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草业学报 ›› 2022, Vol. 31 ›› Issue (12): 146-157.DOI: 10.11686/cyxb2021478

• 研究论文 • 上一篇    

基于全长转录组的盐生草WRKY基因家族的鉴定及其盐胁迫响应模式分析

陈倩1,2(), 徐晓芸1,2, 汪军成1,2, 姚立蓉1,2, 司二静1,2, 杨轲1,2, 韦晓玲1,2, 马小乐1,2, 李葆春1,3, 尚勋武2, 孟亚雄1,2(), 王化俊1,2()   

  1. 1.省部共建干旱生境作物学国家重点实验室,甘肃省作物遗传改良与种质创新重点实验室,甘肃 兰州 730070
    2.甘肃农业大学农学院,甘肃 兰州 730070
    3.甘肃农业大学生命科学技术学院,甘肃 兰州 730070
  • 收稿日期:2021-12-23 修回日期:2022-02-24 出版日期:2022-12-20 发布日期:2022-10-17
  • 通讯作者: 孟亚雄,王化俊
  • 作者简介:E-mail: huajunwang@sina.com
    E-mail: yxmeng1@163.com
    陈倩(1998-),女,甘肃庆阳人,在读硕士。E-mail: 2393484372@qq.com
  • 基金资助:
    国家自然科学基金项目(31960072);财政部和农业农村部:国家现代农业产业技术体系项目(CARS-05-04B-2);甘肃省教育厅:产业支撑计划项目(2021CYZC-12);甘肃省自然基金项目(20JR10RA507);甘肃农业大学伏羲青年英才计划(Ganfx-03Y06);甘肃农业大学国家级大学生创新创业训练计划重点支持领域项目(202110733001)

Identification of a WRKY gene family based on full-length transcriptome sequences and analysis of response patterns under salt stress in Halogeton glomeratus

Qian CHEN1,2(), Xiao-yun XU1,2, Jun-cheng WANG1,2, Li-rong YAO1,2, Er-jing SI1,2, Ke YANG1,2, Xiao-ling WEI1,2, Xiao-le MA1,2, Bao-chun LI1,3, Xun-wu SHANG2, Ya-xiong MENG1,2(), Hua-jun WANG1,2()   

  1. 1.State Key Laboratory of Aridland Crop Science,Gansu Key Laboratory of Crop Improvement and Germplasm Enhancement,Lanzhou 730070,China
    2.College of Agronomy,Gansu Agricultural University,Lanzhou 730070,China
    3.College of Life Sciences and Technology,Gansu Agricultural University,Lanzhou 730070,China
  • Received:2021-12-23 Revised:2022-02-24 Online:2022-12-20 Published:2022-10-17
  • Contact: Ya-xiong MENG,Hua-jun WANG

摘要:

WRKY是一类植物特有的转录因子,尤其在植物生长、发育和逆境过程中起到至关重要的作用。为了鉴定盐生草WRKY基因家族成员、揭示其进化关系及探究HgWRKYs基因在盐胁迫下的表达响应模式,本研究基于盐生草全长转录组数据,利用生物信息学方法对其WRKY转录因子进行鉴定与分析,并通过qRT-PCR方法检测了HgWRKYs家族成员在盐胁迫下的相对表达量。结果表明,在盐生草转录组中共鉴定出31个HgWRKYs,氨基酸数介于220~611 aa,其蛋白N端均具有完整的WRKYGQK保守序列;除HgWRKY20HgWRKY21同时分布于细胞核和线粒体外,其余HgWRKY基因均位于细胞核;系统进化分析将其分为Ⅰ(8个)、Ⅱ(22个)、Ⅲ(1个)三类,第Ⅱ类进一步分为Ⅱ-a(1个)、Ⅱ-b(2个)、Ⅱ-c(1个)、Ⅱ-d(15个)和Ⅱ-e(3个)5个亚类;Motif 1和Motif 2是HgWRKYs基因家族的特征基序,在进化过程中较为保守。qRT-PCR结果表明,HgWRKYs主要在根系中表达,具有组织特异性,其中HgWRKY19HgWRKY23HgWRKY27基因在24 h的表达量最高,将其作为盐胁迫响应候选基因,以期为后续研究盐生草WRKY基因响应盐胁迫的分子机制提供依据。

关键词: 盐生草, WRKY基因家族, 盐胁迫, 生物信息学

Abstract:

WRKY is a plant-specific transcription factor that plays an important role in plant growth, plant development and plant stress responses. This research aimed to identify members of the WRKY gene family in Halogeton glomeratus, explore their evolutionary relationships and elucidate the expression response pattern of HgWRKY genes under salt stress. From the full-length transcriptome data of H. glomeratus, the WRKY gene family members were identified and analyzed by bioinformatics methods, and their relative expression levels under salt stress were detected by qRT-PCR. A total of 31 HgWRKY genes were identified from H. glomeratuswith the coded amino acid number of identified HgWRKYs ranging from 220 to 611. All HgWRKY proteins had complete conserved WRKYGQK sequences at the N-terminal. Subcellularly, all HgWRKYs were located in the nucleus, and HgWRKY20 and HgWRKY21 were also located in mitochondria. Based on phylogenetic analysis, the 31 detected HgWRKYs can be divided into three groups: Ⅰ (8 members), Ⅱ (22 members), Ⅲ (1 member). Group Ⅱ contained five subgroups: Ⅱ-a (1 member), Ⅱ-b (2 members), Ⅱ-c (1 member), Ⅱ-d (15 members) and Ⅱ-e (3 members). Motif 1 and Motif 2 were the characteristic motifs of the HgWRKYs family, and these were found to be relatively conserved in the evolutionary process. The qRT-PCR results indicated that HgWRKYs were mainly expressed in roots with tissue specificity, and the expression levels of HgWRKY19HgWRKY23 and HgWRKY27 genes were the highest at 24 h. These three were therefore selected as candidate genes for salt stress response, in order to provide a basis for further study of the molecular mechanisms of HgWRKY genes in response to salt stress.

Key words: Halogeton glomeratus, WRKY gene family, salt stress, bioinformatics analysis