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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 hypocotyls, leaves and cotyledons of five varieties were used as explants to study the effects of different media, different hormone types and ratios, and proline, glutathione and sucrose on the embryogenic callus and embryoid body of alfalfa Induction, seedling emergence and transplanting. The results showed that the embryogenic callus induction rate and embryoid induction rate were the highest in Zhongmusi No.1 explants. The best embryogenic callus induction medium was modified SH + 2,4-D4.0mg · L-1 + BA0.5mg · L-1; the best embryoid body induction and seedling culture medium were MSO; proline had no effect on embryoid body induction; glutathione Peptide 15mg · L-1, 2% sucrose when induction rate. The embryos of five genotypes were induced from 58% to 77%, average number of embryoids from 25 to 60, embryo regeneration rate from 16% to 72% and rooting rate of 93% The rate is 94%. In addition, alfalfa also established a high-frequency regeneration tissue culture system for the future gene transformation to improve the characteristics of alfalfa laid the foundation.