Bacillus subtilis NCD-2 demonstrates exceptional biocontrol potential against cotton Verticillium wilt. While previous studies have established its direct antifungal activity (e.g., inhibiting Verticillium dahliae 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 V. dahliae 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 V. dahliae. 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., NPR1, ICS1, COI1, and LOX1) 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.

Bacillus subtilis strain NCD-2 is a promising biocontrol agent for soil-borne plant diseases and shows potential for promoting the growth of some crops. The purposes of this study were to analyze the colonization ability of strain NCD-2 in different crops and reveal the plant growth promotion mechanism of strain NCD-2 by rhizosphere microbiome analysis. qRT-PCR was used to determine the populations of strain NCD-2, and microbial communities’ structures were analyzed through amplicon sequencing after application of strain NCD-2. Results demonstrated that strain NCD-2 had a good growth promotion effect on tomato, eggplant and pepper, and it was the most abundant in eggplant rhizosphere soil. There were significantly differences in the types of beneficial microorganisms recruited for different crops after application of strain NCD-2. PICRUSt analysis showed that the relative abundances of functional genes for amino acid transport and metabolism, coenzyme transport and metabolism, lipid transport and metabolism, inorganic ion transport and metabolism, and defense mechanisms were enriched in the rhizospheres of pepper and eggplant more than in the rhizospheres of cotton, tomato and maize after application of strain NCD-2. In summary, the colonization ability of strain NCD-2 for five plants was different. There were differences in microbial communities’ structure in rhizosphere of different plants after application of strain NCD-2. Based on the results obtained in this study, it was concluded that the growth promoting ability of strain NCD-2 were correlated with its colonization quantity and the microbial species it recruited.

Plant Growth Promoting Rhizobacteria, PGPR, can protect plants against soil-borne diseases and abiotic stress conditions. The primary objective of this study was to evaluate the effects of different PGPRs (TF1, TF2, TF3, and TF4) on the rhizosphere microbial community of silage maize in a saline–alkaline field via Illumina MiSeq high-throughput sequencing technology. Results demonstrated that different PGPRs significantly increased the harvest density (by 21.31–45.16%), plant height (by 9.12–19.98%), stem diameter (by 30.07–45.78%), and biomass (by 33.20–65.36%) of silage maize, TF3 treatment significantly increased the fresh weight (by 32.50%), while the other treatments could increase the fresh weight but not significantly. Four microbial agents significantly reduced the contents of soil available phosphorus (AP), electrical conductivity (EC), and neutral phosphatase activity (NPA), while significantly increasing the contents of available potassium (AK), ammonium nitrogen (NH₄⁺-N), nitrate nitrogen (NO₃⁻-N), chitinase activity (ChtA), and urease activity (UA). Specifically, TF2 and TF3 treatments significantly decreased the soil pH value, while not for TF1 and TF4. Microbiome analysis showed that four microbial agents significantly increased the relative abundances of beneficial microorganisms, such as Arthrobacter, Blastococcus, MNDI, Chaetomidium, Alternaria, Sarocladium, Acremonium, and Clonostachys, and significantly decreased the relative abundances of Gibberella and Fusarium. Mental analysis showed that the soil bacterial community structure did not significantly correlate with soil biochemical properties, while the soil fungal community structure significantly and positively correlated with pH. Maize yield significantly and positively correlated with NH₄⁺-N, OM, AP, EC, UA, ChtA, and NPA.

Given the scarcity of known virulence factors in Verticillium dahliae , a fungus that causes severe agricultural losses worldwide, identifying novel virulence genes is essential for understanding its pathogenic mechanisms and ultimately for developing effective disease control strategies. Gamma-glutamyl cyclotransferase (GGCT) has been recognized as a promising drug target, yet its biological functions and regulatory roles in plant-pathogenic fungi remain largely unexplored. Here, we demonstrate that knockout of the VdGGCT gene severely attenuates the development and virulence of V. dahliae and increases resistance to diamide (Dia)-induced oxidative stress. Functional assays using the organic sulfur auxotrophic yeast mutant ABC1723 revealed that despite belonging to the GGCT superfamily, VdGGCT does not exhibit detectable GGCT activity towards GSH, γ-Glu-Met, or γ-Glu-Cys. Integrative transcriptomic and metabolomic analyses revealed that the deletion of VdGGCT significantly downregulated the expression of glutamate and γ-glutamyl amino acids in the γ-glutamyl cycle, with concordant dysregulation of arginine/proline and beta-alanine metabolism at both levels, indicating that VdGGCT modulates the expression and abundance of genes related to pathogenicity, especially in the γ-glutamyl cycle and amino acid metabolism pathways. Collectively, these findings highlight the importance of VdGGCT in the development, oxidative stress response, and virulence of V. dahliae , identifying it as a novel potential target for designing innovative fungicides.

Microplastics have become a global environmental concern, but the impact of microplastics on plant disease remains poorly understood. The primary objective