茶树WRKY转录因子CsWRKY17的克隆与表达分析

刘苗苗; 臧连生; 孙晓玲; 周忠实; 叶萌

doi:10.13305/j.cnki.jts.2021.05.001

茶叶科学 >

2021 , Vol. 41 >Issue 5: 631 - 642

DOI: https://doi.org/10.13305/j.cnki.jts.2021.05.001

研究报告

茶树WRKY转录因子CsWRKY17的克隆与表达分析

刘苗苗 ,
臧连生 ,
孙晓玲 ,
周忠实 ,
叶萌

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1.吉林农业大学生物防治研究所,吉林长春 130118;
2.中国农业科学院茶叶研究所,浙江杭州 310008;
3.中国农业科学院植物保护研究所,北京 100094

刘苗苗,女,硕士研究生,主要从事植物与昆虫互作研究。

收稿日期: 2021-04-30

修回日期: 2021-06-03

网络出版日期: 2021-10-12

基金资助

国家自然科学基金（31901898）、中央级公益性科研院所基本科研业务费（1610212019001）

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Cloning and Expression Analysis of CsWRKY17 Transcription Factor in Tea Plants

LIU Miaomiao ,
ZANG Liansheng ,
SUN Xiaoling ,
ZHOU Zhongshi ,
YE Meng

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1. Institute of Biological Control, Jilin Agricultural University, Changchun 130118, China;
2. Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
3. Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China

Received date: 2021-04-30

Revised date: 2021-06-03

Online published: 2021-10-12

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摘要

WRKY转录因子是植物中最大的转录因子家族之一,在调节植物的抗虫防御反应中发挥关键作用。然而,目前抗虫相关WRKY转录因子的研究还主要集中于草本植物中,木本植物中的研究还十分滞后,仍有大量WRKY未被发掘与鉴定。以茶树龙井43为试验材料,克隆了1个WRKY转录因子基因,命名为CsWRKY17。研究发现,CsWRKY17全长序列为1 141 bp,包含1个987 bp的开放阅读框,编码328个氨基酸。结构域分析表明,CsWRKY17基因具有1个典型的WRKY结构域和1个C₂H₂型锌指结构,属于第Ⅱ亚家族。同源比对及系统发育树分析发现,CsWRKY17与拟南芥的AtWKRY11和AtWRKY17同源关系最近。此外,CsWRKY17具有明显的组织表达特异性,并可被机械损伤、茶尺蠖取食或模拟取食、茉莉酸（JA）等外用激素处理诱导表达。亚细胞定位试验证实CsWRKY17定位在细胞核中。综上结果表明,CsWRKY17在细胞核中发挥功能,并很可能通过调节JA、脱落酸（ABA）、油菜素内酯（BR）和赤霉素（GA）等信号通路来调控茶树对植食性昆虫的诱导防御反应。研究结果为深入解析WRKY转录因子在茶树虫害相关信号通路中的作用打下基础,同时为茶树抗虫基因挖掘和抗虫品种选育提供了理论依据和参考。

关键词： 茶树; WRKY转录因子; CsWRKY17; 表达分析; 抗虫防御

本文引用格式

刘苗苗 , 臧连生 , 孙晓玲 , 周忠实 , 叶萌 . 茶树WRKY转录因子CsWRKY17的克隆与表达分析[J]. 茶叶科学, 2021 , 41(5) : 631 -642 . DOI: 10.13305/j.cnki.jts.2021.05.001

Abstract

WRKY transcription factors, a super family of plant transcription factors, play an essential role in the regulation of plant defense responses to herbivores. While the roles of herbivore-related WRKY transcription factors are well established in grass plants, their roles in woody plants are still largely unknown. Here, we cloned a WRKY transcription factor, named CsWRKY17. CsWRKY17 has a full length of 1 141 bp, contains a 987 bp open reading frame, and encodes 328 amino acids. Based on the conserved domain analysis, CsWRKY17 belongs to the WRKY Ⅱ subfamily, containing one conserved WRKY domain and a typical C₂H₂-type zinc finger motif. Homology alignment and phylogenetic tree analysis show that CsWRKY17 has the closest relationship with AtWKRY11 and AtWRKY17 in Arabidopsis thaliana. Moreover, CsWRKY17 exhibited a tissue specific expression, and was also induced by mechanical wounding, tea geometrid (Ectropis oblique) attack, simulated herbivory, and exogenous phytohormone treatments like JA. Transient expression experiments indicate that it might play a role in the nucleus. Taken together, we proposed that CsWRKY17 is a potential regulator of herbivore-induced defense responses against herbivores in tea plants through JA, ABA, GA and BR signaling. Our study paved the way for molecular analysis of herbivore-related WRKY genes in tea plants, and provided a good genetic resource and theoretical basis for future studies of pest-resistant genes and breeding of tea plants.

Key words： Camellia sinensis; WRKY transcription factors; CsWRKY17; expression analysis; herbivore resistance

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