Comparative Proteomics Analysis of Phosphine-Resistant and Phosphine-Susceptible Sitophilus oryzae (Coleoptera: Curculionidae)

Round 1

Reviewer 1 Report

The present paper is focused to determine potential molecular markers to evaluate the rice weevil phosphine resistance. The authors determined the differences in protein expression of resistance and susceptible strains of Sitophilus oryzae by using 2D PAGE gel electrophoresis and MALDI-TOF/TOF.

Overall, this is a well-performed study that I consider is important and represents a new strategy to obtain rapid results on the evaluation of the rice weevil phosphine resistance.

The authors need to address the below comments to strengthen the quality of the manuscript:

1. Please include in the Introduction information related to a similar study published by Víctor A. Levy-De la Torre et al, Mitochondrial response of the lesser grain borer Rhyzopertha dominica (F.) to modified atmospheres, Journal of Stored Products Research 83 (2019) 338e346 , https://doi.org/10.1016/j.jspr.2019.08.005. .Also in the Results and Discussion part add discussion related to the results obtained in the above-mentioned article and your results.
2. Please include in the Conclusions the information regarding the novelty of the results obtained in the present study and the recommendation of the possible applications of this strategy to evaluate the rice weevil phosphine resistance.

Author Response

We appreciate the consideration of the two anonymous reviewers and the final correction. All suggestions were accepted, and the appropriate revisions were made. 

Thank you so much for your kind comments.

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript by Koo et al. describes a proteomic analysis between the expression profiles of phosphine-resistant and -susceptible strains of Sitophilus oryzae using 2D-PAGE and MALDI-time of flight MS. My comments and suggestions:

1. Although the aim of the study is the comparison and the differences in the total proteins (as it is written in line 50), the authors identified only a short number of protein spots (19 spots).
2. Why they did not select a LC-based analysis (in solutions) instead of gel-based approach?
3. Characteristic MS/MS spectra must be included in the text or should be given Supporting Information.
4. The introduction should be strengthened and a significant number of previous studies and MS-methodologies in the field should be included.
5. Section 3 describes only Results. A different section with Discussion should be included.
6. Conclusions could be re-organized increasing this paragraph as three lines are not enough (lines 171-174).

Author Response

We appreciate the consideration of the two anonymous reviewers and the final correction. All suggestions were accepted, and the appropriate revisions were made. 

Thank you so much for your kind comments.

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

The authors did not address all my comments and questions.

My comment #1: The authors report that total proteome is only 19 proteins? Although they stress the weakness of the 2D-based approach against the LC-MS/MS analysis (reply in my 2^nd comment), they still present the study as "…..the comparison and the differences in the total proteins’’ (line 59 and line 196).

My comment #3: Where are the MS/MS spectra? I can not see them in the main draft as written results. The authors gave a powerpoint file that no one can read it! Please improve the quality of this file, increase all the numbers in the axes and explain in details both in the manuscript and in the Supporting file what you want to present. The authors should also add in page 7: Supporting Information: MS/MS spectra of ......

Comment #4: I suggested the addition of a significant number of previous studies and comparison with different MS-methodologies methodologies. The authors added only 2 studies!

Comment #5: Where is the extra Discussion section? If the authors would like to present Section 3 as Results and Discussion together, their results are only paragraph 3.1;; Limited results part which could be improved.

Comment #6. Please improve your conclusion remarks.

Author Response

Thank you so much for your kind comments.

When we first submitted the manuscript, we received the following notification from the assistant editor: We found that the manuscript is quite short, discussion is only 2 pages and conclusions 4 lines. Our academic editor suggested that paper type should be changed in “communication". Would it be possible for you to change it to communication type?

So we accepted. Therefore, a revision was created without being affected by the length of the content. However, some modifications were made according to the reviewer's request.

Author Response File: Author Response.docx

Round 3

Reviewer 2 Report

The authors have addressed my questions in the last revised version.