Drought is a significant abiotic stressor affecting crops globally. Beneficial microorganisms, such as rhizobia, have been shown to enhance crop resilience to such stresses. In this study, we isolated a highly efficient rhizobacterial strain (Rhizobium sp. PV-6) from the root system of Phaseolus vulgaris and systematically investigated the phenotypic and physiological responses of the plants across seven growth stages under four treatments: W-NO (watering without inoculation of rhizobium), W-RHI (watering with inoculation of rhizobium), D-NO (drought without inoculation of rhizobium), and D-RHI (drought with inoculation of rhizobium). We also examined the variation in microbial communities in rhizosphere and root compartments. Physiological analyses revealed that rhizobium inoculation significantly enhanced plant height, fresh weight and dry weight, root length, lateral root number, and nodule number of red kidney beans. Alpha diversity analysis suggested that the microbial communities in the roots and rhizosphere of red kidney beans show different variant distributions. Beta diversity and species difference analysis revealed that drought treatments (D-NO, D-RHI) recruit Shinella, Nocardioides, Agromyces, Pseudomonas, and Ensifer at rhizosphere compartments, while D-RHI enrich Pseudomonas, Sphingobacterium, Paenibacillus, Bacillus, Massilia, and Lysobacter at root compartments in the T5 stage. Further, transcriptomic analysis revealed that PV-6 enhances drought tolerance in red kidney beans by modulating the expression of genes associated with abiotic stress-related genes. Our findings highlight the potential of Rhizobium sp. PV-6 as a bioinoculant for improving drought tolerance in red kidney beans (Phaseolus vulgaris), providing a foundation for designing synthetic microbial communities for crop stress resilience.

Histone modifications and chromatin‐binding proteins play crucial roles in regulating gene expression in eukaryotes, with significant implications for fungal pathogenicity and development. However, profiling these modifications or proteins across the genome in fungi remains challenging due to the technical limitations of the traditional, widely used Chromatin Immunoprecipitation‐Sequencing (ChIP‐Seq) method. Here, we present an optimized fungal Cleavage Under Targets and Tagmentation‐Sequencing (fCUT&Tag‐Seq) protocol specifically designed for filamentous fungi and dimorphic fungi. Our approach involves the preparation of protoplasts and nuclear extraction to enhance antibody accessibility, along with formaldehyde crosslinking to improve protein‐DNA binding efficiency. We then successfully applied fCUT&Tag‐Seq to accurately profile multiple histone modifications like H3K9me3, H3K27me3, H3K4me3, and H3K18ac, across different plant pathogenic or model fungal species, including Verticillium dahliae , Neurospora crassa, Fusarium graminearum , and Sporisorium scitamineum , showing good signal‐to‐noise ratios, reproducibility, and detection sensitivity. Furthermore, we extended this method to profile chromatin‐binding proteins, such as the histone acetyltransferase Gcn5. This study establishes fCUT&Tag‐Seq as a robust and useful tool for fungal epigenetic research, enabling detailed exploration of chromatin dynamics and advancing our understanding of fungal gene regulation, development, and pathogenicity.

Histone modifications and chromatin-binding proteins play crucial roles in regulating gene expression in eukaryotes, with significant implications for fungal pathogenicity and development. However, profiling these modifications or proteins across the genome in fungi remains challenging due to the technical limitations of the traditional, widely used ChIP-Seq method. Here, we present an optimized CUT&Tag-Seq protocol (fCUT&Tag-Seq) specifically designed for filamentous fungi and dimorphic fungi. Our approach involves the preparation of protoplasts and nuclear extraction to enhance antibody accessibility, along with formaldehyde crosslinking to improve protein-DNA binding efficiency. We then successfully applied fCUT&Tag-Seq to accurately profile multiple histone modifications like H3K9me3, H3K27me3, H3K4me3, and H3K18ac, across different plant pathogenic or model fungal species, including Verticillium dahliae , Neurospora crassa , Fusarium graminearum , and Sporisorium scitamineum , showing good signal-to-noise ratios, reproducibility, and detection sensitivity. Furthermore, we extended this method to profile chromatin-binding proteins, such as the histone acetyltransferase Gcn5. This study establishes fCUT&Tag-Seq as a robust and useful tool for fungal epigenetic research, enabling detailed exploration of chromatin dynamics and advancing our understanding of fungal gene regulation, development, and pathogenicity. Impact Statement: We developed a faster, lowLJinput method to study how genes are turned on and off in fungi, even in tough species that are difficult to analyze with standard methods. Our new approach, named fCUT&Tag-Seq, requires only 10,000 cells and can be completed in just two days. It delivers clearer, more reliable results and has already been successfully applied to multiple fungi, including crop pathogens. By revealing the molecular “control switches” that govern fungal development and virulence, we expect it will accelerate basic research and help identify new targets for controlling destructive plant diseases.

Corrigendum: A Weighted Gene Co-Expression Network Analysis–Derived Prognostic Model for Predicting Prognosis and Immune Infiltration in Gastric Cancer