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Cotton Verticillium wilt, caused by the soil-borne fungal pathogen Verticillium dahliae, is a devastating disease that severely impacts global cotton production. [Objective] To investigate the biocontrol potential and mechanism of endophytic Pseudomonas sp. NWSUAF303 against cotton Verticillium wilt and provide novel microbial resources for managing soil-borne diseases in cotton. [Methods] The strain was identified by phylogenetic analysis based on 16S rRNA gene sequences and phenotypic characterization. Its antifungal spectrum was evaluated via dual-culture and volatile organic compounds (VOCs) inhibition assays. VOCs were detected by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Pot experiments were carried out to assess the disease control efficacy of the strain. RT-qPCR and enzymatic activity assays were employed to elucidate the resistance mechanism of the strain against cotton Verticillium wilt. [Results] Strain NWSUAF303 was identified as Pseudomonas alvandae, exhibiting plant growth-promoting properties including nitrogen fixation, phosphate solubilization, and indole-3-acetic acid (IAA) production. Its non-volatile metabolites inhibited six phytopathogenic fungi, whereas VOCs demonstrated broader antifungal spectrum against seven pathogens, showing the inhibition rates >95% against Sclerotinia sclerotiorum and of 89.27% against V. dahliae 592. The VOCs of this strain downregulated the expression of virulence genes (VdPR1, Vdpf, and VdGAL4) in V. dahliae (P<0.05). Three key antifungal VOCs were identified, including 2,3-butanedione, 2-nonanol, and 6-methyl-2-heptanol, with the inhibitory effect of 2,3-butanedione on V. dahliae being first reported. Pot experiments revealed the control efficacy of 54.40% against Verticillium wilt, which was comparable to that of carbendazim. Strain NWSUAF303 activated the salicylic acid/jasmonic acid (SA/JA) signaling pathway, upregulating the expression of defense-related genes GhPAL, Gh4CL, and GhCHI (P<0.01), while enhancing the activities of peroxidase (POD), polyphenol oxidase (PPO), and superoxide dismutase (SOD). [Conclusion] P. alvandae NWSUAF303 combats Verticillium wilt through dual mechanisms: producing novel antifungal VOCs and activating systemic resistance via SA/JA signaling and defense enzyme coordination. With broad-spectrum antifungal activity and plant growth-promoting properties, this strain represents a promising biocontrol agent for sustainable management of cotton Verticillium wilt.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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