云杉未成熟合子胚诱导体细胞胚胎发生

摘要

Immature zygotic embryos of Picea asperata were used as explants to investigate effects of cone collection dates,genotypes,different concentration of auxin and cytokinin on the induction of embryogenic callus. At the same time, the key factors affecting the differentiation and maturation of somatic embryos were also comprehensively investigated. The results showed that the optimal medium for induction and proliferation of embryogenic callus was 1/2 LM + 2. 2 mg·L -1 2,4-D + 1. 1 mg·L -16-BA + 10 g·L -1 sucrose + 2 g·L -1 gellan gum + 1g·L -1 casein hydrolysate. The best medium for differentiation of somatic embryos was 1/2 LM + 24 mg·L -1 ABA + 50 g·L -1 PEG4000 + 30 g·L -1 sucrose + 1 g· L -1 activated charcoal+ 4 g·L -1 gellan gum + 1 g·L -1 casein hydrolysate. The suitable medium of germination was 1/4 LM + 20 g·L -1 sucrose + 1 g·L -1 AC + 4 g·L -1 gellan gum + 1 g·L -1 casein hydrolysate. All medium was supplemented with 0. 5 g·L -1 L-glutamine,and adjusted to pH 5. 8.%以云杉未成熟合子胚为外植体，研究球果采集日期、基因型、植物生长素和细胞分裂素对愈伤组织诱导的影响，同时对影响体细胞胚分化和成熟的关键因素进行探讨。结果表明：合适的胚性愈伤组织诱导和增殖培养基为1/2 LM +2，4-D 2.2 mg·L -1+6-BA 1.1 mg·L -1+蔗糖10 g·L -1+凝胶2 g·L -1+酶解酪蛋白1 g·L -1；合适的分化培养基为1/2 LM + ABA 24 mg·L -1+ PEG400050 g·L -1+蔗糖30 g·L -1+活性炭1 g·L -1+凝胶4g·L -1+水解酪蛋白1g·L -1；合适的萌发培养基为1/4LM +蔗糖20g·L -1+活性炭1g·L -1+凝胶4g·L -1+水解酪蛋白1 g·L -1。以上培养基中均附加谷氨酰胺0.5 g·L -1，pH5.8。