cong nghe Viet Nam
T O M T A T
NhQng yeu ta b4t Ipi tii moi tnrcmg nhu han, m?tn, iing, nhiet dg cao thucmg lam mit can bSng vS 4p suit thim tiiau gay anh hucmg nghiem trong den sinh truong, phat tri^n va nang suit cay trSng. Tao cay tr6ng biSn dfii gen cai thi€n kha nang chong chju v6i cac dieu kien bit lpi ciia moi truong dang ia mot huong nghifin ctiu trpi^ tam cua c6ng nghfi sinh hoc thuc vit hien nay. Gen codA t6ng hpp ohan lao ma hoa cho choline oxidase tham gia vio sinh tong hop glycine betaine da duoc chuyfin thinh cfing vao cay Xoan ta thoi^ qua Agrobaclerium tumefaciens.
Choi va cay Xoan ta chuyen gen codA nhan dupc trfin mfii truong chon loc bo sung kanamycin duoc khing djnh bing PCR nhin gen codA. £>anh gia 68 dfing Xoan ta chuy&n gen d difiu kien han nhin tao thu dugrc 5 dong chong chju han tot hon so voi dong dfii chiing khdng chuyen gen. Him lugmg glycine betaine tich luy trong la ciia cac dfing Xoan ta chuyfin tir 2,1 - 4,5 mM/g la tuoi cao hem nhifiu so vfii dong Xoan ta doi chiing khong chuyen gen (0,7 mM/g la tuoi). Cac dong Xoan ta chuyen gen cai tliien dupc kha ning chfing ctdu voi difiu kien khfi h^in. E>Sy
\k CO so de ung dung cfing nghe chuyfin gen vao cai thien kha ning chiu han 6 ciy lim nghifip noi chung v i cay Xoan ta nfii rieng.
Tir khfia: Chiu han, chuyen gen, gen codA, glycine betaine, Xoan ta
I. D ^ T VAN D £
Nhung yeu to bat loi tir moi truong nhu han, man, iing, nhiet do cao thucmg lam mat can bang ve ap suat tham thau gay anh huong nghiem ttgng den sinh tnrong, phat trien va nang suat cay ttong (Boyer, 1982; Wang et al., 2001). Mot ttong nhung phan ung thudng gap nhat khi cay chiu cac dieu kien b i t lgi ve nuoc la tang cucmg tong hop va tich liiy cac chat hoa tan tuong thich nhu cac loai duong tan, glycine betaine, proline, v.v. de duy tri ap sudt thim thdu (Hasegava, Bressan, 2000). Glycine betaine (GB) dugc biet dku la mpt ttong nhung chat dong vai tto quan ttgng trong qua trinh dieu chinh ap suEit tham thiu noi bao khi thuc vat sdng ttong cac dieu kien moi trucmg bS.t lgi (Khan et al., 2009). GB la chit hoa tan tucmg thich hieu qua nhat va dugc tim thiy ttong mpt ph^m vi r6ng 6 cac loai dgng vat, vi khuan va mgt s6 loM thuc vat hat kin chiu han han va m§n (Rhodes, Hanson, 1993; Chen, Murata, 2002). GB bao ve cay bang each duy tti can bang nuoc gitia te bao voi mdi truong, 6n dinfa
cac dai phan tii, dieu chinh va duy tri mang thylacoid, dam bao bieu qua quang hgp duoi dieu kien kho han va nong do muoi cao (Yang et al., 2003). Trong tu nfaieu, rau bina, ngo, cii cai duong va liia mach, nhanh chong tich liiy GB khi cay bi tac dgng boi moi truong han faan, nong do muoi cao va nfaiet do thap (Bohnert etal., 1995).
Den nay, cac con duong U8n quan den sinfa tong hgp GB 6 sinh vat da dugc Iam sang to. (J mgt s6 loai thuc vat bac cao GB dugc t6ng hgp til choline thong qua hai phan iing Uen tiep.
Dau tien choline dugc chuyen thanh betaine aldehyde nho sir xiic tac cua choline monooxygenase va tiep theo betaine aldehyde dugc chuyen hoa tfaanfa GB duoi xiic tac ciia betaine aldehyde dehydrogenase (Rathinasbapathi et al., 1997). Trong khi do, con duong sinh tong hgp GB 6 vi khuan Arthrobacter globiformis lai rat don gian, tii choline chuyen hoa thanh san phim true tiep la GB cfai cin xiic tac boi choUne oxidase (Tkuta et al., 1977). Gen codA ma hoa choline oxidase phan lap tir Arthrobacter globiformis, dS TAP CHi KHOA H o r VA rcwtr. N G H £ LAM NGHl£P SO 2 - 2013
Cong nahe sinh hoc & Gidng cav trong cfauyen tfaanh cong vao nhieu ioai cay ttSng va chiing minh cay chuyen gen tang cuong diigc kha nang cfaong chiu cac dieu kien moi truong bit lgi nhu cay Arabidopsis thaliana chuySn gen codA tang cuong kha nang chiu lanfa, nhiet va bang gia (Alia et al., 1998; Sakamoto, Murata, 2000), cay cai be, bach dan, ca cfaua cfauyen gen codA cung tang cuong kha nang chiu man va oxy faoa (Prasad et al., 2000;
Ahmad et al., 2008; Yu et al., 2009).
Vi$t Nam la mpt trong nhung qu6c gia bj tic d6ng manh me cita viec bien d6i khi hiu.
Han faan va dat nfaiem mSn dang ngay m6t gia tang, day se la mgt frong nhiing nguyen nhan chinh lim giim ning suit va chit lugng ciy tr6ng. Vi?c nghien ciiu tao ra cic gidng cay trdng ndng Iim nghiep c6 kha ning chiu han va man cao dang ttd thinh mdt ttong nhiing hudng ngfaien ciiu ttpng tam faien nay. Cac nghien cihi ve chuyen gen vio ciy trong nang cao tinh chdng chju digu kien mdi truong bit lgi mdi chu yiu tip trung vio cac loai ciy ndng nghiep; ddi vdi cic loii cay lam nghiep cdn rit han ch€. Xoan ta Ii loai cay gd ldn, nhiSu uu diem: gd nhe, cd van thd dep, kha bSn, kfad bj mdi mgt, nen dugc diing ttong xiy dung, trang tri npi thit va dieu khic, la lam phin xanh, hat ep liy dau. Vi vay, ciy Xoan ta dugc dinh gia l i mgt ttong nhung loii ciy trong quan trgng ttong chiSn lugc phit tri^n
Iam nghiep d Viet Nam. Vdi gia tri kinfa lk ciia cay Xoan ta, viec iing dung cong nghe chuyen gen dk cai tfaien gidng la can thiet. Trong bai bao nay, tic gia trinfa bay kit qua ngfaien ciiu cfauyin gen codA v i o cay Xoan ta va dinh gii khi nang chju h?ii ciia cay Xoan ta chuyen gen. Kit qua la co sd de tao gidng cay trdng Iam nghiep chuyen gen tang cudng tinh cfadng chju khd han.
KV^^TLlElJVAraOtrONG PHAP NGHIEN c u t L Vat Uu
Hat gidng Xoan ta dugc Vien Cdng nghe sinh hgc Lim nghiep, Dai hgc Lim nghiSp cung cap.
Chiing vi khuan Agrobaclerium tumefaciens LBA4404 mang vector chuyin gen nhi thi pBI121 chira gen codA ma fada cho choline oxidase (gen cocU ting hgp nfain tao dua tten trmh tu cd m i sd AY304485 ttong Ngin hang gen NCBI, dat hang Epoch Life Science, Inc, Hoa Ky tdng hgp) va gen nptll ma hda cho neomycin phospfaottans-ferase H. Gen codA dugc thiet k i them vao diu 5' mdt doan DNA ma hda cfao doan peptide van chuyin vao luc lap - TP (transit peptide sequences of the Rubisco small subunit of tobacco). So dd vector cfauyin gen pBI121 chira gen codA va /iprllnhuHinhOl.
RB NOS-Pro N P n i (kanR) NOS-ter 3SS-Pro
^'•"Clll Xbal sac\
Hinh OL Sa dd vector chuyin gen pBUll chira gen codA vd nptU 2. Phirong phap nghien cirn
2.1. Pbmmgpbip tao ciy Xoan ta chuyin gen Quy trinh chuyjn gen vao ciy Xoan ta thong qua ^i khuin Agrobaclerium tumefaciens duoc tiin hanh nhu sau: Hat Xoan
ta duoc khir triing (theo phuong phap cua Biii Van ThSng va ddng tac gia, 2007), cly 16n moi trudng nay m&n MS b6 sung 3 % sucrose va 7,5 g/l agar. Sau 2 tuin, than mim ciia cay Xoan ta tii sinh tir hat dugc cit thanh nhimg doan nho co kich thuoc 1 cm, nhiing doan than TAP CHi KHOA HOC VA CONG N G H | LAM NGHffiP S 6 2 - 2013
Cong nghe sinh hoc & Giona cav trong liy dugc nhiim vdi dich huyen phii vi khuan
igrobactehum tumefaciens mang vector
;huyin gen (ndng dd vi khuin ODgoo = 0,5) rong 30 phiit mau dugc thim khd va nudi tten ndi trudng ddng nudi ciy MS bd sung I mg/l 3AP + 0,2 mg/l NAA + 200 ^M acetosringon irong 48 gid (nudi trong tdi). Sau tfadi gian 'ldng nudi ciy, m l u dugc nia loai bo vi khuin Jing dung dich cefotaxime 500 mg/l v i ciy i:huyen sang mdi trudng tii sinh chgn lgc SM:
vis bd sung 0,5 mg/I BAP + 0,1 mg/I kinetin + L50 mg/l kanamycin + 300 mg/l cefotaxime, lau 20-25 ngay nudi ciy cac chdi bit diu tii linh. Chgn lgc cac chdi cd tfaan, la mau xanh Iam cay chuyen sang mdi trudng ra re R: MS y 0,3 mg/l IBA + 50 mg/l kanamycin. Cay '"Coan ta chuyen gen sau khi ra r l dugc chuyen '•a ttdng ttong nha ludi.
1.2. Phdn tich su cd mat cua gen codA d cdy :huyen gen
DNA tdng sd dugc tach tir l i xoan theo ijhuong pfaip cua Xavier, Karine (2000). Su
;;6 mat cua gen codA ttong cay chuyen gen lugc kiem tta bing pfain iing PCR vdi cip ndi die hieu: F l : 5'-GCT CTA GAA TGG CAC AAA TTA ACA-3' va R l : 5*-CGA J C T C T C A A T T C A GAT CCT CTT C- J'nfain gen codA v i doan TP cd kich thudc dioang 1,9 kb tfaeo chu trinh nhiet: 94°C/3 jhiit, 30 chu ki [94''C/1 phiit, 50^050 giiy,
?2°C/1 phiit 30 giay], 72°C/10 phiit va kit faiic pfaan img d 4°C. Sin pfaim PCR dugc dem tra bing phuong phip dien di tten gel igarose 0,8%, nhuOm bing ethidium bromide, ioi dudi den UV va chup anfa.
?.i. Phdn tich biiu hien c&a gen codA d mirc td phiin md
RNA tong sd dugc tach tir cac miu la ciia
;ay chuyen gen bing TRIzol (Invittogen, My), 3ieu hien cua gen codA a muc dg phien ma lugc kiem tra bing k^ thuat RT-PCR, sii dung )0 kit Superscript™ One-Step RT-PCR system with Platinum Taq High Fidelity Invitrogen, Hoa Ky) vdi cap mdi PCR die
hieu F2: 5'-GAC TAC ATT GTT GTT GGA ( J G T G-3' va R2: 5'-GTA GAA AGT ACG ACT TCG TTT CTA G-3' nfain mgt doan gen codA CO kich thudc 760 bp theo chu trinh nfaiet: 94''C/3 phut, 30 chu ki [94°C/1 phiit, 56''C/50 giiy, 72°C/1 phiit], 7 2 ' ' a i 0 phiit va kit thiic phan iing d 4°C.
2.4. Bdnh gid tinh chiu han cua cdc ddng Xoan ta chuyin gen d muc dp nhd ludi
Danh gia nhanh khi nang chiu han ciia cic ddng Xoan ta chuyfen gen codA dugc tien hinh d giai doan ciy con (1 thing tudi). Cic ddng Xoan ta chuyen gen va ddi chiing khdng chuyin gen dugc xii ly khd han (khong tudi nuac ), nudi ttong budng sinh trudng vdi di6u kien nhiet do 27±2°C, do i m 60%, cudng do chiiu sang 4.000-5.000 lux, thdi gian chieu sang 14 h/ngiy. Kfai nang chiu han ciia cic dong ciy Xoan ta chuyen gen dugc danfa gia tfadng qua binh thai va phan tich ham lugng glycine betaine ticfa liiy. Pfauong phap tich chiet v i xic dinh ham lugng glycine betaine ttong l i cay Xoan ta dugc thuc hien theo Grieve va Grattau (1983).
m K l ^ QUA NGHIEN CUtJ VA THAO LUAN 1. Tao cac dong cay Xoan ta chuyen gen codA
Tfai nghidm cfauyen gen codA vio Xoan ta dugc chia lam 3 Id vdi tdng sd miu bien nap 448 doan thin mam. Ket qua chuySn gen tfau dugc trinfa bay d bang 01 cho thiy, vdi 448 the nhan gen dugc xam nhiem v i ddng nudi cay vdi chiing Agrobaclerium tumefaciens LBA4404 mang vector chuyen gen pBI121 chiia gen codA sau 5 tuin nudi cay tten mdi tnidng tai sinfa chdi chgn lpc (MS + 0,5 mg/l BAP + 0,1 mg/l kinetin + 30 g/l sucrose + 8 g/l agar + 150 mg/l kanamycin f 300 mg/l cefotaxime) tai sinh dugc 197 chdi. Cac chdi dugc cay chuyen sang mdi trudng ra rl chgn lgc (MS + 0,3mg/I IBA + 20 g/l sucrose + 8,0 g/l agar + 50 mg/l kanamycin), sau 3 tuin cd 68 cfadi ra re, chiem 15,2% so vdi sd miu bien
TAP c m KHOA H Q C VA CONG NGHE LAM NGHIEP S 6 2 - 2013
Cong nghe sinh hoc & Giong cav trong nap. Theo cac nghien cuu cdng bd trudc day cfao thay cic chdi Xoan ta ra re dugc tten mdi trudng chgn lgc 50 mg/l kanamycin la chdi chuyin gen (Biii Van Thing et al.. 2012). Cac ddng ciy Xoan ta cfauyen gen niy dugc chuyen ra ttdng trong nfaa ludi. Kfai cay sinh trudng binfa thudng (ra l i mdi), Ii ciia ddng ciy Xoan ta chuyen gen v i dong ciy ddi chiing khdng chuyen gen (WT) dugc tfau va tich chiit DNA
tong sd, su dung lam kfaudn cho phan ung PCR vdi cap mdi die faieu nfaan gen codA. Ket qua cfao thiy, ca 68 ddng cay Xoan ta kiem tta ddu duong tinh vdi phin iing PCR, cac ddng chuyin gen diu xuit faien mdt bang DNA c6 kich tfaudc' 1,9 kb dung vdi kich thirdc ciia doan gen codA cfauyin vao Xoan ta. DiSu niy cfaiing td cac ddng Xoan ta niy da dugc cfauyin ciu tnic gen codA (Hinh 02f).
Bang 01. Tdng hap ket qud chuyin gen codA vdo Xoan ta Lathi
nghiem 1 2 3 Tong sS
So man bien Dap
150 WO 158 448
So choi/Mol tririTig 150 mg/I Kan
54 68 75 197
SM So choi ra rl/Moi ti-irong R SO mg/l Kan
22 19 27 68
S« chdi dinmg tinfa voi PCR
22 19 27
«8
Hinh 02. Cdc ddng cayXoan ta chuyin gen coda
Ghi cku: mnh a va b. chdi chuySn gen tren moi trucmg tdi sinh co 150 mg/l kanamycin Hinh c-Mi chuyen gen trin mdi truang ra re co SO mg/I kanamycin; Hinh d: chdi khong chuyin gen T i T T r T ' T ™ * " 1"°. '"• "f '"'""^""^ "'•"'' ' ' ^ '^"i '^^^ S^" 'ring i nha luM:
Hinhf: PCR nhan gen cod.4 tu cdc dong chuyin gen.
TAP CHi KHOA HQC VA CONG NGH? LAM NGHffiP S 6 2 - 2013
Cong nghe sinh hoc & Giong cav tronc I. Banh gia kha nang chiu faan cua cac dong
KosLn ta cfauyen gen codA
C i 68 ddng Xoan ta cfauyen gen dugc
;huyin ra ttdng ttong biu dit. Ciy 1 thing tudi lugc tien hanh xir ly han (kfadng tudi nudc).
Sau 10 ngiy xir ly faan, cac ddng cay Xoan ta
;huyen gen va ddng ciy ddi chiing khdng
;huyen gen (WT) cd phan iing khac nfaau vdi liiu kien faan; Trong 68 ddng chuyin gen xir ly ian tfau dugc 5 ddng (TX4, TX12, TX27, 1X28 va TX54) cd khi nang chiu han tdt nfait, :iy sinh trudng v i phat triin hinh thudng ttong lidu kien xir ly fagn 10 ngiy, cdn cac ddng
;huyin gen khac v i ddng WT l i bi heo ni.
rung v i ngimg sinfa tnrdng (Jlinh 03). Die biet, keo dai thdi gian xir iy han nhan tao 20 ngay, 5 ddng cfauyin gen (TX4, TX12, TX27, TX28 va TX54) van sdng nfaung smfa tnidng cfaam lai so vdi ciy ttdng d dieu kien binh thudng. Ngugc lai, cac ddng chuyen gen khic va ddng WT bj chet. Cac ddng cay Xoan ta chuyen gen codA sinh trudng dugc ttong dieu kien mdi tnidng bj kfad ban cd the la do gen codA cfauyen vio hoat ddng sinh tdng hgp choline oxidase giup ciy tang cudng sinh tdng hgp glycine betaine, tir dd duy tri can bing nudc giua cac te bao va mdi tnidng, dn fljnh cac dai pfaan tit, duy tri hieu qua quang fagp dudi diiu kien kfad faan.
^nh 03. Dong Xoan ta chuyen gen (TX4) vd ddi chirng (W7) sau xdr ly han 10 ngdy
Phan tich su bieu hien ciia gen codA a miic 1$ phien ma d 5 ddng Xoan ta chuyen gen
; T X 4 , TX12, TX27, TX28 va TX54) bing k?
iiuat RT-PCR. Kit qui cho thiy c i 5 ddng :huyen gen codA dudi su dieu khien ciia Dromoter 353 ddu cd sir bieu hien manfa d mirc lp mRNA (Hinh 04). Ddng thdi, d i xac dinh l i m lugng glycine betaine tich luy ttong la cua 5 ddng chuyen gen nay va ddng WT sau xit ly ian 10 ngay. Kit qua pfaan ticfa cho thiy, cic
*ddng cfauyen gen cd su tich liiy ham Iugng
•glycine betaine ttong khoang 2,1-4,5 mM/g Ii cuoi, cao facm nfaiiu so vdi ddng ddi chitng khdng chuyen gen (0,7 mM/g ta tuoi) (Hinh 05). Kit qua niy hoin toin pfaii hgp vdi sir
bieu hidn ve hinh thai, ve kha nang chiu khd faan ciia cic dong Xoan ta chuyen gen va ddng ddi chiing sau 10 ngiy khdng tudi nudc. Ddng TX4 cd ham lugng glycine betaine tich lUy cao nfait 4,5 mM/g, tiip theo ddng TX28 l i 4,2 mM/g, ddng TX27 la 3.7 mM/g, ddng TX12 la 2,8 mM/g va thip nfait la ddng TX54 la 2,1 mM/g. Mac dii giiia cac ddng Xoan ta chuyen gen cd sir tich liiy glycine betaine khic nfaau nhung lugng tich luy da dam bio cho cic ddng chuyen gen cai thien dugc kha nang chdng chiu vdi diiu kien khd han. Ket qui tucmg tu cung da dugc cdng bd d ciy hdng, ngd, bdng chuyen gen (Huang et al., 2000: Quan et al., 2004; Lv et al., 2007).
T . ^ CHi KHOA HOC VA CONG NGHE LAM NGHIEP SO 2 - 2013
Contj nghe sinh noc & Giorc csv tronp
Hinh 04. Ste bieu hien cua gen ^j-l-i trong cdc dong cay Xoan ta chuyen gen bang phan ung RT'PCR TX4, TX12, TX27, TX2S, TX54 la dong chuyin gen vd WTla ddng ddi chiing khong chuyin gen
Hinh 05. Ndng d^ GB tinh luy trong Id cua cdc ddng Xoan la chuyin gen vd ddi chung Glycine betaine tich liiy 6 ndng do cao Idiong
gay can tro chiic nang cua tS bio chit ddng thoi CO tic dung dam bao tinh on dinh ciu tnic vi chiic nSng cua cac dai phan tii. Miic dp tich luy glycine betaine tuong quan voi kha nSng ch6ng chju di6u kien bit loi (Yang et al., 2003). Nhiiu loai cay tr6ng chuyen gen codA tang cuimg sinh t6ng hc?p va n'ch luy glycine betaine trong t i bao chong chiu tot hon voi dieu kien moi tnrong b§t lpi (han, man, lanh, bang gia, nhiet va oxy hoa) cung (H duoc bao cao nhir cay Arabidopsis thaliana (Aha et al., 1998; Sakamoto, Murata, 2000), cay hdng (Huang ct .a., 2000), liia (Sawahel, 2003), cai be (Waditee et al., 2005), ca chua (Park et al., 2007; Ahmad et al., 2008), b ^ h dan (Yu et al., 2009). Tuong tu, kit qua nghi6n ciiu chuyen gen codi vao Xoan ta cho thiy cac dong chuyin "gen rich luy glycine betaine cao hon so voi d3i chiing khong chuyin
gen vk cac dong chuyen gen cai thien duoc kha nSng chdng chiu voi diiu kien kho han.
r v . K £ T LVAIS
Da chuyen thanh cdng gen codA ma hda cho choEne oxidase tham gia sinh tdng hop glycme betaine vao cay Xoan ta. Banh gia 68 dong Xoan ta chuyen gen codi it diiu kien h£ln nhan Kio thu duoc 5 dong cay chdng chiu han tdt, la cac ddng 1X4, TX12, TX27, •rX28 va TX54. Ham lupng glycine betaie tich liiy trong la ciia cac ddng Xoan ta chuyin gen (2,1-4,5 mM/g la tuoi) cao hon nhieu so vdi dong ddi chiing khdng chuyin gen (0,7 mM/g la tuoi) din din cac ddng Xoan ta chuyen gen cd kha nang chju han tdt hon so vdi cay ddi chiing khdng chuyin gen. Bay la co sd de ling dung cdng nghe chuyin gen vao cai thiSn kha ning chiu han d cay lam nghidp ndi chung va cAy Xoan ta ndi rieng.
TAP c m KHOA HQC VA CONG NGHf LAM N G H I f P S 6 2 - 2 0 1 3
Cong nghe sinh hoc & Giono cav trong FAX LIEU THAM KHAO
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TAP CHI KHOA HQC VA CONG NGHE LAM NGHEEP S6 2 - 2013
Cong nghe sinh hoc & Gidng cav trong
TRANSFORMATION OF CODA GENE ENCODING CHOLINE OXIDASE INTO Melia azedarach L. ENHANCES DROUGHT TOLERANCE
Bui Van Thang, Le Van Son, Chu Hoang Ha SUMMARY
Drought is one of the environmental stress factors that affects seriously to the growth and development of plants. Recently, application of transgenic plants which are tolerant to the environmental stress conditions is a new approach in plant biotechnology. The codA gene coding for choline oxidase, an enzyme entering in Uie biosynthesis of glycinebetaine, was used to transform into Melia azedarach L. through Agrobacterium tumefaciens mediated transformation. The putative transformed shoots obtamed on selected medium supplying 150 mg/L kanamycin were successfiilly confirmed by PCR analysis using the specific codA primer pair. The results of drought treatment showed that among 68 transgenic lines, five of these were grown better than diat of the wild type. The accumulation of glycine betaine in leaves (2.1 - 4.5 mM g"' fw) of these transgenic Lines was higher than that of the wild type (0.7 mM g"' fw). These results demonstrated tiiat the introduction of a biosynthetic pathway for glycinebetaine into Melia azedarach L. enhanced significantly their drought tolerance.
Key words: Coda gene, drought tolerance, glycine betaine, Melia azedarach L^ ir-ansformation Ngudiphdn biin: TS. Lam Dai Nhan
Ngdy nhdn bdi: BO/4/2013 Ngdy phdn bien: 24/5/2013 Ngdy quyit dinh ddng- 07/6/2013
T < ^ c m KHOA H Q C VA C 6 N G N G H I LAM N G H I | : P S 6 2 - 2013
