'pt^n VI: CdNG NGH$ T^ B A O THgC VAT ..^
Sir PHAT SINH CO QUAN TlT L O P MONG TE BAO CUA GIONG DIEU (Anacardium occidentale L.) CAO SAN BOI
NUOI CAY in vitro
Huynh Hiru Dure, Nguyen Dinh Sy, Nguyen Thi Quynh Phdng Cdng ngh? te bdo thuc vgt, Vi?n Sinh hgc Nhi?t d&i
MdDAU
Cay dieu (Anacardium occidentale L.) Id cay cdng nghiep dem lai hieu qud kinh tl cao d mgt so nude thudc virng nhiet ddi Chdu A vd Chdu Phi do nhan hat dieu cd gid tri cao trong xuat khau. Thdng thudng cdy dugc nhdn gidng tir hat, nhung phuong phdp nay dem lai tinh khdng ddng nhdt ve di truyin (Philip vd Unni, 1994). Cdc phuong phdp nhdn giong vd tinh truyin thdng nhu gidm cdnh, gh^p thudng dugc sir dung de nhdn cdc ddng dilu cd ndng sudt cao, tuy nhien he sd nhdn gidng khdng dap ung nhu cdu. Do phuong phdp nhdn gidng in vitro dugc sir dung vd thdnh cdng tren nhieu lodi cay dn trdi gdn vdi cdy dilu (Barghchi &
Alderson, 1983; Litz vd cgng sy, 1984) nen nhdn gidng in vitro cdy dieu cd tinh khd thi (Vu Nggc Phugng vd cs, 2003).
' Nudi cay Idp mdng te bdo la mgt phuang phdp cho nhieu uu the hon so vdi nhimg phuong phap nhdn gidng in vitro tmyen thdng khdc vd dugc ung dung thdnh cdng tren nhilu lodi cdy khdc nhau (Duong Tdn Nhyt vd cs, 2003). Tuy nhien, viec itng dyng phuong phap nudi cay Idp mdng te bdo d cay thdn gd chua dugc cdng bd nhieu.
Trong bdi ndy chung tdi trinh bdy mdt sd kit qud nghien cuu ve nudi cdy Idp mdng tl bdo tit ddt than mdm vd ddt tir di?p ciia cdy dilu.
VAT LIEU VA PHU'ONG PHAP
Mau nudi cdy Id hat tmdng thdnh cua gidng dilu cao sdn BOI thu dugc tir vudn dau ddng cua Trung tdm Hung Ldc (Vien Khoa hgc Ky thuat Ndng nghiep Mien Nam), Ddng Nai. Hat dilu cd 2 Idp vd, vd cinig ben ngodi vd vd lua ben frong. Hat dugc khir triing bdng dung dich NaOCl (Co sd Vdn Phuoiig, Qudn 11,
|Tp. Ho Chi Minh) vdi ndng dg 1% (w/v) trong 24 gid, sau dd rira lai bdng nude cdt
|v6 triing 3 lan.
Cay mam phdt triln tir hat nudi cdy tren mdi trudng khodng MS khdng cd dudng vd vitamin sau 2 tudn dugc dung ldm nguyen lieu cho thi nghiem. Ldp mdng cat
416 HQI nghi KHOA HOC VA CONG NGH£ 20m
ngang tir 2 loai vat lieu la ddt tu diep vd ddt than cua cdy mam (cd be day khodng 0,3-0,5nim) dugc cdy tren cdc dia petri (O = 10cm) chiia 20ml mdi trudng.
Mdi trudng nudi cdy la mdi trudng MS (Murashige va Skoog, 1962) cd bl sung nude dira 10% (v/v), adenine sulphate (Sigma Chemical Co.,USA) 40mg/L, saccharose (Cty Dudng Bien Hod, Dong Nai) 20g/L, maltose (Sigma Chemical Co., Missouri, USA) lOg/L, agar (Cty Cd phdn Do hop Ha Long, Quang Ninh) 9g/L, than boat tinh 3g/L. Mdi trudng dugc bd sung cdc chat dieu hod sinh trudng thyc vat la 6-benzyladenine (BA), kinetin (KN) vd naphthalene-1-acetic acid (NAA) d cac ndng do khdc nhau. pH cua mdi trudng trudc khi khu trimg la 5,9.
Mdi trudng dugc khir triing d 121 °C, 1 atm trong 20 phut. Phdng nudi cay cd nhiet do 25 ± 2°C, do dm 60 ± 5%. DTa petri dyng mdu dugc che tdi trong 3 ngdy ddu, sau dd dugc ddt dudi cudng do anh sdng 40-50 pmol m"^ s'' vdi thdi gian chieu sdng 12 gid/ngdy.
Mdi dia petri cd 24 mdu cay cua moi loai vat lieu (dot than mam hodc dot tu diep). Thi nghiem cd 3 yeu td Id 3 chdt dilu hod tdng trudng thyc vat, mdi ylu to cd hai muc do (Bang 1). Thi nghiem dugc bd tri hodn todn ngau nhien va moi nghiem thirc gdm 4 dia petri lap lai 3 Idn. Sd lieu dugc phan tich thdng ke bdng phdn mim MSTATC (Dai hgc Michigan, Michigan, USA). Thi nghiem dugc theo ddi trong 28 ngdy.
Bang 1: Md ta thi nghiem (chung cho 2 Ic^i vft ligu)
Nghifm thCpc 1 2 3 4 5 6 7 8
BA (mg/L) 5 5 5 5 10 10 10 10
KN (mg/L) 1 1 0 0 1 1 0 0
NAA (mg/L) 1 0,5
1 0.5
1 0,5
1 0,5
- • *f.
,/f
•s'fa
KET QUA VA THAO LUAN
Ket qud thi nghiem sau 28 ngdy nudi cdy cho thdy cdc Idt mdng d virng dll than chi cho mdt chdi cdn d viing ddt tir diep thi cho nhilu chdi hon. Dilu nay cho thay la do vimg sinh md chd hien dien chung quanh ddt tir diep de bi kich ho^t dudi tac ddng cua chdt dieu hod sinh trudng thyc vat (K. Trdn Thanh Van, 2003)- (Hinh 1).
Phin Vl: CdNG NGHg T £ BAO THUC VAT 417
dot tur dif p
Hinh 1. Ch6i hinh thdnh tip Idp mdng ddt than mim vd ddt tip di?p sau 28 ngdy.
Sd chdi cao nhdt (5, 7 chdi) tii ldp mdng d dot tir diep thudc nghiem thuc 6 ed bd sung lOmg/1 BA, lmg/1 KN vd 0,5mg/l NAA. Ldp mdng tao choi thdp nhdt (3 chdi) khi dugc nudi fren mdi timdng cd bd sung 5mg/l BA vd 0,5mg/l NAA.
Anh hudng cua BA vd KN len sy tao chdi cua ldp mdng vimg dot tir diep thdy rd nhat khi nong do BA lOmg/L vd KN Img/L, frong khi dnh hudng ciia NAA khdng thdy rd ldm trong thi nghiem nay. Ddi vdi ddt than mdm, ndng do cua cdc chdt dieu hoa sinh tmdng thyc vat dugc sir dung trong thi nghiem nay khdng dem lai sy khac biet ve sd choi hinh thanh d cd 8 nghiem thuc (Hinh 2).
S
6 5 4 3 2 1 0
I
i
4
3 4 8
Nghiem thufc
Dot than a O o t t u f d i e p
Hinh 2. Sd chdi tao thdnh tip lat mong ddt than mdm va ddt tip di?p d ngdy thu' 28.
418 HQi nghi KHOA HOC VA C O N G NGHE 2007
Cdc chdi hinh thdnh tii ldp mdng dugc tilp tiic cdy chuyin sang mdi tiirdng MS co bd sung lmg/1 BA vd lmg/1 KN dk phat triln thdnh cay vdi sd lin cdy chuyin la 2 tudn/ldn. Chilu cao cay trung binh dat 3-4cm sau 2 thdng cdy chuyen (Hinh 3).
Hinh 3. Cum chdi cay d i l u phat trien sau 2 thdng cdy chuyin.
KET LUAN
Phuong phdp nudi cay ldp mdng tl bdo cd thi dugc sir dung dk tao chdi tiirc tilp tilr tiyc thugng diep cua hat dilu ndy mdm. Sd chdi dugc tao tryc tilp tir ldp mdng ddt tir diep Id cao nhdt khi mdi tiirdiig cd bd sung lOmg/1 BA, lmg/1 KN vd 0,5mg/l NAA. Doi vdi nudi cdy ldp mdng ciia ddt tiian mdm thi chdi cd till phdt friln tren mdi frudng c6 BA 5mg/L va cd hoac khdng cd kinetin va NAA. Cdn tilp tuc nghien cim dk tim moi tmdng tiiich hgp cho sy phat tiiln chdi tii nudi cdy ldp mdng ddt tiian mdm.
TAI LIEU THAM KHAO I 1. Barghchi M, Alderson PG (1983). hi vifro production of Pistacia species. Acta
Hort. 131,49-60.
Duong Tdn Nhyt, Da Silva JAT, Biii Van Le, Kiem Trdn Thanh Van (2003). Thin cell layer (TGL) morphogenesis as a powerful tool in woody plant and fruit crop mircopropagation and biotechnology, floral genetics andgenetic fransformation. In:
Jain SM & Ishii K (eds.) Micropropagation of woody frees and fruits, pp. 783-814,
Kluwer Academic Publishers, Dordretch, The Netheriands. ' i Litz RE, Knight Jr RJ, Gazit S (1984). hi vifro somatic embryogenesis from
Mangifera indica L. callus. Sci. Hort. 22, 233-240. ^ "
2.
Phin VI: CdNG NGHg T £ I 419 Philip VJ & Urn
In: Bhaskara Ri pp.77-82, CPCR Tran Thanh Vai Bui, Tran Thai regeneration ai Publishers, Dorc
ition of cashew for crop improvement.
Cashew Research and Development, Dncept. hi: Duong Tan Nhut, Van Le .) Thin cell layer culture systerm:
ions. pp. 1-16, Kluwer Academic
li.
T h e 01
(Anacardiun
«^.,
t.r-
Rw
cell layers cashew
e cultivar BOI cultured
, Nguyen Thi Quynh iiology
Cashew (Anacar countiies due to the <
thin cell layer (TCL;
approach to produce BOI were surface s Epicotyls of seedling:
medium (Murashige different concenfratic explants affected the i
TCLS derived from t nodal positions of ef mg/L, respectively, w
)fitable cash crop of several fropical idy on the in vifro organogenesis via . was carried out for an appropriate e scale. Mature seeds of the cultivar '/v) on the MS sugar-free medium.
I;L culture and put on a modified MS lented with BA, KN and NAA at
maltose. The TCL positions on the nber of shoots. Shoot formation from ion was greater than that from other A, KN and NAA of 10, 1, and 0.5 )t formation of the cultivar BOI.