枯草芽孢杆菌J-15次生代谢产物对棉田土壤细菌群落的影响
Effects of Secondary Metabolites of Bacillus subtilis J-15 on Soil Bacterial Community in Cotton Fields
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摘要: 棉花黄萎病是导致棉花减产的主要土传病害之一,在前期工作中筛选到1株对棉花黄萎病病原菌——大丽轮枝菌(Verticillium dahliae)具有较强拮抗作用的枯草芽孢杆菌J-15菌株。为了揭示施用枯草芽孢杆菌J-15次生代谢产物(secondary metabolites of Bacillus subtilis J-15, SMs)对棉田土壤细菌群落多样性的影响规律。将含有大丽轮枝菌的盆栽分为2组,室温黑暗环境下进行培养,模拟该代谢产物施入土壤的情况,其中实验组为添加0.33 mg·mL-1SMs至土壤最大含水量,对照组为添加等量的去离子水。分别在0、10、30和60 d取样,采用Soil DNA Kit试剂盒提取土壤微生物的总DNA,利用高通量测序技术分析棉田土壤细菌群落结构。根据Alpha多样性分析可知,SMs处理土壤后的Shannon、Simpson、Chao1和Ace指数与对照组均无显著差异,说明SMs对土壤中细菌群落的丰富度和多样性无明显影响。根据群落结构分析可知,SMs处理土壤10、30和60 d后,变形菌门和硬壁菌门的相对丰度略微上升,但差异均不显著。属水平上,SMs处理土壤10 d和30 d后,假单胞菌属和芽孢杆菌属的相对丰度略有上升,但差异均不显著。这说明在土壤中施用SMs虽在一定程度上改变了土壤中细菌群落结构但影响却并不显著。由此推断SMs对棉田土壤细菌群落结构及多样性无显著影响,为开发利用枯草芽孢杆菌J-15进行农业上的生物防治提供了依据。Abstract: Cotton verticillium wilt is one of the leading soil-borne diseases that reduce cotton yield. In the previous work, a strain of Bacillus subtilis J-15 with a strong antagonistic effect against Verticillium dahliae, the pathogen of cotton verticillium wilt, was screened to reveal the effect of the application of secondary metabolites of Bacillus subtilis J-15 (SMs) on the diversity of the bacterial community in cotton field soil. To simulate the application of this metabolite into the soil, pot plants containing Verticillium dahliae were divided into two groups and cultured at room temperature in the dark. 0.33 mg·mL-1 SMs were added to the experimental group to maximize soil water content, while the same amount of deionized water was added to the control group. Samples were taken on days 0, 10, 30, and 60, respectively. The total DNA of soil microorganisms was extracted using the Conway Century's Soil DNA Kit kit. The bacterial community structure of cotton field soil was analyzed by high-throughput sequencing technology. According to the Alpha diversity analysis, the Shannon index, Simpson index, Chao1, and Ace index of SMs-treated soil were not significantly different from the control group, indicating that SMs had no significant impact on the richness and diversity of the bacterial community in soil. According to community structure analysis, the relative abundance of Proteobacteria and Scleroderma increased slightly after 10, 30, and 60 d of soil treatment by SMs. However, the difference is insignificant. At the genus level, the relative abundance of Pseudomonas and Bacillus increased slightly after 10 and 30 d of SMs treatment. However, the difference is negligible. The results demonstrated that although the application of SMs in soil changed the bacterial community structure in the soil to a certain extent, the effect was insignificant. On this basis, it is concluded that SMs have no substantial impact on the structure and diversity of the soil bacterial community in cotton fields, providing a basis for the development and utilization of Bacillus subtilis J-15 in agricultural biological control.
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