苜蓿外植体再生系统的建立研究

Establishment of Regeneration System for Explants of Alfalfa

以5个品种的下胚轴、叶片和子叶为外植体研究了不同培养基、不同激素种类和配比以及脯氨酸、谷胱甘肽和蔗糖对苜蓿胚性愈伤组织和胚状体诱导、成苗和移栽的影响,结果表明:中苜1号叶片外植体胚性愈伤组织诱导率和胚状体诱导率最高;最佳胚性愈伤组织诱导培养基为改良SH+2,4-D4.0mg·L-1+BA0.5mg·L-1;最佳胚状体诱导和成苗培养基均为MSO;脯氨酸对胚状体诱导无明显作用;谷胱甘肽为15mg·L-1、蔗糖为2%时诱导率最大。筛选出的5个基因型的胚状体诱导率为58%～77%,平均胚状体数目25～60个,胚状体再生率16%～72%,生根率为93%,移栽成活率为94%。另外,还建立了苜蓿高频再生组织培养体系,为今后的基因转化改良苜蓿性状打下了基础。

The hypocotyl,leaf and cotyledon of five varieties of alfalfa (M.sativa)were used to study the effect of different basic mediums,hormones in different concentrations,proline,GSH and sucrose concentrations on embryogenic callus and embryoid induction,plant development and transplant of alfalfa(M.sativa).The results showed that the leaf embryogenic callus and embryoid induction rate of Zhongmu 1 variety was the highest;the best medium for embryogenic callus induction was the modified SH+2,4-D 4.0 mg·L-1+BA 0.5 mg·L-1,for embryoid induction and plant development was MSO;proline have no effect on embryoid; the highest embryoid induction rate for GSH was 15 mg·L-1and for sucrose was 2%.The embryoid induction rates for the five selected genotypes were 58%～77%,the average embryoid number were 20～60,the embryogenic regeneration rates were 14%～68%,the highest rooting rate was 93% and the transplant survived rate was 94%.By the research the high frequency regeneration system for tissue culture was established and it is benefit to gene transformation of alfalfa.