丁香酚对青枯雷尔氏菌的抑菌效果及作用机制  

The antibacterial effect and action mechanism of eugenol on bacterial wilt pathogen Ralstonia solanacearum

作　　者：刘泽鑫 常晓涵 许汝冰 许雯慧 杨勇[1] 向海波 黎妍妍 Liu Zexin;Chang Xiaohan;Xu Rubing;Xu Wenhui;Yang Yong;Xiang Haibo;Li Yanyan(State Key Laboratory of Biocatalysis and Enzyme Engineering,College of Life Sciences,Hubei University,Wuhan 430062,Hubei Province,China;Tobacco Research Institute of Hubei Province,Wuhan 430030,Hubei Province,China)

机构地区：[1]湖北大学生命科学学院,省部共建生物催化与酶工程国家重点实验室,武汉430062 [2]湖北省烟草科学研究院,武汉430030

出　　处：《植物保护学报》2025年第1期211-221,共11页Journal of Plant Protection

基　　金：中国烟草总公司重大专项项目(110202101047(LS-07),110202201019(LS-03));湖北省烟草公司重点科技项目(027Y2022-023)。

摘　　要：为探究丁香酚对青枯病致病菌的抑菌活性和作用机制,以青枯雷尔氏菌Ralstonia solanacearum为研究对象,采用微量二倍稀释法、琼脂扩散法、毛细管法和结晶紫染色法等方法测定丁香酚对青枯雷尔氏菌最小抑菌浓度(minimal inhibitory concentration,MIC)、最低杀菌浓度(minimum bactericidal concentration,MBC)、运动性、趋化性和生物被膜的影响;对丁香酚处理与否的青枯雷尔氏菌进行转录组分析,并使用实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)技术对几个关键基因表达量进行测定。结果表明:丁香酚对青枯雷尔氏菌的MIC为125μg/mL,MBC为500μg/mL。当丁香酚浓度大于62.5μg/mL时,能著抑制青枯雷尔氏菌的生长、运动性和生物被膜的形成;当丁香酚浓度大于125μg/mL时,能显著促进青枯雷尔氏菌的趋避性。转录组分析结果显示,丁香酚处理的青枯雷尔氏菌中有174个差异表达基因(differentially expressed gene,DEG)上调,347个DEG下调;GO分析结果表明DEG主要富集在前体代谢物与能量的生成、非膜结合的细胞器和RNA结合等生物学过程中;KEGG分析表明DEG主要富集在碳代谢、氧化磷酸化和核糖体等信号通路中。To investigate the antibacterial activity and action mechanism of eugenol against the patho‐gens causing bacterial wilt,Ralstonia solanacearum was used as the target bacterium.Approaches such as the microdilution method,agar diffusion assay,capillary assay,and crystal violet staining were em‐ployed to determine the minimal inhibitory concentration(MIC),minimum bactericidal concentration(MBC),motility,chemotaxis,and biofilm formation of R.solanacearum under treatment with eugenol.Transcriptome analysis was performed on R.solanacearum with and without eugenol treatment,and the expression levels of several key genes were measured using real-time quantitative PCR(RT-qPCR).The results showed that the MIC of eugenol against R.solanacearum was 125μg/mL,and the MBC was 500μg/mL.When the concentration of eugenol exceeded 62.5μg/mL,it significantly inhibited the growth,motility,and biofilm formation of R.solanacearum.At concentrations above 125μg/mL,eugenol significantly promoted the repellent behavior of R.solanacearum.Transcriptome analysis revealed 174 upregulated differentially expressed genes(DEGs)and 347 downregulated DEGs in eugenol-treated R.solanacearum.Gene ontology(GO)analysis indicated that the DEGs were primarily enriched in meta‐bolic processes such as generation of precursor metabolites and energy,non-membrane-bound organ‐elle,and RNA binding.Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis showed that the DEGs were mainly enriched in pathways such as carbon metabolism,oxidative phosphoryla‐tion,and ribosome.

关 键 词：丁香酚 青枯雷尔氏菌 青枯病 转录组 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]