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Archives of Microbiology

, Volume 144, Issue 2, pp 142–146 | Cite as

Dicarboxylic acid transport in Rhizobium meliloti: isolation of mutants and cloning of dicarboxylic acid transport genes

  • Eugene Bolton
  • Brian Higgisson
  • Aiden Harrington
  • Fergal O'Gara
Original Papers

Abstract

The role of the dicarboxylic acid transport (dct) system in the Rhizobium meliloti-Alfalfa symbiosis was investigated. Mutants of R. meliloti CM2 unable to grow on medium containing succinate as the sole carbon source were isolated following chemical and transposon mutagenesis. These mutants were also unable to utilize malate or fumarate as the sole source of carbon. Transport studies with 14C-labelled succinate showed that the mutants were specifically defective in succinate transport. Revertants of both chemical and transposon mutants were obtained at a frequency of 10-5–10-6. The R. meliloti dct mutants were able to nodulate Alfalfa plants but the nodules formed were unable to fix nitrogen. Revertants of the mutants were fully effective on plants. The mutants unable to transport succinate were used to isolate dct genes from a R. meliloti gene bank. Two plasmids containing a common 26.5 Mdal insert were found to complement some of the mutants. The presence of this DNA insert in the complementing mutant strains restored their effectivenss of plants. This DNA fragment encoding succinate transport function(s) was used to produce genetically engineered R. meliloti strains with an increased rate of succinate uptake.

Key words

Rhizobium meliloti Succinate transport Dicarboxylic acids Mutant complementation Nitrogen-fixation 

Abbreviation

dct

dicarboxylic acid transport

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Eugene Bolton
    • 1
  • Brian Higgisson
    • 1
  • Aiden Harrington
    • 1
  • Fergal O'Gara
    • 1
  1. 1.Department of MicrobiologyUniversity CollegeCorkIreland

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