根癌农杆菌介导的g10h基因遗传转化长春花初步研究  

Agrobacterium tumefaciens- Mediated Genetic Transformation of Catharanthus roseus with g10h Gene

作　　者：袁芳[1] 王荃[1] 潘琪芳[1] 田跃胜[1] 赵静雅[1] 唐克轩[1] 

机构地区：[1]上海交通大学农业与生物学院植物生物技术研究中心,上海200240

出　　处：《上海交通大学学报（农业科学版）》2009年第6期606-610,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：863计划(2007AA10Z189);上海市农委项目(沪农科攻字(2005)第1-1-5号);上海市重点学科建设项目(B209)

摘　　要：以长春花下胚轴为基因转化的受体材料,利用根癌农杆菌EHA105介导,将g10h基因转化到3种不同品系的长春花中。结果表明,附加100μmol·L^(-1)乙酰丁香酮的根癌农杆菌悬液在OD_(600)=0.5～0.6时侵染效果较好;PCR分子检测转基因植株部分呈现阳性,证明g10h基因已整合到长春花基因组中,3种不同品系的长春花的再生率和阳性率存在差异;采用Real timePCR技术进一步检测g10h基因表达情况,结果表明转基因长春花植株中g10h mRNA表达量比非转基因对照高1.62～3.79倍。By using the hypocotyls of C roseus as the gene transformation acceptor. The gl0h gene was transferred into three varieties of C. roseus mediated by Agrobacterium tumefaciens EHA105. The results showed that the best infected effect was determined when the suspension liquid medium of Agrobacterium tumefaciens contained 100 μmol·L^-1 acetosyringone, and OD600=0.5-0.6. The PCR screening showed that some transgenic plants were positive, which indicated the gl0h gene was seemed to integrate into genome of C. roseus. And the rate of regeneration and transformation of C. roseus were different in three genotypes. Following analysis of Real time RT-PCR results showed that the expression level of glOh mRNA in the transgenic C. roseus plants was 1.62-3.79 fold higher than that of control （non-transgenie C. roseus plants）.

关 键 词：长春花 根癌农杆菌EHA105 g10h基因 遗传转化 

分 类 号：Q786[生物学—分子生物学] S336[农业科学—作物遗传育种]