普通油茶体胚再生体系研究

Study on somatic embryogenesis regeneration system in Camellia oleifera

为构建农杆菌介导的油茶遗传转化受体系统,选择普通油茶‘长林’4号、53号和166号的幼胚为外植体材料,进行多因素多水平的组织培养试验。结果表明:培养基的p H对愈伤组织诱导率影响不显著,愈伤组织的诱导率受油茶品种影响极显著;‘长林’53号的幼胚极易实现胚性愈伤组织诱导和完整植株再生,MS培养基是最佳培养基,2,4-D和TDZ为幼胚较敏感的植物激素,对愈伤组织诱导影响最大;当MS+2 mg/L 2,4-D+0.5mg/L TDZ处理时,‘长林’53号体胚诱导率为98%;MS+1 mg/L 2,4-D+2 mg/L KT+500 mg/L CH处理时,‘长林’53号体胚诱导率为89.6%。在不同2,4-D、KT、TDZ和添加物水平的L9(34)正交试验中,除3个处理外,其他处理组合的‘长林’53号体胚诱导率都达到100%。已分化的胚性愈伤组织在不含任何激素的MS培养基上,可以实现胚状体的萌发、生长及完整植株再生。

In order to establish genetic transformation receptor system for oil camellia plant via agrobacterium-mediated,‘Changlin＇4,53 and 166 immature zygotic embryos were used as explants to carry out multi-level and multi-factor tissue culture experiments,the results showed that： the effect of culture medium p H on the callus induction rate was not significant,but varieties had a significant effect on callus induction rate. Somatic embryo regeneration was observed from‘Changlin＇53. MS and 2,4- D and TDZ were suitable for the embryo regeneration,the somatic embryogenesis induction rate was 98%,using MS ＋ 2 mg / L 2,4- D ＋ 0. 5 mg / L TDZ,the somatic embryo induction rate was 89. 6% based on MS ＋ 1 mg / L 2,4- D ＋ 2 mg / L KT ＋ 500 mg / L CH,and in addition 2,4-D,KT,TDZ and additives experiments,the somatic embryogenesis induction rate of‘Changlin＇53 was 100%. In hormone-free MS medium,the differentiation embryos could regenerate to plant.