根癌农杆菌介导sHSP基因对半夏的遗传转化  

Genetic transformation of Pinellia ternata with Agrobacterium tumefaciens-mediated sHSP genes

作　　者：郭朝阳[1] 崔婷婷[1] 薛建平[1] 朱艳芳[1] 张爱民[1] 盛玮[1] 滕井通[1] 

机构地区：[1]淮北师范大学生命科学学院资源植物生物学安徽省重点实验室,安徽淮北235000

出　　处：《中国中药杂志》2012年第24期3758-3762,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30973963);安徽省自然科学基金项目(090413252);安徽高校省级自然科学研究重点项目(KJ2009A160)

摘　　要：目的:建立高效的半夏遗传转化体系。方法:以半夏试管苗的叶柄为外植体,采用根癌农杆菌介导法,探索了酚类物质、农杆菌菌液浓度、侵染时间、预培养时间及共培养时间等对半夏遗传转化效率的影响。结果与结论:不经过预培养阶段,用含AS 40 mg.L-1的根癌农杆菌菌液侵染15 min后共培养3 d时可有效提高遗传转化效率。将转化后的叶柄接到含有100 mg.L-1Kan和350 mg.L-1Carb的分化培养基上进行筛选培养,30 d左右在叶柄的两端分化出抗性的试管小块茎。对抗性转化植株进行Gus染色和PCR检测,结果表明外源基因sHSP已经整合到半夏基因组中。Objective： To establish an efficient genetic transformation system of Pinellia ternata. Method ： With petioles from test-tube seedlings of P. ternata as explants, Agrobacterium tunwfaciens mediation method was adopted to explore the effect of phenolic substances, A. tumefaciens＇s concentration, infection time, pre-incubation time and co-cultivation time on genetic transtbrmation efficiency of P. ternata. Result and Conclusion： The genetic transformation efficiency could be effectively enhanced by infecting in A. tumefi^cienseulture containing AS 40 mg·L^-1 for 15 rain for three days. The petioles were put into the differentiation medium containing 150 mg ·L^-1 Kan and 350 mg·L^-1 Carb to screening and cultivation. After around 30 days, microtubers could be observed at both sides ot the petioles. Gus staining and PCR verification on the regenerated plants showed that the exogenous gene sHSP had been integrated into genome of P. ternata.

关 键 词：半夏 遗传转化 SHSP 农杆菌介导 

分 类 号：S567.239[农业科学—中草药栽培]