桃转录因子PpWRKY11的克隆及表达分析  

Cloning and expression analysis of peach transcription factor PpWRKY11

作　　者：王庆杰[1] 张泽杰 高彦刚 陈修德[1] 肖伟[1] 付喜玲[1] 李玲[1] 李冬梅[1] 高东升[1] WANG Qing-Jie;ZHANG Ze-Jie;GAO Yan-Gang;CHEN Xiu-De;XIAO Wei;FU Xi-Ling;LI Ling;LI Dong-Mei;GAO Dong-Sheng(State Key Laboratory of Crop Biology,Shandong Collaborative Innovation Center for Fruit and Vegetable Production withHigh Quality and Efficiency,College of Horticulture Science and Engineering,Shandong Agricultural University,Taian,Shandong 271018,China)

机构地区：[1]作物生物学国家重点实验室山东果蔬优质高效生产协同创新中心山东农业大学园艺科学与工程学院,山东泰安271018

出　　处：《植物生理学报》2019年第4期503-510,共8页Plant Physiology Journal

基　　金：国家自然科学基金(31872041);山东省自然科学基金(ZR2018MC023和ZR201702200074);山东省现代农业产业技术体系果品创新团队项目(SDAIT-06-01)~~

摘　　要：WRKY转录因子能够响应生物及非生物胁迫。本研究基于前期实验室桃花芽休眠解除前后表达量明显下调的Prupe.1G071400基因进行研究,通过定量PCR表明此结果与前期实验结果相符合,证明该基因可能与桃花芽休眠解除密切相关。从桃‘中油四号’叶片克隆了Prupe.1G071400基因,通过生物信息学分析,命名此基因为PpWRKY11; PpWRKY11蛋白编码282个氨基酸,包含特有WRKY结构域。进化树分析发现PpWRKY11与蔷薇科苹果和梨等亲缘关系最为相近;亚细胞定位显示PpWRKY11定位于细胞核中;酵母双杂显示PpWRKY11蛋白没有自主激活活性,进一步对PpWRKY11进行筛库,筛选出互作的蛋白;构建PGEX-PpWRKY11原核表达载体,转化大肠杆菌Rosetta(DE3)诱导出符合大小的蛋白,发现PpWRKY11蛋白主要在沉淀中。WRKY transcription factors are capable of responding to plant biotic and abiotic stresses. This study was based on the Prupe.1G071400 gene whose expression was significantly down-regulated in the peach flower bud dormancy. The results were consistent with the previous experimental results by quantitative PCR, which proved that the gene may be closely related to the dormancy release of peach flower bud. The Prupe.1G071400 gene was cloned from the leaves of the peach ‘Zhong You 4’, and the gene was named PpWRKY11 by bioinformatics analysis. The PpWRKY11 protein encodes 282 amino acids, including the unique WRKY domain. Phylogenetic tree analysis showed that PpWRKY11 was most closely related to Rosaceae apples and pears. Subcellular localization showed that PpWRKY11 was localized in the nucleus;yeast double hybrid showed that PpWRKY11 protein had no autonomous activation activity, and further utilize PpWRKY11 to screen for interaction proteins;The construct PGEX-Pp WRKY11 prokaryotic expression vector transform E. coli Rosetta(DE3)to induce size-matched protein, and found that PpWRKY11 protein is mainly in precipitation. This study laid the foundation for further exploration of the function of the PpWRKY11 gene.

关 键 词：桃 PpWRKY11 芽休眠 基因克隆 亚细胞定位 基因表达 

分 类 号：Q943.2[生物学—植物学] S662.1[农业科学—果树学]