西北植物学报2024，Vol.44Issue(8)：1229-1238,10.DOI:10.7606/j.issn.1000-4025.20230439
不结球白菜BcCHC1蛋白参与调控TuMV侵染的初步研究
Preliminary studies on the involvement of BcCHC1 in the regulation of TuMV infection in non-heading Chinese cabbage
摘要
Abstract
[Objective]To enhance the resistance of non-heading Chinese cabbage to turnip mosaic virus(TuMV),this study preliminarily explored the interaction mechanism between BcCHC1 protein of non-heading Chinese cabbage and TuMV.[Methods]The study firstly identified members of the heavy chain clathrin(CHC)gene family,BcCHCs,from non-heading Chinese cabbage,and then cloned CHC1 gene.BcCHC1 subcellular localization was analyzed,and bimolecular fluorescence complementation(BiFC)as-say was used to verify the interaction between TuMV and BcCHC1.Finally,virus-induced gene silencing(VIGS)was employed to silence BcCHC1 expression and its function was assessed.[Results]BcCHC1 gene from non-heading Chinese cabbage was cloned,with a coding sequence length of 5 124 bp,encoding 1 708 amino acids.qRT-PCR results showed a significant decrease in the relative expression level of Bc-CHC1 gene in non-heading Chinese cabbage infected with TuMV after 30 days.Subcellular localization re-vealed that BcCHC1 protein was located in the cell membrane and nucleus of tobacco epidermal cells.BiFC assay analysis confirmed that BcCHC1 interacted with CI and 6K2:Interaction with CI was mainly occur-ring in the nucleus,while interaction with 6K2 mainly on the cell membrane.Observations of BcCHC1-si-lenced lines showed that silencing BcCHC1 gene led to plant death.[Conclusion]BcCHC1 regulates the infection of non-heading Chinese cabbage by TuMV through interaction with CI and 6K2,thereby affecting the clathrin-dependent endocytosis pathway and virus replication.However,the mechanism by which Bc-CHC1 regulates TuMV infection in non-heading Chinese cabbage requires further study.关键词
不结球白菜/BcCHC1/TuMV/互作蛋白/VIGSKey words
non-heading Chinese cabbage[Brassica campestris(syn.Brassica rapa)ssp.chinensis]/TuMV/BcCHC1/protein interaction/VIGS分类
生物科学引用本文复制引用
刘芳,吕善武,袁梦果,杨紫蕊,侯喜林,张昌伟..不结球白菜BcCHC1蛋白参与调控TuMV侵染的初步研究[J].西北植物学报,2024,44(8):1229-1238,10.基金项目
江苏省自然科学基金项目(BK20200560) （BK20200560）
江苏省农业科技自主创新资金项目[CX(19)3038] （19）
