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Plant Physiol, November 2000, Vol. 124, pp. 1059-1068
Oxygen Regulation of a Nodule-Located Carbonic Anhydrase in
Alfalfa1
Susana
Gálvez,
Ann M.
Hirsch,
Keith L.
Wycoff,2
Stephen
Hunt,
David B.
Layzell,
Adam
Kondorosi, and
Martin
Crespi*
Institut des Sciences Végétales, Centre National de la
Recherche Scientifique, F-91198 Gif-sur-Yvette cédex, France
(S.G., A.K., M.C.); Department of Molecular, Cell, and Developmental
Biology, University of California, 405 Hilgard Avenue, Los Angeles,
California 90095-1606 (A.M.H., K.L.W.); Queen's University, Kingston,
Ontario, Canada K7L 3N6 (S.H., D.B.L.); and Institute of Genetics,
Biological Research Center, Hungarian Academy of Sciences, Szeged, P.O.
Box 521, H-6701 Szeged, Hungary (A.K.)
Control of the permeability to oxygen is critical for the function
of symbiotic nitrogen fixation in legume nodules. The inner cortex (IC)
seems to be a primary site for this regulation. In alfalfa
(Medicago sativa) nodules, expression of the
Msca1 gene encoding a carbonic anhydrase (CA) was
previously found to be restricted to the IC. We have now raised
antibodies against recombinant Msca1 protein and used
them, together with antibodies raised against potato leaf CA, to
demonstrate the presence of two forms of CA in mature nodules. Each
antibody recognizes a different CA isoform in nodule tissues.
Immunolocalization revealed that leaf-related CAs were localized
primarily in the nitrogen-fixing zone, whereas the Msca1
protein was restricted exclusively to the IC region, in indeterminate
and determinate nodules. In alfalfa nodules grown at various
O2 concentrations, an inverse correlation was observed between the external oxygen pressure and Msca1 protein
content in the IC, the site of the putative diffusion barrier. Thus
Msca1 is a molecular target of physiological processes
occurring in the IC cells involved in gas exchange in the nodule.
1
This work was supported by the European Union
Biotechnology program FIXNET (no. PL962319 to S.G.) and by the U.S.
Department of Agriculture-National Research Initiation Competitive
Grants Program (no. 92-37305-2366 to K.L.W.). Additional support came from the Natural Sciences and Engineering Research Council (Canada to
D.B.L.) and from the National Science Foundation (no. 90-23888 to
A.M.H.).
2
Present address: Planet Biotechnology, 2461 Wyandotte Street, Mountain View, CA 94043.
*
Corresponding author; e-mail crespi{at}isv.cnrs-gif.fr; fax
33-1-69823695.
© 2000 American Society of Plant Physiologists
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