1887

Abstract

Rhizobia are a disparate collection of soil bacteria capable of reducing atmospheric nitrogen in symbiosis with legumes (Fix phenotype). Synthesis of the nitrogenase and its accessory components is under the transcriptional control of the key regulator NifA and is generally restricted to the endosymbiotic forms of rhizobia known as bacteroids. Amongst studied rhizobia, strain NGR234 has the remarkable ability to fix nitrogen in association with more than 130 species in 73 legume genera that form either determinate, indeterminate or aeschynomenoid nodules. Hence, NGR234 is a model organism to study nitrogen fixation in association with a variety of legumes. The symbiotic plasmid pSfrNGR234a carries more than 50 genes that are under the transcriptional control of NifA. To facilitate the functional analysis of NifA-regulated genes a new transposable element, Tn was constructed. This transposon combines the advantages of mutagenesis of cloned DNA fragments with a conditional read-out promoter from NGR234 (PwA) that reinitiates NifA-dependent transcription downstream of transposition sites. To test the characteristics of the new transposon, the y4vGHIJ operon was mutated using either the Omega interposon or Tn The symbiotic phenotypes on various hosts as well as the transcriptional characteristics of these mutants were analysed in detail and compared with the ineffective (Fix) phenotype of strain NGRΔ, which lacks a functional copy of . transcription from inserted copies of Tn inside bacteroids was confirmed by qRT-PCR. Unexpectedly, polar mutants in and were Fix on all of the hosts tested, indicating that none of the six genes of the operon of NGR234 is essential for symbiotic nitrogen fixation on plants that form nodules of either determinate or indeterminate types.

Funding
This study was supported by the:
  • University of Geneva
  • Swiss National Science Foundation (Award 31003A-116591)
  • Society of Physics and Natural History of Geneva
  • German Research Council (Award SFB395)
  • Alexander von Humboldt Foundation
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2011-10-01
2024-03-29
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