中国红豆杉中WRKY转录因子基因克隆及其对紫杉烷合成的调控  

Molecular Cloning of Transcription Factors WRKY in Taxus chinensis and Its Regulation on Taxanes biosynthesis

作　　者：肖江蓉 刘建强 黄顺 方银娣 彭洪翠 李慎瑰 张小菊 Xiao Jiangrong;Liu Jianqiang;Huang Shun;Fang Yindi;Peng Hongcui;Li Shengui;Zhang Xiaoju(College of Urban Construction,Wuchang Shouyi University,Wuhan,430070;Xiangyang Product Quality Supervision and Inspection Institute,Xi-angyang,441000;Jingshan County Public Inspection and Testing Center,Jingmen,448000)

机构地区：[1]武昌首义学院城市建设学院,武汉430070 [2]襄阳市产品质量监督检验所,襄阳4000 [3]京山县公共检验检测中心,荆门448000

出　　处：《分子植物育种》2020年第13期4324-4330,共7页Molecular Plant Breeding

基　　金：技术服务战略合作协议项目(wcsy2018-01);国检中心技术开发项目(wcsy2018-02)共同资助。

摘　　要：WRKY是一类专一结合W-box序列的转录因子,它对植物抗病、损伤、衰老等生物学过程起到调控作用,尤其是在植物次级代谢物质合成中起到了重要调控作用。本研究采用中国红豆杉细胞转录组数据库序列设计引物,从中国红豆杉基因组中克隆转录因子基因TcWRKY26,该基因大小为2 202 bp;构建pBI121超表达载体,转化农杆菌LBA4404,并通过农杆菌介导转化红豆杉细胞,继而基于RT-qPCR技术对表达体系进行基因表达量的检测分析,发现转录因子基因明显超表达,相对表达量提高了2.92~3.49倍,紫杉烷合成相关酶dbat基因也有明显的超表达,相对表达量提高了1.78~2.03倍,说明TcWRKY26基因对dbat基因有明显的上调作用,继而为构建高产、稳定的红豆杉基因改良细胞株提供参考依据。WRKY is a class of transcriptional factors which specifically bind to W-box sequence.They involve in the regulation on disease-resistant,defence,caducity and some other biological process.They are important to secondary metabolism of plants.This article designed the primers and then cloned TcWRKY26 gene base on the transcriptome date base of Taxus Chinensis.The size of TcWRKY26 gene is 2202 bp.This work constructed the hyper expression vector,pBI121 and transformed the bacillus LBA4404.Then it was transferred into Taxus cell via Agrobacterium-mediated method.The result reflects that the expression of TcWRKY26 gene were 2.92~3.49 times higher than that of the negative control,the expression of dbat gene were 1.78~2.03 times higher than that of the negative control.It shows that overexpression of TcWRKY26 gene may enhance dbat gene expression in T.chinensis suspension cells.So this work can provide reference for the high-producing and stable gene motificated taxus chinensis clone.

关 键 词：红豆杉细胞 紫杉醇 转录因子 TcWRKY26 RT-QPCR 

分 类 号：S791.49[农业科学—林木遗传育种]