葡萄WRKY54基因克隆及表达特性分析

发布时间：2019-06-24 16:48

【摘要】：为了探究葡萄WRKY54基因的功能,以抗盐葡萄品种Vidal Blanc为材料,采用同源克隆法克隆得到Vv WRKY54基因,并对其进行生物信息学和表达特性分析。结果表明,Vv WRKY54基因c DNA序列为942 bp,编码313个氨基酸。生物信息学分析结果表明,Vv WRKY54蛋白分子量约为35.3091 k Da,等电点为5.45,不稳定系数为55.03,推测其为不稳定蛋白,与已知毛果杨及拟南芥WRKY54蛋白高度同源;亚细胞定位预测结果显示其主要存在于细胞核中。实时荧光定量PCR分析表明,Vv WRKY54在葡萄不同组织中均有表达,其中在梢尖中表达量最高;盐和低温等逆境胁迫因子均能诱导Vv WRKY54上调表达;此外,Vv WRKY54受水杨酸和一氧化氮诱导上调表达,其中水杨酸诱导Vv WRKY54相对表达量在12 h达到最大值,约为对照的50倍。推测Vv WRKY54在植物发育和抵御逆境胁迫中起着重要作用,这为进一步阐明Vv WRKY54的功能及作用机制奠定了分子基础。
[Abstract]:In order to study the function of the WRKY54 gene of the grape, the Vidal Blanc was used as the material to clone the Vv WRKY54 gene by using the homologous cloning method, and the bioinformatics and the expression characteristics of the Vv WRKY54 gene were analyzed. The results showed that the c-DNA sequence of the Vv WRKY54 gene was 942 bp, encoding 313 amino acids. The results of the bioinformatics analysis show that the molecular weight of the Vv WRKY54 protein is about 35.3091 k Da, the isoelectric point is 5.45, the instability coefficient is 55.03, it is presumed that it is an unstable protein, and is highly homologous to the known wild fruit Yang and the Arabidopsis thaliana WRKY54 protein; and the subcellular localization prediction result shows that it is mainly present in the nucleus. The real-time fluorescence quantitative PCR analysis showed that Vv WRKY54 was expressed in different tissues of the grape, in which the expression of Vv WRKY54 was the highest, and the stress-stress factors such as salt and low temperature could induce the up-regulation of Vv WRKY54. In addition, Vv WRKY54 was upregulated by salicylic acid and nitric oxide. In which the relative expression of the salicylic acid-induced Vv WRKY54 reached a maximum value at 12 h, about 50 times that of the control. It is suggested that Vv WRKY54 plays an important role in the development of plant and the resistance to stress, which lays a molecular basis for further elucidating the function and mechanism of Vv WRKY54.
【作者单位】：青岛农业大学生命科学学院/山东省高校植物生物技术重点实验室;
【基金】：国家自然科学基金(31572107、31401844) 青岛农业大学高层次人才科研基金(111339)
【分类号】：S663.1

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