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  

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

作　　者：陈倩 徐晓芸 汪军成[1,2] 姚立蓉 司二静[1,2] 杨轲[1,2] 韦晓玲 马小乐[1,2] 李葆春[1,3] 尚勋武[2] 孟亚雄[1,2] 王化俊[1,2] CHEN Qian;XU Xiao-yun;WANG Jun-cheng;YAO Li-rong;SI Er-jing;YANG Ke;WEI Xiao-ling;MA Xiao-le;LI Bao-chun;SHANG Xun-wu;MENG Ya-xiong;WANG Hua-jun(State Key Laboratory of Aridland Crop Science,Gansu Key Laboratory of Crop Improvement and Germplasm Enhancement,Lanzhou 730070,China;College of Agronomy,Gansu Agricultural University,Lanzhou 730070,China;College of Life Sciences and Technology,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[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

出　　处：《草业学报》2022年第12期146-157,共12页Acta Prataculturae Sinica

基　　金：国家自然科学基金项目(31960072,32001514);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)和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)资助。

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HgWRKY20和HgWRKY21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HgWRKY19、HgWRKY23和HgWRKY27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。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.glomeratus,with 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,the31 detected HgWRKYs can be divided into three groups:Ⅰ(8 members),Ⅱ(22 members),Ⅲ(1 member).GroupⅡcontained five subgroups:Ⅱ-a(1 member),Ⅱ-b(2 members),Ⅱ-c(1 member),Ⅱ-d(15 members)andⅡ-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 HgWRKY19,HgWRKY23 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.

关 键 词：盐生草 WRKY基因家族 盐胁迫 生物信息学 

分 类 号：Q943.2[生物学—植物学]