以刺槐不同胚龄的未成熟合子胚为外植体,采用混合正交试验设计,研究幼胚胚龄和不同外源激素种类及质量浓度对刺槐胚性愈伤组织的诱导和体细胞胚分化、萌发的影响。结果表明:开花后8周(55天左右)是胚性愈伤组织和体胚诱导的最佳外植体取材时期;MS+2,4-D5.0mg·L-1+BA0.5mg·L-1是诱导胚性愈伤组织的最佳培养基,出愈率最高为95.42%±0.02%;MS+NAA0.5mg·L-1+BA0.5mg·L-1+谷氨酰胺250mg·L-1+水解酪蛋白500mg·L-1是体细胞胚诱导和分化的最佳培养基,直接体细胞胚发生率最高为92.40%±0.12%,通过愈伤组织诱导体细胞胚发生的频率最高为90.73%±0.49%。一旦形成球形胚,将培养物转接到不含任何生长调节剂的MS培养基上,体细胞胚经成熟萌发可进一步形成完整小植株。
Immature embryos of Black Locust(Robinia pseudoacacia) were used as explants to study effects of immature zygotic embryo age, as well as the kind and concentration of plant growth regulators in culture media on callus and somatic embryo induction with an orthogonal experiment. The results showed that seeds collected 8 weeks(about 55 days) post-anthesis had the highest ability to initiate embryogenic cultures. The best callus initiation medium was MS+2,4-D 5.0 mg·L-1+ BA 0.5 mg·L-1, with which the highest callus induction rate, 95.42%±0.02%, was obtained. The best somatic embryogenesis medium was MS+NAA 0.5 mg·L-1+BA 0.5mg·L-1+Glutamine 250 mg·L-1+Casein acid hydrolysate 500 mg·L-1, and produced the highest direct and indirect somatic embryogenic frequency of 92.20%±0.12% and 90.73%±0.49%, respectively. Once globular-stage embryo formed, somatic embryos then matured and germinated to develop plantlets by transferring to the same medium without plant growth regulators.