Combined Transcriptome and Proteome Analysis of the Protein Composition of the Brochosomes of the Leafhopper Nephotettix cincticeps
Round 1
Reviewer 1 Report
In this study, the authors performed transcriptomic and proteomic analyses to investigate the protein composition of brochosomes in N. cincticeps. Transcriptomic data were generated from the salivary gland, midgut, malpighian tube, and residual body. Through this analysis, the authors identified 106 malpighian tube-specific genes. Using two different methods, brochosomes were collected from the forewings, resulting in the discovery of 815 common proteins. Based on these two approaches, the authors identified 22 candidate brochosome proteins. With these candidates, the authors conducted motif analysis and analyzed taxonomic distribution. This data would provide valuable insights into understanding brochosomes. Overall, the manuscript is well written and clear, and easy to follow. However, there are some parts that need modification and further explanation.
Major points:
1. How many replicate samples were used for transcriptome analysis? If possible, confirm the expression of several Mt-specific genes using RT-qPCR to enhance the reliability of the transcriptome analysis data.
2. Please make some revisions in cases where the information mentioned in the Method section is duplicated in the figure legend or Results section to avoid redundancy.
Minor points:
1. Please clearly specify whether the author used p-values or q-values to identify Mt-specific genes. It is mentioned as a p-value in the Method section and a q-value in the Results section.
2. Figure S1: Provide the information about A, B, C, … and Z in Figure S1
3. Figure 3E: Provide the information about I, II, III, IV, and V
4. The information mentioned in the Method section differs from that in the Results section. Which one is accurate, the 10% SDS-PAGE gel or the 12% SDS-PAGE gel?
5. If possible, please also include BSM/BSAP in Figure 4.
6. Please use the consistent format or terminology in the manuscript
a. p value / p-value
b. min / minute
c. 1000 g / 1000 ×g
d. 1,195 proteins / 9562 genes (comma)
Author Response
Dear Reviewer,
I sincerely appreciate your time and effort in reviewing our manuscript and providing valuable feedback. Your expertise and insightful comments are highly regarded and will undoubtedly contribute to enhancing the quality of our research.
In response to your valuable suggestions, we have given careful consideration to each of your comments and made revisions to the manuscript. Please find attached a point-by-point response document as well as the revised manuscript.
We believe that these modifications address the concerns you raised and improve the overall quality of the manuscript. Once again, we express our gratitude for your thorough feedback, which is invaluable in refining our work.
Thank you for your continued support and guidance throughout the review process. We look forward to receiving further feedback from you and hope that these revisions meet your expectations.
Sincerely,
Wei Wu
Author Response File: 
Author Response.docx
Reviewer 2 Report
Please, as requested by the journal, differentiate more the simple summary and the abstract which seem to contain the same information.
Keywords: please change these keywords as are already in the title “Brochosome; Leafhopper; Transcriptome; Nephotettix cincticeps”
Please delete this section “0. How to Use This Template”
INTRODUCTION
Please add references to these sentences
- In Cicadellidae, Malpighian tubule is divided into proximal, distal, and terminal segments.
- The lumen of distal segment is surrounded by a single layer of secretory cells with large, spherical nuclei and an extended, rough endoplasmic reticulum.
MATERIAL AND METHODS
- Sample preparation and Illumina sequencing
Why was RNA not verified on agarose gel?
- Brochosomes collection
Line 135: At what stage of their life cycle were the specimens used for the work? Was the sample homogeneous or heterogeneous? Was the presence of brochosomes also verified in the eggs?
- Bioinformatics analysis
Line 201: italics for N. cincticeps
RESULTS
- Nephotettix cincticeps transcriptome sequencing
Some sentences are related to material and method section exclusively, please delete them.
- The authors wrote about the signal peptide sequence, but in the material and method section it was not reported.
FIGURE CAPTIONS
Please enrich all the figure captions; moreover, all letters should be explained.
English language is fine
Author Response
Dear Reviewer,
I sincerely appreciate your time and effort in reviewing our manuscript and providing valuable feedback. Your expertise and insightful comments are highly regarded and will undoubtedly contribute to enhancing the quality of our research.
In response to your valuable suggestions, we have given careful consideration to each of your comments and made revisions to the manuscript. Please find attached a point-by-point response document as well as the revised manuscript.
We believe that these modifications address the concerns you raised and improve the overall quality of the manuscript. Once again, we express our gratitude for your thorough feedback, which is invaluable in refining our work.
Thank you for your continued support and guidance throughout the review process. We look forward to receiving further feedback from you and hope that these revisions meet your expectations.
Sincerely,
Wei Wu
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
The author has addressed all of the reviewer's points in the revised manuscript, and I have no further comments. Therefore, I recommend the publication of the revised manuscript.
