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The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment
BACKGROUND: The mosquito gut harbors a variety of bacteria that are dynamically associated with mosquitoes in various contexts. However, little is known about bacterial factors that affect bacterial inhabitation in the gut microbial community. Enterobacter sp. Ag1 is a predominant Gram negative bact...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4549878/ https://www.ncbi.nlm.nih.gov/pubmed/26306887 http://dx.doi.org/10.1186/s13071-015-1049-1 |
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author | Pei, Dong Jiang, Jinjin Yu, Wanqin Kukutla, Phanidhar Uentillie, Alejandro Xu, Jiannong |
author_facet | Pei, Dong Jiang, Jinjin Yu, Wanqin Kukutla, Phanidhar Uentillie, Alejandro Xu, Jiannong |
author_sort | Pei, Dong |
collection | PubMed |
description | BACKGROUND: The mosquito gut harbors a variety of bacteria that are dynamically associated with mosquitoes in various contexts. However, little is known about bacterial factors that affect bacterial inhabitation in the gut microbial community. Enterobacter sp. Ag1 is a predominant Gram negative bacterium in the mosquito midgut. METHODS: In a mutant library that was generated using transposon Tn5-mediated mutagenesis, a mutant was identified, in which the gene waaL was disrupted by the Tn5 insertion. The waaL encodes O antigen ligase, which is required for the attachment of O antigen to the outer core oligosaccharide of the lipopolysaccharide (LPS). RESULTS: The waaL(−) mutation caused the O antigen repeat missing in the LPS. The normal LPS structure was restored when the mutant was complemented with a plasmid containing waaL gene. The waaL(−) mutation did not affect bacterial proliferation in LB culture, the mutant cells grew at a rate the same as the wildtype (wt) cells. However, when waaL(−) strain were co-cultured with the wt strain or complemented strain, the mutant cells proliferated with a slower rate, indicating that the mutants were less competitive than wt cells in a community setting. Similarly, in a co-feeding assay, when fluorescently tagged wt strain and waaL(−) strain were orally co-introduced into the gut of Anopheles stephensi mosquitoes, the mutant cells were less prevalent in both sugar-fed and blood-fed guts. The data suggest that the mutation compromised the bacterial inhabitation in the gut community. Besides, the mutant was more sensitive to oxidative stress, demonstrated by lower survival rate upon exposure to 20 mM H(2)O(2). CONCLUSION: Lack of the O antigen structure in LPS of Enterobacter compromised the effective growth in co-culture and co-feeding assays. In addition, O-antigen was involved in protection against oxidative stress. The findings suggest that intact LPS is crucial for the bacteria to steadily stay in the gut microbial community. |
format | Online Article Text |
id | pubmed-4549878 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-45498782015-08-27 The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment Pei, Dong Jiang, Jinjin Yu, Wanqin Kukutla, Phanidhar Uentillie, Alejandro Xu, Jiannong Parasit Vectors Research BACKGROUND: The mosquito gut harbors a variety of bacteria that are dynamically associated with mosquitoes in various contexts. However, little is known about bacterial factors that affect bacterial inhabitation in the gut microbial community. Enterobacter sp. Ag1 is a predominant Gram negative bacterium in the mosquito midgut. METHODS: In a mutant library that was generated using transposon Tn5-mediated mutagenesis, a mutant was identified, in which the gene waaL was disrupted by the Tn5 insertion. The waaL encodes O antigen ligase, which is required for the attachment of O antigen to the outer core oligosaccharide of the lipopolysaccharide (LPS). RESULTS: The waaL(−) mutation caused the O antigen repeat missing in the LPS. The normal LPS structure was restored when the mutant was complemented with a plasmid containing waaL gene. The waaL(−) mutation did not affect bacterial proliferation in LB culture, the mutant cells grew at a rate the same as the wildtype (wt) cells. However, when waaL(−) strain were co-cultured with the wt strain or complemented strain, the mutant cells proliferated with a slower rate, indicating that the mutants were less competitive than wt cells in a community setting. Similarly, in a co-feeding assay, when fluorescently tagged wt strain and waaL(−) strain were orally co-introduced into the gut of Anopheles stephensi mosquitoes, the mutant cells were less prevalent in both sugar-fed and blood-fed guts. The data suggest that the mutation compromised the bacterial inhabitation in the gut community. Besides, the mutant was more sensitive to oxidative stress, demonstrated by lower survival rate upon exposure to 20 mM H(2)O(2). CONCLUSION: Lack of the O antigen structure in LPS of Enterobacter compromised the effective growth in co-culture and co-feeding assays. In addition, O-antigen was involved in protection against oxidative stress. The findings suggest that intact LPS is crucial for the bacteria to steadily stay in the gut microbial community. BioMed Central 2015-08-27 /pmc/articles/PMC4549878/ /pubmed/26306887 http://dx.doi.org/10.1186/s13071-015-1049-1 Text en © Pei et al. 2015 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Pei, Dong Jiang, Jinjin Yu, Wanqin Kukutla, Phanidhar Uentillie, Alejandro Xu, Jiannong The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment |
title | The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment |
title_full | The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment |
title_fullStr | The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment |
title_full_unstemmed | The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment |
title_short | The waaL gene mutation compromised the inhabitation of Enterobacter sp. Ag1 in the mosquito gut environment |
title_sort | waal gene mutation compromised the inhabitation of enterobacter sp. ag1 in the mosquito gut environment |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4549878/ https://www.ncbi.nlm.nih.gov/pubmed/26306887 http://dx.doi.org/10.1186/s13071-015-1049-1 |
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