乌拉尔甘草转鲨烯合成酶基因毛状根系研究

Culture of transgenic Glycyrrhiza uralensis hairy root with licorice squalene synthase(SQS) gene

目的:为了研究甘草酸的合成调控途径及提高甘草酸合成水平。方法:将已克隆的乌拉尔甘草鲨烯合成酶基因(GuSQS1,GenBank登录号为:AM182329)通过发根农杆菌Agrobacterium rhizogenes A4的介导,转化甘草下胚轴并诱导毛状根的形成。建立的毛状根系经0.8mg.L-1的除草剂(PPT)抗性筛选、PCR和Southern blotting检测分析后,得到的转基因毛状根系经HPLC检测甘草酸(GA)含量。结果与结论:转基因毛状根中甘草酸的最高含量约为野生型毛状根的3.6倍。

A squalene synthase gene cloned （GuSQS1, accession number in GenBank database： AM182329） from Glycyrrhiza uralensis was transferred into G. uralensis via Agrobacterium rhizogenes A4 for investigating biosynthesis pathway and enhancing synthesis of glycyrrhizic acid （GA）. Hypocotyl explants from G. uralensis were infected with A. rhizogenes A4 containing GuSQS1 gene to induce the hairy roots. The hairy root lines established were selected in medium containing 0. 8 mg·L^-1 phosphinothricin （PPT） and analyzed by PCR and southern blotting. The transgenic hairy roots were cultured in liquid MS medium. GA contents in transgenic hairy roots were detected by HPLC. Results showed that maximal GA content in transgenic hairy root lines was 3.6 times as high as in wild type hairy roots.