Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions

Nitrogen is the second most critical factor for crop production after water. In this study, the beneficial rhizobacterium Pseudomonas protegens Pf-5 was genetically modified to fix nitrogen using the genes encoding the nitrogenase of Pseudomonas stutzeri A1501 via the X940 cosmid. Pf-5 X940 was able...

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Autores principales: Setten, Lorena, Soto, Gabriela Cinthia, Mozzicafreddo, Matteo, Fox, Ana Romina, Lisi, Christian Daniel, Cuccioloni, Massimiliano, Angeletti, Mauro, Pagano, Elba Maria, Diaz Paleo, Antonio Horacio, Ayub, Nicolás Daniel
Formato: Artículo
Lenguaje:Inglés
Publicado: 2017
Materias:
Acceso en línea:http://hdl.handle.net/20.500.12123/1450
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063666
https://doi.org/10.1371/journal.pone.0063666
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author Setten, Lorena
Soto, Gabriela Cinthia
Mozzicafreddo, Matteo
Fox, Ana Romina
Lisi, Christian Daniel
Cuccioloni, Massimiliano
Angeletti, Mauro
Pagano, Elba Maria
Diaz Paleo, Antonio Horacio
Ayub, Nicolás Daniel
author_browse Angeletti, Mauro
Ayub, Nicolás Daniel
Cuccioloni, Massimiliano
Diaz Paleo, Antonio Horacio
Fox, Ana Romina
Lisi, Christian Daniel
Mozzicafreddo, Matteo
Pagano, Elba Maria
Setten, Lorena
Soto, Gabriela Cinthia
author_facet Setten, Lorena
Soto, Gabriela Cinthia
Mozzicafreddo, Matteo
Fox, Ana Romina
Lisi, Christian Daniel
Cuccioloni, Massimiliano
Angeletti, Mauro
Pagano, Elba Maria
Diaz Paleo, Antonio Horacio
Ayub, Nicolás Daniel
author_sort Setten, Lorena
collection INTA Digital
description Nitrogen is the second most critical factor for crop production after water. In this study, the beneficial rhizobacterium Pseudomonas protegens Pf-5 was genetically modified to fix nitrogen using the genes encoding the nitrogenase of Pseudomonas stutzeri A1501 via the X940 cosmid. Pf-5 X940 was able to grow in L medium without nitrogen, displayed high nitrogenase activity and released significant quantities of ammonium to the medium. Pf-5 X940 also showed constitutive expression and enzymatic activity of nitrogenase in ammonium medium or in nitrogen-free medium, suggesting a constitutive nitrogen fixation. Similar to Pseudomonas protegens Pf-5, Pseudomonas putida, Pseudomonas veronii and Pseudomonas taetrolens but not Pseudomonas balearica and Pseudomonas stutzeri transformed with cosmid X940 showed constitutive nitrogenase activity and high ammonium production, suggesting that this phenotype depends on the genome context and that this technology to obtain nitrogen-fixing bacteria is not restricted to Pf-5. Interestingly, inoculation of Arabidopsis, alfalfa, tall fescue and maize with Pf-5 X940 increased the ammonium concentration in soil and plant productivity under nitrogen-deficient conditions. In conclusion, these results open the way to the production of effective recombinant inoculants for nitrogen fixation on a wide range of crops.
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spelling INTA14502017-12-27T14:10:52Z Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions Setten, Lorena Soto, Gabriela Cinthia Mozzicafreddo, Matteo Fox, Ana Romina Lisi, Christian Daniel Cuccioloni, Massimiliano Angeletti, Mauro Pagano, Elba Maria Diaz Paleo, Antonio Horacio Ayub, Nicolás Daniel Pseudomonas Microorganismos Organismos Modificados Genéticamente Bacteria Fijadora del Nitrógeno Fijación del Nitrógeno Fijación del Amonio Nitrogenasa Rizosfera Nitrógeno Asimilable Inoculación Inoculation Arabidopsis Biotecnología Vegetal Sustancias de Crecimiento Vegetal Deficiencias del Suelo Microorganisms Genetically Modified Organisms Nitrogen Fixing Bacteria Nitrogen Fixation Ammonium Fixation Nitrogenase Rhizosphere Digestible Nitrogen Plant Biotechnology Soil Deficiencies Argentina Diazotrofos Endofitos Bacterias Recombinantes Pseudomonas Protogens Pf5 Fijación Biológica del Nitrógeno Deficiencias de Nitrógeno Nitrogen is the second most critical factor for crop production after water. In this study, the beneficial rhizobacterium Pseudomonas protegens Pf-5 was genetically modified to fix nitrogen using the genes encoding the nitrogenase of Pseudomonas stutzeri A1501 via the X940 cosmid. Pf-5 X940 was able to grow in L medium without nitrogen, displayed high nitrogenase activity and released significant quantities of ammonium to the medium. Pf-5 X940 also showed constitutive expression and enzymatic activity of nitrogenase in ammonium medium or in nitrogen-free medium, suggesting a constitutive nitrogen fixation. Similar to Pseudomonas protegens Pf-5, Pseudomonas putida, Pseudomonas veronii and Pseudomonas taetrolens but not Pseudomonas balearica and Pseudomonas stutzeri transformed with cosmid X940 showed constitutive nitrogenase activity and high ammonium production, suggesting that this phenotype depends on the genome context and that this technology to obtain nitrogen-fixing bacteria is not restricted to Pf-5. Interestingly, inoculation of Arabidopsis, alfalfa, tall fescue and maize with Pf-5 X940 increased the ammonium concentration in soil and plant productivity under nitrogen-deficient conditions. In conclusion, these results open the way to the production of effective recombinant inoculants for nitrogen fixation on a wide range of crops. Fil: Setten, Lorena. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina Fil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Mozzicafreddo, Matteo. Università di Camerino. School of Biosciences and Biotechnology; Italia Fil: Fox, Ana Romina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lisi, Christian Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina Fil: Cuccioloni, Massimiliano. Università di Camerino. School of Biosciences and Biotechnology; Italia Fil: Angeletti, Mauro. Università di Camerino. School of Biosciences and Biotechnology; Italia Fil: Pagano, Elba Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina Fil: Diaz Paleo, Antonio Horacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina Fil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2017-10-10T14:32:22Z 2017-10-10T14:32:22Z 2013-05-13 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/1450 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063666 1932-6203 https://doi.org/10.1371/journal.pone.0063666 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 PLoS ONE 8 (5) : e63666. (2013)
spellingShingle Pseudomonas
Microorganismos
Organismos Modificados Genéticamente
Bacteria Fijadora del Nitrógeno
Fijación del Nitrógeno
Fijación del Amonio
Nitrogenasa
Rizosfera
Nitrógeno Asimilable
Inoculación
Inoculation
Arabidopsis
Biotecnología Vegetal
Sustancias de Crecimiento Vegetal
Deficiencias del Suelo
Microorganisms
Genetically Modified Organisms
Nitrogen Fixing Bacteria
Nitrogen Fixation
Ammonium Fixation
Nitrogenase
Rhizosphere
Digestible Nitrogen
Plant Biotechnology
Soil Deficiencies
Argentina
Diazotrofos Endofitos
Bacterias Recombinantes
Pseudomonas Protogens Pf5
Fijación Biológica del Nitrógeno
Deficiencias de Nitrógeno
Setten, Lorena
Soto, Gabriela Cinthia
Mozzicafreddo, Matteo
Fox, Ana Romina
Lisi, Christian Daniel
Cuccioloni, Massimiliano
Angeletti, Mauro
Pagano, Elba Maria
Diaz Paleo, Antonio Horacio
Ayub, Nicolás Daniel
Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions
title Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions
title_full Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions
title_fullStr Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions
title_full_unstemmed Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions
title_short Engineering Pseudomonas protegens Pf-5 for nitrogen fixation and its application to improve plant growth under nitrogen-deficient conditions
title_sort engineering pseudomonas protegens pf 5 for nitrogen fixation and its application to improve plant growth under nitrogen deficient conditions
topic Pseudomonas
Microorganismos
Organismos Modificados Genéticamente
Bacteria Fijadora del Nitrógeno
Fijación del Nitrógeno
Fijación del Amonio
Nitrogenasa
Rizosfera
Nitrógeno Asimilable
Inoculación
Inoculation
Arabidopsis
Biotecnología Vegetal
Sustancias de Crecimiento Vegetal
Deficiencias del Suelo
Microorganisms
Genetically Modified Organisms
Nitrogen Fixing Bacteria
Nitrogen Fixation
Ammonium Fixation
Nitrogenase
Rhizosphere
Digestible Nitrogen
Plant Biotechnology
Soil Deficiencies
Argentina
Diazotrofos Endofitos
Bacterias Recombinantes
Pseudomonas Protogens Pf5
Fijación Biológica del Nitrógeno
Deficiencias de Nitrógeno
url http://hdl.handle.net/20.500.12123/1450
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063666
https://doi.org/10.1371/journal.pone.0063666
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