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Plant Microbiology

Enhanced Nitrogen Fixation in a Rhizobium etli ntrC Mutant That Overproduces the Bradyrhizobium japonicum Symbiotic Terminal Oxidasecbb3

Mario Soberón, Oswaldo López, Claudia Morera, Maria de Lourdes Girard, Maria Luisa Tabche, Juan Miranda
Mario Soberón
Departamento de Biologı́a Molecular de Plantas, Instituto de Biotecnologı́a, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62271, and
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Oswaldo López
Departamento de Biologı́a Molecular de Plantas, Instituto de Biotecnologı́a, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62271, and
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Claudia Morera
Departamento de Biologı́a Molecular de Plantas, Instituto de Biotecnologı́a, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62271, and
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Maria de Lourdes Girard
Centro de Investigación sobre Fijación de Nitrógeno, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México
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Maria Luisa Tabche
Departamento de Biologı́a Molecular de Plantas, Instituto de Biotecnologı́a, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62271, and
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Juan Miranda
Departamento de Biologı́a Molecular de Plantas, Instituto de Biotecnologı́a, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62271, and
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DOI: 10.1128/AEM.65.5.2015-2019.1999
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ABSTRACT

The ntrC gene codes for a transcriptional activator protein that modulates gene expression in response to nitrogen. The cytochrome production pattern of a Rhizobium etli ntrCmutant (CFN2012) was studied. CO difference spectral analysis of membranes showed that CFN2012 produced a terminal oxidase similar to the symbiotic terminal oxidase of bacteroids in free-living cells under aerobic conditions, with a characteristic trough at 553 nm. CFN2012 produced two c-type cytochromes with molecular masses of 27 and 32 kDa, in contrast with the wild-type strain, which produced only a 32-kDa c-type cytochrome. The expression levels of theR. etli fixNOQP operon, which codes for terminal oxidasecbb3, were not affected by the ntrCmutation. However, the production levels of the two c-type cytochromes (27 and 32 kDa) were enhanced at least eightfold when theBradyrhizobium japonicum fixNOQP operon was expressed in CFN2012 from the nptII promoter (pMSfixc), suggesting that these proteins are subunits FixO (27 kDa) and FixP (32 kDa) of cbb3 and that CFN2012/pMSfixc overproduced this terminal oxidase. CFN2012/pMSfixc showed a significant increase in its symbiotic performance as judged by the determination of nitrogenase activities of plants inoculated with this strain, suggesting that the overproduction of cbb3 terminal oxidase correlates with an enhancement in symbiotic nitrogen fixation.

  • Copyright © 1999 American Society for Microbiology
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Enhanced Nitrogen Fixation in a Rhizobium etli ntrC Mutant That Overproduces the Bradyrhizobium japonicum Symbiotic Terminal Oxidasecbb3
Mario Soberón, Oswaldo López, Claudia Morera, Maria de Lourdes Girard, Maria Luisa Tabche, Juan Miranda
Applied and Environmental Microbiology May 1999, 65 (5) 2015-2019; DOI: 10.1128/AEM.65.5.2015-2019.1999

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Enhanced Nitrogen Fixation in a Rhizobium etli ntrC Mutant That Overproduces the Bradyrhizobium japonicum Symbiotic Terminal Oxidasecbb3
Mario Soberón, Oswaldo López, Claudia Morera, Maria de Lourdes Girard, Maria Luisa Tabche, Juan Miranda
Applied and Environmental Microbiology May 1999, 65 (5) 2015-2019; DOI: 10.1128/AEM.65.5.2015-2019.1999
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