HOl NGHj KHOA HOC CONG NGHE SINH HQC TOAN QUOC 2013
LTNG DUNG KY THUAT NUOI CAY LAT MONG Tt B A O BE NHAN GI6NG IN
VITRO CAY HOA D 6 N G T I 6 N (GERBERA JAMESONII BOLUS)Nguyin Thi Ttiu Hing', VC Thi Hue', Nguyin Nhui Ngoc' 'T/udngBaingcLamr^ttiep ViitNam
TOM T A T
Dl xay dmiB dwc he tho.g .i. s.nh,, Jn, o y hoa Dong Ben top » | ^ ^ |^ ^._ b.„ di„ li ei hoa noo diroc nhjp ngi til Tnmg Quoc; h.en lal duoc ua chuong va phu hgp voi dicu nen mien „ m J - 0 5 mm) duoc cay sau J ^ U h i n g s . dugc ci.d,iuh«m»g*»du.nn^^^^^^^^^
len moi iniong cam img tao mo. seo (MS + 1,0 mgfl BAP + 0,i ' " S , ' ' . , ; " . • y^y, .ufct „ „ „h6, soma vi tai slnh ehoi. Moi chuyin sa.£ .mdi tnmng MS bo sung BAP, Kineun, NAA n e ^ I ' S l ^ h g ty i t m l u t f p ^ s ^ " S »<^ '^'»° "^^
ta * 4 J 3 dial Sau I - 2 niJn huin tayen, eiy con in rim dugc cay vao luong dat eo ty le phoi tron « » = t i * " 8 " » ™ T ' " ' S i ^ ' i f e i t S " hm, xo du. (da dugc u chg hoal muc). Tlianh phin gia the cho ty 16 eay song cao nhat { n » 90%) s.u 2 - 3 wan m S n t M% d ? 2 ri. hw vd! 1% * va 4% xo diia. i eay eon da bin rl va 6„ dinh tbi tien hMi bfag timg ca, trong vao luong san xuat.
Tirkhoa: Dfi hoa non. hoa Ddng ti£n, nu6i ciy in vitro, mo seo, lit cSt mong. phoi soma, tai sinh MdroAU
esng ti^n (Gertiera Jamesonli Bolus) la mot trong nhung loSi tioa c6 mau sac dep, tipai sang, da dgng ve chOng loai voi diy du cac logi mau tip do. cam, vang, tring, phan hSng, tfm va c6 gia tn kinh te Idn. Tren b6ng hoa c6 th& co mpt mau dtrn ho?c nhiiu mau xen ke, kich thiroc tjong to va dai nen thich hc^j de tgo thanh bo, Iam lang hoac cam hoa nghe thuat. Mpt trong nhifng iru diem cua hoa dong liin la kha l§u tan, neu frflng trong chgu ftcri gian hoa no- Mr 20 ngay den 30 ngdy, dac bi§t khi diiu kien mfii tnrcrng thit* hcrp, hoa co the ra quanh nam. Dong tien la mot trong 10 loai hoa dygc tring pho biin a nhieu nifcrc, dirgrc ira chuong va tieu thy nhiiu nhat trSn the giwi.
Hoa ding tien co the nhan giong bang nhiiu phucng ph^p nhif: nh^n bing gieo frong hgt, tach cay, nuoi cay in vitro.
Trong do, phuong phap nuoi cly in vitm trong mfii truang nhan tao dug'c i>ng dung rgng rai vi c6 h# s6 nhan^ral cao (tip mgt bO phln nho cua cay sau mot thoi gian ngan c6 the tgo ra hang van cay gifing dap iing nhu d u san xuat) va tranh dugc tinh trang thoai hoa giflng (bang Van Oong, Dinh The Loc, 2003).
Tremg cac loai hoa dfing tiin da va dang ducrc trong tgi Vi?t Nam tfii hoa dong tiin k6p nhap ngi (chu yeu nhgp tit Trung Quoc, Ha Lan, Dai Loan) die hifu qua kinh t i cao nhlt. Cac giing ding tien kep nay co nhiiu uu diem: hoa to. canh day, gom nhiiu tang xip lai voi nhau, mku sac phong phu, hinh ding hoa can dfii, r l t dep, nang suit cao, tuy nhien lai cfi nhucrc diem la thudng bj bit due nen khfing the nhan giflng bang hgt. Giai phap t i l nhlt cho nhan cac giong dflng tien kep nhap nfii la ung dung ky thuat nufli d y mfl - l i bao d l tai sinh cSy in vitro tir ngufin vat lifiu la cac mo lay tir cay ngoai tg nhifin. sau dfi nhan nhanh de tgo ra lugng lan cSy giing co chat luang cao.
Theo Teixeira da Silva (2003), he thflng lap mong t l bao (TCL- thin cell layer) bao gom cac mlu d y co kicti thu6c nho dugc d t ra to cdc bfi phSn khae nhau ci^a friuc vat (than, la, r l . cacbo phgn cua hoa...).Cac lat mong dirge tgo tir cac l^t d t theo chieu dgc dugc goi la iTCL, dugc tao tip lal clt theo chieu ngang ggi la tTCL. Cac iTCL chi chua mflt loai mfi nhu Iflp dan cua t l bao biiu bi hoac m$t vai Ifi'p ( 3 - 6 Ifi'p) cua t i hko vo, ngugc Igl cac tTCL bao gom cac t§ bao Ihugc , cac mfl khae nhau (mfi biiu bi, mfi vo, mfi rugt hay le bao nhu mfi), nhung deu cfi die diem chung la mong, so lugng t i bao giam, d l phan ung mfll each dfing nhat vai mfii tarang. Nhung dac diem nay cfi y nghTa quan trgng trong qua trinh kich thich mau d y phat trifin, phin hfia hinh thii va biet hoa tgo mfl, ca quan, ngoai ra con co the rut nglin thoi gian cam ung phat sinh hinh thai.
Tren the gioi da cfi nhiiu nghifin cuu v i nhan giong, tai sinh In vitro hoa ding tiin li!r mfi sgo co nguon goc tir la, thin, cuong hoa... (Chenna e( aL, 2002; Nafees et al.. 2009; Nazma et aL. 2012). Tuy nhien, nhO'ng nghifin cu'u v i tai sinh chfii hoa ding Sin thfing qua phfii soma tao tij lat mong t l bao con hgn chl. Trong bai bao bay, chiinq tai trinh bay nhirng kit qua nghien cuu v i nhan gifing in vitro cac ciy hoa ding tiin kep bit dgc nhap ngi tir Trung Qufic vai vai lieu- lat d t mfing d i hoa non.
VAT LIEU VA PHU'aNG PHAP Vat lif u
D i hoa non (dirang kinh 1,5 - 3,0 cm) lly ti> cay hoa Dflng tiin kep miu vang phay, hing, dfi nhi den. nhSp nfii tip Tnjng Quic, dugc cung d p bai Cong ty Co phln dau tu xiy dgng Long Vifit, Me Linh, Ha Noi.
Phinmg phap
D i hoa non cua cac cSy dfing tiin kep dirge ri>a duai vol nuac chiy manh trong 15 phut, ngim trong nuac x^ phflng 20 phiiL n>a Igi dufii voi nuoc va trang 5 lin vdi nuoc d t . Miu d i hoa tilp tyc dugc xir ly vdi ethanol 70% trong 30 giay va khu trung tilng HgCl2 0.1% trong 10 phut, trang lgi bing nuac vfi trting 4 - 6 lin.
HQI NGHi KHOA HOC CONG NGHE SINH HOC TOAN Q U 6 c 2013
Cic d i hoa dugc clt theo chilu ngang tao cic lap mong lfi bao - cac ITCL (0,3 - 0,5 mm) - va cay Ifin mfli truong cam ung tao mfl sgo (mfli tnjgng MS b i sung i mg/l BAP va 0,2 mg/llAA), nufii trong 2 tuan. Miu d i cam img Igo mo s$o -duouxdvjhij^ln lfin mni trii-ftng kich thich tgo phfii^soma va tai sinh chfii (mni tripryng MS bo Bimi] RAP, Kinetin. NAA rieng le hay ket hgp). Cac choi mang la thgt dugc d y chuyin sang mfii truang MS bo sung 0,2 mg/l NAA kit hgp vol BAP hoac Kinelin a cac nong dg khic nhau d i nhin nhanh, nufli trong 3 - 4 tuan. Cic chii dil tieu chuan (cao 2,5 - 4-"
an) dugc tach khoi aim chfli va d y chuyin sang mfii trufi'ng tao re l i mfii truang MS bfi sung NAA hogo-IBA. Saul - 2 tuin huin luyen, cay con in vitro dugc d y v i o luflng dit co ty 16 phoi trgn khae nhau giira cac thanh phin dit, cat, trau hun, xa dipa. Khi cay con da bin r i va on djnh thi tien hinh birng tirng cay trflng vio luong san xuaL
Mfll truong ca bin su- dung trong thi nghifim la mfli tnffi'ng MS (Murashige & Skoog, 1962), b i sung 3% sucrose, 0,7%
agar. Mfli tarang nufli d y dugc chinh pH din 5,8 tnrdc khi Hiu tning 6 nhiet dfi 11 r ' c trong 15 phut. Dilu kign nuoi d y In vitm dugc duy tri a nhifit dg 25 ± 2''C; chl do chifiu sang 3000 lux; thai gian chilu sang 16 gio'/ngay.
KET QUA VA T H A O L U A N
Kfch thich tai sinh c h i i tir cac tTCL da cam ipng tao mo seo
Khi cac tTCL dugc nufii d y trfin mfii truang cam ung tgo mfi seo, chi sau 3 - 4 ngiy hinh thai mlu d y cfl su chuyen biin rfi- cac tTCL cong phfing len va xult hien mfi seo miu trlng 6- mep ngoii, khoi mfi seo tilp tyc phat triln v i chuyin din sang miu xanh Tren mfli trufi'ng MS b i sung 1 mg/l BAP v i 0,2 mg/l IAA, 83.67% mlu cay tao mfi s?o.
Cic tTCL d i c^m irng tgo mfl see dugc cly chuyin sang mfii buang tai smh chii (bo sung 0,1 - 2,0 mg/l BAP kit hgp vai 0 - 0,5 mg/l Kinelin v i 0,2 mg/l IAA ho|c NAA).
Sau khoang J - 2 tuan nufii tren rrioi tnrimg tki sinh ehoi, tir cac khii mfi seo bit dau xuit hign mam mong cua phfii soma. Mfit s i logi hinh dgng chil yiu ciia phfli soma hoa ding tiin quan sat dugc du6i kinh hiln vi soi nil l i hinh d u , hinh In^, hinh tim...Cac phfii thuang cfi miu xanh va cfi kha nang tii sinh chii cao tren mfii trud'ng nufli ciy thich hgp.
Ben cgnh do, cic p^hfli cfi thi nh"ah len tgo curn^ptifll soma vai kich thuoc ling din va cac cym phfli nay khi dugc tach nho ra va cay chuyen sang mfli tru-gng nufli d y mai thi tilp tyc cfi sg gia tang ve s6 lugng phfli trong cum.
Ketqua ^Bgng 1) cho thiy kha nang tai sinh choi tir cic miu d y la khae nhau khi duac nuoi trfin cac cfing thirc mfli ' tru&ng 50 sung cic t i hgp chit ffliu hfla slnh trufi'nepth'ac nhau vfi'i ning do khic nhau. Kht ning do BAH trong cac to hgp chit diiu hfia sinh taro'ng bo sung vio mfli trudng d muc thip (0,1 mg/l) ho|e cao (2 mg/l) cho khi ning t i i sinh Choi thap (0 - 23,4% miu d y tai sinh chit). Nfing dO BAP Irong cie to hgp chit dieu hfia sinh trufl'ng fl' khoang 0,3 - 0,5 mg/l eho hi§u qua tai sinh chil eao hon ca (45 - 73.3%).
Cflng thirc mfli trufi'ng tfit nhat cho cac tTCL da cim ung tgo mfi sgo tii sinh chii la mfli trudng MS b i sung 0,3 mg/l BAP kit hgp vol 0.5 mgli Kjnetin va 0,2 mg/l l/kA. Trfin mfli tarang nay, ty l§ miu tao phfli soma va tai sinh chii dat 73,3% vfi'i 4,12 chfii/mlu. Kit qua nay eung tuang duang voi kit qua tii sinh cay hoa dong tiin tir mfl sgo (co nguon gic tu cac mfi la) trang bio cSo cua Surinder va cfing su (2007).
Bang 1. Anh hud'ng cua t6 h g p chSt Siku h6a ainh trud'ng d i n s y tai sinh choi (sau 4 tuan nuoi)
Cong ttiuc
1 2"
3 4
,. _^,
5 8 9"10 11
" .,2 """"
13 14
"15 16 18 19 20 21
Chat dieu hoa sinh tnrong (mg/l) BAP
0.1 0,3 0,5 1
Kinetin
2 ^ 0,1 0,3 0,5 1 2 0.1 0,3 0,5 1 0.1 0,3 0,5 2
0,5 IAA
0.2 0.2 0 2
^""0^2" "
0 , 2 " "
0,2"
NAA
0,5 : 0,2 ' 0.5 ;_ 0,2 0,5 r 0,2 6,5 1 0,2 ' .
• " . " T"" 0.2 i - " : "0.2 1 - : 0.2 - .1. • . I .0?.
0.2 0,5 i - 1 0.2 0,5 : 0,5 J 0.5 0,5
0,2 0,2 "
0,2""
0 2 S i m l u
cay 45 45 45 45 45 45 45 45
" " 4 5 45 45 45 1 '•5 J 45 45 45 45 45 45 45 45
T^ 1$ m l u tai smh choi 1
(%) 1
0
0 : 69,2 ab i
53,6 c ! 40,7 d i 6 T 11.7f 1 73.3 a - ; 65,9 b 53.3 c 1
0 1 9 ^ f
66,7 b 47,5 d j 47,2 d ;
23,4 e 1 0 1
53,3 c 45,0 d \ 38,9 d ;
1
So ch6i/mlu
0 0 3.77 a 3,50 a 3^67 a 0 1,68 d 4,12 a- 3,88 a 2,85 c 0 1,1 d 3,58 a 4,03 a 3,21b 0 1.59 d 3.38 b 2,49 c 2.16 c 0 C i c chu cSi khic nhau trang cung mgi c§l chi s g khae biet co y nghTa b mi>c a = 0,05 theo phuong phkp ttiong ke Duncan Nhin nhanh choT
Cic chii tai sinh tir mfl sgo. khi da cfi 3 - 4 la, cao 1 - 3 cm, la xanh non hogc xanh thim dugc cay chuyin sang mfii tru&ng nhin nhanh.
HOI NGH! KHOA HQC CONG NGHE SINH HOC T O A N QUOC 2013
i^t ., , , * * . , > - ' . , - .,nn im*ng (BAP, Kinelin) lang len tir 0,3 - 2 mg/l del vfti BAP va 0,5 - 5 mg/l * i ;f Khi liam lu-cmg chat g " J ; " J ^ * g 7 „ a „ U sv gia tang h a m V n g BAP hoac KineHn. nhL,ng chSt lirftng ch& I* g'i»"
Kmeln, he so " l l ! " * J „ " ^ " i ' ^ h ^Sdng qua MO, hiiu hiSn IS than oh& manh. han c h i v6 chlcj cao, co thS xuit hien T s ^ d ^ S c ^ * « n " c r c S l V k h i c , cic chSi tao »,anh tren moi l h , 6 j g r f sungBAP hoac K.n^«n " " ^ ^ r ^ r ^ a o k h l S v chuvin sang moi tnjtrng tao rS thi kha nang choi cam ung ra r i rat hap, chat luqmg re kem hon na ,
^ v l i i c c s f ^ f S h ^ n mS .„«,n'g b6 sung chil aiSu hoa ^nh tru.6.n9 vfti nong dp «,4p hoac vfta^ a.^ Mo,
^ g b i sung BAP hoac KineBn vfti ning So 1 - 3 mg/l cho he so "han nhanh choi cao nhimg chiau cao choi han c h . cong aufto chi ro trong nghien ciju cua Duftng Tan Nhirt va cgng sir (2007).
o , „ „„»n sai va thing kS kit qua Irong qua Irlnh thi nghiem, ching toi nhjn thiy cong IhCrc moi m/ftng vfta cho he so
* a n chi^ S l i o v ^ i^io Chit l i K m g " i i l i t la MS b i sung 0,5 mg/l BAP va 0,2 mgfl NAA. Tren moi th,»ng nay h , s i nhfn n h a n h l h i i ^ t 6,18 lin, c h i i i cao chil Sat 3,3 cm, chat Iucrng choi 161, co tiSm nang ra m t,o cay con hoa,.
d i i n h .
B^ng 2. Anh h u o n g cua chM dleu hoa slnh f r u ^ g d i n kha nang nhan nhanh chm (sau 4 t u i n nuoi) Chai dieu tioa sinh tiudng
(mg/l)
BAP
Kinetin
0.3 1.0 2
^ "" 0.5 1.6
< 5,0 ' So choi/mlu
4,33 e 6,18 d
"8.95 c "
2.77 f 3,65 e 5,33 d 8,60 c
Chieu cao choi (cm) 1 3,5 a 1 3.3 a
1,9 b 1.1 c 3.6 a 3,2 a 3,1 a 1,5 b
Than choi m§p, cao m#p,cao l ^ a manh mSnh, thko m l D.cao m^p, cao thgp CSc chir cai kh^c nhau trong ciing mot c6t chi sir k h i c bf§t co y nghTa = 0,05 theo phuong phap th6ng ke Duncan K i c h t h i c h c h o i ra re
Trfin thgc t l , dong ti^n la logi cay co b$ re chum nen de dam bao cay con sinh truang va phat trifin tflt ngoai tg nhien, Vlfc nghifin ciru tao bfi rfi khfie, cung c^p rfit quan trong.Trong nghien ciru tgo re cho chfii, chiing toi sii' dung moi _ , truflng MS bfi sung mfit trong hai loai chit dilu hoa sinh trufi'ng thuoc nhflm auxin la NAA va IBA. C^c choi co chifiu cao tCr 2,5 - 4cm dugc tdch khoi cym cly vko mfii trufi'ng tao re.
Kll quS {Bang 3) cho thiy khi si> dung NAA bo sung vao mfii tm-flng kich thich ra re cho hifiu qua tit hon IBA. Hieu qua tgo re tot nhlt dgt duge fi' cflng thi>c mfii truong b l sung 0,5 mg/l NAA vfl'i 96,67% chfii co re; s l r l trung binh cua chfli dgt 4.33 r l ; r l kha dai; chit lugng re tot, re mgp.
Being 3. Anh h u v n g cua NAA va IBA d§n khs nang kich thich ch6i ra re (s ChSl dleu h6a sinh topfinq (mq/1)
NAA
IBA 0 0,3 0,5 1.0 0,3 0 5 1.0
Ty 1$ ch6i ra r l (%) 4,45 f 75.24 b 96,67 a 63,33 c 29.82 e 46,67 d 60.00 c
S6 r l TB/ch6i 1.13 e 3,68 b 4.33 a 3,67 b 1,60 d 1.95 d 2,67 c
au 4 tuan) Chilu dai r l (cm)
0,67 c 2,55 b 3,67 a 4.10 a 1,88 b 2.00 b 2.17 b
Tinh trgnq rfi
; R l tranq, manh, n q l n I R§ tranq. m^p _^ R l trgng, map ' R l trSnq. mSp 1 Re tranq, manh '< R l t r i n q . manh 1 R l trang. manh cac chO c^i kh^c nhau trong ciing m^t cgt chi sg khae biet co y nghTa d mirc a = 0,05 theo phucrng phap thing ke Duncan
^ h huong cua loai gia the den sinh truwng, phat t r i l n cua cay in vitro gial doan v u * n y c m
Cay con in vitm sau khi huin luyfin dgt cac tifiu chufin: chieu cao cay 3,5 - 5 cm, cfiy co 4 - 5 la va 3 - 5 re, re dai 2 - 4 cm dugc n>a sgch agar bam r l , xu ly^vfi-i dung djdi thufic digt nam VibenC 0,05% trong 2 - 5 phiit, tr^ng bing nufi'c sgch, de rao nuoc roi dem gifim vao luong dit efi cac th&nh phln phfii trfin kh^c nhau giua dat, xo dira, trau hun vk eat.
Sau 2 - 3 tuan eac cay mim bfin re vfi fin ffinh thi tiln hanh b^ng tirng cfiy trong vao luong san xult. T i t ca cac cong thtre dfiu tuoi phun su0ng 2 - 3 lan/ngfiy vol lugng nude tudi IS 5 - 6 l/m7ngay, che 50% anh skng trong 1 tufin dau.
Ket qua duge ting hgp trong Bang 4. - ^ _ Bang 4. Anh hud-ng cOa lo^i gia the d i n sinh tru'd'ng, phat t r i l n cCIa c<iy Jn Wtro
Thdnh p h l n gia the Tong s l cSy dem trong
Ty le cSy s i n g
m
Chieu cao cSy TB , S l l a T B / c S y (ia) -
Tmh hinh slnh truong
9 5 % a l t a A n - 1 % c A t + 4% xo dira 0 95%ddt(lii + 1%cai +
c a y cuTig ckp, sinh t r u i n g tot, _a6ng ahu c a y ci>ng cap, sinh trufitig t i l . dftng deu c a y cOng c g p , sinh trudng t i t , d i n g d i u cac chl> cai khae nhau Irong cung m$l c$t chi s u khae bift c6 y nghia d mdc a - 0,05 Uieo photmg p h ^ thing k# Duncan
HOI NGHI KHOA HOC C 6 N G NGHE SINH HOC TOAN QU6c 2013
Kfit q u a n h $ n d u g c bifiu thj thanh p h l n gia t h i khae nhau co a n h hufl'ng khae nhau d i n kha nang sfing cOa cay gial dogn s a u nufli c l y in vitro, f r o n g cac loai Sianh p h l n gia ths thi cfing thu c co 95%.dSt dfli + 1 % cat + 4 % xo dira u i o ra Stiich hcrp h o n ca v f l i tv Ifi sflng trunp binh sau 4 tuan dat cao n h l i 95,67%, cay c a o 8,67 c m , c u n g c a p v o l bifiu hifin la cay x a n h b f i n g . kha day, sfl la trung binh dat 6,67 la/cay.
'ife-I#-
Kinh 1 . Mot so hlnh anh mo ta cac giai doan ciJa qua trinh nghien c u u nhan giong in vitro hoa d i n g t i l n tir v3t li^u lat cat mong de hoa non
a, b. Phoi soma va choi hinh thanh li> cac tTCL da cam i>ng tao mo seo; c: t(io da c h l i hoa dong tiln tr§n moi truang nhan nhanh;
d cky in vitro fioan chinh, a, f' Cay t i n g d nha i^rdi sau 4 t u i n (e) va sau 10 tuan (f).
K E T L U A N
' D a x i y d u n g d u g c quy trinh nhfin g i l n g in vitm ckc cky d i n g t i l n kep b i t due tir lat mong t l bao d l hoa non (tTCL).
Cac t T C L s a u khi d a c a m Crng tgo m f l s?o d u g c c l y chuyen len mfli t a r o n g tfii sinh chfli eho ty le m a u tgo phfli soma va tai sinh choi dgt 73.3%; s 6 phfli soma trung binh dat 4 , 1 2 phfii/mau c l y . Tren mfli t r u o n g nhan n h a n h . chfii cfi h e sfi niifin dgt 6.18 Ian/4 tuan, von c h i l u cao dgt 3,3cm; T y le c h o i r a r l dgt 96,67%. Cky con In vitm sau khi h u a n l u y f n d u g c c l y vfio lufing cfl t y l e 9 5 % d i t dfli k i t h g p v f i i 1 % cat va 4 % x o dtra cho 9 5 . 6 7 % cay s i n g sau 4 tuan.
T A I L i e u T H A M K H A O
Chenna RA, Mewa LC (2002). Rapid plant regeneration from Gerberayamesom; Bolus callus cultures. Acta Bot Croa( 81(2): 125-134.
Nhul DT. An TTT. Huong NTD. Don NT, Hai NT, Thien NQ, Vu NH (2007). Effect of genotype, explant size, position, and culture medium on shoot generaton of Gert)efa;amesonM by receptacle transverse thin cell layer culture Scienba Hortcuiturae 111(24) 146- figng Van Dflng. Oinh T h I L^c (2003). Hoa d i n g tiln. Nha xuit ban Lao Ddng-Xa Hdi, tr 33-38
Nafees A, /VbduIRK, Uaqat A Inksar AB (2009). Tissue culture of Gertjera. Pak J eof 41(1). 7-10.
Nazma A M I. Hoqueand R. H. Sarker (2012)./nv/fro propagation in three varieties of Gert)era(Geri)era;amesonn Bolus) from flower bud and flower stalk explanls, Planl tissue cult & biotech 22(2). 143-152.
Murashige T. SkoogF(1962), A reused meiJium for rapid growth and bioassays vwfh tobacco tissue cultures. P/jyB/o/P/an( 15:473-497.
Surinder K, Jitender \(K (2007), Plant regenetaton from cell suspensions in Gerfcera jamesonli Bolus. Journal of Fruit and Omamenlal Plant Research 15:157-166.
Teixeira DS (2003). Thin c ^ l layer technology In ornamental [ SjoIsc/tnoJOgy.
: miax>propagation and biotechnology. African Joumal of
IN VITRO PROPAGATION OF GERBERA JAMESONII BOLUS USING
TRANSVERSE THIN CELL LAYER CULTURE
HOI NGH! KHOA HOC C 6 N G NGHE SINH HOC TOAN Q U 6 c 2013
Nguyen Thi Thu Hang', Vu Thi Hue\ Nguyen Nhu Ngoc^
^Vietnam Forestry Unrversity
SUMMARY
Using the flower buds as initial materia], the high efficient protocol for regeneration of i/erftera/aAte^onii B. through transversclhin cell layers (tTCLs) culture was established. The tTCLs (0,3 - 0,5 mm) were cultured on induced medium, which include M;;
supplemented with 1,0 mg/l BAP and 0,2 mg/l IAA, After 2 weeks, explants were transferred to medium with combining or separate addition of BAP, Kinetin, NAA. for en^iyogenesis or shoot regeneration. The highest rate of embryogenesis and shoot formation was 73,3%, with 4,12 shoots per explant afler4 weeks cultured on MS supplemented with 0.3 mg/I BAP, 0,5 mg/I Kinetin and 0.2 n ^ IAA. In vilro shoots were multiplied on MS containing 0,5 mg/l BAP, 0,2 mg/I NAA. The regenerated buds were cultured on MS supplemented with 0,5 mg/l NAA for root generation. After 2 weeks training, the complete plantlets were transferred to nurseiy garden and planted into die comply base (complex of soil, sand, burnt hook and coconut decomposed case). The suitable complex with hi^iest survival ratio is 95% of mountain soil combined with 1% sand and 4% of coconut decon^osed case. When the plantlets begin rooting, they are transferred into field for production.
Keywords: callus, flower buds, GerberaJamesoniiBolas, in vitro culture, regeneration, somatic embryos, transverse thin cell layer.
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