葡萄细胞分裂素响应调节因子VvRR2启动子的克隆与活性分析

Cloning and Activity Analysis of the Promoter of Grapevine Cytokinin Response Regulator VvRR2

细胞分裂素在植物果实生长发育进程中具有重要作用。前期我们对葡萄细胞分裂素响应调节因子基因VvRR2进行了研究,结果发现VvRR2转录本受到多种因素的诱导调节。本研究在此基础上采用同源克隆方法在葡萄中克隆了细胞分裂素响应调节因子基因VvRR2启动子,生物信息学预测分析发现VvRR2启动子序列富含CAAT-box和TATA-box基本元件,还有与光响应、激素应答、组织特异和逆境相关的顺式作用元件。VvRR2启动子连接至pC0390GUS载体,转化烟草叶片,GUS酶活性分析显示VvRR2启动子具有活性。用细胞分裂素和脱落酸处理转化烟草叶片后VvRR2启动子活性没有改变;用生长素、赤霉素、茉莉酸甲酯和水杨酸处理后VvRR2启动子活性显著提高。

The cytokinins play the important role in the plant fruit growth and development. In previous, we studied the response regulator gene VvRR2 of grapevine, and the results showed that VvRR2 transcription was regulated by many factors. On the basis of this study, we cloned the promoter of the cytokinin response regulator in grapevine using homolog cloning method. Bioinformatics prediction analysis of cis-elements found that the promoter sequence was rich in CAAT-box and TATA-box basic elements, and contain the elements that involved in light response, hormone response, tissue specific and stress. VvRR2 promoter connected to p C0390 GUS vector,and the recombinant vector transformed into tobacco leaf. GUS enzyme activity analysis showed VvRR2 promoter had activity. Transformed tobacco leaves treated with cytokinin and abscisic acid, the VvRR2 promoter activity was not altered. When the transformed tobacco leaves treated with auxin, gibberellin, methyl jasmonate and salicylic acid, VvRR2 promoter activity was significantly increased.