枯草芽胞杆菌GXHZ16抑制烟草疫霉病菌的表达谱分析

doi:10.16409/j.cnki.2095-039x.2023.11.022

摘要/Abstract

摘要： 烟草黑胫病由烟草疫霉菌Phytophthora nicotianae侵染引起，是烟草生产上一种分布广泛、为害严重的土传病害。本实验室前期从广西贺州富川烟区健康烟株根际土壤中分离筛选获得一株枯草芽胞杆菌Bacillus subtilis GXHZ16，对峙培养发现该菌株对烟草疫霉菌具有良好的拮抗活性（抑制率65.79%）。为探究菌株GXHZ16对烟草疫霉菌的可能抑制分子机理，本研究分别针对枯草芽胞杆菌GXHZ16与烟草疫霉菌对峙培养36 h（处理组）和0 h（对照组）的混合样品，采用高通量转录组测序（RNA-Seq）技术结合生物信息学分析对峙培养36 h后枯草芽胞杆菌的基因差异表达谱及所涉及的信号通路变化。结果发现：对峙培养36 h后枯草芽胞杆菌GXHZ16有1607个基因发生了显著差异表达，包括810个上调表达基因和797个下调表达基因；其中筛选获得的显著表达基因特别是上调表达的基因（包括鞭毛装配、细菌趋化性相关基因以及与表面活性素、丰原素和制磷脂菌素合成相关的10个相关基因）可能均涉及枯草芽胞杆菌对烟草疫霉菌的抑制作用。进一步的GO富集分析显示，共有1343个差异表达基因被注释到不同的功能亚类中，涉及差异基因最多的为细胞组分类别中的细胞解剖实体。特别地，KEGG富集分析显示，共有551个差异表达基因归到数百个Pathway中，其中涉及差异表达基因最多的是微生物在不同环境中的代谢途径；其他主要代谢途径有鞭毛装配和淀粉蔗糖代谢等；此外，细菌趋化性通路和单环菌素生物合成通路也高度富集。总之，本研究从转录组水平分析获得了枯草芽胞杆菌（与烟草疫霉对峙培养后）的基因差异表达图谱及相关调控代谢途径；为进一步阐明芽胞杆菌抑制植物病原真菌的生防机制并提高枯草芽胞杆菌的生防效果奠定了基础。

关键词: 枯草芽胞杆菌, 烟草疫霉菌, 转录组分析, 差异表达基因

Abstract: Tobacco black shank caused by Phytophthora nicotianae, is a soil-borne disease with wide distribution and serious damage in tobacco production. Bacillus subtilis (GXHZ16 strain) in our laboratory, which exhibited high antagonistic activity against P. nicotiana (inhibition rate: 65.79%), was previously isolated and identified from the rhizosphere soil of healthy tobacco in Fuchuan County, Hezhou, Guangxi Province. To explore the possible inhibitory molecular mechanism of GXHZ16 strain against P. nicotiana, the mixed samples of B. subtilis GXHZ16 andP. nicotianacultured for 36 h (treatment group) and 0 h (control group) were collected, respectively, and then the differentially expressed genes and related signaling pathways in B. subtilis GXHZ16 (cultured for 36 h against P. nicotiana) were investigated by high-throughput transcriptome sequencing (RNA-Seq) and bioinformatics analysis. Our results showed that 1607 genes of B. subtilisGXHZ16 were differentially expressed after 36 h of confrontation culture, including 810 up-regulated genes and 797 down-regulated genes. These differentially expressed genes with up-regulation, including genes related to Flagellar assembly and Bacterial chemotaxis, as well as ten related genes associated with the synthesis of surfactin, fengycin and plipastatin, may be involved in the inhibition of B. subtilisagainst P. nicotiana.Furthermore, GO enrichment analysis showed that a total of 1343 differentially expressed genes have been annotated into different functional subclasses, and the cellular anatomical entity in cell component category was the one with the most differentially expressed genes. More importantly, KEGG enrichment analysis showed that 551 differentially expressed genes were classified into hundreds of pathways. The most differentially expressed genes were involved in the pathway of Microbial metabolism in diverse environments. Other major metabolic pathways included Flagellar assembly and Starch and sucrose metabolism. Moreover, Bacterial chemotaxis pathway and Monobactam biosynthesis pathway were highly enriched. In conclusion, gene expression profiles of B. subtilis (after confrontation culture with P. nicotiana) and related regulatory metabolic pathways were acquired based on transcriptomic analysis in this study, which will provide a basis for further elucidate the biocontrol mechanism of Bacillus to inhibit plant pathogenic fungi and improve the biocontrol efficiency of B. subtilis.

Key words: Bacillus subtilis, Phytophthora nicotianae, transcriptomic analysis, differentially expressed genes

中图分类号:

S476

胡亚杰, 范啟铃, 李群岭, 周效峰, 丁婷, 汪章勋, 吴峰. 枯草芽胞杆菌GXHZ16抑制烟草疫霉病菌的表达谱分析[J]. 中国生物防治学报, 2024, 40(3): 608-619.

HU Yajie, FAN Qiling, Li Qunling, ZHOU Xiaofeng, DING Ting, WANG Zhangxun, WU Feng. Expression Profiles of Bacillus subtilis GXHZ16 against Phytophthora nicotiana[J]. Chinese Journal of Biological Control, 2024, 40(3): 608-619.

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