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Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches

To detect Xanthomonas arboricola pv. pruni, a loop-mediated isothermal amplification (LAMP) detection method were developed. The LAMP assay was designed to test crude plant tissue without pre-extraction, or heating incubation, and without advanced analysis equipment. The LAMP primers were designed b...

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Autores principales: Li, Weilan, Lee, Seung-Yeol, Back, Chang-Gi, Ten, Leonid N., Jung, Hee-Young
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society of Plant Pathology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6901255/
https://www.ncbi.nlm.nih.gov/pubmed/31832043
http://dx.doi.org/10.5423/PPJ.OA.07.2019.0197
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author Li, Weilan
Lee, Seung-Yeol
Back, Chang-Gi
Ten, Leonid N.
Jung, Hee-Young
author_facet Li, Weilan
Lee, Seung-Yeol
Back, Chang-Gi
Ten, Leonid N.
Jung, Hee-Young
author_sort Li, Weilan
collection PubMed
description To detect Xanthomonas arboricola pv. pruni, a loop-mediated isothermal amplification (LAMP) detection method were developed. The LAMP assay was designed to test crude plant tissue without pre-extraction, or heating incubation, and without advanced analysis equipment. The LAMP primers were designed by targeting an ABC transporter ATP-binding protein, this primer set was tested using the genomic DNA of Xanthomonas and non-Xanthomonas strains, and a ladder product was generated from the genomic DNA of X. arboricola pv. pruni strain but not from 12 other Xanthomonas species strains and 6 strains of other genera. The LAMP conditions were checked with the healthy leaves of 31 peach varieties, and no reaction was detected using either the peach leaves or the peach DNA as a template. Furthermore, the high diagnostic accuracy of the LAMP method was confirmed with 13 X. arboricola pv. pruni strains isolated from various regions in Korea, with all samples exhibiting a positive reaction in LAMP assays. In particular, the LAMP method successfully detected the pathogen in diseased peach leaves and fruit in the field, and the LAMP conditions were proven to be a reliable diagnostic method for the specific detection and identification of X. arboricola pv. pruni in peach orchards.
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spelling pubmed-69012552019-12-12 Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches Li, Weilan Lee, Seung-Yeol Back, Chang-Gi Ten, Leonid N. Jung, Hee-Young Plant Pathol J Research Article To detect Xanthomonas arboricola pv. pruni, a loop-mediated isothermal amplification (LAMP) detection method were developed. The LAMP assay was designed to test crude plant tissue without pre-extraction, or heating incubation, and without advanced analysis equipment. The LAMP primers were designed by targeting an ABC transporter ATP-binding protein, this primer set was tested using the genomic DNA of Xanthomonas and non-Xanthomonas strains, and a ladder product was generated from the genomic DNA of X. arboricola pv. pruni strain but not from 12 other Xanthomonas species strains and 6 strains of other genera. The LAMP conditions were checked with the healthy leaves of 31 peach varieties, and no reaction was detected using either the peach leaves or the peach DNA as a template. Furthermore, the high diagnostic accuracy of the LAMP method was confirmed with 13 X. arboricola pv. pruni strains isolated from various regions in Korea, with all samples exhibiting a positive reaction in LAMP assays. In particular, the LAMP method successfully detected the pathogen in diseased peach leaves and fruit in the field, and the LAMP conditions were proven to be a reliable diagnostic method for the specific detection and identification of X. arboricola pv. pruni in peach orchards. Korean Society of Plant Pathology 2019-12 2019-12-12 /pmc/articles/PMC6901255/ /pubmed/31832043 http://dx.doi.org/10.5423/PPJ.OA.07.2019.0197 Text en © The Korean Society of Plant Pathology This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Li, Weilan
Lee, Seung-Yeol
Back, Chang-Gi
Ten, Leonid N.
Jung, Hee-Young
Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches
title Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches
title_full Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches
title_fullStr Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches
title_full_unstemmed Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches
title_short Loop-Mediated Isothermal Amplification for the Detection of Xanthomonas arboricola pv. pruni in Peaches
title_sort loop-mediated isothermal amplification for the detection of xanthomonas arboricola pv. pruni in peaches
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6901255/
https://www.ncbi.nlm.nih.gov/pubmed/31832043
http://dx.doi.org/10.5423/PPJ.OA.07.2019.0197
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