摘要
目的:在建立灰毡毛忍冬再生体系的基础上,建立其遗传转化体系。方法:组织培养方法培养幼苗,根癌农杆菌介导方法转化外植体,gus染色和PCR检测报告基因。结果:侵染时间8 min能获得较理想的转化效果。35 mg/L的卡那霉素和600 mg/L头孢霉素能得到很好的筛选效果,起主导作用的因素是卡那霉素浓度,其不同的浓度间抗性芽诱导率达到了极显著差异。对转基因植株进行gus染色和PCR检测,结果表明gus基因已整合到灰毡毛忍冬基因组中。结论:首次建立了根癌农杆菌(Agrobacterium tumefaciens,EHA105菌株)介导的叶片为受体的遗传转化体系。
Objective： Based on the system of regeneration,the genetic transformation system of Lonicera macranthoides was established. Methods： Tissue culture method of seedlings,Agrobacterium tumefaciens mediated transformation method of explants,report gene was detected by gus staining and PCR. Results： The efficient transformation time was 8 minutes of infection. The good transformation rate was gained with the kanamycin 35 mg / L and cefotaxime 600 mg / L. The concentration of kanamycin had a leading effect on bud differentiation between two antibiotics,and bud induction rate reached extremely significant difference. Results of gus staining and PCR proved that the gus gene was integrated into Lonicera macranthoides genome. Conclusion： The genetic transformation system of Lonicera macranthoides leaves mediated by Agrobacterium tumefaciens EHA105 was established for the first time.
出处
《中药材》
CAS
CSCD
北大核心
2013年第12期1904-1907,共4页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(31200512)
重庆市自然科学基金(cstc2012jjA80018)
重庆文理学院特色林木种质资源创新重点实验室平台建设项目
重庆文理学院校级科研项目(Z2012LX07)
关键词
灰毡毛忍冬
根癌农杆菌
遗传转化
PCR
Lonicera macranthoides Hand.-Mazz.
Agrobacterium tumefaciens
Genetic transformation
PCR
