甘草根特异性过表达HMGR基因毛状根的诱导

Induction and Culture of Glycyrrhiza uralensis Hairy Roots Overexpressing 3-Hydroxy-3-Methylglutaryl-Co A Reductase Gene

目的:构建甘草根特异性过表达3-羟基-3-甲基戊二酰Co A还原酶(HMGR)基因毛状根培养体系。方法:利用根特异性过表达甘草HMGR基因的植物双元表达载体,通过发根农杆菌ACCC10060介导,转化甘草外植体并诱导毛状根的形成,利用PCR法及测序法对转基因毛状根进行验证。结果:诱导得到大量生长良好的甘草根特异性过表达HMGR基因毛状根。结论:为进一步研究功能基因HMGR与甘草酸次生代谢的相关性,以及提高甘草毛状根中的甘草酸含量奠定了良好的实验基础。

Objective： To induce and culture Glycyrrhiza uralensis hairy roots overexpressing 3-hydroxy-3-methyl- glutaryl-CoA reductase gene（HMGR）. Methods： The root specific expression vector containing HMGR gene was transferred into disarmed Agrobacterium rhizogenes ACCC10060, which was then selected on YEB medium supple- mented with 50 mg/L kanamycin sulfate and 20 mg/L hygromycin and validated strictly by PCR and sequencing. Then hypocotyls of 7-day-old seedlings were collected and infected by the above recombinant A.rhizogenes ACCC10060. And hairy roots were tested and verified by PCR and sequencing. Results： A lot of well-grown G. uralensis hairy roots containing root specific over-expressing HMGR genes were obtained. Conclusion： This re- search is significant for further investigations concerned with the correlations between functional gene HMGR and glycyrrhizic acid. It will also lay a good experimental foundation of increasing the content of glycyrrhizic acid in G. uralensis hairy roots.