葡萄WRKY54基因克隆及表达特性分析  

Gene Cloning and Expression Analysis of WRKY54 in Grape

作　　者：肖培连 吕晓彤[1] 侯丽霞[1] 马倩[1] 郝杰[1] 刘新[1] 

机构地区：[1]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,山东青岛266109

出　　处：《核农学报》2017年第1期21-28,共8页Journal of Nuclear Agricultural Sciences

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)

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。In order to explore the function of Vv WRKY54 in plants,the study isolated Vv WRKY54 from the resistant variety Vidal Blanc of grape by homologous cloning,analyzed the sequences using bioinformatics approaches and the expression pattern by Real- Time PCR. The results indicated that Vv WRKY54 was 942 bp in length,encoding a protein of 313 amino acids. The molecular weight of Vv WRKY54 was 35. 309 k Da,isoelectric point was 5. 45 and instability coefficient was 55. 03, speculating it was unstable protein. It shared high homology with WRKY54 in Populus trichocarpa and in Arabidopsis thaliana. Subcellular localization analysis indicated that it was located in nucleus. Real-Time PCR analysis indicated that Vv WRKY54 expressed in different tissues,with the highest expression in grape shoots tip. Vv WRKY54 expression was up- regulated by low temperature,salt stress,and other stress factors. Meantime,Vv WRKY54 was also induced by salicylic acid（ SA） and nitric oxide（ NO）,and the expression induced by SA reached the highest at 12 h,which up to 50 times than the control. These results indicated that Vv WRKY54 has an important role in plant development and stress resistance, which contributes for further illustrating its function and molecular mechanism.

关 键 词：葡萄 WRKY54 基因克隆 表达特性 

分 类 号：S663.1[农业科学—果树学]