KHOA HOC CONG NGHE
IMGHIEIV C i i u IVHAIV GIOIXIG LAIM PHI OIEP TIM [Dendrabium anasmum Lindi.) BAMG KY THUAT
IMUdi CAY /yi/ VITRO
Dang Thi Thu m ' , Pham Thu Ha'
T6MTAT
Lan Phi diep tim {Dendrobium anosmum Lindl.) thuoc chi Dendrobium la loai lan quy. co gia tri Itinh t^
cao. hoa co mau tim. rat da dang va co huong thom de chiu. Hien nay. nhu cau choi lan ngay cang nhieu.
nhung vol ky thuat trOng lan tmyen thong thi khong the nao dap ung dii nhu cau cua con ngudi. Nen viec ap dung phuong phap nhan gidng Uen Uen ddi voi Lan Phi diep tim de dioa man nhu cau cua khach hang v^
thu lai loi ich kinh te la mot viec lam can thiet Ket qua nghien ciiu nhan giong Lan Phi diep tim bang ky thuat nuoi cay in vitio cho thay cong thiic khu triing qua lan thich hop nhat la nhung qua lan vao con 96% va dot 30 giay tren ngon lira den con Oap lai 3 lan) cho ty le mSu sach va ty le mau sach nay mam cao (98.61%).
Moi tiirdng MS co bd sung 0.3 mg/1 BAP. 0.2 mg/1 kinetine. 0.1 mg/l NAA va bd sung 60 g/l khoai tay la thich hop nhat cho nhan nhanh the chOi Lan Phi diep tim voi he sd nhan the choi 11.21 l^n. Moi trudng tdi ini cho su hinh thanh va sinh tinrdng cua rd Lan Phi diep tim la mdi tmdng MS + 0.5 mg/i NAA+40 g/1 dudng sucrose -i 6.5 g/1 agar Cay duoc huan luyen d gia the SOJ^ xo dira + 50% vd thdng cd ty le cay sdng dat cao nhat 93.33%. chat luong cay khde. than Cling, re bam gia the tdt. gia thanh nudi cay md 1 cSy md dai 5 - 7 cm chi 5.000 - 7.000 ddng.
Tiikhda: Lan Phi diep tim, nudi cay in vitro, lua chon moi trudng, tao cay hoan chinh.
1 . DAT VAN BE
Phi diep tim {Dendrobium anosmuni} thudc chi Dendrobium la loai lan quy. cd gia hi kmh te cao.
hoa CO mau hm, rat da dang va cd huong thom dd chiu. Khdng chi cd vai fro lam canh. h-ang tri ma nd con CO the chiia benh. lam huong lidu. Do vay. loai Lan Phi diep tun nay hi lau da la ddi tupng suu tam ciia nhieu ngudi chm lan va nudi trdng lan. Trong tu nhien quan the lan sau khi bi khai thac vSn cd the tai tao toy nhien mat mdt thdi gian rat dai. ti Id sdng thap. Vi vay, vdi sir phat trien cua khoa hpc cdng nghe ndi chung va cdng nghe te bao thirc vat ndi ridng. vide nhan gidng bang ky thuat nudi cay in vitio tao ra sd lupng cay con ldn, on dinh ve mat di Qniyen la giai phap cho su khan hiem ngudn lan gidng va bao tdn loai cd nguy co tuyet chung cao. Xuat phat tir viln de thuc hdn frdn. vide ap dung ky thuat nudi csiy ui vitio de bao tdn va phat frien loai lan quy hiem nay la vd cting can thiet. Tir nhung ly do fren da tidn hanh "Nghien cuu nhan gidng Lan Phi didp tim (Dendrobium anosmum UnAl) bing ky thuat nudi cay in ntio" nhSm xay dung duoc quy frinh nhan gidng in vitio loii Lan Phi diep tim.
2. VAT UEU, m niwi VA PHUDIW PHAP iwflra cuu 2.1. VJt hju nghidn cihi
Loai: Lan Phi didp tim {Dendrobium .mosmum Undl.).
' Tnidng Dai hoc Nong Lam. Dai hpc Thai Nguye
Hinh 1. C4y Lan Phi di$p tim Vat heu nudi ciy; Hat tir qua ciJa Lan Phi didp khde manh. khdng sau henh. cd ngudn gdc dugc nudi dudng tai cac hp gia dinh d huyen Vd Nhai va Dai Tir. tinh Thai Nguyen.
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2.2. Phuong ph4p nghidn ciiu 2.2.1. Phuong phap bd tii tin nghidm
2.2.1.1. Phuong phap tao miu sach Lan Phi diep tim tu vat hdu qua
Xuly qua lan ben ngoai tii cay. Qua lan chin sinh ly (khpang 6 - 8 thang sau khi thu phan) dupc cat va dem ve phdng thi nghiem. Qua Ian phai dupc dam bao cdn nguyen ven khdng hi mit vd. Sau dd duoc khi triing so bd bang cdn 70° nhe nhang lau sach bd mat qua lan.
Khu triing qua ben tiong tii cay vd trung: Qua lan sau khi dupc khii frung ben ngoai se duoc dua vao ben trpng box cay vd frung. Nhung qua lan vap con 96" ddt tren ngon liia den cdn (30 giay) tir 1 den 4 lan.
Cay vao mdi tmdng. Tach phdi hat tir qua lan da dupc khir frung va dua vao mdi frudng nudi cay. Mdi frudng dupc sir dung vao miu gom: MS -i 30 g/1 sucrose -i- 6.5 g/1 agar; pH mdi tttrdng 5.7 ± 0.2. Ket qua duoc danh gia sau 8 hian nudi cay.
Chi heu theo ddi: Sd binh miu sach, sd hinh miu sach nay chdi. sd hinh miu hi nhidm.
2.2.1.2 Phuong phap nghien cuu anh hudng cua chat dieu hda sinh trudng den nhan nhanh the chdh The chdi Lan Phi diep tim thu duoc thi nghiem fren dupc cay vao mdi frtrdng cd thanh phan dinh dudng CO ban theo cdng thiic MS -i- 30 g/1 sucrose -i- 6.5g/l agar hd sung them cac chit didu hda sinh trudng nhu d bang 1. Kdt qua dupc theo ddi sau 8 hiin nudi cay.
Bang 1. Bd tri thi nghidm va thu thap so lidu anh hirong cda chat didu hda smh trudng ddn kha nang nhan nhanh thd chdi Lan Phi didp tim
Cdng thiic thi nghiem
DC CTI CT2 CTS CT4 CTS CT6 CT7 CT8
Chat dieu hda sinh tnrdng BAP (mg/1)
0 0.1 0.3 0.5 0.7 0.1 0.3 0.5 0.7
NAA (mg/1) 0
0.1
0.3
Kmetin (mg/1) 0
0.2
He sd nhan nhanh Gan)
Dac didm the choi
Mdi thinghiem duoc bd tri3 lan lap.
2.2.1.3. Phuungphap nghien cuu anh hudng cua cac chat hdu caden suslnh trudng va phat tnen chdi Phi diep tim
Moi trudng nhan nhanh Lan Phi diep tim dupc str dung cho cac thi nghidm nay gom: MS + 30 g/1 sucrose -i- 5.5 g/1 agar va ngudn huu co bo sung (khoai tay va chudi xanh) vdi ham lupng 0, 30. 60. 90.
120.150.180, 210 g/1. pH mdi frudng 5.7 - 5.8.
Cach lay dich khoai tay; cii khoai tay dupc nia sach. gpt bo vd. dupc cSt nho va can d cac ti Id khac nhau. Sau dd dupc nau chm va diing may xay sinh td de xay thanh dung dich, 100 g hrong ling vdi 100 ml.
Cach lay dich chudi xanh: chudi xanh dupc got bd vd. cit nho va can d cac ti le khac nhau. Sau dd dupc nau chin va diing may xay sinh td de xay thanh dung dich (100 g tiiong ling vdi 100 ml).
Ngudn vat hdu dupc sir dung: The chdi thu dupc tii thi nghiem frdn sau khoang 8 hian frong dieu kidn anh sang phii hpp de tao chdi lan. Nhiing choi lan nay se dupc chpn lam ngudn vat lieu de tien hanh thi nghidm nhan nhanh chdi. Dac diem chdi ban diu:
Chdi cd chat lupng tdt. ddng ddu. co 2 - 3 la. chidu cao 1,5 cm. The chdi duoc cay frong binh tam giic 250 ml. mdi binh 1 thia ca phe the chdi ban dau. mdi cdng thtrc 15 binh. Thoi gian thu sd lieu: Sau s'tuin cay chuyen
Chi Udu theo ddi; He sd nhan nhanh the chdi, chat lupng the chdi.
2.2.1.4. Phuong phap nghien cuu anh hudng cua chat dieu hda sinh tivdng den kha nang bmh thinh va sinh tivdng cua ri Lan Phi diep tim
Chdi Lan Phi diep tim du Ueu chuin vd kich thudc va sd la (chdi dd tao rd phai dam bao yeu '
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co: 3 - 4 ddt than, 3 • 4 la, cao 3 • 4 cm). Chdi Lan Phi hda sinh trudng (NAA BAP) d cac ndng dp khac di$p tim duoc cay vao mdi tnrdng dinh dtidng MS -i- nhau nhu 6 bang 2. Chdi dupc ciy frong binh tam giic 30 g/1 sucrose* 6.0 g/1 agar -r bd sung cac chat dieu 250 ml: 5 -7 cay/ binh.
Bing 2. Bd tri thi nghidm va thu thap sd U^u anh hudng ciia chat didu hda sinh trudng ddn k h i nang tao va phat tridn dia r^ Lan Phi didp tiro m vitro
Cdng thiic thi nghiem
DC RRl RR2 RR3 RR4 RR5 RR6 RR7 RR8
Chat didu hda smh trudng (mg/1) NAA
0 0.1 0.3 0.5 0.7 0.1 0,3 0,5 0,7
BAP 0 0,05
0
Ti Id chdi ra rd(%)
Sd rd TB/chdi
Chieu dai rd TB/chdi (cm)
Dac diem ctiard
Mdi thi nghidm dupc bdtil3 lan lap, 30 cbdi/lan.
2.2.1.5. Phuongphap dim cay ra gia the tidng va chSm sdc dnha ludi
Cay con hoan chinh co chi&u cao 4 - 5 cm, rk map, 5 - 6 rfe/cay huan luyen duoi anh sang tan xa khoang 1 - 2 tuan h-uoc khi ra cay. Sau do cay con duoc ma sach thach bam b rk va cay ra cac loai gia th^ khac nhau bao g6m: 100% xa dira, 50% xo dua + 50% vo thong va 100% dan h-dng. Gia the duac dat vao cac khay nhua chia 16 san loai kich thuoc 48 l6/khay (kich thuoc 15: 3 x 3 cm). M&i 16 cay 1 cay. Cay trong
trong cac loai gia the thi nghiem duoc d^t trong nha luoi duoc che sang 50% bang luoi den. M6i ngay tuoi phun suong cho cay lan 2 lan, sau 8 tuan cham soc, thu thap so heu
2.2.2. Phuongphap thu tiiSp va xuli sdlieu Thu thap va xu ly ket qua bang phan mem thong ke toan hoc Microsoft office Excel 7.0 va phdn mem SPSS.
* Tinh toan cac chi tieu thu thap
+ Ty + Ty le binh mhx sach tai sinh (%)
+ He s6 nhan nhanh the choi Oan)
• H6 so nhan choi (ldn) f Chieu dai TB cua rk (era)
So binh mau sach Tfingso binh mau cay
Tongsothechbi T6ng so the choi ban dau S6 Choi tao thanh Tong so mSu cay Tdng chieu dai cua re
Tdng so r i
xlOO XlOO 3. KET QUA NGHIBU CUU VA THAO LUAN
3.1. K^t qua tao mSu sgch Lan Phi diep tim Tao mhx sach la dieu kien tien quyet co y nghia vo ciing quan trpng trong su thanh cong ciia mot quy trinh nhan gidng m viti-o doi voi bat ki doi hrong nghien cuu nao, bai cac thi nghiem co the tiep tuc duoc thuc hien khi da tao ra. Doi voi Lan Phi ditP tira, vai vat lieu la qua, viec khu tning va gieo hat
giiip hat nay mam hieu qua la mpt hong nhung chi tieu quan trong ciia thi nghiem nay. Ket qua tao mdu sach duoc the hien h-ong bang 3.
Bang 3 cho thdy, khi tang so lan dot qua tir 1 - 4 lan da lam cho ty le mdu sach tang len dang ke {tu 61,76% den 100%). Tuy nhien, a cac cong thuc CTI d6n CTS dat 100% s6 mdu sach co kha nang nay mdm, trong khi CT4 (4 lan dot) chi dat 92,8,)% vi co
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mdt sd hinh miu sach hat hi chuyen man den va khdng cd kha nang nay mam. Didu nay cd the do ddt den lan thu 4 lam cho nhiing phdi nim sat vd hi ndng va mat siic nay mam. So sanh vdi ket qua khu hiing qua lan mot sd loai thudc chi Dendrobium cda mdt sd nghidn ciiu di trudc cho thay: Nguydn Quynh Trang va cdng sii (2013) [2). Nguydn Van Viet (2017)
[4], deu khii hung qua lan bang HgCl^ 0.1%. Nguydn Van Viet cd kdt qua khu Wing hang dung dich HgCl, 0.1% trong 12 phiit va nudi cay trdn mdi h-udng
Knops. cho ty 16 miu sach la 86,7%, ty le mau phat stah thd chdi (protocorm) la 76,7% - thaip hon nhieu so vol CT3 ciia nghien ciiu nay. Mat khac, thuy ngan clorua la mdt hda chat kha ddc. Nen viec sir dung cdn 96% de khu tiling nhu h-ong nghidn ciiu nay viia don gian lai it ddc hai. Nhu vay. vdi qua Lan Phi didp hm cd thd nhung qua vao cdn 96% va ddt 30 giay h-dn ngon lira den cdn (3 lan) cho ly le mSu sach va mau sach nay mam tdt nhat.
Cdng thiic CTI CT2 CT3 CT4
Chat khii trimg
Cdn 96°
B4ng3.Kd Cach khii
triing Ddt 1 lan Ddt 2 lan Ddt 3 lan Ddt 4 lan
t qua mau s ^ h Lan Phi didp tim Sdbmh
mau cay 68 70 72 70
Sd hinh mau sach
42 52 71 70
Tl le mau sach (%) 61.76 74,28 98,61 100
Ti Id mau tai sinh chdi (%)
61,76 74,28 98,61 92,85
Hinh 2. Hgt Lan Phi difip tim phdt tri^n sau 4 tuan nuoi cay (B)
3.2. K^t qua nhan nhanh th^ ch6i Lan Phi di6p tim
3.2.1. Ket qua anh hudng cua mdi trudng dinh dudng CO ban den kha nang nhan nhanh the chdi Bang 4. K^t qua anh huong cua m6i tru6ng duih duong d^n kha nang nhan nhanh th^ ch6i Lan Phi digptim
Viec lira chpn moi truang nuoi cay thich hgp la van de quan trpng anh huong den su thanh cong ciia qua trinh nuoi cay in vitro. Qua viec tham khao mpt so tai beu cho thay moi truang thich hop cho da s6 loai lan dac biet la cac loai lan thupc ho Dendrobium la cac loai moi truang: Knudsons C, Knops, MS, B5,...Trong pham vi nghien cuu ciia ngi dung nay tien hanh thu nghiem 3 loai moi truang la Knudsons C, MS va B5, co bo sung them 0,3 mg/1 BAP + 0,1 mg/1 NAA + 0,2 mg/1 kinetin + 20 g/1 sucrose + 6,5 g/1 agar. K^t qua sau 8 tuan dupc the hi§n a bang 4.
Moi tmdng
MS Knudsons C
B5 Sig.
He sd nhan nhanh th^
choi sau 8 tuan Odn) 6,54+0,17^
5,25+0,13'' 3,24+0,26^
0.0001
Dac diem ciia the ch6i sau 8 tudn
nuoi cay The ch6i xanh tdt, co
long Th^ ch6i hoi vang The chdi nho, yen, ch^t nhieu, hai vang
(Cac sd heu duoc theo sau bdi cac ky Or khac nhau cdy nghia khac biet vdi do tin cay 95%)
Hmh 3. Vnk chdi Lan Phi difp hm h-dn mdi hudng MS
Ket qua d bang 4 cung nhu hinh anh cac mau thi nghiem. cho thSy mdi frudng MS co ban cd anh hudng hch cue nhat den kha nang nhan nhanh chdi Lan Phi diep tim voi he sd nhan 6.54 lan. the chdi cd mau xanh. the chdi khde hinh thanh nhitu Id tien dd cho su phat hien tii the chdi thanh chdi
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3.2.2 Ket qua anh hudng aia chat dieu hda stiih Chat dieu hda stah tiTidng d nhimg ndng dd tmdng ddn nhan nhanh Old choi thich hop cd kha nang Uiuc day qua hinh tang smh the chdi rd ret (cdng thiic thi nghidm so voi ddi chiing).
Bang 5. Kdt qua i n h hudng dia chft di^u hda smh hudng Ain k h i nSng nhin nhanh Cong
thiic thi nghidm
DC CTI CT2 CTS CT4 CTS CT6 CT7 CTS
Chat dieu hda smh trudng BAP
(mg/1) 0 0,1 0,3 0,5 0,7 0,1 0,3 0,5 0,7
NAA (mg/1) 0
0,1
0,3 Sig
KmeUn (mg/1) 0
0,2
Hd sd nhan nhanh the chdi (lan) 2.33+0.04' 5.44+0.07"
9.44±0.11' 7.22±0.14'' 6.04+0. U ' 5.22+0.13'' 3.87±0.05' 3.17+0.06' 3.15+0.22' 0.0001
Ty Id mlu phat trien thanh chdi
(%)
0 100 100 100 100 100 100 100 100
sd thd chdi phat trien thanh
chdi 0 16.97+2.02'^
23.53+1.96' 19.63+0.77' 14.7+0.15' 12.53±0.87'' 8.43+0.42'
9.4+0.76' 9.07±0.16'
0.0001
Dac didm thd chdi
+ + +++
++
++
++
+ + + Ghi chu: (+): The chdi it, phat trien kem, vang; (++): The chdi va chdi tnmg binh, xanh; (+++): Thd chdi va chdi nhiiu, phat trien manh.
d cong thiic CT2 (0,3 mg/1 BAP ket hap vai 0,2 mg/1 kinetine va 0,1 mg/I NAA) cho ket qua t6t nhat voi h^ s6 nhan th^ choi la 9,44 lan, so the ch6i phat tnen thanh choi ion. Tuy nhien, O nong do cao (CT7, CT8) lai CO nhimg anh huong khong tot den kha nang nhan nhanh the ch6i, lam cho hieu qua nhan the ch6i giam ro ret chi con hon 3 lan.
3.2.3. Kit qua anh hudng cua khoai tay den kha nang nhan nhanh the chdi Lan Phi diep tim
Khoai tay c6 chua cac yeu to vi luong va da lugng nhu Fe, Ca, P... va mpt so vitamin. Khoai tay duac su dung lam nguon carbohydrate b6 sung b^n canh thanh phdn duong saccharose cho choi va cay trong di^u kien in vitio. Trong 100 g khoai tay co chiia 17,47 g carbohydrate; 0,78 g duong, cac thanh phan vi lupng nhu Ca, Fe, Mg, Zn, vitamin B6...(theo United States Department of Agriculture - 2014). Sau 8 tuan nuoi cay, cac the choi Lan Phi diep tim tren moi trudng co bo sung dich khoai tay a cac ti le khac nhau cho thay co su khac bi^t dang ke so VOI ddi chiing (0% khoai tay). Thi nghidm duac tien hanh va thu thap so lieu sau 8 tuan va Hinh 4. I h ^ ch6i Phi di^p tim trfin moi tni6ng MS + duac the hien a bang 6.
0^ mg/1 BAP + 0,2 mg/1 kmetine + 0,1 mg/1 NAA
Bang 6. K^t qua anh huong cua khoai tay d^n kha nang nhan nhanh t h i ch6i Cong thuc
thi nghiem DC KTl KT2
Khoai tay (g/0
30
So mau cay
He s6 nhan nhanh the
ch6i 2,33±0.04^
Ty le mdu phat trien thanh choi
(%)
0
So the choi phat tri^n thanh choi
54,86+0,86'
Dac diem the ch6i
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KT4 KT5 KT6 KT7 _j
90 120 150 180 210
30 30 30 30 30 Sig
8.84+0.04°
7.7+0.09' 6.75+0.03°
5.7+0.02' 4.32±0.02' 0,0001
100 100 100 100 100
42.9±1.20°
30.2±0.57' 20.4±0.47'' 17.13+0.19' 11.27±0.29' 0.0001
+++
++
++
++
+
Ghi chu: (+): Thi chdi it, phat tiien kim, vang; (++): The chdi va chdi trung bmh, xanh; (+++): ThS chdi va chdi nhiiu, phat tiien manh.
nghidn cihi cua Vu Ngoc Lan (2013), khi da dua ra quy trinh nhan gidng in vitio gidng lan Thach hoc {D. nobile Lindl): moi tnrong nhan cum protoconm CO bd sung 60 g/1 khoai tay cho he sd nhan nhanh 4,47 ldn sau 8 tuan nuoi cay (Vu Nggc Lan, 2013) [1].
So sanh voi ddi chiing cho thay mdi truong co bd sung chat hiru co gitip tang manh qua trinh nhan nhanh the choi, ddng thdi giiip the chdi phat trien thanh chdi khoe manh, sd lupng vAnkxx. La ca sd tao ngudn vat li^u cho nhan nhanh chdi Ian sau nay. Nhu vay, dua vao ket qua phan tich ta thdy moi tnrdng K2:
MS + 30 g/1 sucrose + 5,5 g/1 agar + 60 g/1 khoai tay la thinh hgp nhat cho nhan nhanh the chdi Lan Phi diep tim.
3.2.4. Kit qua anh hudng cua chudi xanh den kha ndng nhin nhanh the chdi Ian Phi diep tim
Sau 8 tuan nudi cay, cac the chdi Lan Phi diep tim tren mdi trudng MS + 30 g/1 sucrose + 5,5 g/1 agar cd bd sung them ham lupng chudi xanh d cac ti le khac nhau: 0, 30, 60, 90,120,150,180, 210 g/1. Mt qua duac the hien d bang 7.
A B Hinh 5. T h i chdi Lan Phi diep tim sau 8 tudn nuoi
c^y h ^ n mdi huemg KT2 (A) va KT7 (B) Bang 6 cho thay khi tang ham lupng khoai tay tir 0 den 60 g/1 thi cd su gia tang ro r6t v6 cac chi tieu theo doi va dat cao nhat d thi nghiem cd bd sung 60 g/1 khoai tay, cho he sd nhan nhanh the chdi cao nhat la 11,28 lan, trong dd th^ choi phat trien thanh chdi trung binh la 54,86 tren mot cum mdu ban ddu.
Tuy nhi^n, khi tiep tuc tang ham lupng khoai tay thi he sd nhan nhanh the choi va su phat trien cua the chdi thanh chdi deu bi giam. Mpt so man cd hien tuong ch6t the choi. Ket qua nay cung phii hpp vdi
Bang 7. K^t qua anh huong ciia chuoi xanh d^n kha nflng nhdn nhanh 1h^ chdi Cdng thiic
thi nghiem BC
Cl C2 C3 C4 C5 C6 C7
Chuoi xanh (g/1)
0 30 60 90 120 150 180 210
Sd mau cay
30 30 30 30 30 30 30 30 Sig
He sd nhan nhanh the choi (Ian)
2.33+0.04' 3.69+0.04d 5.05+0.07' 6.06±0.03' 4.38+0.02' 3.82+0.01'' 2 74+0.03' 2.1+0.08' 0.0001
Ty le miu phat trien thinh chdi
(%)
0 ^ 100
100 100 100 100 100 100
sd the chdi phat trien thinh chdi
0 7.77+1.71'' 11.77+1.15'^
20.83+0.75"
13.7+0.15'"
9.53±0.62°' ' 6.6±0.I7' 2.67+0.15^" '
0.0001
Dac diem the
chdi + ++
+++
++
+ + + Chichu: (Cac sd heu duoc theo sau bdi cac ky tir khac nhau cd Ji nghia khac biet vdi dd tm cay 95% r i.
The chdi it, phat tiien kem, vang: (++): The choi va chdi tivng binh, xanh: (+++): The chdi va chdi nhieu phi trien manh, xanh.
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KHOA HOC CONG NGHi
Nhung ket qua nghien ciiu vdi loai Lan Phi di$p tim lai cho thay mdi trudng cd bd sung 90 g/1 chudi xanh cd th^ chdi phat tridn nhieu nhat. Trong cdng thiic nay cho thay su tang sinh ve sd lupng the chdi kem nhung lai kich thich the chdi mpc thanh chdi nhidu, chdi khde va phat trien ddng deu. Khac vdi nghien ciiu ciia Saranjeet Kaur, K. K Bhutani (2012) [4] khi nhan nhanh lan Cymbidium pendulum (Roxb.) thi phai d ndng dp 150 g/1 trd len thi he sd nhan chdi giam di va cd hi^n tuang chet.
Nhu vay, dua vao ket qua phan tich cho thay moi trudng C3: MS + 30 g/1 sucrose + 5,5 g/I agar + 90 g/1 chudi xanh la thich hop nhat cho nhan nhanh the chdi Lan Phi diep tim.
3.3. K^t qud anh hudng ciJa ch^t d i ^ hda sinh trudng d^n kha nang hinh th^nh vd sinh tnrdng rk.
Lan Phi di$p tim
Bang 8. K^t qua anh hudng cua ch^t di^u hda sinh tnrdng d^n kha nang tao vd phdt tri^n ciia rk Lan Phi di^p timiD vitro
A B Hinh 6. Th^ chdi Lan Phi di^p tim sau 8 hidn nudi
c^y tren mdi tivdng C3 ( ^ vd C7 (B) Ket qua nghifen cim nhan nhanh lan Cymbidium pendulum (Roxb.) cua Saranjeet Kaur, K. K Bhutani (2012) da cho kdt qua d ham lupng chudi tren (75 g/1) bat lgi cho su sdng ciia protocorm; cac co quan gidng nhu protocorm cd hien tugng chet.
Cong thiix:
thi iighiem
DC KRl RR2 RES RR4 RR5 RR6 RR7 RR8 _ Sig.
Chat dieu hda sinh trudng
(mg/1) NAA
0 0,1 0,3 0.5 0.7 0.1 0.3 0,5 0,7
BAP 0
0,05
0 Tyle chdi ra
rd(%) 100 100 100 100 100 100 100 100 100 0,0001
sd rd hung hinh/chdi I.66±0.05' 1.86±0.11'^
6.96+0.06'"
4.26±0.05' 3.08+0.04' 2.85+0.04' 4.58+0.05"' 7.64+0.09' 5.12+0.03' 0,0001
Chidu dii rd tmng hinh/chdi
(cm) 1.85+0.01' 2.72+0.08"
2.13+0.05'' 4.18+0.10' 3.54±0.06'"' 3.74±0.05'"
3.36+0.12' 3.73+0.09'"' 2.23+0.07' 0.0001
Dac diem cua rd
Re ngin. manh. cay thap Rd map. ngin vi it. ciy to map. li to dai xanh dam
Rd mip. nhieu rd. rd ngin. ciy thap. moc deu de chdi nhidu Rd map, dai, ciy l i to vi day nhat Re mip, dai, cay cao map, la xanh dii to
Rd mip, dii, ciy cao to, l i to vi dii, ciy xanh tdt
Rd map, dii. ciy rit cao to. la to vi dii xanh dim
Rd map, dai, miu xanh dim, nhieu ciy cao to, li to v i dii, cay xanh tdt Rd map. nhidu rd. rd ngin. cay thip mip. l i to nhimg ngin
(Cac sdheu duoc theo sau bdi cac ky tii khac nhau Cdng doan cudi ciing cua qua trinh nudi cay In vitio la hinh thanh rh de tao cay hoan chinh. Giai doan nay dam bao chdi ra rk ddng deu va dat ti le s6ng cao khi dem ra Q-dng. Chdi sii dung cho thi nghiem nay dam bao yeu cdu: cd 3 - 4 ddt than, 3 - 4
CO y nghia khac biet vdi do tin cay 95%)
la, cao 3 - 4 cm). Vai trd ctia chat dieu hda sinh trudng mang tinh chat quyet djnh la auxin. Auxin la nhdm chat cd tac dung len tang phat smh ciia mau nudi cdy va dSc biet dgiai doan nay co tac dung hinh thanh rk. Chdi I-an Phi diep tim dupc cay vao mdi
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 7/2020
107KHOA HOC C O N G NGHE
trudng MS +40 g/1 duong sucrose + 6.5 g/1 agar + hd sung cic chat didu hda smh Unidng (NAA, BAP) d cac ndng dp khic nhau. Ket q u i sau 8 hiin duoc the hidn trong bang 8.
Ket qua phin tich phuong sai mdt nhin td cua k h i n i n g ra rd, dp dii rd, kha nang smh hiidng cua chdi deu cd sig - 0,0001 < 0,05. Dieu dd chiing td h i m luong cic chat dieu hda sinh hudng khic nhau se anh hudng rd ret ddn k h i ning ra rd. chidu dai rd v i k h i nang smh hudng cua chdi Lan Phi diep tim.
Hinh 7. Chdi Lan Phi didp tun ciy trdn mdi budng ra r5RR7
Khi bd sung BAP d ndng dd thap (0,05 mg/1) ket hpp void ndng do NAA thap (0,1 mg/l), sd rd it (1,86 rd/chdi), rd khdng ddng ddu v i h i n g Khi tang ndng do NAA len (0,3 mg/I) sd rd r i t nhieu (6,96 rd/chdi) nhung rd khdng ddng deu va ciy rat t h i p (chidu cao chdi ting them 0,66 cm). Khi tiep tac ling ndng do NAA len (0.5 mg/l v i 0.7 mg/l) sd rd giim dan (theo thii hr 4.26 rd/chdi va 3.08 rd/chdi). Vi vay. bd sung BAP d ndng dp 0,05 mg/l li khdng can thiet a thi nghidm tao v i phit trien rd. Nhu vay. mdi tnrdng tdi uu cho chdi Lan Phi didp hm ra rd la RR3: MS + 0,5 mg/l NAA+40 g/1 dudng sucrose + 6,5 g/1 agar. Khi so sinh ket qui trong cdng thiic ra rd ciia nghidn cihi niy vdi mdt sd nghien ciiu trudc dd vdi nhirng loii lan thupc chi Dendrobium cho thay:
+ Vdi cdng trinh nghien ciiu ciia Nguydn Quynh Trang v i cdng su (2013) [21, cdng thiic bd sung 0,5 mg/l IBA v i cdng thiic 0,3 mg/l 1BA+ 0,1 mg/l NAA cho ty le choi ra rd dat 98%, sd rd trung hinh dat trdn 3 rd/chdi. Vdi ket qua nghien ciiu niy cho sd rd li 7.64 rd/chdi, rd khde manh mau xanh dim, ddng deu v i rd dii li 3,73 cm.
NAA cd anh hudng ldn den kha nang ra rd cua chdi Lan Phi didp tim nudi cay in vitio. Cic chi tieu theo ddi deu cd su sai khac co y nghia giira cic cdng thiic thi nghiem. O ndng dp NAA thap (0.1 mg/l) sd rd trung hmh/chdi thap (2.85 rd/chdi), cic rd khdng ddng deu. Khi tang NAA ldn 0,3 mg/l sd lupng rd cao ban (3,36 rd/chdi), rd dai deu. Khi ting NAA ldn 0.5 mg/l sd lugng rd thu duoc cao nhat (7.64 rd/chdi), rd dii ddu (3,73 cm) va cd miu xanh dim. Tiep tuc tang NAA len 0.7 mg/l k h i nang ra rd, die diem rd deu kem hon so vdi cdng thiic chiia 0.5 mg/l NAA.
Bang 9. Kdt qua huan luydn ciy Phi digp ra gii the v i chim sdc trong n h i ludi
+ Vdi nghien ciiu cua Selvaraju Parthibhan v i cs (2015) [5], nudi cay m vitiota protocorms a Dendrobium aqueumLindley, choi nudi ciy trdn mdi truong bd sung 1/2 MS vdi IBA 5 mg/l tao ra hning binh 8.75 rd/ciy, my nhien rS dii nhat (1.48 cm) d ndng do NAA 7 mg/l. Nhung cdng thiic nay cho s6 rd nhidu nhung rd khdng dii.
3.4. Kdt q u i huin luy$n ciy Lan Phi didp tun ra gii the v i chim sdc trong n h i ludi
STT Loai gia the
50% xa diia + 50% vd thdng 100% ddn h-ang
Sd miu cay (ciy)
Sig
Ty le ciy
song'"' Chat luong cay Ciy ydu. rd mdi bim gia the Ciy khde. thin cung. rd bam gia the tdt Ciy khde. thin ciing. xuit hi6iiiri;;j;^7j"j~- gii the tdt
Bang 9 cho thay ciy duoc huin luyen d gii the 50% xa dita + 50% vd thdng cd ty le ciy sdng dat cao nhat 93.33%, chat lugng ciy khde, thin ciing, rd ham g i i the tdt.
Ket q u i nghidn ciiu cung cho thay khi nudi ciy md: gia thanh giam do hd sd nhin ciia qua hinh nudi ciy md cao, do dd khi nudi ciy md Lan Phi didp tim
bang nudi cay md mdt ciy md dii 5 - 7 cm chi 5 000 - 7.000 ddng (neu mua sd lugng ldn cd the chi cdn 4.000 ddng/ciy). Nguoc lai ciy dugc nhan bing kei vdi kich thudc tuong ta phii 50 - 80 nghm dong/civ (theo g i i thi mrdng). Mat khic. de tao ngudn gidng ldn thi vide nhin b i n g kei gap „i,ii„" ."f ^ khin. '™ ^ °
KHOA HOC C O N G N G H £
mdt sd hidn phip ky thuit nudi trdng hai loii Ian ban dia {Dendrobium nobile Lindl., Dendrobium chiysantiium LmdI.) tai H i Ndi. 2013. Thu vien Qudc gia Viet Nam.
2. Nguydn Quynh Trang. Vu Thi Hue. Khuit Thi Hii Nmh. Nguydn Thi Tho (2013). Nhin gidng m vitio Lan Phi diep tim {Dendrobium anosmum). Tap chi Khoa hpc vi Cdng nghd Lim nghiep, sd 3 - 2013.
3. Nguydn Van Viet (2017). Ung dung ky thuit nudi ciy m vitio tiong nhin gidng Lan Hoing thio ken {Dendrobium htuidorum Lindley). Tap chi Khoa hoc vi Cdng nghe Lim nghidp. so 4. 2017.
4. Kaur Saranjeet., Bhuhmi K. K. (2012). Organic growth supplement stimulants for m vitio multiphcation of Cymbidium pendulum (Roxb.) Sw.
Hort. Sci. (Prague), 39:47 - 52.
5. Selvaraju Parthibhan. Mandah Venkateswara Rao. Thiruppathi Senthd Kumar (2015). In vitio regeneration from protocorms in Dendrobium aqueum lindley - An unperiled orchid. Asian Pacdic Joumal of Tropical Biomedicine. December 2015, Pages 227-233.
4.linUMN
Dd tdi uu qua tiinh tai smh chdi cay Lan Phi didp dm, quy tiinh nhin gidng nen duoc thuc hien tidn mdi tiirdng MS. Cdng thiic khil tiling q u i lan Uiich hgp nhat Ii nhiing q u i Ian vao cdn 96% vi ddt 30 giay tren ngpn lua ddn cdn flip lai 3 lin) cho ty Id miu sach vi ty le miu sach nay mim cao (98.61%).
Mdi tiTidng MS cd hd sung 0.3 mg/l BAP. 0.2 mg/l kinetin, 0,1 mg/l NAA va bd sung 60 g/I khoai tay li tiiich hgp nhit cho nhan nhanh the chdi Lan Phi diep tim vdi hd sd nhin thd chdi 11,21 lin.
Mdi tiTidng tdi uu cho su hinh thinh v i stah tmdng cua rd Lan Phi didp tim li mdi tindng MS + 0.5 mg/l NAA+40 g/I dudng sucrose + 6.5 g/I agar.
Ciy dugc huin luydn d gia the trdng 50% xa diia + 50% vd thdng v i cham sdc trong nha ludi cd ty Id ciy sdng dat cao nhat 93.33%. chat lugng cay khoe.
thin Cling, rd ham gii the tdt gii thinh nudi cay md 1 ciy md dii 5 - 7 cm. chi 5.000 - 7.000 ddng/ ciy.
TAI UEU THAM KHAO
1. Vu Nggc Lan (2013). Nghien ciiu nhin gidng in vitio khdng su dung chat dieu tidt sinh trudng vi
STUDYING ON IN W75?0 PROPAGATION OF Dendrobium anosmum LmdL Dang Thi Thu Ha, Pham Ttm Ha Summary
Dendrobium anosmum of the genus Dendrobium is a precious orchid species with high economic value.
The flowers of Dendrobium anosmum has a vary range of purple and has a pleasant Iragrance. Currendy.
with the increasing demand for orchids, bui with tradiUonal orchid cultivation techniques, it is impossible to meet ihe needs of people. The applicahon of advanced breeding methods for orchid Dendrobium anosmum to satisfy the needs of customers and recover economic benefits is necessary. The results of propagation of Dendrobium anosmum by in vitio culture technique showed thai the orchid sterilizaUon formula is most suitable to immerse the fruit of Dendrobium anosmum in 96% alcohol and bum 30 seconds on die flame of alcohol lamp (repeated 3 Umes) gave a processed sample proportion and a high proportion of germination (98.61%). MS environment supplemented with 0.3 mg/l BAP. 0.2 mg/l kinetine. 0.1 mg/l NAA and additional 60 g potato is most suitable for rapid multiplication of buds with a coefficient of 11.21 times. The optimal environment for the formation and growth of Dendrobium anosmum roots is MS + 0.5 mg/l NAA + 40 g/I sucrose + 6.5 g/1 agar. Trees were Uained in the subsh-ates which include 50% Coconut fiber and 50%
pine bark to have the highest survival rate up to 93.33%. Trees have high quality which cost of only 5000 - 7000 VND/ti-ee for its height from 5 - 7 cm.
Keywords: Dendrobium anosmum, in vitro culture, environmental selection, create completed Uee.
