Fusarium pseudograminearum causes destructive crown disease in wheat. The velvet protein family is a crucial regulator in development, virulence, and secondary metabolism of fungi. We conducted a functional analysis of FpVelB using a gene replacement strategy. The deletion of FpVelB decreased radial growth and enhanced conidial production compared to that of wild type. Furthermore, FpVelB modulates the fungal responses to abiotic stress through diverse mechanisms. Significantly, virulence decreased after the deletion of FpVelB in both the stem base and head of wheat. Genome-wide gene expression profiling revealed that the regulation of genes by FpVelB is associated with several processes related to the aforementioned phenotype, including “immune”, “membrane”, and “antioxidant activity”, particularly with regard to secondary metabolites. Most importantly, we demonstrated that FpVelB regulates pathogen virulence by influencing deoxynivalenol production and modulating the expression of the PKS11 gene. In conclusion, FpVelB is crucial for plant growth, asexual development, and abiotic stress response and is essential for full virulence via secondary metabolism in F. pseudograminearum.

The soil-borne pathogen Fusarium pseudograminearum is capable of causing a highly destructive crown disease in wheat. The purpose of this study was to characterize the biological functions, such as of virulence and secondary metabolites (SMs), of a putative RNA-Pol II transcription elongation factor, FpRtfA, in F. pseudograminearum. The current study revealed that the deletion of FpRtfA reduced radial growth compared to wild type in F. pseudograminearum. In addition, FpRtfA was found important to conidiation and response to metal ions and oxidative stress. More importantly, the virulence to the wheat stem base or head was decreased when FpRtfA was deleted. Using genome-wide gene expression profiling, FpRtfA was found to regulate several processes related to the above phenotype, such as the carbohydrate metabolic process, and the response to oxidative stress and oxidoreductase activity, especially for SMs. Further, we proved that FpRtfA exerts its regulatory effect on the virulence of pathogens by modulating the expression of the PKS gene, but not the generation of DON. In conclusion, FpRtfA has important roles in growth, asexual development, and the response to metal ions and oxidative stress. More importantly, FpRtfA is required for SMs and full virulence in F. pseudograminearum.

Rapid phenotyping of water- and salt-soluble wheat protein fractions can support grain quality evaluation and breeding. Whole-wheat flour from water-nitrogen and water-fertilizer trials was used to predict measured albumin and globulin contents and their derived sum, termed the metabolic-protein sum (albumin + globulin). After systematically comparing various preprocessing, wavelength selection, and regression approaches, a framework combining order-optimized derivative preprocessing (ODP), a hybrid feature-selection approach based on minimum redundancy maximum relevance (MRMR) and recursive feature elimination (RFE) and a tabular prior-data fitted network (TabPFN) regressor provided consistently strong test-set performance. The optimal derivative orders were 1.0 for albumin and the metabolic-protein sum and 0.8 for globulin. The ODP-MRMR-RFE-TabPFN framework retained 147, 46, and 70 wavelengths for albumin, globulin, and the metabolic-protein sum, yielding test set coefficient of determination (R²) values of 0.847, 0.693, and 0.850; root mean square error (RMSE) values of 3.073, 0.951, and 3.368 mg g⁻¹; and residual prediction deviation (RPD) values of 2.554, 1.804, and 2.586, respectively. Leave-one-cultivar-out validation revealed substantial variation in predictive performance across held-out cultivars, with R² and RPD ranging from 0.282 to 0.736 and from 1.180 to 1.945, respectively, for albumin, and from 0.416 to 0.750 and from 1.308 to 2.001, respectively, for globulin. Thus, although albumin achieved quantitative-level performance under the random train-test split, this performance was not consistently maintained for unseen cultivars, whereas globulin was generally more suitable for screening. SHapley additive explanations (SHAP) and partial dependence plots (PDPs) revealed that visible wavelengths contributed strongly to all models, with near-infrared (NIR) wavelengths supplying complementary information for globulin and the metabolic-protein sum. Overall, this framework offers rapid, interpretable flour-based estimation of metabolically active wheat protein fractions.