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Assessment of SCAR markers to design real-time PCR primers for rhizosphere quantification of Azospirillum brasilense phytostimulatory inoculants of maize.
Couillerot, O; Poirier, M-A; Prigent-Combaret, C; Mavingui, P; Caballero-Mellado, J; Moënne-Loccoz, Y.
Afiliación
  • Couillerot O; Université de Lyon, F-69622, Lyon, France.
  • Poirier MA; Université Lyon 1, Villeurbanne, France.
  • Prigent-Combaret C; CNRS, UMR5557, Ecologie Microbienne, Villeurbanne, France.
  • Mavingui P; Université de Lyon, F-69622, Lyon, France.
  • Caballero-Mellado J; Université Lyon 1, Villeurbanne, France.
  • Moënne-Loccoz Y; CNRS, UMR5557, Ecologie Microbienne, Villeurbanne, France.
J Appl Microbiol ; 109(2): 528-538, 2010 Aug.
Article en En | MEDLINE | ID: mdl-20141548
AIMS: To assess the applicability of sequence characterized amplified region (SCAR) markers obtained from BOX, ERIC and RAPD fragments to design primers for real-time PCR quantification of the phytostimulatory maize inoculants Azospirillum brasilense UAP-154 and CFN-535 in the rhizosphere. METHODS AND RESULTS: Primers were designed based on strain-specific SCAR markers and were screened for successful amplification of target strain and absence of cross-reaction with other Azospirillum strains. The specificity of primers thus selected was verified under real-time PCR conditions using genomic DNA from strain collection and DNA from rhizosphere samples. The detection limit was 60 fg DNA with pure cultures and 4 x 10(3) (for UAP-154) and 4 x 10(4) CFU g(-1) (for CFN-535) in the maize rhizosphere. Inoculant quantification was effective from 10(4) to 10(8) CFU g(-1) soil. CONCLUSION: BOX-based SCAR markers were useful to find primers for strain-specific real-time PCR quantification of each A. brasilense inoculant in the maize rhizosphere. SIGNIFICANCE AND IMPACT OF THE STUDY: Effective root colonization is a prerequisite for successful Azospirillum phytostimulation, but cultivation-independent monitoring methods were lacking. The real-time PCR methods developed here will help understand the effect of environmental conditions on root colonization and phytostimulation by A. brasilense UAP-154 and CFN-535.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Azospirillum brasilense / Reacción en Cadena de la Polimerasa / Cartilla de ADN / Zea mays País/Región como asunto: America do sul / Brasil Idioma: En Revista: J Appl Microbiol Asunto de la revista: MICROBIOLOGIA Año: 2010 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Azospirillum brasilense / Reacción en Cadena de la Polimerasa / Cartilla de ADN / Zea mays País/Región como asunto: America do sul / Brasil Idioma: En Revista: J Appl Microbiol Asunto de la revista: MICROBIOLOGIA Año: 2010 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Reino Unido