小麦赤霉病拮抗菌JB7的生防特性

doi:10.13560/j.cnki.biotech.bull.1985.2024-1187

摘要/Abstract

摘要：

目的 Bacillus methylotrophicus JB7可以抑制Fusarium graminearum（Fg）的生长并能控制小麦赤霉病，但其抑菌及控病机制并不明确，探究其抑菌及控病机制，为其应用提供理论依据。方法通过对禾谷镰刀菌孢子萌发、抑菌能力和抗氧化酶活性测定等方法，研究JB7无菌上清液（cell-free supernatants, CFS）对Fg孢子数量、菌丝生长和抗氧化酶活性的影响，利用转录组学测序技术分析差异表达基因（DEGs），扩增子测序技术评估喷施JB7后对麦穗毒素含量和微生物群落结构的影响。结果菌株JB7的CFS对Fg菌丝生长和分生孢子萌发具有较强的抑制作用，同时CFS引起Fg菌丝超氧化物歧化酶（superoxide dismutase, SOD）、过氧化氢酶（catalase, CAT）活性上升，过氧化物酶（peroxidase, POD）和谷胱甘肽还原酶（glutathione reductase, GR）活性下降。转录组学分析表明，菌株JB7 CFS影响了与氧化应激和毒素有关基因的表达，喷施JB7改变了麦穗中微生物的群落结构，降低了麦穗中Fg的密度和毒素含量。结论菌株JB7的CFS通过触发氧化应激和降低毒素水平展现出对Fg的抗真菌作用，且菌株JB7能改变麦穗微生物群落结构，降低Fg的密度和毒素含量。

关键词: 小麦赤霉病, 禾谷镰刀菌, 甲基营养型芽胞杆菌, 转录组学, 扩增子测序

Abstract:

Objective Bacillus methylotrophicus JB7 may inhibit Fusarium graminearum (Fg) growth and control Fusarium head blight in wheat; however, the mechanisms underlying its antifungal activity and disease management remain unclear. In this study, the antifungal and disease control mechanisms of B. methylotrophicus were explored, which may provide a theoretical basis for its further application. Method The effects of cell-free supernatants (CFS) from strain JB7 on the number of Fg spores, mycelium growth and antioxidant enzyme activity of Fg were studied through assessment of spore germination, antifungal activity and antioxidant enzyme activity. Transcriptomic sequencing was employed to analyze differentially expressed genes (DEGs), while amplicon sequencing was used to evaluate the effects of JB7 spraying on wheat ears mycotoxin content and microbial community structure. Result The CFS from strain JB7 showed a strong inhibitory effect on mycelial growth and germination of Fg. Additionally, the CFS led to an increase in the activity of superoxide dismutase (SOD) and catalase (CAT) in Fg mycelium, while causing a reduction in peroxidase (POD) and glutathione reductase (GR) activities. Transcriptome analysis indicates that the CFS from strain JB7 affected the expression of genes related to oxidative stress and toxin production. The application of JB7 resulted in an alteration of microbial community structure in wheat ears, leading to a decrease in Fg density and mycotoxin content. Conclusion The CFS from strain JB7 shows antifungal effects on Fg by triggering oxidative stress and reducing mycotoxin levels, and strain JB7 may change the microbial community structure in wheat ears and reduce the density and mycotoxin content of Fg.

Key words: Fusarium head blight, Fusarium graminearum, Bacillus methylotrophicus, transcriptome, amplicon sequencing

张越, 毕钰, 慕雪男, 郑子薇, 王志刚, 徐伟慧. 小麦赤霉病拮抗菌JB7的生防特性[J]. 生物技术通报, 2025, 41(7): 261-271.

ZHANG Yue, BI Yu, MU Xue-nan, ZHENG Zi-wei, WANG Zhi-gang, XU Wei-hui. Biocontrol Characteristics of Strain JB7 against Fusarium graminearum[J]. Biotechnology Bulletin, 2025, 41(7): 261-271.

图/表 6

图1 菌株JB7对禾谷镰刀菌（Fg）的抑制效果A：菌株JB7不同浓度CFS对孢子萌发的影响；B：菌株JB7中挥发物质对Fg的抑菌效果；C：菌株JB7中挥发物质对Fg的抑菌效果平板图。不同小写字母表示处理间差异显著（P<0.05）。****表示在P<0.000 1水平存在显著差异。下同

Fig. 1 Inhibitory effect of strain JB7 on Fusarium graminearum (Fg)A: The effect of different concentrations of CFS from strain JB7 on spore germination. B: The inhibitory effect of volatile compounds from strain JB7 on Fg. C: Plate of the inhibitory effect of volatile compounds from strain JB7 on Fg. The different lowercase letters indicate significant differences among treatments (P<0.05). **** indicates significant differences at P<0.000 1 . The same below

图2 菌株JB7 CFS对Fg菌丝抗氧化酶活性的影响*、**、***分别表示在P<0.05、P<0.01、P<0.001水平存在显著差异。下同

Fig. 2 Effects of CFS from strain JB7 on the antioxidant enzyme activities of Fg mycelia*, **, and *** indicate significant differences at P<0.05, P<0.01, and P<0.001, respectively. The same below

图3 Fg差异表达基因富集分析A：差异基因的GO富集分析；B：差异基因的KEGG富集分析

Fig. 3 Enrichment analysis of DEGs in FgA: GO enrichment analysis of DEGs; B: KEGG enrichment analysis of DEGs

图4 与氧化应激和毒素有关基因的表达分析A：与氧化应激有关基因的聚类热图；B：与毒素有关基因的聚类热图；C：DON的生物合成；D：利用RT-qPCR检测与氧化应激和毒力因子有关基因的表达

Fig. 4 Expression analysis of genes related to oxidative stress and mycotoxin in FgA: Clustering heatmap of genes related to reactive oxygen species. B: Clustering heatmap of genes related to mycotoxin. C: Biosynthesis of deoxynivalenol. D: Gene expression of genes related to oxidative stress and mycotoxin using RT-qPCR

图5 Fg在麦穗上定殖及其分泌毒素的量A：JB7组麦穗颖片剖面图；B：JB7组组织内部剖面图；C：W组麦穗颖片剖面图；D：W组组织内部剖面图；E：麦穗上真菌毒素含量；箭头表示Fg菌丝分布

Fig. 5 Colonization of Fg in wheat ears and its mycotoxin secretionA: Profile map of glumes wheat ears in JB7 treatment. B: Profile map of inside tissues in JB7 treatment. C: Profile map of glumes wheat ears in water treatment. D: Profile map of inside tissues in water treatment. E: Mycotoxin content in wheat ears. The arrows indicate the distribution of the Fg mycelium

图6 菌株JB7对麦穗中微生物群落结构的影响A：基于Bray-Curtis距离的细菌群落的主坐标分析；B：基于Bray-Curtis距离的真菌群落的主坐标分析；C：不同处理中细菌属水平的相对丰度；D：不同处理中真菌属水平的相对丰度；E：麦穗中Bacillus的绝对丰度；F：麦穗中Fg的相对丰度

Fig. 6 Effects of strain JB7 on microbial community structure in wheat earsA: Principal coordinate analysis of bacterial community based on Bray-Curtis distance. B: Principal coordinate analysis of fungal community based on Bray-Curtis distance. C: Relative abundance of bacterial genera in different treatments. D: Relative abundance of fungal genera in different treatments. E: Absolute abundance of Bacillus in wheat ears. F: Relative abundance of Fg in wheat ears

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