Cong nghe sinh hpc & Giong cdy trong
vilTNG DUNG KY THUAT NUOI CAY IN VITRO TRONG NHAN GIONG QUE LAN HI/ONG (AerUes odorata Lour.)
Nguyen Van Viet', Biii Van T h a n g \ Nguyen Thj H i r ^ g ' , Nguyen Thi Thu HSng '•^•^Trtfdmg Dai hpc Ldm nghiep
''Trung tdm phdt triin Ldm nghiep Hd Npi
T O M T A T
Vi nhan gidng cay Qui lan huong bSng ky thuat nuoi ciy in vitro da duac nghien ciru thanh cong. Kit qua nghien ciiu cho thiy sat khuin be mat qua bang ethanol 70% trong 2 phut, khii trimg bang dung dich HgCl2 0,1% trong 15 phiit va nuoi cay tren moi trucmg Knops, cho ty le mau sach 94,33% va 88,67 % miu tai sinh thi ch6i sau 5 toan. Moi truang Knops bo sung BAP 0,5 mg/l, Kinetin 0,2 mg/l va NAA 0,3 mg/l cho kha nang tao th£ ch6i tot nhit, cac the chdi to, map, xanh thim va he so tao thi choi dat 4,78 lan sau 4 tuin. Nhan nhanh choi teen moi truang khoang ca ban Knops bo sung BAP 0,8 mg/l, Kinetin 0,4 mg/l, NAA 0,1 mg/l, nude dira 100 ml/1, dich chilt khoai tay 100 ml/1, sucrose 30 g/l va agar 4 g/l, chdi phat triin nhanh, map, mau xanh dim va he s6 nhan chdi dat 9,67 lin sau 5 tuin. Ty le chdi ra rl d?t 96,67%, so re trung binh 3,46 rl/cay vdi chieu dai re ttung binh 3,83 cm khi nuoi tren moi truang Knops b6 sung NAA 0,3 mg/l, nude dira 100 ml/1, djch chiet khoai tay 100 ml/1, sucrose 20 g/l va agar 4 g/l sau 5 tuan nuoi cay. Cay con hoan chinh dupe huan luyen va chuyen ra trong tten gia the ddn trang, xu ly gia the 2 ngay trong nude, cho ti 1? cay song 93,33%. Quy trinh vi nhan giong nay co the ap dung de san xuat hang loat cay gidng Qui lan huang chit lupng tdt, dap ling nhu cau nguon cay gidng Que lan huomg hien nay.
Tir khoa: Aerides odorata Lour., in vitro, knops, nhan gi6ng, Que lan hucng, ra r l , the choi.
I. DAT VAN DE
Que lan huang {Aerides odorata Lour.) la mgt ddng Giang huang thudc hg phong lan {Orchidaceae)- mgt loai hoa dang rat dugc quan tam va ua chudng vi hoa cd kilu dang, mau sac dep, tda ra mui huong thom va la loai hoa quy hiim cua rimg Hda Binh. Ngoai gia tri tham my thi Que lan huang cdn cd gia tri ve kinh te rat ldn. Do la loai hoa cd huang tham nen cdn dugc su dung d l tach chilt liy tinh dau phuc vu cho nganh my phim, tao huang nude hoa... Hien tai loai hoa nay cimg dang dugc nghien ciiu ling dung trong nganh y hgc...
Cay mgc nhieu thanh bui, cd nhiiu bui cao den 1 met, loai lan thudng mgc trong rimg, bam tren cay go, dudi tan cay thua... Than cay manh, cay bieu sinh, than mgc khde va map.
La mim, mau luc nhat, xoin d d4u, dai tdi 25 cm, la dai tdi 35 cm, rong khoang 4 cm, mat tren mau xanh, mat dudi cd mau hoi tim nhat.
La xep deu ra hai ben, hai cong xudng, d dau
la cd vet khuyet khdng deu.
Hoa mgc thanh chiun thdng xudng, dai khoang 40 cm. Hoa nhd, cau tnic nac, mau trang cd dinh tit ca cac phien mau hdng va mgt dai tia d giiia canh mdi. Canh mdi cudn lai, hinh dng rong, bao phan cd md dai. Hoa nd phan ra bdn phia trdng rat hai hda, canh hoa budng xudng thanh chum. Hoa mgc thanh chiim, cd mau dam d dau canh va mdi hoa.
Hoa to khoang 2 - 3 cm, cd huomg tham. Hoa thudng thay cd hai mau chinh la trang va mau tim hdng. Qua nang hinh trai xoan, cd cudng kha dai. Nd ra theo 3 - 6 dudng mit dgc, khi chin qua nd ra va manh vd cdn dinh lai vdi nhau d phia dinh va phia gdc. Hat rat nhieu nhung nhd li ti, xop, chiia diy khdng khi. Hat trudng thanh sau 2 - 1 8 thang.
Mac dii Que lan huang cd khu vuc phan bd rong, tuy nhien hien nay loai lan nay dang bi khai thac can kiet do nhu cau ldn ciia con ngudi cdng them mdi trudng sdng hi thu hep 162 TAP CHI KHOA HOC v A CONG NGHE L A M NGHIEP SO 6-2016
do nan pha riing, chay rung ngay cang nhieu nen lan rimg khong con noi cu mi. Mat khac, trong tu nhien ngi nhfi trong hat lan thudng bi tieu giam, mudn nay mam phai co sir cgng sinh cua n i m Rhiroctonia nen ty le tai sinh b§ng hat rat kem, ben canh do nhan giong tir cac ca quan sinh dudng thudng rat cham, he so nhan gidng thip va anh hudng den cay me.
Da cd mgt sd tac gia bao cao xay dung thanh cdng quy trinh nhan giong Que lan huang (Aerides odorata Loiu.) tii cac minh la, my nhien quy trinh thudng keo dai do phai tao chdi thong qua gjai doan tao caUus. Hien nay, ky thuat nhan gidng in vitro vai he sd nhan tir mgt qua lan la rat ldn. Do do, nhan gidng in vitro hoa Ian hi phdi hat trong ong nghiem Idia hoan hao va thudng dugc sti dung trong nhan gidng Ian Dendrobium.
Bai bao nay thdng bao kit qua nghien cuu nhan gidng thanh cdng cay Que Ian huong bing ky thuat nudi cay in vitro dat hieu qua cao.
n. VAT UEU, PHUONG PBAP NGHIEN C L t 2.1. Vat lieu nghien ciru
Vat Ueu nudi cay la hat cd ngudn gdc tir qua Que Ian huong dugc myen chgn tu nhiing cay cd tinh trang tdt tai \-udn lan thudc xa Huang Ngai, Thach Thit, Ha Ndi.
Cac loai mdi trudng nudi cay dugc ghi d bang 1.
Hda chat dimg de khu trimg mau bao gdm:
edianol 70%, HgCL 0,1% va Javen 6%.
2.2. Phuorng p h a p nghien cuu
Nuoi cay khai dpng: Mau qua Que lan huong nia sach bing nude may, ngam mau trong dimg djch xa i^Kng loang 5 - 1 0 phiit, nia lai dudi \di nude may 2 - 3 lan, trang lai bing nude cit vd triing 2 - 3 Ian. Sat khuan be mat bing ethanol 70% trong 2 phut, trang lai bing nude cit vd trimg 3 - 4 lin. Khu ttimg man bing HgCh 0,1% va
Javen 6% trong cac khoang thdi gian khac nhau, nia lai mau bing nude cat vd triing sau moi lan khu trung. Tach vd qua, trai hat len mdi tiudng nudi cay khdi dgng (MTKD). Sau 5 6 tuin hat bat diu tai sinh the chdi (protocorm) la nguyen lieu cho cac nghien curu tiq> tfaeo.
Nlian nhanh the choi: Mau hat Que Ian huong dugc nudi cay tren mdi tnidng tao the chdi (Ti.g) gdm mdi trudng chit khoang co ban Knops bd simg cac chit dieu hda sinh tnidng (DHST) cd ndng do khac nhau, dudi anh sang gian den neon. Sau 4 tuan nudi cay, mau tao the chdi, thdng ke sd the chdi trong binh va tinh he sd nhan.
NJian nhanh clioL Cac choi huu hieu dugc nudi cay tren mdi trudng nhan nhanh chdi (Ni-g). Mdi trudng nudi cay sii dvmg la Knops cd bd sung cac chat DHST vdi ndng do khac nhau. Ket qua thi ngfaiem dugc theo ddi trong 5 tuin, thong ke he sd nhan chdi va dac diem cua cum chdL
Tgo cay hoan dunk: Chgn cum chdi phat trien ddng deu, dung keo hoac dao sac tach cac chdi hiru hieu va cay len mdi trudng cam ling ra re (R1-9) de binh nudi dudi anh sang gian den neon, sau 5 tuan chdi ra re tao cay con hoan chinh, thdng ke sd re tien chdi, do chieu dai re va danh gia chi sd ra re.
Huan tuyen vd ra ngdi: Cac cay con Que lan huang in vitro trong binh tir thi nghiem tren dugc huan lu\'en trong nha ludi 1 tuan.
Sau dd lay cay ra khdi binh va nia sach mdi trudng dudi vdi nude chay va cay vao cac khay da chuan bj gia the cd thanh phan khac nhau. Dat cac khay cay trong nha ludi, neu trdi nang can che 70° 0 anh sang bang ludi den, tudi nude .^ - 3 laiLngay. Thdng ke ty le cay sdng chet ^ a danh gia chat lugng cay sau 6 tuan.
T.\P CHI KHOA HOC v A CONG NGHE LAM N G H I E P S 6 6-2016 163
Cong nghe sinh hpc & Gidng cdy trong
Bang 1. Thanh phin cac loai moi tnrorng nuoi cay Que Ian hwmg in vitro
„ . . . .. i Ky hieu moi
Giai doan nuoi cay ^ ^ ^ ^ ^ Thanh phan moi truomg nuoi cay Nudi cay khdi ddng MTKD
Moi trudng nhan .p thi chdi '"*
Nhan nhanh chdi Ra rl tao cay hoan
chinh Huan luyen va ra
ngoi
N i .
Rl-9
Knops bd sung sucrose 30 g/l, agar 7 g/l.
Knops bd sung BAP 0,1 - 0,5 mg/l, Kinetm 0,1 - 0,2 mg/l, NAA 0,1 - 0,3 mg/l, nude dira 100 ml/I, dich chiet khoai tay 100 ml/1, sucrose 30 g/l, agar 4 g/l.
Knops bd sung BAP 0,2 - 1,6 mg/l, Kinetin 0,2 - 0,4 mg/l, NAA 0,1 - 0,2 mg/l, nude dira 100 mI/1, dich chilt khoai tay
100 ml/I, sucrose 30 g/l, agar 4 g/l.
Knops bd sung IBA 0,1- 0,3 mg/l, NAA 0,1 0,3 mg/l, sucrose 20 g/l, dich chilt khoai tay 100 ml/1, agar 4 g/I.
Cac loai gia thi (ddn tring, vd thdng, xa dira) thdi gian ngam cac loai gia the (2 - 6 ngay).
Cac thi nghiem dugc bd tri trong cac binh thuy tmh hinh tru (5 mau/binh), moi cdng thiic thi nghiem lap lai 3 lan. Sd Ueu dugc xii ly bang phan mem Excel.
Dieu kien nudi cay: Cudng do chieu sang 2000 lux; thdi gian chilu sang: 14 gid/ngay;
nhiet do phdng nudi: 25 ± 2 C.
Cac loai mdi trudng nudi cay trong nghien ciiu dua tren mdi trudng dinh duong ca ban Knops. Cac mdi trudng nudi cay dugc chuan do pH = 5,8; khii trimg d 1 IS^C, ap suit latm trong 17 phiit.
III. KET QUA v A T H A O L U A N
3.1. Tao mau sach va tai sinh the choi in vitro
Hien nay da cd nhieu cdng trinh cdng bd ve ket qua nghien ciiu khii trimg mau qua ciia cac gidng lan. Ke thira cac ket qua dd, mau qua Que lan huang sau khi thu hai ve dugc nia sach, khii trimg be mat bang ethanol 70% trong 2 phut. Tiep theo, tien hanh kbit trimg mau bing HgCla 0,1% (8 17 phiit) va Javen 6%
(10 - 17 phiit), cac cdng thiic thi nghiem dugc thilt k l nhu bang 2. Mau sau khi khu trimg dugc nudi ciy tren mdi trudng nudi cay khdi dgng.
Sau thdi gian theo ddi 5 - 6 man, ket qua thi nghiem kbit triing tao mau sach cho thay (bang 2), khi su dung cac loai hda chat khii trung HgCl2 0,1% va Javen 6%, vdi thdi gian khii trimg 8 - 1 7 phiit cho ty le mau sach Ian lugt la 60 - 100% va 50,33 - 83,33%. Trong dd, dung HgCh 0,1% khii trimg trong 15 phiit dat ty le mlu sach 94,33% (hinh la), ty le mau tai sinh thi chdi cao nhit dat 88,67% sau 40 ngay. Tuy nhien cd the keo dai them thdi gian khu trimg den 17 phiit va thu dugc ket qua cao hon (100%), nhung kha nang tai sinh the chdi thap (70%). Ben canh dd, su dung Javen 6% cho ty le mau sach va mau tai sinh thap hom so vdi HgCh, do dd lua chgn chat khu triing la HgCla 0,1%, thdi gian khii triing la 15 phiit se hieu qua nhat de tao m l u sach va tai sinh the chdi in vitro (hinh la-b).
Bang 2. Anh hudng cua loai hoa chat, thdi gian khir triing den kha nang tao mau sach va tai sinh the choi
Loai hoa chit
HgCl2 0,l%
Thtri gian (phiit)
8 10 12 15 17
Ty le mau sach (%) 60,00 76,67 87,00 94,33 100
Ty If mau tai sinh (%) 58,67 75,33 83,00 88,67 70,00
Thdi gian tai sinh (ngay)
38 39 39 40 41 164 TAP CHI KHOA HOC v A CONG NGHE LAM N G H I E P SO 6-2016
T . . . .X Thoi gian Loai hoa chat
(phut)
Ty 1^ mlu sach Ty If miu tai sinh Thdi gian tai sinh
(%) (%) (ngiy) Javen 6%
(NaClO)
10 12 15 17
50,33 60,67 66,67 83,33
46,67 56,67 63,33 60,67
39 40 41 41 3.2. Nhan nhanh the ch^i Que lan hucrng
The chdi tai sinh tir hat lan trong mdi trudng nudi cay khdi ddng dugc cay chuyen sang mdi trudng nhan nhanh the chdi. Thi nghiem dugc bd tri gdm 8 cdng thiic cd sir dimg cac chat
dieu hda sinh trudng thuc vat vdi ndng do khac nhau va 1 cdng thiic ddi chiing khdng bd sung chat dieu hda sinh trudng thuc vat (bang 3).
Sau 4 tuan theo ddi va thu thap sd lieu ve he sd tao the chdi, dac diem the chdi.
Bang 3. Anh hirdng cua nong do BAP, Kinetin, NAA den kha nang nhan nhanh the choi CTTN
DC Tl T2 T3 T4 T5 T6 T, Ts
( BAP
0,2 0,3 0,5 1 0,2 0,3 0,5 1
Chat DHST (mg/l)
Kinetin NAA 0,1
0,1 0,1 0,1 0,2 0,2 0,2 0,2
0,1 0,1 0,1 0,1 0,3 0,3 0,3 0,3
He so nhan nhanh (lan)
1,44 2,78 3,78 4,55 3,33 3,78 4,11 4,78 3,89
Dac diem the choi The chdi it, mau nau xam The chdi nhieu, map, xanh nhat The chdi nhieu, map, xanh The chdi rat nhieu, map, xanh The choi nhieu, xanh nhat The chdi nhieu, map, xanh nhat The chdi nhieu, map, xanh The chdi nhieu, map, xanh dam The chdi nhieu, xanh nhat Ket qua cho thay (bang 3), nudi cay the chdi
tren mdi trudng Knops cd bd stmg cac chat dieu hda sinh trudng thuc vat khac nhau cd anh hudng rd ret den qua trinh tao the chdi in vitro.
O cdng thiic ddi chiing khdng bd stmg chat dieu hda sinh trudng thuc vat, he sd nhan the chdi thap (1,44 lan), chat lugng the chdi kem, kich thudc nhd. Dac biet cdng thiic T7 da cho he sd tao thi chdi cao nhat dat 4,78 va the chdi nhiiu, map, mau xanh dam (hinh le). He sd tao the chdi Que lan huang tren mdi trudng chiia BAP 0,5 mg/l, Kinetin 0,2 mg/l, NAA 0,3 mg/l cao ban so vdi bao cao ciia Nguyen Thi Som va cs (2012) khi nhan nhanh the choi Dendrobium
fimbriatum Hook trong 8 man chi dat 4,63 lan.
3.3. Nhan nhanh choi Que lan huonng Nhan nhanh chdi la cdng doan cd y nghia het siic quan trgng ddi vdi viec tao ra sd lugng ldn cay con in vitro. Cac chat dieu hda sinh trudng thuc vat Kinetin, BAP va NAA thudng dugc bd stmg ket hgp de lam cho he sd nhan cua chdi tang len rd ret. Trong cac thi nghiem nay, su dung cac chat dieu hda sinh trudng thuc vat cd ndng do khac nhau de kich thich tao nhieu chdi nham dap ling cac yeu cau tao ra he so nhan cao, chdi sinh trudng, phat trien tdt. Cac the chdi lan dugc tao ra tir thi nghiem tren dugc cay vao mdi trudng nhan nhanh de tao cum chdi.
Ket qua cho thay (bang 4), khi sit dung TAP CHI KHOA HOC v A CONG NGHE L A M N G H I E P S 6 6-2016 165
Cong nghe sinh hpc & Gidng cdy trong cdng thiic mdi tnrong nudi ciy khac nhau cd anh hudng rd ret din kha nhan nhanh chdi Qui Ian huang. Sd chdi TB/mlu ciy ban diu
dat 4,67 - 9,67 Ian, chilu cao trung binh ciia chdi dat 2,2 - 2,7 cm va so la tnmg binh cua mgt chdi dat 2 - 3,5.
Bang 4. Anh hudng ciia ning do BAP, Kinetin, NAA den kha nang nhan nhanh choi CTTN
DC Ni Na
N j N4 N5 Ne N , Ng
Chat DHST (mg/l) BAP
0,2 0,4 0,6 0,8 1,0 1,2 1,4 1,6
Kinetin 0,4 0,4 0,4 0,4 0,2 0,2 0,2 0,2
NAA 0,1 0,1 0,1 0,1 0,2 0,2 0,2 0,2
Kha S i choi T B / m l u
2,67 5,33 6,00 6,67 9,67 6,33 5,67 5,00 4,67
nang tai sinh ciia Chieu cao TB
choi (cm) 1,7 2,2 2,4 2,6 2,7 2,5 2,4 2,6 2,3
chdi S o l a TB/choi
1,6 2,0 2,1 2,2 3,5 2,5 2,4 2,6 2,4 6 cdng thiic N M ndng do BAP tang (0,2 -
0,8 mg/I), Kinetin 0,4mg/l va NAA 0,1 mg/l din den sd choi TB/mau tang 5,33 - 9,67 trong khi tang nong do chit DHST thi so chdi TB/mau lai giam xuong. Kit qua nghien ciiu cho thay, cdng thiic mdi trudng N4 (mdi trudng khoang ca ban Knops bo sung BAP 0,8 mg/l va Kinetin 0,4 mg/l, NAA 0,1 mg/l) cho ket qua tdt nhit, vdi gia tri trung binh: sd chdi dat 9,67 chdi/mlu cay ban dau, chieu cao dat 2,7 cm va sd la 3,5 (hinh Id).
3.4. Kich thich ra re tao cay hoan chinh Tao cay con hoan chinh la giai doan cudi cimg ciia qua trinh nhan gidng bang phuang phap nudi cay md te bao. Mdi trudng nudi cay cd bd sung chat DHST thuc vat vdi cac ndng do khac nhau cho ket qua thu dugc d bang 5.
Ket qua thi nghiem cho thay (bang 5) d cdng thiic ddi chiing chi dat ty le choi ra re 53,33% va sd re trung binh 0,98 va chieu dai
tnmg binh re dat 1,73 cm, chi sd ra r l 1,70 vdi chit lugng r l ngan, xiu. Cac cdng thiic R1.3 co ndng do IBA 0,1 - 0,3 mg/I ty le chdi ra r l thap 56,67 - 70%, cdng thiic R4-6 cd nong do NAA 0,1 0,3 mg/l ty le choi ra re cao hom 80 96,67%, trong khi phdi hgp ndng do cua NAA va IBA nhu d cac cdng thiic R7.9 thi ty le ra rl giam (83,33 - 70%). Do dd lira chgn cdng thiic R^ vdi thanh phan la mdi trudng khoang ca ban knops bd sung NAA 0,3 mg/l cho ty le chdi ra r l cao nhat dat 96,67%, sd re TB/cay 3,46 va chieu dai TB/rl dat 3,83 cm, chi sd ra r l 13,25 (hmh le). Kit qua dat dugc ciing tuomg tu nhu bao cao cua Huidrom S. Devi et al.
(2013) cho ring mdi trudng V2 MS bd simg NAA 0,5 mg/l thich hgp nhat de kich thich ra re tao cay con hoan chmh loai Aerides odorata Lour, trong khi Hongthongkham J. et al. (2014) bao cao moi trudng chiia BA 5 mg/l cho ty le ra re cao nhat dat 96%.
Bang S. Anh hudng cua nong do IBA va NAA den kha nang ra r l CTTN
DC Rl R2 R3 R4
C h i t DHST (mg/I) IBA NAA
0,1 0,2 0,3
0,1
Ty le choi ra re (%) 53,33 56,67 60,00 70,00 80,00
So re TB/cay
0,98 1,08 1,10 1,28 2,67
Chieu dai TB/re (cm)
1,73 1,83 2,47 2,93 3,13
Chi so ra re 1,70 1,98 2,72 3,75 8,36
166 TAP CHI KHOA HOC vA CONG NGHE LAM NGHIEP SO 6-2016
CTTN
R5 R6 R7 R8
R,
Chit DHST (mg/l) IBA NAA
0,1 0,2 0,3
0,2 0,3 0,3 0,2 0,1
Ty le chii ra re (%) 93,33 96,67 83,33 80,00 70,00
So re TB/cay
2,73 3,46 2,51 2,29 2,02
Chilu dai TB/re (cm)
3,63 3,83 3,40 2,97 2,83
Chi so ra re 9,91 13,25
8,53 6,80 5,72 3.5. Huan luyfn va ra ngdi
Cac binh cay con dii tieu chuan chieu cao >
2,5 cm, cd 2 - 4 re va 3 - 4 la dugc huin luyen cay trong thdi gian 1 tuan d nha ludi cd mai che (can che them bang ludi den de tranh anh sang true xa). Sau do lay cay ra khdi binh, nia sach mdi trudng nudi cay dudi vdi nude may.
tiep tuc trdng cay vao khay vdi cac gia the khac nhau. Thi nghiem dugc bd tri vdi 9 cdng thiic, gdm 3 loai gia the (xa dira, ddn trang, vd thdng) dugc xu ly bang each ngam trong nude vdi thdi gian khac nhau, tudi nude 3 lan/ngay dam bao vira dii do am, chi tieu theo ddi la ty le song (%) trong 6 tuin.
Bang 6. Anh hudng cua loai gia the va thoi gian xu ly den ty If song Loai gia the Thoi gian xu ly
gid the (ngiy)
Ty le song (%)
Chat Ivorng cay
Xadita
Ddn trang
V6 thdng
30,00 36,66 60,00 93,33 83,67 73,33 43,33 46,66 80,00
Kem Kem Trung binh
Tdt Tot Tnmg binh
Kem Kem Tmng binh Ket qua thi nghiem cho thiy (bang 6), khi
dimg ddn trang da dugc xii ly 2 ngay trong nude lam gia the trdng Qui lan huang cho ty le sdng cao nhit dat tdi 93,33% sau 6 tuin theo ddi. Vao thdi diem nay cay Que lan huomg da bat dau hinh thanh la mdi va re vin tiep tuc dugc keo dai (hinh If). Nhu vay, vdi thdi gian xii ly 2 ngay gia the ddn dii mem cd kha nang gili nude tdt, du do xdp, giiip r l cay giu im tdt, phii hgp vdi viec nudi trdng Que lan huang.
IV. KET LUAN
Mau qua Qui lan huomg dugc sat khuan b l mat bing ethanol 70% trong 2 phiit, khu triing
bang HgCb 0,1% trong 15 phiit. Cay hat tren mdi trudng khoang ca ban Knops bd sung sucrose 30 g/l, agar 7 g/l, ty le mlu sach dat 94,33% va mlu tai sinh thi chdi 88,67%. Nhan nhanh the chdi Que lan huomg tren mdi trudng Knops bd sung BAP 0,5 mg/l, Kinetin 0,2 mg/l, NAA 0,3 mg/l, nude dira 100 ml/1, dich chiet khoai tay 100 ml/1, sucrose 30 g/l, agar 4 g/l cho he sd nhan the chdi dat 4,78 lan. Nhan nhanh chdi Que lan huomg tren mdi trudng Knops bd stmg BAP 0,8 mg/l, Kinetin 0,4 mg/l, NAA 0,1 mg/l, nude dira 100 ml/1, dich chilt khoai tay 100 ml/1, sucrose 30 g/l va agar 4 g/l TAP C H i KHOA HOC v A CONG NGHE L A M NGHIEP SO 6-2016 167
Cong nghe sinh hpc & Giong cay trong
dat kit qua 9,67 chdi TB/mlu, chieu cao TB/chdi 2,7 cm, sd la TB/chdi 3,5 va chdi phat triin tot. Tao cay Qui lan huomg hoan chinh tren mdi trudng Knops bd sung NAA 0,3 mg/l, nude dira 100 ml/1, dich chilt khoai tay 100 ml/l, sucrose 20 g/I va agar 4 g/l dat ty le ra rl
96,67%, so re TB/cay 3,46 va chilu dai TB/re 3,83 cm, chi so ra rl 13,25. Cay Qui Ian huang hoan chinh dugc huin luyen 1 tuan trong nha ludi va trdng vao khay cd gia the ddn trang da dugc xu ly 2 ngay trong nude, ty le cay sdng dat 93,33%.
Hinh 1. Cac giai doan trong quy trinh nhan giong cay lan Que lan hirong
Ghi chd: a) Mdu sach; b) Mdu sach tdi sink; c) Tao thi choi; d) Nhan nhanh chdi;
e) Cdy hoan chinh; f) Cdy trong trong dim trdng
T A I L I E U T H A M
KHAO
1. Hongthongkham J., Bunnag S. (2014). In vitro propagation and cryopreservation of Aerides odorata Lour. (Orchidaceae). Pakistan journal of biological sciences, 17(5): 608 - 618.
2. Huidrom S. Devi, Sanglakpam I. Devi, Thingbaijam D. Singh (2013). High frequency plant regeneration system of Aerides odorata Lour, through foliar and shoot tip culture. Not Bot Horti Agrobo, 41(1):169-176.
3. Nguyen Quynh Trang, Vu Thi Hue, Khuit Thi Hai Nmh, Nguyen Thi Tha (2013). Nhan gidng in vitro lan Phi diep tim. Tap chi Khoa hoc vd Cong nghe Ldm nghiep, 3(\y. 1 6 - 2 1 .
4. Nguyin Thi San, Nguyin Thi Ly Anh, Vfl Ngoc
Lan, Tran The Mai (2011). Nhan giong in vitro loai lan Dendrobium fimbriatum Hook (Hoang thao long nhan).
Tap chi Khoa hpc va Phdt triin, 10(2): 263 - 271.
5. Nguyen Van Song (2011). Nhan giong in vitro lan Kim diep (Dendrobium chrysotoxuom) mot loai lan rtmg CO nguy ca myet chung. Tap chi Khoa hoc Dai hoc Hui, 64: 127-136.
6. Pham Hoang Ho (2003). Cdy co Viet Nam Quyin in. NXB Tre.
7. Trin Hgp (1989). Phong lan Viet Nam. NXB Nong nghiep.
8. Tran Van Minh, Nguyen Van Uyin (2001). Vi nhan giong Phong lan nhdm Dendrobium tten quy mo cong nghiep. Tap chi Khoa hoc vd Cong nghe, 1: 1-9.
168 TAP CHI KHOA HOC vA CONG NGHE LAM NGHIEP SO 6-2016
Cong nghe sinh hpc & Giong cdy trong
USING IN VITRO CULTURE TECHNIQUE FOR PROPAGATION OF AERIDES ODORATA LOUR
Nguyen Van Viet', Nguyen Thi Huong^, Bui Van Thang^, Nguyen Thi Thu Hang ''^'^Vietnam National University of Forestry
^Forestry Development Center in Hanoi
S U M M A R Y
Micropropagation of Aerides odorata Lour, by in vitro cultural technique has been successfully stodied.
The results showed that the optimal method for bud sterilization was soaked in 70% ethanol for 2 minutes, in HgC12 0.1% for 15 minutes and then cultoring the samples with Knops medium, provided the proportion of purified samples was 94.33% and the regeneration rate was 88.67%. The most knops basal medium supplemented with 6-benzylaminopurine (BAP) 0.5 mg/l, Kinetin (Ki) 0.2 mg/l, a-naphthalene acetic acid (NAA) 0.3 mg/l, the protocorm were big, plump, dark green and the coefficient of formed protocorm reached 4.78. Propagating the buds in Knops with BAP 0.8, Kinetin 0.4 mg/l, NAA 0.1 mg/l, coconut water 100 ml/1, potatoes extract lOOml/1, sucrose 30 g/l, agar 4 g/l, the buds developed rapidly and were dark green, the coefficient of formed buds was 9.67. The rooted shoots 96.67%, the average number of roots was 3.46 per individual and the average length of roots was 3.83 cm when cultured after 5 weeks in Knops medium supplemented with NAA 0.3 mg/l, coconut water 100 ml/1, potatoes extract 100 ml/1, sucrose 20 g/l, agar 4 g/l. The completed nurslings were trained and planted on the sphagnum moss immersed in 2 days got the rate of living was 93.33%. The plantlets were perfectly acclimatized under greenhouse condition with high humidity before transferred to field. This procedure can be applied for mass production of Aerides odorata Lour, to meet the commercial demand.
Keywords: Aerides odorata Lour., in vitro, Knops, propagation, protocorm, rooting.
Ngvoi phan bien Ngay nhan bai Ngay phan bien Ngay quyet d|nh dang
PGS.TS. Ha Van Huan 10/9/2016
05/10/2016 25/10/2016
