ScRpb4, Encoding an RNA Polymerase Subunit from Sugarcane, Is Ubiquitously Expressed and Resilient to Changes in Response to Stress Conditions
Round 1
Reviewer 1 Report
This manuscript presents interesting results on the transcriptional machinery in sugarcane under stressful conditions. However, it requires major revisions before further processing.
- The abstract should be rewritten. The language of the article should be passive. The use of subject pronouns in the article, especially in the abstract, materials and methods, and results is not correct. For example:
We performed an…
We show that …
We observed that
This statement is not corrected, please correct them as passive
I strongly recommend that the Language should be checked by an English professional.
- The challenges in this experiment that have been answered in the results should be explained in detail in the discussion section. In my opinion, the discussion is poorly written. The results are very good, but it is better to spend more time discussing these results with new references to better interpret that the reader can better understand the relationship between the results of this study and previous research results.
Because of the novelty of this research and very little works that have been done in this regard, I think it is better not to change the title and it is appropriate
Suggested comments for correcting the abstract section:
1- Please do not use "we" especially in the abstract section. For example
Line 16- 17: Please replace "We show that, evolutionarily, ScRpb4 is conserved across kingdoms." With "The results demonstrate that ScRpb4 is evolutionarily conserved across kingdoms."
Line 18: please delete "we observed that"
Line 20-21: please replace "We also identified optimal conditions to express ScRpb4 in vitro for further studies" with "Optimal conditions to express ScRpb4 in vitro for further studies were identified"
2 In one sentence, state your general conclusion from doing this research at the end of the abstract
3 The materials and methods section is well written and does not need to be corrected
4 The results section is also accompanied by very good data and is very well presented, just, it would be better if the authors could interpret each of the results obtained in the discussion section based on previously published articles, although I know that very little work has been done in this area.
Author Response
Manuscript: Agriculture-1515747
Title: ScRpb4, encoding an RNA polymerase subunit from sugarcane, is ubiquitously expressed and resilient to changes in response to stress conditions
We thank the anonymous reviewers for their careful reading of our manuscript and their insightful comments and suggestions. In addition to our responses to each reviewer’s comment (below – responses in green) we are also providing a revised manuscript that reflects their suggestions and comments. We feel that this has resulted in an improved version of our original manuscript.
Reviewers' comments:
Reviewer #1
This manuscript presents interesting results on the transcriptional machinery in sugarcane under stressful conditions. However, it requires major revisions before further processing.
- The abstract should be rewritten. The language of the article should be passive. The use of subject pronouns in the article, especially in the abstract, materials and methods, and results is not correct. For example:
We performed an…
We showed that …
We observed that
This statement is not corrected, please correct them as passive
I strongly recommend that the Language should be checked by an English professional.
Response: We thank reviewer #1 for the insightful comments. We have modified the abstract as suggested. However, we would like to point out that the use of subject pronouns is common and widely accepted in the scientific community. There are countless examples in many top journals in different areas. Please see below some instances:
- https://www.nature.com/articles/s41477-021-01031-8
- https://www.nature.com/articles/s41477-021-01008-7#Abs1
- https://www.science.org/doi/10.1126/sciadv.abh2434
- https://www.science.org/doi/10.1126/science.abg0929
- https://www.pnas.org/content/118/43/e2112970118
- https://academic.oup.com/plcell/advance-article/doi/10.1093/plcell/koab317/6484645?searchresult=1
- https://www.mdpi.com/2077-0472/11/11/1091
- https://www.mdpi.com/2073-4395/11/10/1957
- https://www.cell.com/molecular-plant/fulltext/S1674-2052(21)00488-3#relatedArticles
- https://www.cell.com/molecular-plant/fulltext/S1674-2052(19)30398-3
- The challenges in this experiment that have been answered in the results should be explained in detail in the discussion section. In my opinion, the discussion is poorly written. The results are very good, but it is better to spend more time discussing these results with new references to better interpret that the reader can better understand the relationship between the results of this study and previous research results.
Response: We thank reviewer #1 for his suggestions. We have expanded our discussion and included new references. Due to the very little work that has been done in this area, we have included papers in plants when available. New paragraphs have been included in the improved discussion [Lines 344-348, 356-359, 385-392, 401-403, 411-414].
Because of the novelty of this research and very little works that have been done in this regard, I think it is better not to change the title and it is appropriate
Response: We have kept the title as suggested by the reviewer.
Suggested comments for correcting the abstract section:
1- Please do not use "we" especially in the abstract section. For example
Line 16- 17: Please replace "We show that, evolutionarily, ScRpb4 is conserved across kingdoms." With "The results demonstrate that ScRpb4 is evolutionarily conserved across kingdoms."
Response: We have changed the sentence as suggested by the reviewer.
Line 18: please delete "we observed that"
Response: We have changed the sentence as suggested by the reviewer.
Line 20-21: please replace "We also identified optimal conditions to express ScRpb4 in vitro for further studies" with "Optimal conditions to express ScRpb4 in vitro for further studies were identified"
Response: We have changed the sentence as suggested by the reviewer.
2 In one sentence, state your general conclusion from doing this research at the end of the abstract
Response: We thank reviewer #1 for bringing this point into discussion. Thus, based on the reviewer’s suggestions we have added a paragraph in the abstract and the introduction on the reason of performing this research.
The abstract [Lines 21-24] paragraph can be read as: “In this study, we present an initial characterization of the sugarcane polymerase II subunit IV. Our results open the window to more specific experiments to study ScRpb4 function, for instance, crystal structure determination and pull-down assays as well as their function under biotic and abiotic stresses.”
3 The materials and methods section is well written and does not need to be corrected
Response: We thank reviewer #1 for the positive comments.
4 The results section is also accompanied by very good data and is very well presented, just, it would be better if the authors could interpret each of the results obtained in the discussion section based on previously published articles, although I know that very little work has been done in this area.
Response: We thank reviewer #1 for the positive comments. We have also improved the discussion using previously published articles.
Reviewer 2 Report
Kim et al. have provided a good study based on the characterization of the sugarcane RNA polymerase subunit IV (ScRpb4). The experiment is well planned and executed and most of the necessary information is provided in the manuscript. However, I suggest authors to add a few lines about the benefit of the manuscript/work in the abstract. They can add that how the presented data will be beneficial for further research work in this direction. The novelty of this manuscript should be highlighted in both the abstract and introduction. One more point, PCR conditions can be added to the manuscript. From Line 310-313, delete the lines ‘Authors should discuss the results and how they can be interpreted from the perspective of previous studies and of the working hypotheses. The findings and their implications should be discussed in the broadest context possible. Future research directions may also be highlighted.’ I think these lines were mistakenly left.
Regards,
Author Response
Manuscript: Agriculture-1515747
Title: ScRpb4, encoding an RNA polymerase subunit from sugarcane, is ubiquitously expressed and resilient to changes in response to stress conditions
We thank the anonymous reviewers for their careful reading of our manuscript and their insightful comments and suggestions. In addition to our responses to each reviewer’s comment (below – responses in green) we are also providing a revised manuscript that reflects their suggestions and comments. We feel that this has resulted in an improved version of our original manuscript.
Reviewers' comments:
Reviewer #2
Kim et al. have provided a good study based on the characterization of the sugarcane RNA polymerase subunit IV (ScRpb4). The experiment is well planned and executed and most of the necessary information is provided in the manuscript.
Response: We thank reviewer #2 for the positive comments.
However, I suggest authors to add a few lines about the benefit of the manuscript/work in the abstract. They can add that how the presented data will be beneficial for further research work in this direction. The novelty of this manuscript should be highlighted in both the abstract and introduction.
Response: We thank reviewer #2 for bringing this point into discussion. Thus, based on the reviewer’s suggestions we have added a paragraph on the abstract and the introduction strategy.
The abstract [Lines 21-24] paragraph can be read as: “In this study, we present an initial characterization of the sugarcane polymerase II subunit IV. Our results open the window to more specific experiments to study ScRpb4 function, for instance, crystal structure determination and pull-down assays as well as their function under biotic and abiotic stresses.”
The new paragraph in the introduction [Lines 78-82] can be read as: “Here, we present an initial characterization of the sugarcane polymerase II subunit IV (ScRpb4). Our results open the window to expand the knowledge of ScRpb4 function. Determination of ScRpb4 crystal structure, protein–protein interactions and its response under biotic and abiotic stresses will improve our understanding of the complex sugarcane gene expression machinery.”
One more point, PCR conditions can be added to the manuscript.
Response: We have added PCR conditions in the material and methods section [Lines 109-116]. It can be read now as: “PCR conditions were as follows: a final volume of 25 µL containing 50 ng of plasmid DNA, 0.20 µmol L−1 of each primer, 1.5 mmol L−1 MgCl2, 0.2 mmol L−1 of each dNTP, 1X reaction buffer, and 1 U Phusion high-fidelity DNA polymerase (#F-530S, ThermoFisher Scientific, Waltham, MA) using a ProFlex PCR System (ThermoFisher Scientific, Waltham, MA). Amplifications were performed using the following program: denaturation step of 4 min at 94°C, followed by 35 cycles of a denaturation step at 94°C for 45 s, an annealing step at 60°C for 45 s, an extension step at 72°C for 1.5 min, and then a final extension step at 72°C for 5 min.”
From Line 310-313, delete the lines: ‘Authors should discuss the results and how they can be interpreted from the perspective of previous studies and of the working hypotheses. The findings and their implications should be discussed in the broadest context possible. Future research directions may also be highlighted.’ I think these lines were mistakenly left.’
Response: We thank reviewer #2 for pointing this out. We have removed this paragraph.
Round 2
Reviewer 1 Report
The authors have used most of the reviewer's comments and corrections to further improve their manuscript.
Reviewer 2 Report
Comments have been addressed by the authors. Accept in the present form
