摘要
艾(Artemisia argyi)是常用的中草药和灸用原料植物,具有重要的药用和经济价值,然而其药用活性成分的合成调控研究鲜有报道。WRKY转录因子家族是植物中最大的转录因子家族之一,在植物生长发育、抗逆以及次生代谢合成调控等方面发挥重要作用。本研究基于转录组数据系统鉴定艾的WRKY家族转录因子,分析其蛋白理化性质、进化关系和保守基序等特性,并研究该家族基因在茉莉酸胁迫下的表达模式。实验结果表明,从转录组水平共鉴定到37个AarWRKY转录因子,其氨基酸数目为119-547个,分子量为13.4-59.8 kDa,理论等电点为4.86-10.03。根据拟南芥WRKY家族分类,34个AarWRKY蛋白归类在I、II、III三组,而3个蛋白未归类。AarWRKY蛋白均具有保守的WRKY结构域,且同一组蛋白具有相似的保守基序。基因表达谱分析以及利用qRT-PCR方法检测基因表达差异发现,外源茉莉酸甲酯处理后,6个AarWRKY基因的表达发生了显著差异,其中3个基因上调表达,3个基因下调表达。进一步蛋白互作网络预测发现这些差异表达的AarWRKY基因可能参与JA响应和萜类化合物的生物合成调控等途径。本研究为艾WRKY家族基因的功能研究及其在艾叶活性成分累积中的调控作用解析奠定基础。
Artemisia argyi is a commonly used Chinese herb and a key raw material for moxibustion, which has important medicinal and economic values. However, there are few reports on the biosynthesis and regulation of active ingredients in A. argyi. The WRKY gene family is one of the largest transcription factors(TFs) in plants and plays major roles in modulating plant growth and developmental processes, stress responses and secondary metabolic. In this study, we systematically identified the WRKY TFs in A. argyi using the RNA-seq data, and analyzed physicochemical properties of proteins, evolutionary relationship, conservative motifs and their expression profiles under jasmonic acid(JA) stress.The results indicated that 37 AarWRKYs were identified from the transcriptome level, the length of amino acids was119-547 aa, molecular weight of proteins was 13.4-59.8 kDa, and the theoretical isoelectric points were 4.86-10.03.According to the classification of WRKY gene family in Arabidopsis thaliana, 34 AarWRKY proteins were classified into groups I, II, and III, but 3 AarWRKY proteins were not classified. AarWRKY proteins have the conserved WRKY domains and proteins in the same group share similar conserved motifs. Gene expression heat map and qRT-PCR results showed that the expression of 6 AarWRKY genes were significantly changed under exogenous methyl jasmonate treatment. Among them, 3 genes were up-regulated and 3 genes were down-regulated. The protein-protein interaction network prediction indicated that these differentially expressed AarWRKY genes might be involved in JA response and the regulation of terpenoids biosynthesis. This study lays the foundation for the functional research of AarWRKY genes and their regulatory role in the accumulation of active ingredients in A. argyi.
作者
易小哲
王梦月
邬兰
刘霞
刘佳伟
农熠瑛
陈士林
师玉华
Yi Xiaozhe;Wang Mengyue;Wu Lan;Liu Xia;Liu Jiawei;Nong Yiying;Chen Shilin;Shi Yuhua(Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine,Artemisinin Research Center,Institute of Chinese Meteria Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Chemical Engineering and Life Sciences,Wuhan University of Technology,Wuhan 430070,China;Hubei Ai Ai Tie Health Technology Co,Ltd.,Huanggang 435300,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第5期1811-1824,共14页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
中国中医科学院科技创新工程(CI2021A04107):黄花蒿等药用植物高效基因编辑体系建立与种质创新研究,负责人:师玉华。
关键词
艾
WRKY转录因子
茉莉酸胁迫
基因表达分析
实时荧光定量PCR
Artemisia argyi
WRKY transcription factors
Jasmonic acid stress
Gene expression analysis
Real-time fluorescence quantitative PCR
