Functional Analysis of GhEXLB2 in Regulating Cotton Resistance to Verticillium Wilt

Verticillium wilt (VW), caused by the soil-borne fungus Verticillium dahliae, is a major disease that markedly compromises both the yield and fiber quality of cotton. In this study, we explored the function and underlying mechanism of the cotton expansin gene GhEXLB2 in response to VW infection. Expression profiling revealed that members of the GhEXL family exhibit distinct patterns across tissues and under various biotic and abiotic stresses. Notably, GhEXLB2, which encodes an extracellular protein, showed the strongest induction following V. dahliae challenge. Ectopic expression of GhEXLB2 in Arabidopsis thaliana promoted root elongation and root hair formation, and was associated with improved resistance to the pathogen. In contrast, silencing GhEXLB2 in cotton via virus-induced gene silencing (VIGS) led to pronounced vascular browning, increased pathogen recovery, and a lower level of disease resistance. In addition, RNA-seq profiling of GhEXLB2-silenced (VIGS) cotton plants revealed that most differentially expressed genes were enriched in pathways related to phytohormone signaling and plant-pathogen interactions, with salicylic acid (SA) signaling and WRKY transcription factors emerging as central regulatory components. Analysis of the GhEXLB2 promoter further identified multiple cis-acting elements associated with stress and hormone responsiveness. When integrated with protein-protein interaction (PPI) prediction data, these results suggest that GhEXLB2 may be modulated by a network of transcription factors and signaling pathways. Collectively, the evidence supports a positive association between GhEXLB2 and VW resistance. This study provides a framework for understanding expansin functions in cotton defense against VW.