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CRISPR loci-PCR as Tool for Tracking Azospirillum sp. Strain B510.
Rilling, Joaquin I; Maruyama, Fumito; Sadowsky, Michael J; Acuña, Jacquelinne J; Jorquera, Milko A.
Afiliação
  • Rilling JI; Applied Microbial Ecology Laboratory (EMAlab), Departamento de Ciencias Químicas y Recursos Naturales, Universidad de La Frontera, Temuco 4780000, Chile.
  • Maruyama F; Center of Plant, Soil Interaction and Natural Resources Biotechnology, Scientific and Technological Bioresource Nucleus (BIOREN), Universidad de La Frontera, Temuco 4780000, Chile.
  • Sadowsky MJ; Office of Industry-Academia-Government and Community Collaboration, Hiroshima University, Hiroshima 739-8511, Japan.
  • Acuña JJ; Department of Soil, Water, and Climate, Department of Plant and Microbial Biology, and BioTechnology Institute, University of Minnesota, St. Paul, MN 55812, USA.
  • Jorquera MA; Applied Microbial Ecology Laboratory (EMAlab), Departamento de Ciencias Químicas y Recursos Naturales, Universidad de La Frontera, Temuco 4780000, Chile.
Microorganisms ; 9(7)2021 Jun 22.
Article em En | MEDLINE | ID: mdl-34206618
Azospirillum-based plant and soil inoculants are widely used in agriculture. The inoculated Azospirillum strains are commonly tracked by both culture-dependent and culture-independent methods, which are time-consuming or expensive. In this context, clustered regularly interspaced short palindromic repeats (CRISPR) loci structure is unique in the bacterial genome, including some Azospirillum species. Here, we investigated the use of CRISPR loci to track specific Azospirillum strains in soils systems by PCR. Primer sets for Azospirillum sp. strain B510 were designed and evaluated by colony and endpoint PCR. The CRISPRloci-PCR approach was standardized for Azospirillum sp. strain B510, and its specificity was observed by testing against 9 different Azospirillum strains, and 38 strains of diverse bacterial genera isolated from wheat plants. The CRISPRloci-PCR approach was validated in assays with substrate and wheat seedlings. Azospirillum sp. strain B510 was detected after of two weeks of inoculation in both sterile and nonsterile substrates as well as rhizosphere grown in sterile substrate. The CRISPRloci-PCR approach was found to be a useful molecular tool for specific tracking of Azospirillum at the strain level. This technique can be easily adapted to other microbial inoculants carrying CRISPR loci and can be used to complement other microbiological techniques.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Microorganisms Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Chile País de publicação: Suíça

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Microorganisms Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Chile País de publicação: Suíça