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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。
Cytokinin plays an important role in the process of plant fruit growth and development. We studied the cytokinin responsive regulatory factor gene VvRR2 in early prophase and found that VvRR2 transcript was regulated by many factors. Based on this, we cloned the cytokinin response regulator gene VvRR2 promoter by homologous cloning method. Bioinformatics analysis showed that the VvRR2 promoter sequence is rich in CAAT-box and TATA-box elements. There are cis-acting elements that are involved in photoresponsiveness, hormonal responses, tissue-specific and adversity. The VvRR2 promoter was ligated into the pC0390GUS vector and tobacco leaves were transformed. Analysis of GUS enzyme activity revealed that the VvRR2 promoter is active. VvRR2 promoter activity did not change after transformation of tobacco leaves with cytokinin and abscisic acid; VvRR2 promoter activity was significantly increased after treatment with auxin, gibberellin, methyl jasmonate and salicylic acid.