Previous Article | Next Article 
Applied and Environmental Microbiology, October 2000, p. 4292-4299, Vol. 66, No. 10
Laboratorio de Microbiología,
Departamento de Biotecnología, Escuela Técnica Superior
de Ingenieros Agrónomos, Universidad Politécnica de
Madrid,1 and Consejo Superior de
Investigaciones Científicas,2 Ciudad
Universitaria s/n, 28040 Madrid, Spain
Received 29 March 2000/Accepted 17 July 2000
Hydrogen evolution by nitrogenase is a source of inefficiency for
the nitrogen fixation process by the Rhizobium-legume
symbiosis. To develop a strategy to generate rhizobial strains with
H2-recycling ability, we have constructed a Tn5
derivative minitransposon (TnHB100) that contains the ca. 18-kb
H2 uptake (hup) gene cluster from Rhizobium leguminosarum bv. viciae UPM791. Bacteroids from
TnHB100-containing strains of R. leguminosarum bv.
viciae PRE, Bradyrhizobium japonicum, R. etli,
and Mesorhizobium loti expressed high levels of hydrogenase activity that resulted in full recycling of the hydrogen evolved by
nitrogenase in nodules. Efficient processing of the hydrogenase large subunit (HupL) in these strains was shown by immunoblot analysis
of bacteroid extracts. In contrast, Sinorhizobium meliloti, M. ciceri, and R. leguminosarum bv. viciae UML2
strains showed poor expression of the hup system that
resulted in H2-evolving nodules. For the latter group of
strains, no immunoreactive material was detected in bacteroid extracts
using anti-HupL antiserum, suggesting a low level of transcription of
hup genes or HupL instability. A general procedure for the
characterization of the minitransposon insertion site and removal of
antibiotic resistance gene included in TnHB100 has been developed and
used to generate engineered strains suitable for field release.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Generation of New Hydrogen-Recycling
Rhizobiaceae Strains by Introduction of a Novel
hup Minitransposon
*
Corresponding author. Mailing address: Laboratorio de
Microbiología, Escuela Técnica Superior de Ingenieros
Agrónomos, Universidad Politécnica de Madrid, Ciudad
Universitaria s/n, 28040 Madrid, Spain. Phone: 34-91-3365759. Fax:
34-91-3365757. E-mail: jpalacios{at}bit.etsia.upm.es.
Applied and Environmental Microbiology, October 2000, p. 4292-4299, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Maroti, G., Tong, Y., Yooseph, S., Baden-Tillson, H., Smith, H. O., Kovacs, K. L., Frazier, M., Venter, J. C., Xu, Q.
(2009). Discovery of [NiFe] Hydrogenase Genes in Metagenomic DNA: Cloning and Heterologous Expression in Thiocapsa roseopersicina. Appl. Environ. Microbiol.
75: 5821-5830
[Abstract] [Full Text] -
Lenz, O., Gleiche, A., Strack, A., Friedrich, B.
(2005). Requirements for Heterologous Production of a Complex Metalloenzyme: the Membrane-Bound [NiFe] Hydrogenase. J. Bacteriol.
187: 6590-6595
[Abstract] [Full Text] -
Peralta, H., Mora, Y., Salazar, E., Encarnacion, S., Palacios, R., Mora, J.
(2004). Engineering the nifH Promoter Region and Abolishing Poly-{beta}-Hydroxybutyrate Accumulation in Rhizobium etli Enhance Nitrogen Fixation in Symbiosis with Phaseolus vulgaris. Appl. Environ. Microbiol.
70: 3272-3281
[Abstract] [Full Text] -
Baginsky, C., Brito, B., Imperial, J., Palacios, J.-M., Ruiz-Argueso, T.
(2002). Diversity and Evolution of Hydrogenase Systems in Rhizobia. Appl. Environ. Microbiol.
68: 4915-4924
[Abstract] [Full Text] -
Brito, B., Palacios, J. M., Imperial, J., Ruiz-Argueso, T.
(2002). Engineering the Rhizobium leguminosarum bv. viciae Hydrogenase System for Expression in Free-Living Microaerobic Cells and Increased Symbiotic Hydrogenase Activity. Appl. Environ. Microbiol.
68: 2461-2467
[Abstract] [Full Text]
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»