特异性过表达SQS1基因的甘草毛状根培养体系的建立  

Culture System Establishment of Glycyrrhiza uralensis Hairy Roots Over-Expressing Squalene Synthase 1 Gene Root-Specifically

作　　者：周姗[1] 袁伯川 马永生[1] 杨瑞[1] 王礼强[1] 刘颖[1] 

机构地区：[1]北京中医药大学中药学院,北京100102

出　　处：《生物技术通讯》2016年第5期638-642,共5页Letters in Biotechnology

基　　金：国家自然科学基金(81508131);北京中医药大学杰出青年人才项目(2016JYBXJQ002)

摘　　要：目的:建立根特异性过表达鲨烯合酶(SQS)基因的甘草毛状根培养体系。方法:构建根特异性过表达甘草SQS1基因的发根农杆菌ACCC10060工程菌;侵染甘草无菌苗胚轴,共培养48 h,以诱导毛状根的形成;多次除菌后,利用PCR法及测序法对诱导获得的甘草毛状根进行验证。结果:PCR结果显示扩增得到了长度约为730、580和1400 bp的基因片段,分别与烟草根特异性启动子TobRB7、发根农杆菌rolC基因和甘草SQS1基因长度一致;测序结果进一步确定了PCR扩增序列的正确性,从而证明了甘草根特异性过表达SQS1基因毛状根诱导成功。结论:获得了大量生长良好的特异性过表达SQS1基因的甘草毛状根,为研究功能基因SQS1与甘草酸次生代谢的相关性,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 establish the culture system of Glycyrrhiza uralensis hairy roots over-expressing squa-lene synthase 1（SQS1） gene root- specifically. Methods： The recombinant Agrobacterium rhizogenes ACCC10060 with a root-specific promoter TobRB7 and G.uralensis SQS1 gene was constructed, which was used to infect the hy-pocotyls of 7-day-old G.uralensis seedlings for 20 min, and co-cultured for 48 hours. The hairy roots were testedand verified by PCR and sequencing. Results： PCR results showed that a 730 bp, a 580 bp, and a 1400 bp frag-ments were obtained. And the further sequencing results demonstrated that they were tobacco root-specific promot-er TobRB7, A.rhizogenes rol C gene, and G.uralensis SQS1 gene, respectively, which declared that the G.uralensis hairy roots over- expressing SQS1 gene root- specifically were induced successfully. Conclusion： A lot of well-grown G.uralensis hairy roots over-expressing SQS1 root-specifically were obtained. This paper is significant for re-vealing the correlation between functional gene SQS1 and the biosynthesis of glycyrrhizic acid, and developing thecontent of glycyrrhizic acid in G.uralensis hairy roots.

关 键 词：甘草 根特异性过表达 鲨烯合酶1 发根农杆菌 毛状根 

分 类 号：Q943.2[生物学—植物学]