摘要
在本实验室建立的狗蔷薇(Rosa canina)类原球茎(PLB)再生途径基础上,以类原球茎为转化受体,采用根癌农杆菌(Agrobacterium tumefaciens,GV3101菌株)介导法,通过评价类原球茎GUS基因瞬时表达率,采用正交试验设计优化狗蔷薇遗传转化条件,建立遗传转化体系。最佳转化条件为:用OD600值为0.6的农杆菌菌液侵染狗蔷薇类原球茎30min,然后在含有100μmol/L乙酰丁香酮的MS培养基上共培养2d,利用此优化的转化系统,经组织化学染色及PCR检测获得了转GUS基因阳性植株。
Genetic transformation system of Rosa canina was established based on the PLBs （protocorm-like bodies） regeneration protocol developed by our lab. By evaluating transient GUS expression, several factors were studied which may affect genetic transformation mediated by Agrobacterium tumefaciens （GV3101 strain）. Genetic transformation was optimized by inoculation of PLBs. At a density of OD600 = 0.6 for 30min, PLBs were co-cultivated with Agrobacterium tumefaciens on the medium containing l00μmol/L acetosyringone for 2d. Transgenic plants of Rosa canina were identified by GUS histochemical staining and PCR testing.
出处
《核农学报》
CAS
CSCD
北大核心
2012年第2期270-274,共5页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金(31171993)
农业部行业科研专项(200903020)
关键词
狗蔷薇
类原球茎
根癌农杆菌
遗传转化
Rosa canina
protocorm-like bodies
Agrobacterium tumefaciens
genetic transformation
