Applicability of the 16S-23S rDNA internal spacer for PCR detection of the phytostimulatory PGPR inoculant Azospirillum lipoferum CRT1 in field soil.

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  • Additional Information
    • Source:
      Publisher: Oxford University Press Country of Publication: England NLM ID: 9706280 Publication Model: Print Cited Medium: Internet ISSN: 1365-2672 (Electronic) Linking ISSN: 13645072 NLM ISO Abbreviation: J Appl Microbiol Subsets: MEDLINE
    • Publication Information:
      Publication: 2022- : Oxford : Oxford University Press
      Original Publication: Oxford : Published for the Society for Applied Bacteriology by Blackwell Science, c1997-
    • Subject Terms:
      Soil Microbiology*; Azospirillum lipoferum/*genetics ; Azospirillum lipoferum/*isolation & purification ; DNA, Ribosomal Spacer/*genetics ; Polymerase Chain Reaction/*methods; Base Sequence ; Molecular Sequence Data
    • Abstract:
      Aims: To assess the applicability of the 16S-23S rDNA internal spacer regions (ISR) as targets for PCR detection of Azospirillum ssp. and the phytostimulatory plant growth-promoting rhizobacteria seed inoculant Azospirillum lipoferum CRT1 in soil.
      Methods and Results: Primer sets were designed after sequence analysis of the ISR of A. lipoferum CRT1 and Azospirillum brasilense Sp245. The primers fAZO/rAZO targeting the Azospirillum genus successfully yielded PCR amplicons (400-550 bp) from Azospirillum strains but also from certain non-Azospirillum strains in vitro, therefore they were not appropriate to monitor indigenous Azospirillum soil populations. The primers fCRT1/rCRT1 targeting A. lipoferum CRT1 generated a single 249-bp PCR product but could also amplify other strains from the same species. However, with DNA extracts from the rhizosphere of field-grown maize, both fAZO/rAZO and fCRT1/rCRT1 primer sets could be used to evidence strain CRT1 in inoculated plants by nested PCR, after a first ISR amplification with universal ribosomal primers. In soil, a 7-log dynamic range of detection (10(2)-10(8) CFU g(-1) soil) was obtained.
      Conclusions: The PCR primers targeting 16S-23S rDNA ISR sequences enabled detection of the inoculant A. lipoferum CRT1 in field soil.
      Significance and Impact of the Study: Convenient methods to monitor Azospirillum phytostimulators in the soil are lacking. The PCR protocols designed based on ISR sequences will be useful for detection of the crop inoculant A. lipoferum CRT1 under field conditions.
    • Accession Number:
      0 (DNA, Ribosomal Spacer)
    • Publication Date:
      Date Created: 20090709 Date Completed: 20100420 Latest Revision: 20091216
    • Publication Date:
      20231215
    • Accession Number:
      10.1111/j.1365-2672.2009.04393.x
    • Accession Number:
      19583800