《植物生理学报》 2015, 51(10): 1743-1748

牡丹PsWRKY基因的克隆和表达特性分析

石红梅, 战新梅, 管世铭, 盖树鹏, 刘春英, 张玉喜^*

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：张玉喜；E-mail: zhang-yuxi@163.com；Tel: 13793284502

摘　要：

以课题组前期454高通量测序筛选到的类WRKY基因的部分cDNA序列为基础, 通过RACE扩增、测序和序列拼接获得了牡丹1 091 bp PsWRKY基因全长cDNA, 包括5′ UTR区57 bp, 3′ UTR区92 bp, 编码框942 bp, 推测编码313个氨基酸序列。生物信息学分析结果表明PsWRKY基因推测氨基酸序列中不具有信号肽序列, 暗示其不是分泌性蛋白; 在氨基酸序列中存在核定位信号, 与亚细胞定位预测结果相一致, 这与其作为转录因子行使功能相关。与其他已知植物的WRKY蛋白的同源性分析结果表明牡丹PsWRKY与葡萄VvWRKY的同源性最高, 为54%。实时定量PCR分析PsWRKY基因的表达特性结果显示PsWRKY基因在初花期牡丹的叶片和心皮中转录水平较高。在人工低温处理的牡丹花芽的休眠解除过程中, PsWRKY基因受低温诱导, 且转录水平在整个休眠进程中呈上调表达趋势。研究结果为进一步解析牡丹花芽的休眠解除的分子机制提供理论依据。

关键词：PsWRKY; RACE; 生物信息学分析; 表达特性

收稿：2015-06-12   修定：2015-09-10

资助：国家自然科学基金面上项目(31471908和31372104)。

Cloning and Expression Pattern Analysis of PsWRKY in Tree Peony (Paeonia suffruticosa)

SHI Hong-Mei, ZHAN Xin-Mei, GUAN Shi-Ming, GAI Shu-Peng, LIU Chun-Ying, ZHANG Yu-Xi^*

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

Corresponding author: ZHANG Yu-Xi; E-mail: zhang-yuxi@163.com; Tel: 13793284502

Abstract:

In this study, based on the partial cDNA sequence in tree peony obtained by 454 high-throughput sequencing, 1 091 bp PsWRKY full-length cDNA sequence was obtained using RACE amplification, including 57 bp 5′ UTR, 92 bp 3′ UTR. The coding fame of 942 bp encoded 313 amino acids. The results of bioinformatics analysis indicated that PsWRKY has no signal peptide, which suggested that it was not a secreted protein. NLS was found existing in the PsWRKY protein sequence, which was consistent with that of the prediction of subcellular localization. The result of homology analysis indicated that the similarity between the PsWRKY and VvWRKY was highest with that of 54%. The expression patterns were analyzed by using real-time quantitative PCR, and the results showed that the transcript level of PsWRKY at the early stage of flowering in leaf and carpel were the highest. During the whole process of dormancy release after chilling treatments, PsWRKY was induced by low temperature, and its transcript was up-regulated. All results would provide theoretical basis for further analysis the molecular mechanism of dormancy release in tree peony.

Key words: PsWRKY; RACE; bioinformatics analysis; expression pattern