摘要
橡胶树体胚遗传转化体系已成功建立．但抗性胚状体的循环增殖可获得大量抗性胚状体的同时，其后期的继代工作繁重。本文对橡胶树基因组DNA的CTAB提取方法进行了优化，同时筛选出了适合PCR检测的胚块重量。文中建立的胚块DNA的CTAB提取方法及筛选出的1～3mg重的胚块提取的DNA量均可满足早代胚状体PCR分析鉴定，且PCR鉴定结果与胚块的GUS染色结果吻合。该方法可用于橡胶树体胚遗传转化抗性胚状体早代筛选．
Agrobacterium tumefaciens-mediated Hevea somatic embryogenesis transformation system was successfully established. But a large propagation efficiently propagated from the secondary embryogenesis would lead to heavy burden in the following subculture. Thus the analysis of early positive embryo clones is very important. In this paper, the method of DNA extraction was optimized according to the conventional method of CTAB, meanwhile, the suitable weight of embryo used for PCR analysis was chosen. Experiments indicated that the optimized DNA extraction method and the scale of 1-3 mg all could accord with the PCR analysis of early embryos. Moreover, the results of PCR analysis were coincided with GUS histochemical staining. Hence, the method in this paper is available to early embryos screen in somatic embryogenesis transformation.
出处
《热带作物学报》
CSCD
北大核心
2013年第5期866-869,共4页
Chinese Journal of Tropical Crops
基金
国家自然科学基金(No.30960322)
国家自然科学基金(No.31200503)
现代农业产业技术体系建设专项(No.NYCYTX-34)
关键词
橡胶树
遗传转化
次生体胚
PCR
GUS
Rubber tree
Genetic transformation
Secondary embryo
PCR
GUS
