title: Selection of nitrogen-fixing deficient Burkholderia vietnamiensis strains by cystic fibrosis patients: involvement of nif gene deletions and auxotrophic mutations.
abstract: Burkholderia vietnamiensis is the third most prevalent species of the Burkholderia cepacia complex (Bcc) found in cystic fibrosis (CF) patients. Its ability at fixing nitrogen makes it one of the main Bcc species showing strong filiations with environmental reservoirs. In this study, 83% (29 over 35) of the B. vietnamiensis CF isolates and 100% of the environmental ones (over 29) were found expressing the dinitrogenase complex (encoded by the nif cluster) which is essential in N(2) fixation. Among the deficient strains, two were found growing with ammonium chloride suggesting that they were defective in N(2) fixation, and four with amino acids supplements suggesting that they were harbouring auxotrophic mutations. To get insights about the genetic events that led to the emergence of the N(2)-fixing defective strains, a genetic analysis of B. vietnamiensis nitrogen-fixing property was undertaken. A 40-kb-long nif cluster and nif regulatory genes were identified within the B. vietnamiensis strain G4 genome sequence, and analysed. Transposon mutagenesis and nifH genetic marker exchanges showed the nif cluster and several other genes like gltB (encoding a subunit of the glutamate synthase) to play a key role in B. vietnamiensis ability at growing in nitrogen-free media. nif cluster DNA probings of restricted genomic DNA blots showed a full deletion of the nif cluster for one of the N(2)-fixing defective strain while the other one showed a genetic organization similar to the one of the G4 strain. For 17% of B. vietnamiensis clinical strains, CF lungs appeared to have favoured the selection of mutations or deletions leading to N(2)-fixing deficiencies.
authors.affiliation: Research Group on "Bacterial Opportunistic Pathogens and Environment", Université de Lyon, Lyon, F-69003, France.
authors.forename: Aymeric
authors.initials: A
authors.lastname: Menard
authors.forename: Claire
authors.initials: C
authors.lastname: Monnez
authors.forename: Paulina
authors.initials: P
authors.lastname: Estrada de Los Santos
authors.forename: Christine
authors.initials: C
authors.lastname: Segonds
authors.forename: Jesus
authors.initials: J
authors.lastname: Caballero-Mellado
authors.forename: John J
authors.initials: JJ
authors.lastname: Lipuma
authors.forename: Gerard
authors.initials: G
authors.lastname: Chabanon
authors.forename: Benoit
authors.initials: B
authors.lastname: Cournoyer
mesh_terms: D019121:Burkholderia Infections / Q000382:microbiology*; D042602:Burkholderia cepacia complex / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000382:microbiology*; D004269:DNA, Bacterial; D004275:DNA, Ribosomal / Q000145:classification*; D019143:Evolution, Molecular; D017353:Gene Deletion; D006801:Humans; D008168:Lung / Q000382:microbiology / Q000502:physiology; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation / Q000235:genetics* / Q000502:physiology; D009591:Nitrogenase / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D012988:Soil Microbiology
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D009591:Nitrogenase