桑属植物总RNA的提取及花青素合成基因MCHS的鉴定

Extraction of Total RNA and Identification of Anthocyanin Biosynthesis Gene MCHS in Morus Alba

桑属植物细胞中含大量蛋白酶及多糖等生物功能性分子。以新鲜的桑叶、桑果为试验材料,应用Trizol法提取总RNA。试验表明,使用Trizol法配合氯仿—苯酚试剂提取RNA操作简单,且能够有效去除桑叶中多糖和多酚类等次生物质,从而得到高质量的RNA,以获得的RNA为模板,设计简并引物,成功从桑叶中鉴定出花青素合成的关键酶基因MCHS基因。该试验为分子水平研究桑科类黄酮代谢机制、以及桑葚功能性食品的开发等提供了一定的理论依据。

Mulberry cells contain a large number of proteases and polysaccharides and other biological functional molecules,using fresh mulberry leaves,mulberry(fruit),the total RNA was isolated by Trizol combined with Chloroform-phenol.Purity and concentration of isolated RNA indicated that polysaccharide,phenol,and other biological molecules could be effectively eliminated in the mulberry plant cell by this method.Using the high-quality RNA as template,key genes of MCHS for anthocyanin synthesis was successfully identified by degenerate RT-PCR.The experimental results provide an experimental basis for the molecular level research on mechanism of flavonoids and the breeding of new varieties.