麻疯树胚乳愈伤组织诱导及其污染消除

经过比较分析各种消毒方法,获得了较为理想的麻疯树(Jatrophacurcas)胚乳愈伤组织,并动态观察其类型、形态特征、增殖速率和形态发生能力,为麻疯树胚乳植株再生及愈伤组织和细胞培养生产药用成分奠定了基础.以麻疯树成熟胚乳为外植体进行愈伤组织的诱导,测试0.4mg/L的分裂素(BA、KT与TDZ)与2.0mg/L的生长素(IAA、IBA、NAA与2,4D)搭配组合的诱导效果.结果表明,对胚乳愈伤诱导效果而言,2.0mg/L的2,4D效果最好,NAA的效果次之,IBA又次之,IAA的效果最差.BA和KT诱导效果差异不显著,TDZ的添加可促进胚乳愈伤组织的诱导.选择适宜的消毒剂(种类、浓度)和消毒(处理)时间对胚乳外植体的接种成功具有至关重要的作用.在本试验所采取的灭菌方式中,用70%酒精处理30s,然后用10%NaClO处理25min,灭菌效果最佳,同时对胚乳外植体的生长影响最小.图3表1参23

Callus Induction from Jatropha curcas Endosperm and Elimination of Microbial Contamination in Culture

For further investigations on potential production of triploid plantlet and curcin production from in vitro endosperm cultures of physic nut (Jatropha curcas L.), the endosperm calli of J. curcas were obtained and the calli could rapidly proliferate and have morphogenetic potentiality by comparatively analyzing different disinfection methods and hormone combinations. The mature endosperms of J. curcas were taken as explants for callus induction, and 0.4 mg/L of cytokinin(BA, KT and TDZ) and 2.0 mg/L of auxin (IAA, IBA, NAA and 2,4-D) were used to test callus induction efficiencies. The result showed that the efficiencies of BA and KT were not quite different. TDZ could promote the induction of calli more effectively. Among all the hormone combinations, 2.0 mg/L of 2,4-D induced more calli with base effect and 2.0 mg/LNAA did the second. IAA could make endosperm form calli as well, but the color of the calli was slightly lighter than those induced by IBA, NAA and 2,4-D with lowest effect. Appropriate disinfectants and sterilization time were important to successfully inoculate the endosperm explants. In order to fix appropriate disinfectants and sterilization time, the explants were marinated in 70% ethanol for 30 s firstly, and then 0.1% HgCl_ 2 or 10% NaClO was used for disinfection, and finally, the appropriate disinfectant and the optimum sterilization time were found, that is, treating with 70% ethanol for 30 s, then disinfecting with 10% NaClO for 25 min. Fig 3, Tab 1, Ref 23