Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves
In the tobacco phyllosphere, some of the microbes may have detrimental effects on plant health, while many may be neutral or even beneficial. Some cannot be cultivated, so culture-independent methods are needed to explore microbial diversity. In this study, both metagenetic analysis and traditional...
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| Format: | Article |
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.843389/full |
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| author | Li-Gang Xiang Li-Gang Xiang Han-Cheng Wang Feng Wang Liu-Ti Cai Wen-Hong Li Tom Hsiang Zhi-He Yu |
| author_facet | Li-Gang Xiang Li-Gang Xiang Han-Cheng Wang Feng Wang Liu-Ti Cai Wen-Hong Li Tom Hsiang Zhi-He Yu |
| author_sort | Li-Gang Xiang |
| collection | DOAJ |
| description | In the tobacco phyllosphere, some of the microbes may have detrimental effects on plant health, while many may be neutral or even beneficial. Some cannot be cultivated, so culture-independent methods are needed to explore microbial diversity. In this study, both metagenetic analysis and traditional culture-dependent methods were used on asymptomatic healthy leaves and symptomatic diseased leaves of tobacco plants. In the culture-independent analysis, asymptomatic leaves had higher microbial diversity and richness than symptomatic leaves. Both asymptomatic and symptomatic leaves contained several potentially pathogenic bacterial and fungal genera. The putative bacterial pathogens, such as species of Pseudomonas, Pantoea, or Ralstonia, and putative fungal pathogens, such as species of Phoma, Cladosporium, Alternaria, Fusarium, Corynespora, and Epicoccum, had a higher relative abundance in symptomatic leaves than asymptomatic leaves. FUNGuild analysis indicated that the foliar fungal community also included endophytes, saprotrophs, epiphytes, parasites, and endosymbionts. PICRUSt analysis showed that the dominant functions of the bacterial community in a symptomatic leaf were cellular processes and environmental information processing. In the other five foliar samples, the dominant functions of the bacterial community were genetic information processing, metabolism, and organismal systems. In the traditional culture-dependent method, 47 fungal strains were isolated from 60 symptomatic tobacco leaf fragments bearing leaf spots. Among them, 21 strains of Colletotrichum (29%), Xylariaceae (14%), Corynespora (14%), Pestalotiopsis (10%), Alternaria (10%), Epicoccum (10%), Byssosphaeria (5%), Phoma (5%), and Diaporthe (5%) all fulfilled Koch’s postulates and were found to cause disease on detached tobacco leaves in artificial inoculation tests. Symptoms on detached leaves caused by three strains of Corynespora cassiicola in artificial inoculation tests were similar to the original disease symptoms in the tobacco field. This study showed that the combined application of culture-dependent and independent methods could give comprehensive insights into microbial composition that each method alone did not reveal. |
| first_indexed | 2024-12-11T13:16:42Z |
| format | Article |
| id | doaj.art-f9d62575d05844e4adea5f3390acf7be |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-11T13:16:42Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-f9d62575d05844e4adea5f3390acf7be2022-12-22T01:06:03ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-04-011310.3389/fmicb.2022.843389843389Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco LeavesLi-Gang Xiang0Li-Gang Xiang1Han-Cheng Wang2Feng Wang3Liu-Ti Cai4Wen-Hong Li5Tom Hsiang6Zhi-He Yu7College of Agriculture, Yangtze University, Jingzhou, ChinaGuizhou Provincial Academician Workstation of Microbiology and Health, Guizhou Academy of Tobacco Science, Guiyang, ChinaGuizhou Provincial Academician Workstation of Microbiology and Health, Guizhou Academy of Tobacco Science, Guiyang, ChinaGuizhou Provincial Academician Workstation of Microbiology and Health, Guizhou Academy of Tobacco Science, Guiyang, ChinaGuizhou Provincial Academician Workstation of Microbiology and Health, Guizhou Academy of Tobacco Science, Guiyang, ChinaGuizhou Institute of Plant Protection, Guizhou Academy of Agricultural Sciences, Guiyang, ChinaSchool of Environmental Sciences, University of Guelph, Guelph, ON, CanadaCollege of Life Sciences, Yangtze University, Jingzhou, ChinaIn the tobacco phyllosphere, some of the microbes may have detrimental effects on plant health, while many may be neutral or even beneficial. Some cannot be cultivated, so culture-independent methods are needed to explore microbial diversity. In this study, both metagenetic analysis and traditional culture-dependent methods were used on asymptomatic healthy leaves and symptomatic diseased leaves of tobacco plants. In the culture-independent analysis, asymptomatic leaves had higher microbial diversity and richness than symptomatic leaves. Both asymptomatic and symptomatic leaves contained several potentially pathogenic bacterial and fungal genera. The putative bacterial pathogens, such as species of Pseudomonas, Pantoea, or Ralstonia, and putative fungal pathogens, such as species of Phoma, Cladosporium, Alternaria, Fusarium, Corynespora, and Epicoccum, had a higher relative abundance in symptomatic leaves than asymptomatic leaves. FUNGuild analysis indicated that the foliar fungal community also included endophytes, saprotrophs, epiphytes, parasites, and endosymbionts. PICRUSt analysis showed that the dominant functions of the bacterial community in a symptomatic leaf were cellular processes and environmental information processing. In the other five foliar samples, the dominant functions of the bacterial community were genetic information processing, metabolism, and organismal systems. In the traditional culture-dependent method, 47 fungal strains were isolated from 60 symptomatic tobacco leaf fragments bearing leaf spots. Among them, 21 strains of Colletotrichum (29%), Xylariaceae (14%), Corynespora (14%), Pestalotiopsis (10%), Alternaria (10%), Epicoccum (10%), Byssosphaeria (5%), Phoma (5%), and Diaporthe (5%) all fulfilled Koch’s postulates and were found to cause disease on detached tobacco leaves in artificial inoculation tests. Symptoms on detached leaves caused by three strains of Corynespora cassiicola in artificial inoculation tests were similar to the original disease symptoms in the tobacco field. This study showed that the combined application of culture-dependent and independent methods could give comprehensive insights into microbial composition that each method alone did not reveal.https://www.frontiersin.org/articles/10.3389/fmicb.2022.843389/fulltobaccomicrobial diversitypathogenmetagenetic analysisculture-dependent method |
| spellingShingle | Li-Gang Xiang Li-Gang Xiang Han-Cheng Wang Feng Wang Liu-Ti Cai Wen-Hong Li Tom Hsiang Zhi-He Yu Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves Frontiers in Microbiology tobacco microbial diversity pathogen metagenetic analysis culture-dependent method |
| title | Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves |
| title_full | Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves |
| title_fullStr | Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves |
| title_full_unstemmed | Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves |
| title_short | Analysis of Phyllosphere Microorganisms and Potential Pathogens of Tobacco Leaves |
| title_sort | analysis of phyllosphere microorganisms and potential pathogens of tobacco leaves |
| topic | tobacco microbial diversity pathogen metagenetic analysis culture-dependent method |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.843389/full |
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