Journal of Nuclear Agricultural Sciences ›› 2021, Vol. 35 ›› Issue (9): 1987-1993.DOI: 10.11869/j.issn.100-8551.2021.09.1987

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

Cloning and Expression Analysis of Transcription Factor PpWRKY18 in Peach

LI Sen1(), WANG Qingjie1(), CHEN Xiude1, ZHANG Rui1, LI Ling1, DU Guiying2, FU Xiling1,*()   

  1. 1 College of Horticulture Science and Engineering, Shandong Agricultural University/State Key Laboratory of Crop Biology/Shandong Collaborative Innovation Center for Fruit and Vegetable Production with High Quality and Efficiency, Taian, Shandong 271018
    2 Wulian Fruit Tree Station, Rizhao, Shandong 262300
  • Received:2020-05-27 Accepted:2020-08-21 Online:2021-09-10 Published:2021-07-22
  • Contact: FU Xiling

桃WRKY转录因子PpWRKY18克隆及表达分析

李森1(), 王庆杰1(), 陈修德1, 张蕊1, 李玲1, 杜桂英2, 付喜玲1,*()   

  1. 1 山东农业大学园艺科学与工程学院/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 泰安 271018
    2 五莲县果树站,山东 日照 262300
  • 通讯作者: 付喜玲
  • 作者简介:李森,男,主要从事果树分子生物学研究。E-mail: lisensdau@163.com;
    王庆杰,男,主要从事果树分子生物学研究。E-mail: wangqingjie0914@163.com第一联系人:**同为第一作者。
  • 基金资助:
    十三五重点研发计划子课题(2018YFD1000104);山东省现代农业产业技术体系果品创新团队(SDAIT-06-01);山东省自然科学基金(ZR201702200074)

Abstract:

In order to explore the important role of WRKY transcription factors in plant resistance to adversity stress. This study obtained the PpWRKY18 gene in peach, and bioinformatics analysis showed that the PpWRKY18 protein contained a typical WRKY functional domain. Phylogenetic tree analysis showed that PpWRKY18 in peach was more closely related those in to Rosaceae species. PpWRKY18 was predicted to be a nucleolus localization protein, and was confirmed to be located in nucleus using transient expression in tobacco leaf through Agrobacterium tumefaciens injection. Fusion protein PGEX-PpWRKY18 was expressed after induction, and showed that the PpWRKY18 was mainly expressed in the inclusion body. Real-time PCR showed that the expression of PpWRKY18 was induced by drought, and the expression of PpWRKY18 decreased after rehydration. The protein phosphatase protein family (Protein Phosphatase 2C), C2H2 protein family (PpZAT5), ERF protein family (PpERF9), and BHLH protein family (PpBHLH92) were predicted to interact with PpWRKY18, therefore we speculate that PpWRKY18 regulates drought stress by interacting with these proteins, and may provide a theoretical basis for investigating the molecular mechanism of PpWRKY18 response to drought.

Key words: peach, PpWRKY18, bioinformatics analysis, drought

摘要:

为了探究WRKY转录因子在植物抵抗逆境胁迫方面的重要作用,本研究从桃中克隆PpWRKY18基因,通过生物信息学分析PpWRKY18蛋白含有典型WRKY功能结构域。系统进化树分析表明,桃PpWRKY18与蔷薇科物种的亲缘关系最为相近。生物信息学预测PpWRKY18蛋白定位细胞核,并通过农杆菌瞬时注射烟草叶片证实PpWRKY18定位于细胞核。对PGEX-PpWRKY18融合蛋白进行诱导表达,发现PpWRKY18融合蛋白主要在包涵体中表达。荧光定量PCR结果表明,PpWRKY18受干旱诱导表达量上升,复水后PpWRKY18表达量下降。生物信息学预测显示,蛋白磷酸酶蛋白家族(Protein Phosphatase 2C)、C2H2蛋白家族(PpZAT5)、ERF蛋白家族(PpERF9)、BHLH蛋白家族(PpBHLH92)可能与PpWRKY18蛋白互作,推测PpWRKY18蛋白通过与这些蛋白互作协同响应干旱胁迫。本研究为探索PpWRKY18响应干旱的分子机制提供了理论依据。

关键词: 桃, PpWRKY18, 生物信息学分析, 干旱