544 HQI nghi KHOA HQC VA C O N G NGHg 2007
ITNG D U N G
CONG NGHE PHOI soma TRONG BAO TON vA PHAT TRIEN CAY QUE
(Cinnamomum cassia Nees et Eberth)
Bui Thj Tudng Thu, Tran Van Minh '-^ ^ PTNTD ve phia Nam ve CNTBTV. Vien Sinh hgc Nhiet dai
MODAU
Cay que la mdt trong nhinig loai cay dac san d Viet Nam, cd gia tri kinh te cao.
Que dugc trdng nhieu vimg frong nudc tu Bac vao Nam. Tuy nhien, hien nay nhan dan chia ra lam hai loai que: que vd mdng va que vd day. trong dd que vd day cho nhieu tinh dau hon. Cay que dugc du nhap va trdng trpt theo tap quan timg dia phuong, chu yeu tir
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cay thyc sinh bang hat, cd nhieu gidng khac nhau, cay que thoai hda dan, dan den ham lugng tinh dau khai thac thap. Cdng tac gidng que trong thdi gian qua cd gang chpn cay dau ddng de bao tdn va phat trien, nhung nhung ky thuat ddng hda cdn han che nhu giam hom va ghep canh (Gamborg, 1993). Nhung rao can dau tien van la cdng tac gidng. Gidng thuan chimg sinh trudng nhanh va cd nang suat cao dd dugc chpn Ipc. Cay que cd chira nhieu tinh dau, rat khd tai sinh in vitro (Mamood, 1993). U'ng dung cong nghe phdi soma trong bao tdn va ddng hda cay que me dau ddng la cin thiet trong phat frien cac viing chuyen canh cay que.
VAT LIEU VA PHU'ONG PHAP Vat lieu
Mdu nudi cdy: cay que dau dimg dugc thu thap tai Tien Phudc, Quang Nam. Mau dugc vd frnng bing hypochlorite caici (10%) trong 10 phiit, sau dd dugc vd trimg bing HgCb (0,1%) frong 5 phiit. Chdi non doan 1-1,5cm dugc su dyng trong nudi ciy.
Dieu kien nudi cdy: mdi trudng dugc vd trimg d 1 2 r c va lat frong 25 phut. Nhiet dp phdng nudi cay 28+2°C. Cudng dp chilu sang 34,2pmol/m^/s. Thdi gian chilu sang 8gid/ngay.
Mdi truang nudi cdy: mdi trudng dinh dudng khoang co ban MS (Murashige- Skoog, 1962) va WPM (Lloyd and McCown, 1981), dugc bd sung BA (6-benzyl aminopurine), Kinetin (6-furfrirylaminopurine), NAA (a-naphthalene acetic acid), glutamine, glycin, vitamin Bl va nudc dua (CW)
Phuong phap J Thi nghiem dugc bd tri theo CBD don ylu td, 4 lin lap lai, moi lin lap lai nudi cay
5 binh tam giac 300ml, mdi binh tam giac chua 65ml mdi frudng thi nghiem va dupe
Ph^n VI: CdNG NGHg Tg BAO T H I / C VAT 545
ciy 5 mlu. Sd lieu thu thap dugc phan tich thdng ke bing phin mim MSTATC (P=0.05). Ty le mau nudi ciy phat sinh tl bao soma (%), sinh khdi tl bao sau 60 ngay nudi ciy (g), do tang sinh khdi (g), ty le phat sinh phdi (%>).
W&T QUA VA THAO LUAN
;• Nudi cdy phdt sinh te bdo soma: Chdi non chua hda gd cua cay que sau khi vd trimg dugc nudi ciy tren mdi trudng phat sinh te bao soma. Mdi trudng nudi ciy MS, WPM va B5. Kit qua nghien cuu cho thay hau het cac mdi trudng khoang co ban nudi ciy diu cho phat sinh tl bao soma. Trong dd mdi trudng nghiem thirc A2: WPM + BA (lmg/1) + Kin (Img/l) nudi ciy cho phat sinh phdi. Phdi soma que cd mau frang sira va tl bao chit dam dac
Anh hu&ng cua mdu nudi cdy den phdt sinh te bdo soma: Than la re cay que in vitro dugc sir dung trong nudi ciy phat sinh phdi fren mdi trudng nghiem thiic A2. Ket qua nghien cuu cho thiy ca 3 loai mau nudi ciy diu cho phat sinh te bao soma. Trong dd miu nudi ciy than cho phat sinh tl bao soma cao (nghiem thiic B1)
Nudi cdy tdng sinh ti bdo soma tren mdi tru&ng agar: Te bao phdi soma (nghiem thuc B2) dugc nudi ciy tang sinh tren mdi trudng WPM va MS cd bd sung BA, kinetin, 2.4D, glutamine va CW. Kit qua nghien cuu cho thiy mdi trudng nghiem thiic C2:
WPM + BA (lmg/1) + Kin (lmg/1) cho tang sinh tl bao soma manh me (14,333g) sau 15 ngay nudi ciy. T I bao soma cd mau nau ciia nhua cay que sau 45 ngay nudi ciy
Nudi cdy phdt sinh ti bdo phdi soma: Tk bao soma nudi ciy tren mdi trudng nghiem thuc C2 dugc nudi ciy chuyin sang rndi trudng nudi ciy phat sinh phdi. Mdi tiudng nudi ciy phat sinh phdi la WPM cd bd sung BA, kinetin, CW. Ket qua nghien cim cho thiy, tl bao soma phan chia va biet hda manh me sang tl bao phdi soma sau 45 ngay nudi ciy tren mdi trudng nghiem tiiuc D5: WPM + BA (lmg/1) + Kin (lmg/1). Te bao phdi cd dang hinh ciu, tach rdi rac va khdng thanh cum, cd mau frang nau dng anh sang ciia tinh dau que
Anh hu&ng cua sd Idn cdy truyin din phdt sinh phdi soma: T I bao soma dugc nudi cay ' fruyen nhilu lin fren mdi tiirdng nghiem tiiuc D5, tiidi gian giiia hai lin ciy fruyen la 45
ngay, cang vl sau tl bao phdi soma xuit hien cang nhilu tir thi he cay fruyen F5 frd di, den thi he F8 toan bd tl bao phdi soma biet hda tiianh phdi hmh dng cd Id tien smh.
Tdi sinh phdi soma: T I bao phdi soma nudi ciy fruyin tren mdi trudng nghiem thirc D5, duoc nudi ciy tai sinh fren mdi trudng WPM cd bd sung BA, kinetin, CW. Ket qua nghien cuu cho tiiiy, mdi trudng nghiem thirc E3 va E4 cd ty le tai sinh cao (8 va 7 choi / binh). Tren mdi trudng nghiem tiiiic E4: WPM + BA (0,5mg/l) + Kin (lmg/1) + CW (10%) cho phat sinh chdi cd la phat triln
K£TLUAN
Chdi non cay qui tiiu tir rimg phdng hp dugc vd frimg va nudi ciy phat sinh te bao soma in vitro tren mdi frudng WPM + BA (lmg/1) + Kin (lmg/1). Than |a ^f/^^^^ '"^ ^ ^ nang nudi ciy phat sinh tl bao soma. T I bao soma tang sinh manh me (14,3 J3g; sau o
546 H(}i nghj KHOA HQC vA C O N G NGHg 2007
ngay nudi ciy tren mdi trudng WPM + BA (lmg/1) + Kin (lmg/1). T I bao soina cd mau nau cua nhya cay qui sau 45 ngay nudi cay. Te bao soma biet hda thanh te bao phoi soma tren mdi trudng WPM + BA (lmg/1) + Kin (lmg/1). T I bao phdi cd dan^ hinh ciu, tach rdi rac va khdng thanh cum, cd mau frang nau dng anh sang ciia tinh dau que. Tir lin ciy'truyin tiiir 5 frd di, kha nang phat sinh phdi nhieu sau nudi cay. Phdi soma dugc tai sinh tren mdi'trudng WPM + BA (0,5mg/l) + Kin (lmg/1) + CW (10%) cho phat sinh chdi cd la phat triln. U'ng dung cdng nghe phdi soma trong bao tdn va phat friln cay qui chung minh cd hieu qua
TAI LIEU THAM KHAO
1. Gamborg O. L. (1986). Protoplasts and plant regeneration in culmre. In: Demain AL, Solomon NA (eds) Manual of industrial microbiology and biotechnology.
American Society for Microbiology, Washington, DC
2. Gamborg O. L. (1993). Potential of biotechnology in micropropagation and genetic improvement of woody species. In: Proc Application of plant in vifro technology.
16-18 Nov 1993, University of Malaysia, Malaysia
3. Lloyd G. & B. McCown (1980). Commercially feasible micropropagation of laurel, Kahnia latifoUa, by use of shoot tip culture. Comb. Proc. Int. Plant Crop Soc. (30) 421-427 4. Mamood M. (1993). The total phenols and growth of cocoa (Theobroma cacao L.)
tissue culmre after supplemented with phenylalanine in vifro. In: NA Shamaan etal.
(eds) Proc. Application of plant in vitro technology. 16-18 Nov 1993, University of
Malaysia, Malaysia ^ 5. Murashige T. & R. Skoog (1962). A revised medium for rapid growtii and
bioassays with tobacco tissue culmres. Physiol. Plant. (15)431 -497 ~
SUMMARY
Application of somatic embryogenesis culture techniques in conservation and development of
Que (Cinnamomum cassia Nees et Eberth)
Bui Thi Tuong Thu, Tran Van Minh
National Key Lab of Plant Cell Biotechnology. Institute of Tropical Biology Stems were the well explants in cultivation. The explants were culfrue for somatic cell initiation well, for somatic cell mass' propagation, for somatic embryogenesis on medium WPM + BA (lmg/1) + Kin (lmg/1). Regeneration of somatic embryogenesis was culmred on medium WPM + BA (0,5mg/l) + Kin (lmg/1) + CW (10%)). A system of initiation, mass propagation, and regeneration culmre for conservation and cloning to develop the orchard and export planting area were established