枯草芽孢杆菌调控甘蓝根际微生物多样性及根肿病抗性效应研究

doi:10.16420/j.issn.0513-353x.2025-0665

摘要/Abstract

摘要：

根肿病由芸薹根肿菌（Plasmodiophora brassicae，Pb）引起，严重影响甘蓝的产量和品质。枯草芽孢杆菌（Bacillus subtilis，BS）作为生防菌株，其调控根际微生态机制尚不明确。本研究中，通过设置BS、Pb、Pb + BS复合处理及无菌水对照灌根，结合16S rDNA高通量测序与根际代谢组分析，并测定病情指数、生物量及防御酶活性等生理指标，系统揭示了枯草芽孢杆菌BS调控甘蓝根肿病抗性机制。结果表明：与Pb处理相比，Pb + BS复合处理的Shannon指数呈上升趋势，二者根际微生物α多样性差异显著，表明BS处理提高了受Pb胁迫下的微生物群落多样性。与无菌水对照比较，施加BS有509个差异代谢物上调表达；k-means聚类分析显示，其中364种代谢物在特定趋势群集中丰度上调。施加BS处理显著增加了差异代谢物数量，富集在苯丙烷和黄酮类等防御相关代谢通路，并重塑了微生物—代谢物关联网络。关联分析表明，黏球菌（Myxococcota）、硝化螺旋菌（Nitrospirota）和甲基异螺旋菌（Methylomirabilis）的富集与优化根际氮转化效率、降低活性氮胁迫相关。同时，BS诱导防御酶（POD和SOD）时序性激活（POD峰值达对照组的10.43倍），复合处理（Pb + BS）的病情指数显著降低（盆栽降低54.5%），有效抑制了根肿菌增殖。本研究表明，枯草芽孢杆菌通过重塑根际微生物群落结构和阶段性激活植物防御系统，形成协同抗病效应，从而有效抑制甘蓝根肿病，为植物—微生物互作理论及病害生物防治提供重要依据。

关键词: 甘蓝, 根肿病, 枯草芽孢杆菌, 根际微生物, 植物—微生物互作, 代谢组学

Abstract:

Clubroot disease，caused by Plasmodiophora brassicae（Pb），severely impacts the yield and quality of cabbage. The regulatory mechanism of Bacillus subtilis（BS）on the rhizosphere microecology，as a biocontrol strain，remains unclear. In this study，by setting up root irrigation with BS，Pb，Pb + BS combined treatments，and a sterile water control，combined with 16S rDNA high-throughput sequencing and rhizosphere metabolomics analysis，and measuring physiological indices such as disease index，biomass，and defensive enzyme activity，we systematically revealed the mechanism by which Bacillus subtilis（BS）regulates clubroot resistance in cabbage. The results showed that the Shannon index of the Pb + BS combined treatment increased compared with the Pb group，and the rhizosphere microbial α diversity differed significantly between the Pb group and the Pb + BS group，indicating that BS treatment improved the diversity of the microbial community under Pb stress. Compared with the sterile water control，509 differential metabolites were upregulated after BS treatment；k-means clustering analysis further showed that 364 of these metabolites were upregulated in specific trend clusters. BS treatment significantly increased the number of differential metabolites，enriched in defense-related metabolic pathways such as phenylpropane and flavonoids，and reshaped the microbial-metabolite association network. Association analysis showed that the enrichment of Myxococcota，Nitrospirota，and Methylomirabilis was associated with optimized rhizosphere nitrogen transformation efficiency and reduced reactive nitrogen stress. Meanwhile，BS induced the sequential activation of defense enzymes of POD and SOD（POD peak value reached 10.43 times that of the control group），and the disease index of the combined treatment（Pb + BS）was significantly reduced（54.5% reduction in potted plants），effectively inhibiting the proliferation of clubroot bacteria. This study demonstrates that Bacillus subtilis effectively inhibits clubroot disease in cabbage by reshaping the rhizosphere microbial community structure and periodically activating the plant defense system，thereby creating a synergistic disease resistance effect. This provides an important basis for the theory of plant-microbe interactions and the biological control of diseases.

Key words: cabbage, Plasmodiophora brassica, Bacillus subtilis, rhizosphere microbes, plant-microbe interactions, metabolomics

王艳花, 邵伟兴, 安冬燕, 梅雪雯, 李勤菲, 司军. 枯草芽孢杆菌调控甘蓝根际微生物多样性及根肿病抗性效应研究[J]. 园艺学报, 2026, 53(6): 1779-1792.

WANG Yanhua, SHAO Weixing, AN Dongyan, MEI Xuewen, LI Qinfei, SI Jun. Bacillus subtilis Regulates the Diversity of Rhizosphere Microorganisms and the Resistance to Clubroot in Cabbage[J]. Acta Horticulturae Sinica, 2026, 53(6): 1779-1792.

图/表 7

图1 不同处理下甘蓝的病情指数及生长指标 CK：无菌水对照；Pb：根肿菌处理；BS：枯草芽孢杆菌处理；Pb + BS：复合处理。多重比较差异显著性分析，P < 0.05。下同

Fig. 1 Effects of different treatment groups on the incidence index and growth of Brassica oleracea CK：Sterile water control；Pb：Plasmodiophora brassica treatment group；BS：Bacillus subtilis treatment group；Pb + BS：Combined treatment group. Multiple comparisons were performed for significance，P < 0.05. The same below

图2 不同处理下甘蓝叶片POD和SOD活性的动态变化

Fig. 2 Dynamic changes in POD and SOD activities in Brassica oleracea leaves under different treatments

图3 不同处理下甘蓝根际土样微生物群落的多样性与组成 A：PCA；B：Shannon多样性（*、**号标示组间t检验差异显著，α = 0.05，α = 0.01）；C：聚类树—柱状图复合图中，丰度排名前10的物种

Fig. 3 Diversity and composition of rhizosphere microbial communities in Brassica oleracea under different treatments A：PCA；B：Shannon diversity，* and ** indicates the significant difference in the t-test（α = 0.05，α = 0.01）；C：Clustering tree-bar chart，the top 10 species in terms of species abundance

图4 甘蓝根际各处理组与对照组比较的差异代谢物火山图

Fig. 4 Differences in metabolite volcano plots between different treatment groups and the control group in the rhizosphere of Brassica oleracea

图5 甘蓝根际差异代谢物的k-means聚类分析括号中为代谢物数量

Fig. 5 Cluster analysis of differential metabolites in the rhizosphere of Brassica oleracea The number of metabolites is in brackets

图6 甘蓝根际共有差异代谢物KEGG富集图

Fig. 6 KEGG enrichment pathway map of common differential metabolites in the rhizosphere of Brassica oleracea

图7 不同处理下甘蓝根际代谢物聚类群组与微生物门类的相关性热图红色表示正相关，而蓝色表示负相关。*、**和***号表示代谢物簇和微生物之间显著相关（P < 0.05，P < 0.01和P < 0.001）

Fig. 7 Correlation heatmap of rhizosphere metabolite clusters and microbial phyla in Brassica oleracea under different treatments Red indicates a positive correlation，while blue indicates a negative correlation.*，** and *** indicate significant correlations between metabolite clusters and microorganisms（P < 0.05，P < 0.01 and P < 0.001）

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