VIEW ARTICLE | DOI: 10.1094/MPMI-8-0492
Characterization of a Rhizobium melilotiProline Dehydrogenase Mutant Altered in NodulationEfficiency and Competitiveness on Alfalfa Roots. Jose I. Jimenez-Zurdo . Departamento de Microbiologi'a, Estacion Experimental del Zaidfn, CSIC, Profesor Albareda 1, 18008 Granada, Spain. Pieter van Dillewijn(1), Maria J. Soto(1), Maria R. de Felipe(2) ,Jose Olivares(1), and Nicolas Toro(1). (1) Departamento de Microbiologi'a, Estacion Experimental del Zaidfn, CSIC, Profesor Albareda 1, 18008 Granada, Spain (2) Centro de Ciencias Medioambientales, CSIC, Serrano 115 Bis, 28006 Madrid, Spain. MPMI 8:492-498. Accepted 6 March 1995 . Copyright 1995 The American Phytopathological Society.
Additional Keywords: symbiosis
Rhizobium meliloti strain GRM8 is able to transform orni-thine into proline by means of an ornithine cyclodeami-nase and, therefore, has the ability to use either of these amino acids as its sole carbon and nitrogen source. By Tn5 insertion mutagenesis we obtained a GRM8 mutant derivative strain (LM1) unable to catabolize either ornithine or proline. DNA hybridization studies showed that the LM1 mutant carries a single Tn5 insertion within a chromosomally located gene that, as deduced from a partial nucleotide sequence, encodes a proline dehydrogenase (ProDH). Enzymatic assays confirmed the lack of ProDH activity in cell extracts of strain LM1 and revealed that production of this enzyme is inducible in the parental strain by proline and ornithine. Ultrastructural nodule microscopy analysis, acetylene reduction assays, and dry-weight determinations of nodulated alfalfa plants showed no obvious defect in the nitrogen fixation process of the ProDH" mutant LM1. However, nodulation tests and competition assays demonstrated that in R. meliloti ProDH is required for nodulation efficiency and competitiveness on alfalfa roots.