草业学报2016，Vol.25Issue(11)：50-56,7.DOI:10.11686/cyxb2015590
乌拉尔甘草离体再生与遗传转化技术的研究
In vitro research on receptor regeneration and genetic transformation in Glycyrrhiza uralensis
摘要
Abstract
A study has been undertaken to investigate the system of receptor regeneration and genetic transfor-mation of Glycyrrhiza uralensis in vitro ,in order to provide a preliminary experimental basis for breeding G. uralensis via genetic engineering.Using stem segments as explants,tests were done to select the most appro-priate medium formula for adventitious bud differentiation,propagation and elongation.Furthermore,GUS and GFP genes were transferred into G.uralensis through Agrobacterium tumefaciens-mediated transformation methods.The results showed that the optimum formula for bud differentiation and proliferation was a Murash-ing and Skoog (MS)medium supplemented with 0.1 mg/L thidiaxuron,0.1 mg/L 6-benzyladenine and 0.1 mg/L indole-3butyric acid.Using this formula,the differentiation rate of adventitious buds was 38.8% and the rate of multiplication 3.67.The most suitable formula for bud elongation was a MS medium containing 0.5 mg/L α-naphthalene acetic acid.Compared to stems without axillary buds,the differentiation rate of adventi-tious buds reached 100%.Moreover,these buds can survive in antibiotic containers and so become the optimal transformation explants.Stems with axillary buds were pre-cultured for 7 days in the differentiation medium, then infected by an Agrobacterium tumefaciens solution (OD600 =0.6)for 10 min and co-cultured for 3 days, following which the explants were transferred to a selective medium based on the differentiation medium and supplemented with 50 mg/L kanamycin and 250 mg/L carbenicillin.After 20 days,the GFP transient expres-sion rate reached 78.88% and GFP expressed strongly by GFP detection.5 transformed plants were selected randomly for testing and the presence of GUS and GFP genes in the genome were confirmed by polymerase chain reaction,indicating that 5 transformed plantlets had been successfully produced.关键词
乌拉尔甘草/茎段/不定芽分化/遗传转化/绿色荧光蛋白Key words
Glycyrrhiza uralensis/stem segments/adventitious bud differentiation/genetic transformation/green fluorescent protein (GFP )引用本文复制引用
胡海英,张芮,段艳慧,谢应忠..乌拉尔甘草离体再生与遗传转化技术的研究[J].草业学报,2016,25(11):50-56,7.基金项目
宁夏自然科学研究基金项目（NZ13034）资助。 （）
