Defense Responses Stimulated by Bacillus subtilis NCD-2 Through Salicylate- and Jasmonate-Dependent Signaling Pathways Protect Cotton Against Verticillium Wilt.

NCD-2 demonstrates exceptional biocontrol potential against cotton Verticillium wilt. While previous studies have established its direct antifungal activity (e.g., inhibiting mycelial growth and spore germination), our work reveals a novel mechanism: NCD-2 primes systemic resistance in cotton by activating plant immune-signaling pathways. Firstly, transcriptional profiling uncovered that NCD-2 triggers a defense response in roots analogous to infection, allowing cotton to maintain a more balanced state when confronted with pathogen attacks. Meanwhile, the mutant strains ∆fen and ∆srf-defective in lipopeptide synthesis-also improved cotton resistance to Verticillium wilt by activating partially identical defense pathways in cotton roots. Furthermore, the application of lipopeptide compounds derived from NCD-2, particularly surfactin and fengycin, could enhance host resistance to . Using an RT-qPCR approach, we found that numerous resistance-related genes were induced by these lipopeptide compounds. The up-regulation of SA/JA pathway markers (e.g., , , , and ) revealed NCD-2's activation of plant immune signaling. Using virus-induced gene silencing (VIGS), we conclusively linked SA and JA signaling to NCD-2-induced defense priming. Silencing either pathway abolished resistance, highlighting their indispensable coordination. By bridging mechanistic insights and agricultural applicability, our work positions NCD-2 as a sustainable alternative to conventional fungicides, addressing both crop productivity and environmental health.