A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne

The first step in managing herbicide-resistant weeds is to confirm their resistance status. It is, therefore, crucial to have a rapid, reliable and cost-effective technique to assess samples for herbicide resistance. We designed and evaluated three derived cleaved amplified polymorphic sequence (dCA...

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Autores principales: Ghanizadeh, Hossein, Griffiths, Andrew G., Buddenhagen, Christopher E., Anderson, Craig B., Harrington, Kerry C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7853469/
https://www.ncbi.nlm.nih.gov/pubmed/33529261
http://dx.doi.org/10.1371/journal.pone.0246028
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author Ghanizadeh, Hossein
Griffiths, Andrew G.
Buddenhagen, Christopher E.
Anderson, Craig B.
Harrington, Kerry C.
author_facet Ghanizadeh, Hossein
Griffiths, Andrew G.
Buddenhagen, Christopher E.
Anderson, Craig B.
Harrington, Kerry C.
author_sort Ghanizadeh, Hossein
collection PubMed
description The first step in managing herbicide-resistant weeds is to confirm their resistance status. It is, therefore, crucial to have a rapid, reliable and cost-effective technique to assess samples for herbicide resistance. We designed and evaluated three derived cleaved amplified polymorphic sequence (dCAPS) markers for detecting glyphosate resistance in Lolium perenne. conferred by non-synonymous mutations at codon-106 in the enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene. The dCAPS markers involve amplification of the target region, digestion of the amplified products with restriction enzymes and gel-based visualisation of the digested products. The results showed that all three dCAPS markers could successfully detect mutations at codon-106 in the target enzyme. The dCAPS markers can also inform us of the zygosity state of the resistance allele and was confirmed by sequencing the target region of the EPSPS gene. The markers described here are effective quick tests for the monitoring and evaluation of the target-enzyme mechanism of glyphosate resistance in Lolium perenne.
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spelling pubmed-78534692021-02-09 A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne Ghanizadeh, Hossein Griffiths, Andrew G. Buddenhagen, Christopher E. Anderson, Craig B. Harrington, Kerry C. PLoS One Research Article The first step in managing herbicide-resistant weeds is to confirm their resistance status. It is, therefore, crucial to have a rapid, reliable and cost-effective technique to assess samples for herbicide resistance. We designed and evaluated three derived cleaved amplified polymorphic sequence (dCAPS) markers for detecting glyphosate resistance in Lolium perenne. conferred by non-synonymous mutations at codon-106 in the enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene. The dCAPS markers involve amplification of the target region, digestion of the amplified products with restriction enzymes and gel-based visualisation of the digested products. The results showed that all three dCAPS markers could successfully detect mutations at codon-106 in the target enzyme. The dCAPS markers can also inform us of the zygosity state of the resistance allele and was confirmed by sequencing the target region of the EPSPS gene. The markers described here are effective quick tests for the monitoring and evaluation of the target-enzyme mechanism of glyphosate resistance in Lolium perenne. Public Library of Science 2021-02-02 /pmc/articles/PMC7853469/ /pubmed/33529261 http://dx.doi.org/10.1371/journal.pone.0246028 Text en © 2021 Ghanizadeh et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Ghanizadeh, Hossein
Griffiths, Andrew G.
Buddenhagen, Christopher E.
Anderson, Craig B.
Harrington, Kerry C.
A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne
title A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne
title_full A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne
title_fullStr A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne
title_full_unstemmed A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne
title_short A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne
title_sort pcr plus restriction enzyme-based technique for detecting target-enzyme mutations at position pro-106 in glyphosate-resistant lolium perenne
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7853469/
https://www.ncbi.nlm.nih.gov/pubmed/33529261
http://dx.doi.org/10.1371/journal.pone.0246028
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