| 史文炯, 付振超, 曾洪梅*.拟南芥受体激酶AtCERK1胞外结构域互作蛋白的筛选与验证[J].植物保护,2025,51(3):113-121. |
| 拟南芥受体激酶AtCERK1胞外结构域互作蛋白的筛选与验证 |
| Screening and validation of extracellular domain-interacting proteins of Arabidopsis thaliana receptor kinase AtCERK1 |
| 投稿时间：2025-02-17 修订日期：2025-03-20 |
| DOI：10.16688/j.zwbh.2025072 |
| 中文关键词: 拟南芥 几丁质激发子受体激酶胞外结构域 酵母双杂交 互作蛋白 |
| 英文关键词:Arabidopsis thaliana chitin elicitor receptor kinases 1 extracellular domain yeast-two-hybrid interacting protein |
| 基金项目:国家重点研发计划(2017YFD0200900) |
|
| 摘要点击次数: 29 |
| 全文下载次数: 17 |
| 中文摘要: |
| 几丁质激发子受体激酶1(chitin elicitor receptor kinase 1, CERK1)是一种含有溶素基序(lysin motif, LysM)的模式识别受体(pattern recognition receptor, PRR), 在植物免疫响应过程中起重要作用。目前发现CERK1识别的激发子主要为碳水化合物, 是否识别蛋白类配体还不清楚。禾谷镰刀菌Fusarium graminearum是众所周知的真菌病原体, 可以侵染谷物也可以侵染拟南芥。为了确定CERK1的新功能并筛选获得新的激发子, 我们以拟南芥几丁质激发子受体激酶胞外结构域(Arabidopsis thaliana chitin elicitor receptor kinase 1 extra cellular domain, AtCERK1-ECD)为诱饵, 通过酵母双杂交(yeast-two-hybrid, Y2H)筛选禾谷镰刀菌cDNA文库, 获得候选互作蛋白内切葡聚糖酶5(Fg-endoglucanase-5)。内切葡聚糖酶是一类参与植物免疫反应及微生物侵染、定殖、共生的蛋白, Fg-endoglucanase-5是其中的一种。进一步, 通过谷胱甘肽S-转移酶下拉(GST pull-down)和双分子荧光互补(bimolecular fluorescence complementation, BiFC)试验验证了AtCERK1-ECD与Fg-endoglucanase-5的相互作用, 并分析了Fg-endoglucanase-5的功能。本研究获得了AtCERK1-ECD的互作蛋白Fg-endoglucanase-5并阐明了它们在植物-微生物相互作用中的功能, 有助于开发新型激发子并找到植物病害防治的新策略。 |
| 英文摘要: |
| Chitin elicitor receptor kinase 1(CERK1), a lysin motif-containing pattern recognition receptor (PRR), plays an important role in plant immune responses. Currently, known elicitors recognized by CERK1 are primarily carbohydrates, while its potential recognition of protein ligands remains unclear. Fusarium graminearum is a well-known fungal plant pathogen that can infect both cereals and Arabidopsis thaliana. To determine the novel functions of CERK1 and identify new elicitors, we used the extracellular domain of A.thaliana chitin elicitor receptor kinase 1 (AtCERK1-ECD) as bait to screen a F.graminearum cDNA library by yeast-two-hybrid (Y2H) system. This screening identified Fg-endoglucanase-5 as a candidate interacting protein. Endoglucanase are a class of proteins involved in plant immune responses as well as microbial infection, colonization and symbiosis, with Fg-endoglucanase-5 being one such protein. Further validation of the interaction between AtCERK1-ECD and Fg-endoglucanase-5 was performed using glutathione S-transferase pull-down (GST pull-down) and bimolecular fluorescence complementation (BiFC) assays. Additionally, the function of Fg-endoglucanase-5 was analyzed. Identifying AtCERK1-ECD-Fg-endoglucanase-5 interaction and clarifying their roles in plant-microbe interactions may contribute to the development of novel elicitors and new strategies for plant disease management. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
