Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers

Since their discovery, plant-growth promoting rhizobacteria from the genus Azospirillum have been sub-jected to intensive research due to their biotechnological potential as crop inoculants. Phylogeneticanalysis of Azospirillum spp. is carried out by 16S rRNA sequencing almost exclusively, but incon...

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Main Authors: Maroniche, Guillermo Andrés, Garcia, Julia Elena, Salcedo, Florencia, Creus, Cecilia Mónica
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: 2017
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/1032
http://www.sciencedirect.com/science/article/pii/S0944501316307327?via%3Dihub
https://doi.org/10.1016/j.micres.2016.11.009
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author Maroniche, Guillermo Andrés
Garcia, Julia Elena
Salcedo, Florencia
Creus, Cecilia Mónica
author_browse Creus, Cecilia Mónica
Garcia, Julia Elena
Maroniche, Guillermo Andrés
Salcedo, Florencia
author_facet Maroniche, Guillermo Andrés
Garcia, Julia Elena
Salcedo, Florencia
Creus, Cecilia Mónica
author_sort Maroniche, Guillermo Andrés
collection INTA Digital
description Since their discovery, plant-growth promoting rhizobacteria from the genus Azospirillum have been sub-jected to intensive research due to their biotechnological potential as crop inoculants. Phylogeneticanalysis of Azospirillum spp. is carried out by 16S rRNA sequencing almost exclusively, but inconsis-tencies and low confidence often arise when working with close species. In this work, it was observedthat these difficulties might be explained by a high number of rRNA operons with considerable inter-genic variability within Azospirillum genomes. To search for alternative genetic markers from a list ofhousekeeping genes, the correlation between pairwise gene and whole-genome similarities was exam-ined. Due to its good performance, rpoD was selected for further analyses. Genus-specific primers for thePCR-amplification and sequencing of rpoD from Azospirillum spp. were designed and tested on 16 typestrains of different species. The sequences obtained were used for inferring a phylogenetic tree of thegenus, which was in turn used as a reference to successfully identify a collection of 31 azospirilla iso-lated from many different locations of Argentine. In addition, several strains that might represent novelspecies were detected. The results indicate that the sequencing of rpoD is a suitable alternative methodfor a confident molecular identification in Azospirillum spp.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
language Inglés
publishDate 2017
publishDateRange 2017
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spelling INTA10322019-03-22T13:13:37Z Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers Maroniche, Guillermo Andrés Garcia, Julia Elena Salcedo, Florencia Creus, Cecilia Mónica Azospirillum Marcadores Genéticos Rizobacterias Genotipos Genetic Markers Rhizobacteria Genotypes Since their discovery, plant-growth promoting rhizobacteria from the genus Azospirillum have been sub-jected to intensive research due to their biotechnological potential as crop inoculants. Phylogeneticanalysis of Azospirillum spp. is carried out by 16S rRNA sequencing almost exclusively, but inconsis-tencies and low confidence often arise when working with close species. In this work, it was observedthat these difficulties might be explained by a high number of rRNA operons with considerable inter-genic variability within Azospirillum genomes. To search for alternative genetic markers from a list ofhousekeeping genes, the correlation between pairwise gene and whole-genome similarities was exam-ined. Due to its good performance, rpoD was selected for further analyses. Genus-specific primers for thePCR-amplification and sequencing of rpoD from Azospirillum spp. were designed and tested on 16 typestrains of different species. The sequences obtained were used for inferring a phylogenetic tree of thegenus, which was in turn used as a reference to successfully identify a collection of 31 azospirilla iso-lated from many different locations of Argentine. In addition, several strains that might represent novelspecies were detected. The results indicate that the sequencing of rpoD is a suitable alternative methodfor a confident molecular identification in Azospirillum spp. Inst. de Microbiología y Zoología Agrícola IMyZA Fil: Maroniche, Guillermo Andrés. Unidad Integrada Balcarce-Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias-Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Garcia, Julia Elena. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Salcedo, Florencia. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Creus, Cecilia Mónica. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina 2017-08-24T13:50:00Z 2017-08-24T13:50:00Z 2017 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion http://hdl.handle.net/20.500.12123/1032 http://www.sciencedirect.com/science/article/pii/S0944501316307327?via%3Dihub 0944-5013 https://doi.org/10.1016/j.micres.2016.11.009 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Microbiological research 195 : 1-10. (January 2017)
spellingShingle Azospirillum
Marcadores Genéticos
Rizobacterias
Genotipos
Genetic Markers
Rhizobacteria
Genotypes
Maroniche, Guillermo Andrés
Garcia, Julia Elena
Salcedo, Florencia
Creus, Cecilia Mónica
Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers
title Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers
title_full Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers
title_fullStr Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers
title_full_unstemmed Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers
title_short Molecular identification of Azospirillum spp.: limitations of 16S rRNA and qualities of rpoD as genetic markers
title_sort molecular identification of azospirillum spp limitations of 16s rrna and qualities of rpod as genetic markers
topic Azospirillum
Marcadores Genéticos
Rizobacterias
Genotipos
Genetic Markers
Rhizobacteria
Genotypes
url http://hdl.handle.net/20.500.12123/1032
http://www.sciencedirect.com/science/article/pii/S0944501316307327?via%3Dihub
https://doi.org/10.1016/j.micres.2016.11.009
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