南方农业学报2025，Vol.56Issue(3)：840-852,13.DOI:10.3969/j.issn.2095-1191.2025.03.015
一种烟草叶斑类病害的诊断及其叶际微生态特征分析
Diagnosis of a type of tobacco leaf spot disease and its phyllosphere microecological characteristics
摘要
Abstract
[Objective]To diagnose an unknown leaf spot disease of tobacco and study its microecological characteris-tics,which provided scientific basis for the precise prevention and control of this unknown tobacco leaf spot disease.[Method]Using diseased tobacco leaves with unknown leaf spot disease collected from the tobacco growing region of Shibing in Guizhou as materials,the phyllosphere fungi were isolated,purified and identified by tissue culture method.Based on Koch's postulates,screening and verification of pathogenic bacteria was conducted by stab inoculation.High-throughput sequencing technology(ITS and 16S rRNA)was utilized to analyze the phyllosphere microbial communities and diversity.[Result]Four strains of fungi SBR111,SBR121,SBR222 and SBR232 were isolated from the diseased to-bacco leaves.Strains SBR111 and SBR121 were identified as Didymella,and strains SBR222 and SBR23 were identified as Hypoxylon by molecular identification.According to Koch's postulates,strain SBR121 was confirmed as the patho-genic fungus,and morphological and molecular identification classified it as Didymella segeticola.The phyllospheric dominant fungi phyla of diseased and healthy leaves all were Ascomycota,Basidiomycota and Mortierellomycota,and the phyllospheric dominant bacteria phyla of diseased and healthy leaves all were Proteobacteria and Firmicutes.For di-seased leaves,the dominant genera of fungi were Phoma,Alternaria and Boeremia,while those of bacteria were Pseudo-monas,unidentified_Rhizobiaceae,Stenotrophomonas and Sphingomonas.For healthy leaves,the dominant genera of fungi were Alternaria,Phoma,Boeremia and Hannaella,and those of bacteria were Pseudomonas,Sphingomonas,un-identified_Rhizobiaceae and Methylobacterium.For phyllosphere fungi Alpha diversity index of healthy tobacco leaves,except that the coverage index was equal to that of diseased tobacco leaves,diversity index and abundance index were higher than diseased tobacco leaves.The phyllosphere bacteria diversity index and abundance index of healthy tobacco leaves were lower than that of diseased tobacco leaves.The microbial community functional prediction results indicated that phyllosphere fungi in diseased leaves were mainly distributed in plant_pathogens-wood_saprotroph fungi group,while the phyllosphere fungi in healthy leaves were mainly distributed animal_pathogen-endophyte-plant_pathogen-wood_saprotroph and endophyte-plant_pathogen groups;the phyllosphere bacteria of diseased and healthy tobacco leaves were mainly distributed in 3 pathways:metabolism,genetic information processing and environmental information pro-cessing.[Conclusion]Didymella segeticola is the pathogen of tobacco leaf spot in Shibing tobacco growing region of Gui-zhou.There are differences in the community structure and diversity of phyllosphere fungi and bacteria between healthy and diseased tobacco leaves.The diversity and richness indexes of phyllosphere fungi in diseased tobacco leaves are lower than those in healthy tobacco leaves,and the diversity index and richness index of phyllosphere bacteria are higher than those in healthy tobacco leaves.关键词
烟草/亚隔孢壳属/叶斑病/扩增子高通量测序/真菌群落/细菌群落Key words
tobacco/Didymella/leaf spot disease/amplicon high-throughput sequencing/fungi community/bacte-ria community分类
农业科技引用本文复制引用
林丽,雷文洁,汪汉成,陈丽莉,蔡刘体,陈兴江,彭丽娟..一种烟草叶斑类病害的诊断及其叶际微生态特征分析[J].南方农业学报,2025,56(3):840-852,13.基金项目
国家自然科学基金项目(32160522) （32160522）
贵州省"百层次"创新型人才项目(黔科合平台人才-GCC[2022]028-1) （黔科合平台人才-GCC[2022]028-1）
贵州省科技创新人才团队项目(黔科合平台人才-CXTD[2023]021) （黔科合平台人才-CXTD[2023]021）
中国烟草总公司贵州省公司科技项目(2024XM06) National Natural Science Foundation of China(32160522) （2024XM06）
Guizhou"Hundred Layers"Innova-tive Talent Project(QKHPTRC-GCC[2022]028-1) （QKHPTRC-GCC[2022]028-1）
Guizhou Science and Technology Innovation Talent Team Project(QKHPTRC-CXTD[2023]021) （QKHPTRC-CXTD[2023]021）
Guizhou Company Science and Technology Project of China National Tobacco Corpo-ration(2024XM06) （2024XM06）
