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Comparison of loop-mediated isothermal amplification, polymerase chain reaction, and selective isolation assays for detection of Xanthomonas albilineans from sugarcane

Duarte Dias Vanessa, Fernandez Emmanuel, Cunha Marcos G., Pieretti Isabelle, Hincapie Martha, Roumagnac Philippe, Comstock Jack C., Rott Philippe. 2018. Comparison of loop-mediated isothermal amplification, polymerase chain reaction, and selective isolation assays for detection of Xanthomonas albilineans from sugarcane. Tropical Plant Pathology, 43 (4) : pp. 351-359.

Journal article ; Article de recherche ; Article de revue à facteur d'impact Revue en libre accès total
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Abstract : A loop-mediated isothermal amplification (LAMP) assay was developed and compared to polymerase chain reaction (PCR), nested PCR, and selective isolation assays for detection of Xanthomonas albilineans, the causal agent of sugarcane leaf scald. The pathogen was isolated on selective medium from 44 out of 45 (98%) samples taken from symptomatic stalks, and from 44 out of 70 (63%) samples from asymptomatic stalks that were collected from plots with symptomatic stalks. Forty-two (93%), 41 (91%), and 42 (93%) symptomatic samples tested positive by LAMP, PCR and nested PCR, respectively. The pathogen was detected in 19 (27%), 8 (11%), and 25 (36%) of the 70 asymptomatic samples by LAMP, PCR and nested PCR, respectively. Symptomatic stalks were mainly, but not always, associated with high populations of the pathogen (107–109 CFU/ml), and asymptomatic stalks with low populations (<103 CFU/ml) or no bacteria. Although our LAMP and nested PCR methods detected 10 CFU/ml of X. albilineans in suspensions prepared with pure culture, they sometimes failed to detect the pathogen in samples with low pathogen populations. Isolation on selective medium along with another method should therefore be used for detection of the pathogen in asymptomatic stalks, especially in quarantine programs.

Mots-clés géographiques Agrovoc : Floride

Classification Agris : H20 - Plant diseases

Champ stratégique Cirad : Axe 4 (2014-2018) - Santé des animaux et des plantes

Auteurs et affiliations

  • Duarte Dias Vanessa, University of Florida (USA)
  • Fernandez Emmanuel, CIRAD-BIOS-UMR BGPI (FRA)
  • Cunha Marcos G., UFG (BRA)
  • Pieretti Isabelle, CIRAD-BIOS-UMR BGPI (FRA)
  • Hincapie Martha, University of Florida (USA)
  • Roumagnac Philippe, CIRAD-BIOS-UMR BGPI (FRA) ORCID: 0000-0001-5002-6039
  • Comstock Jack C., USDA (USA)
  • Rott Philippe, CIRAD-BIOS-UMR BGPI (FRA) ORCID: 0000-0001-6085-6159 - auteur correspondant

Source : Cirad-Agritrop (https://agritrop.cirad.fr/587708/)

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