发根农杆菌介导的垂序商陆遗传转化体系的建立及毛状根株系生物量分析  

Establishment of Phytolacca Americana L. Genetic Transformation System by Agrobacterium Rhizogenes and Analysis of Hairy Root Strains Biomass

作　　者：姚庆收[1] 单长民[1] 武玉永[1] 于敏[1] 逄高燕 

机构地区：[1]滨州医学院药学院,山东烟台264003

出　　处：《时珍国医国药》2014年第8期1991-1994,共4页Lishizhen Medicine and Materia Medica Research

基　　金：山东省自然科学基金资助项目(No.ZR2010HQ054);农业部橡胶树生物学重点开放实验室/省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室开放课题基金项目资助(No.KLOF1106);山东省高等学校青年骨干教师国内访问学者项目资助

摘　　要：目的建立发根农杆菌介导的垂序商陆遗传转化体系及毛状根株系生物量分析,为利用基因工程技术开发商陆奠定研究基础。方法活化发根农杆菌R1601,利用其不同浓度的菌液对垂序商陆的叶片和茎段进行转化,利用PCR技术鉴定毛状根;随机选取毛状根培养,鉴别不同株系间的生物量差异。结果垂序商陆对发根农杆菌R1601侵染敏感,4个浓度都能高效诱导垂序商陆形成毛状根,其中A600为1.0时,茎段和叶片的遗传转化效率都能达到100%;PCR鉴定表明毛状根中已经整合进了T-DNA序列;不同毛状根株系生物量分析表明毛状根株系生物量呈极显著差异。结论建立了发根农杆菌R1601介导的垂序商陆的高效遗传转化体系,不同毛状根株系间生物量存在极显著差异。Objective To establish the genetic transformation system of Phytolacca americana L. by Agrobacterium rhizogenes mediated and analysis of hairy root strains biomass for the base of Phytolacca americana L. research using genetic engineering technology. Methods Activated Agrobacterium rhizogenes RI601, transforming leaves and stems of Phytolacca americana L. by u- sing different concentrations of Agrobacterium rhizogenes R1601 ; Hairy roots identified by PCR; randomly selected hairy roots to culture,identified the biomass differences of different strains. Results Phytolacca americana L. is sensitive by Agrobacterium rhi- zogenes R1601 infected; Four concentrations can induce hairy roots of Phytolacca americana L. highly efficient, the genetic transformation efficiency of stems and leaves can reach 100 % if A600 is 1.0 ; PCR identification showed the T - DNA sequences had integrated into the hairy roots of Phytolacca americana L. ; The biomass analysis showed the biomass of different hairy root strains was significantly different. Conclusion The genetic transformation system highly efficient of Phytolacca americana L. by Agrobacterium rhizogenes has been established,the biomass of different hairy root strains was significantly different.

关 键 词：发根农杆菌 RI质粒 垂序商陆 毛状根 遗传转化 

分 类 号：R282.71[医药卫生—中药学]