Preliminary studies on the involvement of BcCHC1 in the regulation of TuMV infection in non-heading Chinese cabbage
[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.
non-heading Chinese cabbage[Brassica campestris(syn.Brassica rapa)ssp.chinensis]TuMVBcCHC1protein interactionVIGS
NETL
NSTL
万方数据
