紫萼玉簪查尔酮合酶基因的克隆及功能验证

Cloning and Function Analysis of CHS Gene From Hosta ventricosa

查尔酮合酶是花色素生物合成过程中的关键酶。为研究玉簪查尔酮合酶基因的结构及功能,基于玉簪转录组数据库信息,利用RT-PCR的方法克隆得到紫萼玉簪查尔酮合酶基因,命名为HvCHS,构建pCAMBIA3301-HvCHS载体,农杆菌介导法转化烟草,验证HvCHS基因功能。结果表明:HvCHS基因的ORF全长1 176 bp,编码391个氨基酸,分子质量为42.798 kD,等电点为6.48,是亲水性的同源二聚体蛋白;预测亚细胞定位于细胞质中,与大花君子兰Clivia miniata(ARI70436.1)、水仙Narcissus tazetta var.chinensis(AFM36770.1)的同源性较高;转基因烟草的花瓣中花色苷含量明显高于野生型烟草,说明HvCHS基因的过表达能够促进花色苷积累。

Chalcone synthase is a key enzyme in anthocyanin biosynthesis. In order to study the structure and function of chalcone synthase gene, based on the information of Hosta transcriptome database, the chalcone synthase gene of Hosta ventricosa was cloned by RT-PCR and named HvCHS. The pCAMBIA3301-HvCHS plant vector was constructured to verify HvCHS gene function by tobacco transform. The results show that the open reading fragment size of the HvCHS gene was 1 176 bp, encoding 391 amino acids, its relative molecular weight was about 42.798 kD, and the isoelectric point was 6.48. HvCHS protein was a hydrophilic Homodimer protein. Subcellular localization was predicted to be in cytoplasm and HvCHS protein sequences had high homology with Clivia miniataer(ARI70436.1) and Narcissus tazetta var.chinensis(AFM36770.1). The content of anthocyanin in the petals of transgenic tobacco was significantly higher than that of wild type tobacco, indicating that HvCHS gene can promote the synthesis of anthocyanin.