Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>

<i>Aliarcobacter butzleri</i> is an emergent enteropathogen, showing high genetic diversity, which likely contributes to its adaptive capacity to different environments. Whether natural transformation can be a mechanism that generates genetic diversity in <i>A. butzleri</i> i...

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Main Authors: Marina Bonifácio, Cristiana Mateus, Ana R. Alves, Emanuel Maldonado, Ana P. Duarte, Fernanda Domingues, Mónica Oleastro, Susana Ferreira
Format: Article
Language:English
Published: MDPI AG 2021-07-01
Series:Pathogens
Subjects:
Online Access:https://www.mdpi.com/2076-0817/10/7/909
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author Marina Bonifácio
Cristiana Mateus
Ana R. Alves
Emanuel Maldonado
Ana P. Duarte
Fernanda Domingues
Mónica Oleastro
Susana Ferreira
author_facet Marina Bonifácio
Cristiana Mateus
Ana R. Alves
Emanuel Maldonado
Ana P. Duarte
Fernanda Domingues
Mónica Oleastro
Susana Ferreira
author_sort Marina Bonifácio
collection DOAJ
description <i>Aliarcobacter butzleri</i> is an emergent enteropathogen, showing high genetic diversity, which likely contributes to its adaptive capacity to different environments. Whether natural transformation can be a mechanism that generates genetic diversity in <i>A. butzleri</i> is still unknown. In the present study, we aimed to establish if <i>A. butzleri</i> is naturally competent for transformation and to investigate the factors influencing this process. Two different transformation procedures were tested using exogenous and isogenic DNA containing antibiotic resistance markers, and different external conditions influencing the process were evaluated. The highest number of transformable <i>A. butzleri</i> strains were obtained with the agar transformation method when compared to the biphasic system (65% versus 47%). <i>A. butzleri</i> was able to uptake isogenic chromosomal DNA at different growth phases, and the competence state was maintained from the exponential to the stationary phases. Overall, the optimal conditions for transformation with the biphasic system were the use of 1 μg of isogenic DNA and incubation at 30 °C under a microaerobic atmosphere, resulting in a transformation frequency ~8 × 10<sup>−6</sup> transformants/CFU. We also observed that <i>A. butzleri</i> favored the transformation with the genetic material of its own strain/species, with the DNA incorporation process occurring promptly after the addition of genomic material. In addition, we observed that <i>A. butzleri</i> strains could exchange genetic material in co-culture assays. The presence of homologs of well-known genes involved in the competence in the <i>A. butzleri</i> genome corroborates the natural competence of this species. In conclusion, our results show that <i>A. butzleri</i> is a naturally transformable species, suggesting that horizontal gene transfer mediated by natural transformation is one of the processes contributing to its genetic diversity. In addition, natural transformation can be used as a tool for genetic studies of this species.
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spelling doaj.art-187dff054a8a4856aed4fd0298228db52023-11-22T04:39:23ZengMDPI AGPathogens2076-08172021-07-0110790910.3390/pathogens10070909Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>Marina Bonifácio0Cristiana Mateus1Ana R. Alves2Emanuel Maldonado3Ana P. Duarte4Fernanda Domingues5Mónica Oleastro6Susana Ferreira7CICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, PortugalCICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, PortugalCICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, PortugalC4-UBI-Cloud Computing Competence Centre, University of Beira Interior, 6200-284 Covilhã, PortugalCICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, PortugalCICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, PortugalNational Reference Laboratory for Gastrointestinal Infections, Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, 1649-016 Lisbon, PortugalCICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, Portugal<i>Aliarcobacter butzleri</i> is an emergent enteropathogen, showing high genetic diversity, which likely contributes to its adaptive capacity to different environments. Whether natural transformation can be a mechanism that generates genetic diversity in <i>A. butzleri</i> is still unknown. In the present study, we aimed to establish if <i>A. butzleri</i> is naturally competent for transformation and to investigate the factors influencing this process. Two different transformation procedures were tested using exogenous and isogenic DNA containing antibiotic resistance markers, and different external conditions influencing the process were evaluated. The highest number of transformable <i>A. butzleri</i> strains were obtained with the agar transformation method when compared to the biphasic system (65% versus 47%). <i>A. butzleri</i> was able to uptake isogenic chromosomal DNA at different growth phases, and the competence state was maintained from the exponential to the stationary phases. Overall, the optimal conditions for transformation with the biphasic system were the use of 1 μg of isogenic DNA and incubation at 30 °C under a microaerobic atmosphere, resulting in a transformation frequency ~8 × 10<sup>−6</sup> transformants/CFU. We also observed that <i>A. butzleri</i> favored the transformation with the genetic material of its own strain/species, with the DNA incorporation process occurring promptly after the addition of genomic material. In addition, we observed that <i>A. butzleri</i> strains could exchange genetic material in co-culture assays. The presence of homologs of well-known genes involved in the competence in the <i>A. butzleri</i> genome corroborates the natural competence of this species. In conclusion, our results show that <i>A. butzleri</i> is a naturally transformable species, suggesting that horizontal gene transfer mediated by natural transformation is one of the processes contributing to its genetic diversity. In addition, natural transformation can be used as a tool for genetic studies of this species.https://www.mdpi.com/2076-0817/10/7/909<i>Aliarcobacter butzleri</i>natural competencetransformation
spellingShingle Marina Bonifácio
Cristiana Mateus
Ana R. Alves
Emanuel Maldonado
Ana P. Duarte
Fernanda Domingues
Mónica Oleastro
Susana Ferreira
Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>
Pathogens
<i>Aliarcobacter butzleri</i>
natural competence
transformation
title Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>
title_full Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>
title_fullStr Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>
title_full_unstemmed Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>
title_short Natural Transformation as a Mechanism of Horizontal Gene Transfer in <i>Aliarcobacter butzleri</i>
title_sort natural transformation as a mechanism of horizontal gene transfer in i aliarcobacter butzleri i
topic <i>Aliarcobacter butzleri</i>
natural competence
transformation
url https://www.mdpi.com/2076-0817/10/7/909
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