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

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posted on 2023-05-10, 07:36 authored by Hossein Ghanizadeh, Andrew GriffithsAndrew Griffiths, Chris BuddenhagenChris Buddenhagen, Craig AndersonCraig Anderson, Kerry Harrington
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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Rights statement

© 2021 Ghanizadeh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Language

  • English

Does this contain Māori information or data?

  • No

Publisher

PLOS

Journal title

PLoS One

ISSN

1932-6203

Citation

Ghanizadeh, H., Griffiths, A. G., Buddenhagen, C. E., Anderson, C. B., & Harrington, K. C. (2021). A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne. PLoS One, 16(2), e0246028. doi:10.1371/journal.pone.0246028