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A pyrosequencing-based method to quantify genetic substitutions associated with resistance to succinate dehydrogenase inhibitor fungicides in Botrytis spp. populations

The genetic underlying resistance mechanisms in the population of the phytopathogenic fungus Botrytis cinerea are well documented. Specifically, several genetic substitutions associated with succinate dehydrogenase inhibitor (SDHI)-based fungicide resistance have been identified in the succinate deh... Full description

Journal Title: Pest management science March 2016, Vol.72(3), pp.566-73
Main Author: Gobeil-Richard, Mélanie
Other Authors: Tremblay, David-Mathieu , Beaulieu, Carole , Van Der Heyden, Hervé , Carisse, Odile
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: E-ISSN: 1526-4998 ; PMID: 25900263 Version:1 ; DOI: 10.1002/ps.4026
Link: http://pubmed.gov/25900263
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recordid: medline25900263
title: A pyrosequencing-based method to quantify genetic substitutions associated with resistance to succinate dehydrogenase inhibitor fungicides in Botrytis spp. populations
format: Article
creator:
  • Gobeil-Richard, Mélanie
  • Tremblay, David-Mathieu
  • Beaulieu, Carole
  • Van Der Heyden, Hervé
  • Carisse, Odile
subjects:
  • Botrytis
  • Pyromark Q24
  • Sdhi
  • Fungicide Resistance
  • Pyrosequencing
  • Botrytis -- Drug Effects
  • Drug Resistance -- Genetics
  • Fungicides, Industrial -- Pharmacology
  • Sequence Analysis, DNA -- Methods
  • Succinate Dehydrogenase -- Antagonists & Inhibitors
ispartof: Pest management science, March 2016, Vol.72(3), pp.566-73
description: The genetic underlying resistance mechanisms in the population of the phytopathogenic fungus Botrytis cinerea are well documented. Specifically, several genetic substitutions associated with succinate dehydrogenase inhibitor (SDHI)-based fungicide resistance have been identified in the succinate dehydrogenase gene. The objective of the present work was to develop a molecular tool for accurate quantification of these genetic substitutions within Botrytis populations. A test using the PyroMark Q24 instrument was designed to detect and quantify five genetic substitutions associated with SDHI resistance. The technique is based on sequencing by synthesis, and it generated quantitative and accurate data with a limit of quantification of a minimum of 500 spores. There was a linear relationship between the known and estimated percentages of spores with the targeted genetic substitutions and wild-type strains at ratios of 0-100%, with a 20% increment. With the pyrosequencing assay developed in this study, a large number of Botrytis spp. individuals can be characterised in a timely fashion with greater accuracy than by commonly used methods. Hence, pyrosequencing-based methods will be useful for improving our understanding of fungicide resistance, detecting the arrival of new genetic substitutions, monitoring shifts in fungal populations and assessing the effectiveness of antiresistance strategies, and for routine monitoring of fungicide resistance.
language: eng
source:
identifier: E-ISSN: 1526-4998 ; PMID: 25900263 Version:1 ; DOI: 10.1002/ps.4026
fulltext: fulltext
issn:
  • 15264998
  • 1526-4998
url: Link


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titleA pyrosequencing-based method to quantify genetic substitutions associated with resistance to succinate dehydrogenase inhibitor fungicides in Botrytis spp. populations
creatorGobeil-Richard, Mélanie ; Tremblay, David-Mathieu ; Beaulieu, Carole ; Van Der Heyden, Hervé ; Carisse, Odile
ispartofPest management science, March 2016, Vol.72(3), pp.566-73
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subjectBotrytis ; Pyromark Q24 ; Sdhi ; Fungicide Resistance ; Pyrosequencing ; Botrytis -- Drug Effects ; Drug Resistance -- Genetics ; Fungicides, Industrial -- Pharmacology ; Sequence Analysis, DNA -- Methods ; Succinate Dehydrogenase -- Antagonists & Inhibitors
descriptionThe genetic underlying resistance mechanisms in the population of the phytopathogenic fungus Botrytis cinerea are well documented. Specifically, several genetic substitutions associated with succinate dehydrogenase inhibitor (SDHI)-based fungicide resistance have been identified in the succinate dehydrogenase gene. The objective of the present work was to develop a molecular tool for accurate quantification of these genetic substitutions within Botrytis populations. A test using the PyroMark Q24 instrument was designed to detect and quantify five genetic substitutions associated with SDHI resistance. The technique is based on sequencing by synthesis, and it generated quantitative and accurate data with a limit of quantification of a minimum of 500 spores. There was a linear relationship between the known and estimated percentages of spores with the targeted genetic substitutions and wild-type strains at ratios of 0-100%, with a 20% increment. With the pyrosequencing assay developed in this study, a large number of Botrytis spp. individuals can be characterised in a timely fashion with greater accuracy than by commonly used methods. Hence, pyrosequencing-based methods will be useful for improving our understanding of fungicide resistance, detecting the arrival of new genetic substitutions, monitoring shifts in fungal populations and assessing the effectiveness of antiresistance strategies, and for routine monitoring of fungicide resistance.
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1The technique is based on sequencing by synthesis, and it generated quantitative and accurate data with a limit of quantification of a minimum of 500 spores. There was a linear relationship between the known and estimated percentages of spores with the targeted genetic substitutions and wild-type strains at ratios of 0-100%, with a 20% increment.
2With the pyrosequencing assay developed in this study, a large number of Botrytis spp. individuals can be characterised in a timely fashion with greater accuracy than by commonly used methods. Hence, pyrosequencing-based methods will be useful for improving our understanding of fungicide resistance, detecting the arrival of new genetic substitutions, monitoring shifts in fungal populations and assessing the effectiveness of antiresistance strategies, and for routine monitoring of fungicide resistance.
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abstractThe genetic underlying resistance mechanisms in the population of the phytopathogenic fungus Botrytis cinerea are well documented. Specifically, several genetic substitutions associated with succinate dehydrogenase inhibitor (SDHI)-based fungicide resistance have been identified in the succinate dehydrogenase gene. The objective of the present work was to develop a molecular tool for accurate quantification of these genetic substitutions within Botrytis populations. A test using the PyroMark Q24 instrument was designed to detect and quantify five genetic substitutions associated with SDHI resistance.
doi10.1002/ps.4026
pmid25900263
date2016-03