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

Authors :
Philippe Rott
Martha Hincapie
Jack C. Comstock
Isabelle Pieretti
Vanessa Duarte Dias
Marcos Gomes da Cunha
Emmanuel Fernandez
Philippe Roumagnac
IFAS, Everglades Research & Education Center
University of Florida [Gainesville] (UF)
Universidade Federal de Goiás [Goiânia] (UFG)
Biologie et Génétique des Interactions Plante-Parasite (UMR BGPI)
Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)
Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)
Sugarcane Field Station
USDA-ARS : Agricultural Research Service
Brazil Scientific Mobility Program - CAPES (Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior)
CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico)
Florida Sugar Cane League
USDA National Institute of Food and Agriculture : Hatch/Rott FLA-BGL-005404
Source :
Tropical Plant Pathology, Tropical Plant Pathology, 2018, 43 (4), pp.351-359. ⟨10.1007/s40858-018-0216-2⟩
Publication Year :
2018
Publisher :
HAL CCSD, 2018.

Abstract

International audience; 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 (10(7)-10(9) CFU/ml), and asymptomatic stalks with low populations (< 10(3) 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.

Details

Language :
English
ISSN :
19832052
Database :
OpenAIRE
Journal :
Tropical Plant Pathology, Tropical Plant Pathology, 2018, 43 (4), pp.351-359. ⟨10.1007/s40858-018-0216-2⟩
Accession number :
edsair.doi.dedup.....51baa55fc82870295e011990ec57b8ad
Full Text :
https://doi.org/10.1007/s40858-018-0216-2⟩