《植物生理学报》 2013, 49(3): 289-296

三个葡萄WRKYs基因的克隆及表达特性分析

侯丽霞*, 王文杰*, 郭秀萍, 傅佩宁, 刘新**

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

通信作者：刘新；E-mail: liuxin6080@yahoo.com.cn；Tel: 0532-88030224

摘　要：

以葡萄抗性品种‘左优红’组培苗为材料, 利用同源克隆法克隆VvWRKY13、VvWRKY45和VvWRKY71, 测序结果显示, VvWRKY13扩增片段大小为681 bp, 编码226个氨基酸; VvWRKY45扩增片段大小为549 bp, 编码182个氨基酸; VvWRKY71扩增片段大小为936 bp, 编码311个氨基酸序列。利用生物信息学分析VvWRKY13、VvWRKY45和VvWRKY71序列显示,这3个蛋白均含有保守的WRKY结构域; 分子量分别为25.7、20.8和34.8 kDa, pI分别为9.08、9.41和6.94。实时定量PCR结果表明, 3个WRKYs均在叶片中表达量最高。逆境相关的信号物质, 如水杨酸、脱落酸、茉莉酸和一氧化氮以及高盐、低温和渗透等逆境胁迫均可诱导VvWRKY13、VvWRKY45和VvWRKY71的表达。推测3个基因均参与了葡萄抵御逆境胁迫的过程。

关键词：葡萄; WRKYs; 基因克隆; 表达分析

收稿：2013-01-10   修定：2013-01-22

资助：青岛市自然科学基金[10-3-4-5-5-jch和12-1-4-5-(12)-jch]和青岛农业大学高层次人才启动基金(630722)。

Gene Cloning and Expression Analysis of Three WRKYs in Vitis vinifera L.

HOU Li-Xia*, WANG Wen-Jie*, GUO Xiu-Ping, FU Pei-Ning, LIU Xin**

Key Lab of Plant Biotechnology in Universities of Shandong, College of Life Sciences, Qingdao Agricultural University, Qingdao, Shangdong 266109, China

Corresponding author: HOU Li-Xia; E-mail: liuxin6080@yahoo.com.cn; Tel: 0532-88030224

Abstract:

WRKY is an important transcription factor that can regulate the expression of downstream stress re-sistance genes, it can be widely involved in plant biotic and abiotic stress resistance. Using homology cloning method, the full-length cDNA of VvWRKY13, VvWRKY45 and VvWRKY71 was cloned from leaves of Vitis vin-ifera cultivar ‘Zuoyouhong’ tissue culture seedling. Bioinformatic analysis indicated that VvWRKY13, VvWRKY45 and VvWRKY71 separately consisted of 681, 549, 936 nucleotides encoding 226, 182, 311 amino acid with molecular weight 25.7, 20.8, 34.8 kDa, isoelectric point 9.08, 9.41 and 6.94, respectively. Three VvWRKYs all possessed one conserved WRKY domain. Real-time PCR analysis showed that three VvWRKYs were expressed in all tested tissues, with the highest expression in leaves. VvWRKY13, VvWRKY45 and VvWRKY71 was significantly induced by stress-related signal substances such as salicylic acid (SA), abscisic acid (ABA), jasmonate acid (JA), nitric oxide (NO). Moreover, three VvWRKYs were accumulated in response to salt stress, cold stress and osmotic stress. It suggested these three VvWRKYs were involved in resistance to various stresses in grape.

Key words: Vitis vinifera L.; WRKYs; gene clone; expression analysis