Identification of Burkholderia multivorans ATCC 17616 genetic determinants for fitness in soil by using signature-tagged mutagenesis

Yuji Nagata, Junko Senbongi, Yoko Ishibashi, Rie Sudo, Masatoshi Miyakoshi, Yoshiyuki Ohtsubo, Masataka Tsuda

    Research output: Contribution to journalArticlepeer-review

    11 Citations (Scopus)

    Abstract

    To identify bacterial genetic determinants for fitness in a soil environment, signature-tagged mutagenesis (STM) was applied to a soil bacterium, Burkholderia multivorans ATCC 17616. This strain was randomly mutagenized by each of 36 different signature-tagged plasposons, and 36 mutants with different tags were grouped as a set. A total of 192 sets consisting of 6912 independent mutants were each inoculated into soil and incubated. Two-step STM screening based on quantitative real-time PCR of total DNAs extracted from the resulting soil samples using the tag-specific primers led to the selection of 39 mutant candidates that exhibited a reduction in relative competitive fitness during incubation in the soil, and 32 plasposon-insertion sites were determined. Among them, mutants having plasposon insertion in fur, deaD or hrpA exhibited reduced fitness during incubation in soil when compared with the control strain. The deficiency in the soil fitness of the fur mutant was recovered by the introduction of the wild-type fur gene, indicating that the fur gene is one of the genetic determinants for fitness in the soil.

    Original languageEnglish
    Pages (from-to)883-891
    Number of pages9
    JournalMicrobiology (United Kingdom)
    Volume160
    Issue numberPART 5
    DOIs
    Publication statusPublished - 2014 May

    ASJC Scopus subject areas

    • Microbiology

    Fingerprint Dive into the research topics of 'Identification of Burkholderia multivorans ATCC 17616 genetic determinants for fitness in soil by using signature-tagged mutagenesis'. Together they form a unique fingerprint.

    Cite this