禾谷镰刀菌木聚糖酶FGSG_03624互作蛋白筛选及其与小麦通用应激蛋白TaUSP-1A的互作分析OA

The Fusarium graminearum xylanase FGSG_03624 exerts dual functions during wheat in-fection:it disrupts cell wall integrity and induces cell necrosis to activate plant immune responses.To further clarify the molecular mechanism underlying FGSG_03624-triggered immunity,subcellular lo-calization of FGSG_03624 was analyzed via bioinformatics prediction and transient expression in Nic-otiana benthamiana leaves.Yeast two-hybrid assay was performed using FGSG_03624 as the bait to screen interacting proteins from the wheat(Anong 1589)spike cDNA library constructed after F.graminearum infection.The protein-protein interactions were further verified by Y2H assay,lucifer-ase complementation assay(LCA),and bimolecular fluorescence complementation assay(BiFC).The expression patterns of candidate interacting protein gene TaUSP-1A were analyzed by qRT-PCR,and the disease resistance of transgenic Arabidopsis thaliana was evaluated.Results indicated that FGSG_03624 is a secreted protein localized to the cell wall and/or intercellular spaces.Subsequent identifica-tion of its interacting proteins should be conducted the split-ubiquitin membrane yeast two-hybrid sys-tem.Using pBT3STE-FGSG_03624 as the bait vector,40 potential FGSG_03624-interacting proteins were identified from the wheat cDNA library under the condition of adding 12 mmol·L-1 3-AT.These proteins are involved in transcriptional regulation,signal transduction,stress response,mem-brane transport,and plant-pathogen interaction.Among them,wheat universal stress protein TaUSP-1A,associated with stress response,was confirmed to interact with FGSG_03624.TaUSP-1A expression was upregulated after F.graminearum infection,and its overexpression in Arabidop-sis thaliana enhanced plant resistance to the pathogen.These findings indicate that FGSG_03624 may trigger wheat immune responses through interactions with host proteins such as TaUSP-1A.