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dc.contributor.author
Ambrosio, Rafael
dc.contributor.author
Ortiz Marquez, Juan César Federico
dc.contributor.author
Curatti, Leonardo
dc.date.available
2018-07-13T17:01:13Z
dc.date.issued
2017-03
dc.identifier.citation
Ambrosio, Rafael; Ortiz Marquez, Juan César Federico; Curatti, Leonardo; Metabolic engineering of a diazotrophic bacterium improves ammonium release and biofertilization of plants and microalgae; Academic Press Inc Elsevier Science; Metabolic Engineering; 40; 3-2017; 59-68
dc.identifier.issn
1096-7176
dc.identifier.uri
http://hdl.handle.net/11336/52034
dc.description.abstract
The biological nitrogen fixation carried out by some Bacteria and Archaea is one of the most attractive alternatives to synthetic nitrogen fertilizers. However, with the exception of the symbiotic rhizobia-legumes system, progress towards a more extensive realization of this goal has been slow. In this study we manipulated the endogenous regulation of both nitrogen fixation and assimilation in the aerobic bacterium Azotobacter vinelandii. Substituting an exogenously inducible promoter for the native promoter of glutamine synthetase produced conditional lethal mutant strains unable to grow diazotrophically in the absence of the inducer. This mutant phenotype could be reverted in a double mutant strain bearing a deletion in the nifL gene that resulted in constitutive expression of nif genes and increased production of ammonium. Under GS non-inducing conditions both the single and the double mutant strains consistently released very high levels of ammonium (>20 mM) into the growth medium. The double mutant strain grew and excreted high levels of ammonium under a wider range of concentrations of the inducer than the single mutant strain. Induced mutant cells could be loaded with glutamine synthetase at different levels, which resulted in different patterns of extracellular ammonium accumulation afterwards. Inoculation of the engineered bacteria into a microalgal culture in the absence of sources of C and N other than N2 and CO2 from the air, resulted in a strong proliferation of microalgae that was suppressed upon addition of the inducer. Both single and double mutant strains also promoted growth of cucumber plants in the absence of added N-fertilizer, while this property was only marginal in the parental strain. This study provides a simple synthetic genetic circuit that might inspire engineering of optimized inoculants that efficiently channel N2 from the air into crops.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Academic Press Inc Elsevier Science
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.subject
Ammonium Excretion
dc.subject
Fertilization
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Microalgae
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Nitrogen Assimilation
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Nitrogen Fixation
dc.subject
Plants
dc.subject.classification
Otras Ciencias Biológicas
dc.subject.classification
Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
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Otras Biotecnología Agropecuaria
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Biotecnología Agropecuaria
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CIENCIAS AGRÍCOLAS
dc.title
Metabolic engineering of a diazotrophic bacterium improves ammonium release and biofertilization of plants and microalgae
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-07-11T14:40:49Z
dc.journal.volume
40
dc.journal.pagination
59-68
dc.journal.pais
Estados Unidos
dc.description.fil
Fil: Ambrosio, Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para Investigaciones Biológicas Aplicadas; Argentina
dc.description.fil
Fil: Ortiz Marquez, Juan César Federico. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para Investigaciones Biológicas Aplicadas; Argentina
dc.description.fil
Fil: Curatti, Leonardo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para Investigaciones Biológicas Aplicadas; Argentina
dc.journal.title
Metabolic Engineering
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.ymben.2017.01.002
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S1096717616301112
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