参与连翘苯丙烷合成途径的WRKY转录因子的筛选与鉴定  

Screening and identification of transcription factors WRKY involved in phenylpropane synthesis pathway in Forsythia suspensa

作　　者：张佳琪 刘晓冉 陈佳茜 郭广洋 谭新杰 赵杏利[1,2] 胥华伟 刘浩[1,2] 侯典云 ZHANG Jia-qi;LIU Xiao-ran;CHEN Jia-xi;GUO Guang-yang;TAN Xin-jie;ZHAO Xing-li;XU Hua-wei;LIU Hao;HOU Dian-yun(College of Agricultural,Henan University of Science and Technology,Luoyang 471023,China;Luoyang Engineering Research Center of Breeding and Utilization of Dao-di Herbs,Luoyang 471023,China)

机构地区：[1]河南科技大学农学院,河南洛阳471023 [2]洛阳市道地药材繁育与创新利用工程技术研究中心,河南洛阳471023

出　　处：《中草药》2023年第18期6055-6064,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(U1404829);中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”(2060302);河南省自然科学基金项目(202102110156);河南省中药材产业科技特派员服务团、河南省中药产业技术体系建设专项资金资助。

摘　　要：目的筛选出参与连翘Forsythia suspensa苯丙烷合成途径的WRKY转录因子并进行生物信息学分析。方法通过对从连翘转录组数据中筛选出的62条连翘WRKY基因序列进行鉴定;通过采用蛋白理化性质和基序分析、蛋白结构分析、系统进化分析、功能注释、蛋白互作分析、外源激素处理后实时荧光定量分析等方式进行相关分析。结果最终筛选出52条具有WRKY结构域的连翘WRKY转录因子。WRKY转录因子编码蛋白的氨基酸数目在105~728,相对分子质量在12277.95~80321.99。蛋白基序分析显示其均有WRKY结构域,与拟南芥WRKY转录因子构建系统进化树将连翘52个WRKY转录因子进一步分为3大类;筛选出5个可能参与连翘苯丙烷合成途径的连翘WRKY转录因子,并推测其可能以被MAPK级联反应所调控的方式介导连翘中苯丙烷合成途径。结论通过分析转录组测序结果,从52个连翘WRKY转录因子中筛选出5个可能参与连翘苯丙烷合成途径的转录因子,其表现出组织特异性表达,且对不同浓度茉莉酸甲脂表现出不同的响应,为进一步研究其作用机制提供研究方向。Objective To screen out the WRKY transcription factors involved in the phenylpropane synthesis pathway of Forsythia suspensa and conduct bioinformatics analysis.Methods A total of 62 WRKY gene sequences screened from F.suspensa transcriptome data were identified.Then the correlation analysis was carried out by means of protein physical and chemical properties and motif analysis,protein structure analysis,phylogenetic analysis,functional annotation,protein interaction analysis,real-time fluorescence quantitative analysis after exogenous hormone treatment.Results A total of 52 WRKY transcription factors with WRKY domain were screened.The number of amino acids of the protein encoded by WRKY transcription factor ranged from 105 to 728,and its molecular weight ranged from 12277.95 to 80321.99.The analysis of protein motif showed that all of them had WRKY domains,and the phylogenetic tree was constructed with Arabidopsis WRKY transcription factors to further divide 52 WRKY transcription factors of F.suspensa into three categories;Five Forsythia WRKY transcription factors that may participate in the phenylpropane synthesis pathway of F.suspensa were screened,and it was speculated that they may mediate the phenylpropane synthesis pathway of F.suspensa in a way regulated by MAPK cascade reaction.Conclusion By analyzing the transcriptome sequencing results,this study screened five transcription factors that may participate in the phenylpropane synthesis pathway of F.suspensa WRKY from 52 transcription factors,which showed tissue-specific expression and different responses to different concentrations of methyl jasmonate,providing a research direction for the further study of its mechanism of action.

关 键 词：连翘 次生代谢产物 WRKY 转录因子 苯丙烷合成途径 

分 类 号：R286.12[医药卫生—中药学]