紫娟茶叶片中花青素合成酶基因的克隆及表达分析

Cloning and Expression Analysis of Anthocyanin Synthase(ANS)Gene in Leaves of Zijuan Tea

本研究通过RT-PCR从紫娟茶(Camellia sinensis var.assamica)叶片中克隆花青素合成酶(anthocyanin synthase,ANS)基因,进行生物信息学分析,以实时荧光定量PCR研究ANS基因在紫娟茶顶芽及芽下第一、第二、第三叶中的表达情况。克隆得到了ANS基因的两个cDNA,即ANSa、ANSb,获得GenBank登录号分别为MN266484、MN266485,二者的功能结构域均属于2-酮戊二酸和Fe(Ⅱ)-依赖的氧化酶家族。通过荧光定量PCR发现两个ANS基因在不同幼嫩程度的叶片中表达量存在差异,ANSa在顶芽中表达量最高,ANSb则在芽下第一叶中表达量最高。本研究明确了ANS在紫娟茶同一时期不同幼嫩程度叶片中的表达情况,为进一步研究紫娟茶花青素的合成与积累奠定了基础。

In this study,the ANS gene was cloned from the leaves of Zijuan tea(Camellia sinensis var.assamica)by RT-qPCR and analyzed by bioinformatics.The expressions of two ANS genes in the top buds,the first,the second and the third leaves under the buds of Zijuan tea were studied by real-time fluorescence quantitative PCR.The two cDNAs of ANS genes,named ANSa and ANSb,respectively,were cloned and submitted to GenBank,with the numbers of MN266484 and MN266485.Their functional domains belong to 2-ketoglutarate and Fe(Ⅱ)-dependent oxidase family.The qPCR showed that the expression levels of the two ANS genes were different in different leaves of Zijuan tea.The highest expression of ANSa was in the top buds and ANSb was in the first leaves under the buds.This study clarified the expression of ANS in different leaves of Zijuan tea in the same period,which laid a foundation for further study on the synthesis and accumulation of anthocyanins in Zijuan tea.