PONE-D-20-36361

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

PLOS ONE

Dear Dr. Ghanizadeh,

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Reviewer #1: Yes

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Reviewer #1: Comments:

Manuscript # PONE-D-20-36361

The manuscript by Ghanizadeh et at. describes the technique of detecting mutations on EPSPS at the position Pro-106 for perennial ryegrass (Lolium perenne), which was developed based on the dCAPS method published earlier by Neff et al. (Plant Journal 1998). The primers were designed in a way that the PCR products can only be cut by the enzymes when the codon coding for proline-106 at the first and second nucleobases are not mutated which is susceptible to glyphosate. These dCAPS target-site mutation markers were further tested using progeny from the ‘Blenheim’ population. This approach/technique can detect the target enzyme mutations as well as the zygosity of the mutant alleles. The manuscript generally is well written, and the experimental design is fine, and the experiments were apparently well-executed. However, I have a few comments to the manuscript, listed below.

In table 1, the MscI cut site should be TGG/CCA.

Legends in figures 1 and 3, lines 446 and 467, the term “company” should be specified.

In figure 3, “RR” glyphosate-resistant individuals (lanes 11 and 14) seem to have the cleaved (~180 bp) dCAPS amplicon, although the bands are faint. The authors may want to elaborate it a bit in the text.

There are 4 figures in the manuscript. Figure 4 (chromatogram) did not have legend and was not mentioned in the text.

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Reviewer #1: No

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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.

PONE-D-20-36361R1

Dear Dr. Ghanizadeh,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

David D Fang, Ph.D.

Academic Editor

PLOS ONE

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