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作 者 ：辛肇军,孙晓玲,陈宗懋*

期 刊 ：西北植物学报 2013年 5期 页码：864~871

关键词：茶树；醇脱氢酶基因；克隆；表达；番茄转化；

Keywords:tea, alcohol dehydrogenase gene, clone, expression, tomato transformation,

摘 要 ：根据茶树醇脱氢酶基因(CsiADH1)的cDNA序列设计引物,采用RT-PCR方法从茶树品种‘龙井43’中克隆了CsiADH1序列,分析了CsiADH1在生物和非生物胁迫下的诱导表达情况并转化番茄。结果表明:CsiADH1包含一个1 044 bp的最大开放阅读框,编码347个氨基酸。qRT-PCR分析显示,CsiADH1的表达受到茶尺蠖取食、机械损伤、茉莉酸和水杨酸的诱导;将CsiADH1基因ORF区域克隆进pCAMBIA1301载体中,构建了由CaMV35S启动子驱动的CsiADH1基因植物表达载体 pCAMBIA-ADH,并以农杆菌介导的方法侵染番茄‘中蔬四号’子叶,经PCR鉴定,获得了8个转CsiADH1基因阳性植株。该结果为进一步揭示CsiADH1基因在植物诱导防御反应中的分子机理研究奠定了基础。

Abstract:Primers were designed according to the cDNA sequence of tea alcohol dehydrogenase gene (CsiADH1).RT-PCR method was used to clone CsiADH1 from ‘Longjing 43’.The expression levels of CsiADH1 under biotic and abiotic stress were analyzed by qRT-PCR.The results showed CsiADH1 contains an open reading frame of 1 044 bp which encodes a protein of 347 amino acids.Tea geometrid feeding,wounding,JA and SA treatment up-regulated the expression levels of CsiADH1.Using molecular cloning techniques,the ORF region sequence was cloned into the pCAMBIA1301 vector under the control of a consistent promoter CaMV35S.The recombinant pCAMBIA-ADH was then inserted into tomato cultivar‘Zhongshu 4’using Agrobacterium tumefaciens-mediated transformation.8 transgenic plants were obtained by PCR identification.The results will help to further understand the molecular mechanism of CsiADH1 in plant induced defense response.

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