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 044bp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。
Primers were designed according to the cDNA sequence of tea alcohol dehydrogenase gene ((Tsi- ADH1). RT-PCR method was used to clone CsiADH1 from 'Longjing 43'. The expression levels of Csi- ADH1 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 con- sistent promoter CaMV35S. The recombinant pCAMBIA-ADH was then inserted into tomato cuhivar ' Zhongshu 4 ' using Agrobacterium tumefaciezls-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.