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The Fusarium graminearum xylanase FGSG_03624 exerts dual functions during wheat infection: 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 localization of FGSG_03624 was analyzed via bioinformatics prediction and transient expression in Nicotiana 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, luciferase 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 identification of its interacting proteins should be conducted the split-ubiquitin membrane yeast two-hybrid system. 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, membrane 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 Arabidopsis 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.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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