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Applied and Environmental Microbiology, March 2000, p. 937-942, Vol. 66, No. 3
Laboratorio de Microbiología, 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 5 August 1999/Accepted 29 November 1999
A limited number of Rhizobium and
Bradyrhizobium strains possess a hydrogen uptake (Hup)
system that recycles the hydrogen released from the nitrogen fixation
process in legume nodules. To extend this ability to rhizobia that
nodulate agronomically important crops, we investigated factors that
affect the expression of a cosmid-borne Hup system from Rhizobium
leguminosarum bv. viciae UPM791 in R. leguminosarum
bv. viciae, Rhizobium etli, Mesorhizobium loti,
and Sinorhizobium meliloti Hup
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Nickel Availability and hupSL Activation by
Heterologous Regulators Limit Symbiotic Expression of the
Rhizobium leguminosarum bv. Viciae Hydrogenase System in
Hup
Rhizobia

strains. After
cosmid pAL618 carrying the entire hup system of strain
UPM791 was introduced, all recipient strains acquired the ability to
oxidize H2 in symbioses with their hosts, although the
levels of hydrogenase activity were found to be strain and species
dependent. The levels of hydrogenase activity were correlated with the
levels of nickel-dependent processing of the hydrogenase structural
polypeptides and with transcription of structural genes. Expression of
the NifA-dependent hupSL promoter varied depending on the
genetic background, while the hyp operon, which is
controlled by the FnrN transcriptional regulator, was expressed at
similar levels in all recipient strains. With the exception of the
R. etli-bean symbiosis, the availability of nickel to
bacteroids strongly affected hydrogenase processing and activity in the
systems tested. Our results indicate that efficient transcriptional
activation by heterologous regulators and processing of the hydrogenase
as a function of the availability of nickel to the bacteroid are relevant factors that affect hydrogenase expression in heterologous rhizobia.
*
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: 34-91-3365753. Fax:
34-91-3365757. E-mail: jpalacios{at}bit.etsia.upm.es.
Present address: Departamento de Bioquímica, Facultad de
Agronomía, Universidad de la República, CP 11600 Montevideo, Uruguay.
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