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

Authors
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
Publication Year
2013
Language
English
Format
article
Status
Published version
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.
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
Source
PLoS ONE 8 (5) : e63666. (2013)
Subject
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
Access level
Open access
License
Repository
INTA Digital (INTA)
Institution
Instituto Nacional de Tecnología Agropecuaria
OAI Identifier
oai:localhost:20.500.12123/1450