Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VERTICAL AND TEMPORAL VARIATIONS OF SOIL BACTERIAL AND ARCHAEAL COMMUNITIES IN WHEAT-SOYBEAN ROTATION AGROECOSYSTEM
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript entitled “Vertical and temporal variations of soil bacterial and archaeal
communities in maize-soybean rotation agroecosystem” submitted by Yokota et al. aimed to study the variations in bacterial and archaeal population in surface/middle and deep soil. There is no novelty in this work. There are various corrections listed below.

1.	Raw data for this manuscript have been submitted but it is already published in other paper. Disentangling the role of salinity-sodicity in shaping soil microbiome along a natural saline-sodic gradient DOI:10.1016/j.scitotenv.2020.142738.
2.	Please remove the plagiarised words from the manuscript.
3.	Please change the reference pattern in whole manuscript. It should be year wise first 2006 then 2007 and so on.
4.	Please rewrite the abstract and mention how your study is better than others.
5.	Line 260 improve the sentence; reference have repeated two-time Shade et al. Correct it.
Experimental design: 6.	In line 100 can bacterial population survived at -80℃??
7.	Line 104-105 provide references for estimation of soil parameters (organic carbon, NPK etc.).
8.
Validity of the findings: Discussion part is too short and discuss in details with recent references.
9.	Discuss about the role of different phyla in soil how they involved in nutrient cycle.
10.	Please add these two-reference related to this study in line 263
a) Chaudhary P, Khati P, Chaudhary A, Maithani D, Kumar G, Sharma A (2021d) Cultivable and metagenomic approach to study the combined impact of nanogypsum and Pseudomonas taiwanensis on maize plant health and its rhizospheric microbiome. PLoS ONE 16(4): e0250574. https://doi.org/10.1371/journal. pone.0250574,
b) Chaudhary P, Sharma A, Chaudhary A, Khati P, Gangola S, Maithani D (2021b). Illumina based high throughput analysis of microbial diversity of rhizospheric soil of maize infested with nanocompounds and Bacillus sp. Applied Soil Ecology. 159: 103836. doi: http://doi.org/10.1016/j.apsoil.2020.103836
11.	Add this reference in line 241.
Kukreti B, Sharma A, Chaudhary P, Agri U, Maithani D (2020). Influence of nanosilicon dioxide along with bioinoculants on Zea mays and its rhizospheric soil. 3Biotech. 10: 345. https://doi.org/10.1007/s13205-020-02329-8,
12.	You can add these references in line 283 related to soil fertility.
a)	Kumari S, Sharma A, Chaudhary P, Khati P (2020). Management of plant vigor and soil health using two agriusable nanocompounds and plant growth promotory rhizobacteria in Fenugreek. 3Biotech. 10: 461. https://doi.org/10.1007/s13205-020-02448-2,
b) Chaudhary P, Khati P, Gangola S, Kumar A, Kumar R, Sharma A (2021c) Impact of nanochitosan and Bacillus spp. on health, productivity and defence response in Zea mays under field condition. 3 Biotech. 11: 237. http://doi.org/10.1007/s13205-021-02790-z.
               c) Chaudhary P, Khati P, Chaudhary A, Gangola S, Kumar R, Sharma A (2021a) Bioinoculation using indigenous Bacillus spp. improves growth and yield of Zea mays under the influence of nanozeolite. 3Biotech. 11: 11. https://doi.org/10.1007/s13205-020-02561-2.
13.	Please discuss the outcome and how it benefits for others (its value).
Additional comments: Please check the accession number you have provided because it is already published in another paper. How is it possible to submit the information in two papers.  Please remove the plagiarism in the manuscript.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VERTICAL AND TEMPORAL VARIATIONS OF SOIL BACTERIAL AND ARCHAEAL COMMUNITIES IN WHEAT-SOYBEAN ROTATION AGROECOSYSTEM
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this paper, the authors examined in depth the possible vertical and temporal patterns of soil bacterial and archaeal communities in maize-soybean rotation agroecosystem. In particular, the authors pointed out that the bacterial and archaeal communities are impacted and shaped by the soil depth. Conversely, significant temporal patterns were not observed for both communities. The authors also focused on the rare taxa, identifying the rare bacterial community as the key determinants of the soil fertility index.
I find interesting the general concept of the work that incorporates temporal variation, in addition to vertical variation, as a putative determinant in influencing the distribution of soil microbes. However, I have some comments/suggestions on some points that I believe the authors should clarify and/or improved.
Experimental design: 1) Soil characteristics (as TN, TC, etc.) obtained by the physiochemical analysis are not reported as supplementary results, as well as the methodologies are not explained in Materials and Methods. I think that these results, if available, should be reported as supplementary results. Otherwise, the authors should mention it only in the results sections as “data are not reported”. More important, are these parameters measured for the different sampling times? I think the authors should report this information.
Validity of the findings: 2) The analysis was conducted in a maize-soybean rotation agro-ecosystem over one year. However, it seems to me that the agricultural context in which this analysis is carried out is little emphasized.  For example, in the introduction, as well as in the discussion, the authors do not explicit/clarify why it is important to understand if (and eventually how) the soil depth influences the diversity and composition of soil microbial communities in agricultural soils especially when a crop rotation is performed. The deeper soils are important for crop yield, especially for cereal crops as maize. Therefore, the authors should explain how the information on the vertical patterns can help the sustainability of agricultural systems. Moreover, the soil prokaryotic community compositions are not linked to the agricultural context and the crops present in the different sampling times.
3) The authors should discuss more deeply why they did not detect a temporal variation in this particular crop rotation system. In particular, by Random forest analysis, the authors found the Total Nitrogen as one of the key determinants for bacterial richness and evenness. It is well known that the use of the leguminous plant in a crop rotation is crucial for increasing the total nitrogen in the soil. Concurrently, they found that the bacterial richness did not show temporal variations after the rotation with soybean, which certainly increased the Nitrogen content. I think that the authors should examine in-depth this point. Also, in the discussion section, the authors stated that “The temporal variations of the soil microbial community were attributable to two factors: the shift of soil nutrients and/or the plant growth stage (Yang, 2017)” (line 261-263) and justify their results saying that the soil samples were collected from only two plant individuals. What about the shift of soil nutrients? Again, did the authors verified if soil nutrients contents and, in general, soil properties were different among the sampling times? If not, I think that the phrase “The minimal impact may also be due to no apparent difference in soil environmental conditions among sampling times” (line 265-266) should be removed.
Additional comments: 4) In general, the impression is that the authors glossed over the results, which are sometimes roughly reported. Moreover, It is often not clear to which figures (and to which panels of the figures) the authors were referring. Line 172- 174 for example.
5) The authors have confused Figure 3 with Figure 4.
6) I suggest the author using the Tax4fun software package, which can predict the functional profiles of bacterial and archaeal communities. This might shed a light on the functionality of rare bacterial and archaeal communities the authors found, then greatly improved the paper.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VERTICAL AND TEMPORAL VARIATIONS OF SOIL BACTERIAL AND ARCHAEAL COMMUNITIES IN WHEAT-SOYBEAN ROTATION AGROECOSYSTEM
Review round: 1
Reviewer: 3

Output:
Basic reporting: Overall the manuscript is well written, according to the journal's instructions, except for the reference section. We recommend to remove the initial number from the list of references and adopt the following Journal reference format: List of authors (with initials). Publication year. Full article title. Full title of the Journal, volume: page extents. DOI (if available) (see author’s instructions here https://peerj.com/about/author-instructions/#reference-format for more details)

Literature references are appropriate and figures/tables are well presented.

Regarding the raw data the authors affirmed having deposited the sequences in the NCBI SRA repository (accession No. SRP271579). However, those sequences do not seem to correspond to the sequences of this manuscript, being 121 samples (instead of 72) related to the microbiome of saline-sodic soils in west Songnen Plain. We recommend to carefully check and revise the info.
Experimental design: The authors presented a manuscript aiming to address the vertical and temporal patterns of soil bacterial and archaeal communities in maize-soybean rotation agroecosystem at a scale of millions of sequences per sample. The main goals were to address: (1) Which factor from soil depth and temporal fluctuations is the key determinant in shaping bacterial and archaeal communities; (2) Whether there exist differences in the vertical distribution pattern between soil bacteria and archaea; (3) Whether rare taxa occupy a larger proportion in the surface soil layer than in the deep soil layer.

Although both title and conclusions clearly refer to maize-soybean rotation, in the M&M section the authors described a wheat-soybean experimental design. This has to be carefully revised.

In general, the proposed methods were appropriate for the scope of the work. However, methods used to assess soil physicochemical characteristics were not described and no references were provided.

Considering the relevance of the "rare" bacterial taxa in this work, a better description of the criteria adopted for the selection of the "cut-off" value at 0.1% rather than 0.01% of the reads should be provided and possibly supported by literature or a clear motivation.
Validity of the findings: Overall, the topic of this work is not very innovative and many studies already addressed the distribution of soil microbial communities across soil depth during the last two decades (i.e. Eilers, K. G., Debenport, S., Anderson, S., & Fierer, N. (2012) Digging deeper to find unique microbial communities: the strong effect of depth on the structure of bacterial and archaeal communities in soil. Soil Biology and Biochemistry, 50, 58-65) as well as the role and temporal shifts of rare communities (i.e. Shade, A., Jones, S. E., Caporaso, J. G., Handelsman, J., Knight, R., Fierer, N., & Gilbert, J. A. (2014). Conditionally rare taxa disproportionately contribute to temporal changes in microbial diversity. MBio, 5(4), e01371-14). However, the focus put on the spatial-temporal distribution of rare taxa across soil depth represents an intriguing item to be better explored.

The authors claimed to have addressed a number of soil physiochemical parameters (es. TC, TN, DOC, pH, etc.) but their values were not included in the manuscript nor as results or into the supplemental materials. Considering their relevance for the interpretation of the work and that they have been used for microbial community analysis, it is highly recommended to include them in the main text as additional table.

The authors concluded that "we evaluated the effects of soil depth and sampling time on soil bacterial and archaeal community and co-occurrence networks in maize-soybean rotation agroecosystem at a depth of millions of sequences per sample". However, it's not maize but wheat and they did not get millions of sequences per sample.
Finally, the authors argued that "our study also revealed the potential of rare microbial taxa in soil biogeochemical function" but it was rather speculative and poorly supported by results. The discussion on this point should be significantly improved and supported by literature.
Additional comments: Minor comments:
Line 29: The reference of Caporaso et al.2011 seems to be poorly relevant here.
Line 56: bacterial diversity
Line 99: four sample sites?
Line 114: Qiime or Qiime2?
Lines 124-125: Relative abundances below 0.01% were defined as “rare” but above 0.1% as “abundant”. Are you sure that they were not the same value? Please, check it. In any case,
Line 172: the authors claim that the evenness index was significantly higher in October than in April (Fig.1). However, looking at the values reported in figure 1 (C), it does not seems to be true. Please, check it carefully.
Line 237: the authors affirmed that “we observed that bacterial richness and evenness in the middle soil layer were roughly equal to that in the surface”. However, the values of bacterial richness reported in Fig.1 show very different values between surface and middle soil samples. Please, check it. Moreover, one possible  explanation proposed by the authors rely on the equal distribution of organic carbon in the two layers. However, the values of carbon distribution across soil layers is missing.
Lines 245-247: a comment/discussion on the eveness of archea is needed.
Lines 281-283: the authors claimed “We also found that rare taxa were key determinants of soil fertility, indicating that rare taxa had an over-proportional role in the soil biogeochemical cycling process”. However, it is not clear to me the results upon they based their speculation.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VERTICAL AND TEMPORAL VARIATIONS OF SOIL BACTERIAL AND ARCHAEAL COMMUNITIES IN WHEAT-SOYBEAN ROTATION AGROECOSYSTEM
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am recommended this manuscript for publication.
Experimental design: Well explained
Validity of the findings: Good
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VERTICAL AND TEMPORAL VARIATIONS OF SOIL BACTERIAL AND ARCHAEAL COMMUNITIES IN WHEAT-SOYBEAN ROTATION AGROECOSYSTEM
Review round: 2
Reviewer: 2

Output:
Basic reporting: I appreciated the efforts the authors made in reviewing the manuscript. I think most of the hints were adequately taken into account, as re-uploading the correct sequences, including the soil physiochemical parameters, and discussing more deeply some points. However, I still found the current version superficial in both reporting and deeply analyzing some available data/information.
Experimental design: In my concern, the putative functionality of rare taxa is the most engaging aspect of this manuscript. However, I believe this was not sufficiently explored although was previously suggested. Despite the authors employing the FAPROTAX tool, the functional prediction and, consequently, the distribution of the functional categories from the rare bacterial communities were not found out.
Validity of the findings: The authors reported that “the rare bacterial community was the key determinant of the soil fertility index, as is clearly shown by the supplementary data (figure S2 and 3). It would have been interesting to explore in which specific soil biogeochemical cycling processes (functional categories) are involved the prokaryotic phyla the authors found and their related proportion.
Additional comments: I strongly suggest a revision of the English language.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VERTICAL AND TEMPORAL VARIATIONS OF SOIL BACTERIAL AND ARCHAEAL COMMUNITIES IN WHEAT-SOYBEAN ROTATION AGROECOSYSTEM
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors re-edited the reference format as requested. It is not reported in alphabetic order but the journal will edit it.

The authors re-uploaded the sequences as requested on NCBI SRA repository (accession number No. PRJNA766099). However, in the text (line 117) there is still the wrong number. Please, modify it.
Experimental design: The authors revised the manuscript according with all my previous comments.

Just a last comment: at Line 119, please cite “Bolyen, E., Rideout, J. R., Dillon, M. R., Bokulich, N. A., Abnet, C., Al-Ghalith, G. A., ... & Caporaso, J. G. (2018). QIIME 2: Reproducible, interactive, scalable, and extensible microbiome data science (No. e27295v1). PeerJ Preprints (https://doi.org/10.1038/s41587-019-0209-9) instead of “Caporaso et al., 2010” for QUIIME2.
Validity of the findings: The authors added a table (Table S1), as requested. However, the units of the soil variables and the statistical meaning of the letters was not specified. Please, add them.
Additional comments: No comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE EFFECT OF FARMLAND ON THE SURFACE WATER OF THE ARAL SEA REGION USING MULTI-SOURCE SATELLITE DATA
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. This article conformed to professional standards of expression.
2. This article referenced relevant literatures and provided sufficient background.
3. The structure of the article was reasonable. The content of line 278 (3.3 Accuracy assessment) should be located before 3.1 and 3.2, I think. Only the accuracy assessment reached the satisfied degree, the analysis to the farmland and waters variations were dependable.
Experimental design: Multisource satellite data is being used in many fields, and the utilization in the field of water in the article is a useful attempt. The methods in the article described with sufficient details, and can provide help to the relevant research.
Validity of the findings: The multisource satellite data used in the article are robust, the data processing methods are reasonable, and then the conclusions ate crecdible.
Additional comments: 1.	Examining in the pater title is suggested to delete.
2.	Farmland irrigation in the paper title and abstract is inaccurate. No farmland irrigation information, such as irrigation area, irrigation type, etc.，is provided in the paper. Based on the content (abandoned, replanted farmland) in the paper, land utilization maybe more accurate than farmland irrigation. And the related content in the paper can be adjusted accordingly.
3.	In Line 249, water variations maybe inaccurate, and waters variations maybe more accurate.
4.	Figure 7, different line types are suggested to be used to make the figure more clearly expressed.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE EFFECT OF FARMLAND ON THE SURFACE WATER OF THE ARAL SEA REGION USING MULTI-SOURCE SATELLITE DATA
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: See attached PDF

This paper assess the effect of irrigation on surface water. Multi-source data were employed to extract the variations of water area, water level, water storage in the Aral Sea Region. Using this data, relationship between surface water and irrigation is developed. In general, the manuscript is well explained. However, there are some issues associated with the quality of this work that need to be addressed prior to publication:

1. In line 58, you mentioned that it was verified that the reduction of downstream runoff is not associated with the upstream of the Amu Darya River and the Syr Darya River. It was verified by whom? Kindly give reference here.
2. Kindly consider rewriting sentences like in line 50 to 52 “To explore the reasons for the shrinking of the Aral Sea, numerous researches have been conducted on the rivers’ source, as presented below.”
3. In line 85 “At present, it has become an essential measure of long-term monitoring variations in surface water in large areas (Singh et al., 2012)”. What it indicate in this sentence?
4. Kindly go through your paper and remove all typos and grammar related mistakes.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE EFFECT OF FARMLAND ON THE SURFACE WATER OF THE ARAL SEA REGION USING MULTI-SOURCE SATELLITE DATA
Review round: 1
Reviewer: 3

Output:
Basic reporting: The paper is overall clear and cited relevant references in most places needed. The paper follows a professional article structure. However, there are places that need improvement.

1.	Abstract:
a. “Therefore, the surface water of the Aral Sea continuously deteriorated each year.” How can the increase in drainage deteriorate the surface water? I do not understand how the “therefore” plays a role here. In addition, the “deteriorate” is confusing. It sounds like you are talking about water quality; however, you only consider water quantity.
b. You mentioned irrigation methods/models. This is very confusing. To me, irrigation methods are drip irrigation, sprinkling irrigation, surface irrigation, etc. Apparently, the paper does not consider different types of irrigation methods.
c. I suggest adding implications at the end of the abstract, e.g., who can benefit from your study?
2.	Lines 58-59, reference is needed.
3.	Line 75, what do you mean by “establish a scientific method”? Are the methods in the previous studies not scientific? The statement is too general and arbitrary.
4.	Line 194, I am wondering about the uncertainties of the three datasets about water height. Why is a simple average good enough to be used here? What if one of them has better quality/ less uncertainties?
5.	You calculated water storage using equation (5). Then what about the water storage from GRACE data? It is unclear which data/calculation you used to reflect water storage in later analysis.
6.	Line 202, why do you only validate the 1987 classification results? What about other years?
7.	Lines 219 and 222, what are the xb, xab?
8.	Line 244-245, based on what evidence, did you state that the decrease in runoff is impacted by replanting of the farmland? I thought this is something you need to demonstrate in later analysis.
9.	Line 254, where is the Dike Kokaral dam? Why can it reduce the water flowing into East Aral?
10.	Line 287, availability should be “capability”?
11.	Line 302, wtaer is a typo.
12.	Figure 1, is “Reacher” a typo?
Experimental design: Some details of the methodology are unclear or not robust.

1. I do not think the sample sizes are robust for the correlation analysis (Figs. 8 and 9). There are only seven samples. Would it be possible to conduct the correlation using the values of each grid cell instead of an annual value? In addition, the values shown in Figs. 8 and 9 are based on the classification using random forest, so the correlation is afterthought analysis. Unless you have independent observations or mechanism hypotheses to verify your results, I do not see the reason why the correlation analysis can test the reliability of water extraction results.

2. Details of the random forest are unclear to me. First, how did you select the 200 training samples? Will the different choices of the samples influence the validation results? Second, what is the value for ntree? How are the validation results sensitive to parameter tuning? Third, mtry is the square root value of the number of variables. What are the variables?
Validity of the findings: I do not think the paper has good novelty.

1. The authors highlight that their contribution is establishing the real connection between irrigation and surface water, and the method they use is the entropy method. First, it is unclear what criteria they use to determine that the coupling coordination is high-quality. Second, the coupling degree values quantified in this study can only verify the importance of irrigation on surface water in the Aral Sea Region. However, such importance has been observed and reported by numerous previous studies. To name only a few:
Shibuo, Yoshihiro, Jerker Jarsjö, and Georgia Destouni. "Hydrological responses to climate change and irrigation in the Aral Sea drainage basin." Geophysical Research Letters 34.21 (2007).
Micklin, Philip. "The past, present, and future Aral Sea." Lakes & Reservoirs: Research & Management 15.3 (2010): 193-213.
Cretaux, Jean-François, René Letolle, and Muriel Bergé-Nguyen. "History of Aral Sea level variability and current scientific debates." Global and Planetary Change 110 (2013): 99-113.
I do not see the value of the coupling degree values. For example, how can the coupling degree value you quantified here provide implications to water resources management? I would say more innovative or quantitative research could be to specify the contribution of irrigation to the water fluxes, i.e., how much water decreases are due to irrigation and how such amount varies over time.

2. The discussion is very general, especially in section 4.2. The effect of farmland irrigation on surface water is the contribution of this paper, as the authors highlighted in the introduction. However, the discussion is just a repeat of what they have done. I suggest the authors discuss the influencing mechanism of irrigation on surface water in the Aral Sea. In addition, it is necessary to compare their results with previous literature to highlight their contribution or to verify the reliability of their results.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE EFFECT OF FARMLAND ON THE SURFACE WATER OF THE ARAL SEA REGION USING MULTI-SOURCE SATELLITE DATA
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I am overall satisfied with the authors’ responses. Many thanks to the authors for providing further analysis. I have several additional comments as follows.

Major comments:
1. The mechanisms of abandoned and replanted farmland leading to decreases of the Aral Sea are unclear to me. Abstract mentions the “Two types of farmland changes… led area of the Aral Sea to decrease”. Please clarify the mechanism separately for abandoned farmland and replanted farmland in the discussion section, i.e., how they can both lead to the decreases of the Aral Sea. To my understanding, abandoned and replanted farmland are two opposite ways of farmland utilization. It is unclear why they lead to the effects of the same direction on the Aral Sea. If they both lead to a decrease, which leads to a greater decrease, and why?
I notice that the authors try to discuss it in section 4.2. Unfortunately, this section is confusing. The authors mentioned that “if abandoned farmland were used to plant cotton, the water loss would reach 0.028 km3 every year”, and such an amount of water loss could provide water use for tens of thousands of people. To my understanding, the authors are trying to stress that farmland/cropland leads to a great amount of water loss. In other words, if the abandoned farmland were not used for planting, the water loss would not be that great. However, later in the text, the authors mentioned: “abandoned farmland has aggravated the loss of water”. This is contradictory to the previous evidence. In addition, it is unclear what the “water wasted” the authors refer to, is it the same as water loss?
2. The description of the selection of training samples is still unclear. In line 195, “the training samples were selected according to the high-resolution image of Google Earth (Figure 3).” The “according to” is confusing to me. Does it mean that “the high-resolution image of Google Earth” is a criterion you used to select training samples? If so, based on what criteria did you determine it is high-resolution? Did you randomly choose the 200 training samples from all the high-resolution images of Google Earth?

Minor comments:
Abstract:
Line 17, “unscientific” should be “improper”?
Lines 18 and 36, sentences usually do not start with “And”.
Line 36, “coupling coordination between irrigation and surface water” should be between “surface water” and “farmland”?
Line38, “The findings are contributed for …”, delete “are”. Change “formulate” to “formulating”.
Line41, What “ideas”? This is very ambiguous. Please describe directly what insights or intellectual merits your work provides to the academic community.

Introduction:
Line 133, consider deleting “to address the above issues”.

Results:
Line 287: Consider mentioning the Syr Darya and the Amu Darya in the caption of Figure 3. In addition, what do you mean by “mainly diffused especially around the Amu Darya”? Are you looking at the percentage of abandoned farmland in each Darya River? If so, suggest adding the number or percentage in the text.
Lines 289-291: The sentences are very unclear to me. First, I can’t see why Fig. 4 shows the annual change peaked between 1987 and 1993. To me, the figure shows the area of abandoned farmland, but the colors between the years are very difficult to distinguish. It may be better to indicate the values of abandoned farmland for each year in the label. Second, what do you mean by “abandoned rate”? Is it the rate of change in abandoned farmland between years (e.g., 2002 to 2016), or the percentage of abandoned farmland in a given Darya in each year from 2002 to 2016? Third, “the abandoned rate was elevated by 5.7% in 2017”. What baseline are you compared to when you say “elevated by 5.7%”?
Line 297: Again, I can’t determine the replanted rate for each year from Figure 5, let alone the trend of the rate.
Fig. 8, The figure also shows equivalent water height, but it is not described in the text.

Discussion:
Section 4.1 is like a summary of what you did instead of a real discussion. Discussion should discuss how the results can be interpreted from the perspective of previous studies. For example, do your results contradict or coincide with previous studies, and why?
Line 379, “Most e previous”, “e” is redundant.

Others:
The paper lacks space between words in many places. To name only a few, line 70, “landsto” should be “lands to”. Line 122, “Regionis” should be “Region is”. I suggest authors carefully check the spaces throughout the paper.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE EFFECT OF FARMLAND ON THE SURFACE WATER OF THE ARAL SEA REGION USING MULTI-SOURCE SATELLITE DATA
Review round: 3
Reviewer: 1

Output:
Basic reporting: 1. This article conformed to professional standards of expression.
2. This article referenced relevant literature and provided sufficient background.
3. The structure of the article was reasonable.
Experimental design: Multi-source satellite data is being used in many fields, and the utilization in the field of water area analysis in the article is a useful attempt. The methods in the article described with sufficient details, and can provide help to the relevant research.
The multi-source satellite data used in the article are robust, the data processing methods are reasonable, and then the conclusions ate credible.
Validity of the findings: 1.In line 165，2.3.1 Detection Algorithm of Farmland Change, maybe more accurate.
2.In line 183，2.3.2 Detection Algorithm of waters (or water area) Change, maybe more accurate.   Please pay attention to the difference between water and waters.
3.Line 266, Table should be located behind related text.
In Figure 3, a1,a2, bi,b2,b3, is suggested to be located in the bottom of the figures.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE EFFECT OF FARMLAND ON THE SURFACE WATER OF THE ARAL SEA REGION USING MULTI-SOURCE SATELLITE DATA
Review round: 3
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I am quite pleased with the authors' responses to my comments. I appreciate the authors’ work. The paper is appropriate to publish at PeerJ after addressing the following minor comments.

Line 294, change “by” to “to”.
Lines 331 and 332, please change “Water mass” to lowercase.
Line 362, “schoolers” should be “scholars”.
Line 425, “disturbed” should be “disturbance”.
Lines 428-429 and Line 432, “it is necessary to identify whether or not need to add other indices for improving accuracy.” and “we aim at more complete data to observe the change of the surface water”. Please fix the grammar.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A COMPARATIVE STUDY OF BACTERIAL DIVERSITY BASED ON EFFECTS OF THREE DIFFERENT SHADE SHED TYPES IN THE RHIZOSPHERE OF PANAX QUIQUEFOLIUM L.
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper use clear, unambiguous, technically correct text. Most part of he article conform to professional standards of courtesy and expression，however，you‘d better  find a professional native language  English speakers to help revise the full article.
The paper include sufficient introduction and background to demonstrate how the work fits into the broader field of knowledge.
The structure of the article basically conform to an acceptable format of ‘standard sections’.
Figures in this paper is relevant to the content of the paper, of sufficient resolution, and appropriately described and labeled.
Experimental design: The submission clearly define the research question, which was relevant and meaningful. The knowledge gap being investigated was identified clearly.

Methods described in this article had sufficient information to be reproducible by another investigator.
Validity of the findings: The data on which the conclusions are based  made available in an acceptable discipline-specific repository. The data was robust, statistically sound, and controlled.

Conclusions are well stated, linked to original research question. However, Individual conclusions need to be carefully considered.
Additional comments: Overall, I agree that this article will be published in Peer J unless you modify your paper according to senior editor and my suggestion.   Suggestion will be upload in the specified page.
you‘d better  find a professional native language  English speakers to help revise the full article.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A COMPARATIVE STUDY OF BACTERIAL DIVERSITY BASED ON EFFECTS OF THREE DIFFERENT SHADE SHED TYPES IN THE RHIZOSPHERE OF PANAX QUIQUEFOLIUM L.
Review round: 1
Reviewer: 2

Output:
Basic reporting: The language and grammar of the article including the title requires revisions.
Experimental design: Line 95-102 The description of climate characteristics in Weihai area is too detailed, please simplify it.
Line 114-115 The sampling information is too simple, please describe it in detail. Please specify the number of specific sampling sites.
Line 118 Please describe the chemical analysis in detail and cite related references.
Line 200 Fig. S6C–S6E? Please check it.
Line 234 ginkgo roots?
Validity of the findings: The difference in environmental conditions and fresh weight, disease rate etc. of American ginseng under three different shade sheds should be analyzed in Results and Discussion parts.
Were the soil properties the same in each experiment site before American ginseng was planted? Did the type of shade sheds cause the difference in soil properties? Please explain them in Results and Discussion parts.
The analysis of the reason (how shed type affected soil properties) is lacking in the Discussion part.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A COMPARATIVE STUDY OF BACTERIAL DIVERSITY BASED ON EFFECTS OF THREE DIFFERENT SHADE SHED TYPES IN THE RHIZOSPHERE OF PANAX QUIQUEFOLIUM L.
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: UNRAVELING THE HIDDEN PALEOBIODIVERSITY OF THE MIDDLE DEVONIAN (EMSIAN) CRINOIDS (CRINOIDEA, ECHINODERMATA) FROM POLAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: Dear Ladies and Gentlemen,
Thank you for the opportunity to review the submitted manuscript. It is very well prepared, contains historical, older and current studies and represents a useful contribution of supraregional importance to these.
You can find my suggestions and additions in the attached PDF.
I am available to answer any questions and I consent to my comments and my name being sent to the authors.
Dr. Jan Bohatý
Experimental design: see PDF
Validity of the findings: see PDF
Additional comments: see PDF


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: UNRAVELING THE HIDDEN PALEOBIODIVERSITY OF THE MIDDLE DEVONIAN (EMSIAN) CRINOIDS (CRINOIDEA, ECHINODERMATA) FROM POLAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper is very well organized and clearly written, in good English rendering.  The background/context and references are appropriate.  The paper is well structured, with descriptions following standard format, with data provided.  The paper fulfills the stated aims in providing descriptions of newly discovered specimens of crinoids as complete or partial calyxes,
Experimental design: The paper is highly significant in reporting crinoid taxa known from complete and partial calyxes, heretofore not well represented in the Emsian of this region, despite the fact that crinoid remains were well known from columnals and columnals.  The descriptions are very detailed and thorough, and the illustrations are excellent in quality.
Validity of the findings: The paper provides complete data on the fossils described, and the conclusions are concisely stated and sound.
Additional comments: I found very few minor corrections that are highlighted and explained as notations.  The institution for Dr Ausich is incorrect and noted in the byline at the very beginning of the manuscript (but is OK in the following, line 8) .  There is a section (lines 146-157) enclosed in " marks that should have the source included if it has been taken from another work.  In the caption for Fig. 5 attached to the figure, the explanation of part H appears to be truncated (lines not numbered), but it is OK in the full list of figure captions.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VALIDATION OF A BATTERY OF INHIBITORY CONTROL TASKS REVEALS A MULTIFACETED STRUCTURE IN NON-HUMAN PRIMATES
Review round: 1
Reviewer: 1

Output:
Basic reporting: I want to commend the authors on their thorough and careful reporting. I especially appreciated Figure 1, which really helped to orient me as I was reading through the methods and results.

However, the introduction is rather long. I think that some of the descriptions of content validity, repeatability, and impurity could be cut back considerably (ie lines 92-139).

Second, when introducing your study species (ie lines 147-151) I think it would be worth presenting what’s already known about macaques’ performance in inhibitory control tasks, especially those that you include in your study, to help frame and inform your hypotheses (e.g. Hopper et al., 2021; Lauwereyns et al 2000; Rathke & Fischer, 2020; Reeves et al. 2017; Vardanjani et al., 2021)

Lastly, I found it a little confusing that the analyses description was provided before the detailed task descriptions. I suggest reordering this.

Cited References
Hopper et al 2021. A comparative perspective on three primate species' responses to a pictorial emotional Stroop task. Animals, 11(3), 588

Lauwereyns et al 2000. Interference from Irrelevant Features on Visual Discrimination by Macaques (Macaca fuscata): A Behavioral Analogue of the Human Stroop Effect. J. Exp. Psychol. Anim. Behav. Proc. 26, 352–357

Rathke and Fischer 2020 Differential ageing trajectories in motivation, inhibitory control and cognitive flexibility in Barbary macaques (Macaca sylvanus). Philosophical Transactions of the Royal Society, 375(1811).

Reeves et al. 2017. Within-session reversal learning in rhesus macaques (Macaca mulatta). Animal Cognition 30, 9750983

Vardanjani et al., 2021 The effects of emotional stimuli and oxytocin on inhibition ability and response execution in macaque monkeys. Behavioural Brain Research, 412, 113409.
Experimental design: In this study, the authors report a series of touchscreen tasks presented to macaques to assess inhibitory control. My main concern with this study is that I am not convinced that the first test is actually testing what is claimed. I suggest that this is presented a “training” exercise (i.e. familiarizing the macaques with touching stim on a touchscreen) and that this could even be moved to supplemental materials so that the focus of the analysis can be on the tasks that appear to actually test relevant metrics. In addition to this more fundamental concern, I have some other questions and request for clarifications that I present in the order in which they arose as I read the article.

For the first task, I am not sure what an incorrect response would be given that there was no choice to be made – the monkeys just had to touch a red triangle. Please clarify this. Relatedly, and more importantly, this task is very simple and represents the basic training phase of any touchscreen training so it seems a leap to describe this as an executive function task. What planning was required by the macaques? Also, if the touchscreens were new to the monkeys at the start of this study, why would you expect repeatability in this? Surely there would be a learning curve with time (as measured in decreasing response latencies)? How familiar with the touchscreens were the macaques prior to this and what other stimuli had you trained them on?

With the second task, what happened if the macaque touched the distractor stimuli? And was the distractor presented simultaneously with the target? This was not clear (line 433).

Line 473 – but now the time limit for the response was longer. Have you analyzed their responses to see how many responses were made within the previous trial time limit?

Line 525 – why was the no-go stim presented for such a brief period? Are the monkeys able to respond (incorrectly) this quickly? How does this relate to their average response latencies in other tasks and how does this method compare to other go/no-go tasks used previously? It just seems that it’s somewhat setting the monkeys up for success without allowing them to fail.

Line 552 “there was still a sign that the subjects tried to override their prepotent response” this seems like a big claim. It could just be that the red triangle was more familiar to the monkeys and so they were more likely to reach for it than the unfamiliar green circle, not because they “knew” they shouldn’t respond to that second stim.
Validity of the findings: I have some concerns regarding the presentation and interpretation of task 1 - see my above comments
Additional comments: Minor comments
Line 185 is the “free roaming room” and indoor space or did these macaques have outdoor access?

Line 203 what was the average group size that the monkeys were housed in?

Line 205 “from the higher rank” this is unclear, please clarify. Did you simply classify the macaques as high vs low ranked from the David’s score and 15 were high and 6 were low?

Line 240 “the timer was paused,” is this a built in timer within the software or were you manually recording the times? This makes it a little unclear.

Line 409 but see Hopper et al. 2021 who only found that positive stim interfered with macaques’ responses in a Stroop task.

Line 465 “wasn’t” should be “was not”

Line 670 could these comparisons be presented in a table? I found it hard to parse all these comparisons from the text and I think a summary table would be more accessible.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VALIDATION OF A BATTERY OF INHIBITORY CONTROL TASKS REVEALS A MULTIFACETED STRUCTURE IN NON-HUMAN PRIMATES
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: The tasks used are appropriate and overall the methodology is well explained.
Validity of the findings: Due to the nature of the study, the methods and results sections are very long and a bit difficult to follow. The findings are valid and the conclusions drawn by the authors are in line with their findings. This is useful manuscript for animal cognition researchers and I have relatively minor, specific comments for the authors.
Additional comments: Line 21: is inhibitory control only restricted to unproductive responses? Inhibition may also work on responses that are productive, but potentially less so than delayed responses. I suggest you revise this section of the abstract to be more consistent with your definition of inhibition in the introduction, where productivity is better explained.
Line 66: check typo and add comma to 33,000
Line 204: section on dominance rank is vague. What was the minimum number of interactions used to calculate rank for each animal. What do you mean as ‘higher rank’?
Line 293: how many study subjects did you have in each of the three rank categories?
Line 297: since model selection has been criticised by some authors, wouldn’t it be simpler/better to just compare the full model to a null model containing all your control variables but not the test variables? From the way you describe the aims in the introduction, this model selection approach seems unnecessary. Please clarify point this or revise the analyses presented.
Moreover, it is not clear how many data points were used for these models. This is important to understand whether your sample size could ‘accept’ the number of variables you used in the model.
Due to the nature of the study, the results section is very long and for the reader it’s difficult to keep track of the different findings.  I suggest you add a summary table with the main findings of your study, if space is available.
Discussion: unless I missed it, you don’t mention anywhere in your manuscript that a possible source of variance in the literature on inhibitory control is due to species-specific differences. For example, it is possible that inhibition could be a general construct in some taxa and a family of multi-components in other taxa. It would be useful to briefly discuss this option in your manuscript, especially in relation to whether differences in support of one of these two hypotheses may be related to the species being tested.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: VALIDATION OF A BATTERY OF INHIBITORY CONTROL TASKS REVEALS A MULTIFACETED STRUCTURE IN NON-HUMAN PRIMATES
Review round: 1
Reviewer: 3

Output:
Basic reporting: Review of “Validation of a battery of inhibitory control tasks reveals a multifaceted structure in non-human primates (#65913)” submitted to PeerJ.

This article tries to answer whether Inhibitory control (IC) is a general construct or a family of distinct sub-components, by testing monkeys in touchscreen tasks aimed to target 3 aspects of IC (a new battery of tasks, another goal of the current MS). The authors found some support for a general construct but also some support for distinct sub-components. I found the MS easy to read and generally convincing (not least, as the authors place their findings well into the literature and also do not oversell their results).

Was there “Clear, unambiguous, professional English
language used throughout.”? Yes.

Did “Intro & background show context.”? Yes.

Was the “Literature well referenced & relevant.”? Yes.

Were “Figures relevant, high quality, well
labelled & described.”? Yes.

Was the “Raw data supplied”? The authors claim that this was supplied – but I could not find the data (and neither a functioning link to that data in a repository) myself among the files I received.
Experimental design: Was the “Original primary research within Scope of
the journal.”? Yes.

Was the “Research question well defined, relevant
& meaningful.”? Yes.

“It is stated how the research fills an identified knowledge gap.”? Yes.

Was there “Rigorous investigation performed to a
high technical & ethical standard.”? Yes. I especially liked the “distraction of other group members” aspect. Note that I did not see the ethic board decision letters.

Were “Methods described with sufficient detail &
information to replicate.”? Yes.
Validity of the findings: Impact and novelty: not assessed, as per PeerJ design.

Was “All underlying data provided;
and were they robust, statistically sound, &
controlled.”? Yes.

Were “Conclusions well stated, linked to
original research question & limited to
supporting results.”? Yes.
Additional comments: “to first demonstrate the content validity of a cognitive task seems a crucial step in order to both justify its use and draw evolutive conclusions from it.” I could not agree more. It is nice to see the current efforts to overcome such hurdles.
I wish to congratulate the authors on a nice study and nice paper that goes quite some way in this direction.

-	The authors mention pilot subjects. Could the authors explain whether this data was entered into the main data set? Could they also state whether these subjects were excluded from later testing?

-	Iine 711: word repeat


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CYBER-VICTIMIZATION AND ITS ASSOCIATION WITH DEPRESSION AMONG VIETNAMESE ADOLESCENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: Not appropriate to stated objectives.
Validity of the findings: Valid.
Additional comments: 1.	Major concern I have with the paper is the language that the authors use which overstates the findings of their study. The (cross-sectional) design used by authors cannot support the objectives and conclusions drawn. The objectives are stated thus “to assess the effect of cybervictimization on depression” which implies a causal association. This is not supported by the design. The authors have to redo the language (say “association with” than “effect on”) everywhere in the paper, as appropriate.
2.	Statistical analysis – better to say precision (in lieu of type I error) was set at 5%.
3.	The study was carried out during the first wave of the COVID pandemic and students could have been spending more time on the internet for academic purpose, I would like to know how this was factored in when classifying IA as mild/moderate/severe based on parameters such as time spent using internet.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CYBER-VICTIMIZATION AND ITS ASSOCIATION WITH DEPRESSION AMONG VIETNAMESE ADOLESCENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript is well-structured and in accordance with PeerJ standards. Clear and professional language is used; introduction and background show context with relevant literature being referenced; and figures are relevant, high quality, well-labelled and described.

The results section is very well-structured. First it demonstrates the relationship between symptoms of depression and internet use and internet addiction (Table 1). Second, it shows the association between symptoms of depression and the environment (family, neighborhood, school, and cyber victimization; Table 2). Finally in Table 3 the covariates are adjusted for, and the relationship between symptoms of depression and cyber victimization is reported.

In the supplementary materials Thanh Thai et al., have also included the raw dataset. As observed from the attached raw dataset, personally identifiable information is not present.

Minor comments on basic reporting
	A sentence comparing the method used to measure symptoms of depression with DSM-5 needs to be made. This manuscript measured symptoms of depression using the Center for Epidemiologic Studies – Depression scale (lines 142–143). While the authors mention that this is a well-established assessment test, a sentence comparing/contrasting it with the standard DSM-5 is warranted.

	This manuscript needs to discuss that there might be possible gender difference in self-reporting. That there were more female students volunteering for this study is also noteworthy (Table 1 and line 163). What is particularly interesting to note is that 1.5 times more female students self-reported symptoms of depression. Gender differences in self-reporting for epidemiological studies is well documented. This is particularly true for studies of mental health disorders, and this probable skew needs to be discussed.

	The authors report chi-squared results (Tables 1, 2, and 3). The standard that the authors are following for reporting this statistic is not mentioned. It can be observed that this manuscript is conforming to the APA standard. However, reporting that fact would be ideal.
Experimental design: In this study Thanh Thai et al., aims to understand the relationship between symptoms of depression and cyber victimization of students in Vietnam. This original primary research article is within the scope of PeerJ. The authors showed rigor in defining their research question and designing experiments to address it.

Depression is a multifactorial psychiatric disorder. To control for this, the authors assessed for multiple categories at the level of individual student characteristics, environmental factors (family, neighborhood, school), and the main variables of cyber victimization and symptoms of depression. This shows necessary rigor. This is further highlighted by their sampling approach. Thanh Thai et al. utilized a multi-stage cluster sampling method. The authors selected secondary and higher secondary schools from 3 urban and 1 suburban area (grades 8–12) and invited all students from 40 classes of these schools to participate in this study. The measures used are well-established.

In addition, the authors have also shared the template for the participant information and consent form, questionnaire, and data codebook. This reviewer was able to easily load the provided raw data file and browse through it.
Validity of the findings: The data analysis methods — such as sampling weights — to control for the sampling approach is rigorous. Also, adjustment for confounding covariates was performed.

The authors also show balanced reporting of their conclusions. E.g., pertinent societal/household factors that can confer protection has been mentioned, and their findings are well contextualized with similar studies in other countries. In addition, the authors also dedicated an entire section in discussing the limitations of this study. It demonstrated that Thanh Thai et al., are mindful that this is not a causal study, and have been careful not to imply the same.
Additional comments: I recommend this study by Thanh Thai et al., for publication in PeerJ. Only minor comments on basic reporting.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CYBER-VICTIMIZATION AND ITS ASSOCIATION WITH DEPRESSION AMONG VIETNAMESE ADOLESCENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No concerns
Experimental design: No concerns
Validity of the findings: No concerns
Additional comments: Thank you for making the necessary changes.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CYBER-VICTIMIZATION AND ITS ASSOCIATION WITH DEPRESSION AMONG VIETNAMESE ADOLESCENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed the comments.
Experimental design: NA
Validity of the findings: NA
Additional comments: NA


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ON THE ANDEAN GENUS LESCHENIUS (COLEOPTERA: CURCULIONIDAE: ENTIMINAE): UPDATED PHYLOGENY, WITH A NEW SPECIES FROM ECUADOR, DISCOVERY OF MALES, AND LARVAL DESCRIPTION OF THE POTATO WEEVIL LESCHENIUS VULCANORUM
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an important contribution and update to the knowledge of the Naupactini of the New World, specially for the naupactine fauna of the Andean Region and the South American transition zone.

The taxon sampling in the present contribution follows findings in del Río, Marvaldi & Lanteri (2012) and Lanteri & del Rio (2017). The genus-level relationships depicted here agree with del Río, Marvaldi & Lanteri (2012) but no with those of Lanteri & del Rio (2017). This is a important point of discussion that is missing in the paper (see comments on PDF).
The character statistics (l, ci, and ri) are not mentioned. This can be added in Table 1 (see PDF).
The figures are very helpful. I suggested some small changes in the figure captions (see PDF).
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ON THE ANDEAN GENUS LESCHENIUS (COLEOPTERA: CURCULIONIDAE: ENTIMINAE): UPDATED PHYLOGENY, WITH A NEW SPECIES FROM ECUADOR, DISCOVERY OF MALES, AND LARVAL DESCRIPTION OF THE POTATO WEEVIL LESCHENIUS VULCANORUM
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is clear and well written. It includes relevant references for most of the aspects discussed. My only complain (very minor) is that figure legends were not included in the main word document, but only visible in the final PDF compiled by the journal's platform.
Some of the figures might be improved by increasing the size of the images, reducing blank space around relevant parts.
Experimental design: The manuscript presents new informaion: a new species, lectotype designations, a new synonym, descriptions of males and immature stages.
The methods are clearly presented.
Validity of the findings: The manuscript presents relevant supporting information.
Additional comments: The manuscript is very clear and well supported. I only have a few suggestions marked throughout the text, especially concerning morphological terminology and consistency on the use of that terminology. I also added a few questions in the discussion that the authors might be able to tackle with the information presented. I also suggest to summarize information on ecology/associated plants for each of the treated species, since that information is presented already, but is sort of diluted in the material examined section, and might be relevant to discuss the implications of the newly found phylogenetic relationships. See comments in both the main word document and the PDF (only in the figure captions for this one).


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NONTARGETED AND TARGETED METABOLOMICS APPROACHES REVEAL THE KEY AMINO ACID ALTERATIONS INVOLVED IN MULTIPLE MYELOMA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is written in proper professional English, clear and unambiguous. The introduction describes well enough the background of the study and included sufficient references. However, there is no message of the novelty of the study compared to the similar studies previously published.
Experimental design: - Number of sample is to less for a conclusion. Validation study with higher sample number is recommended.
- The individual stratification is not clearly described in the manuscript.
- There is no information about cofounders, e.g., medication, comorbidity, smoking, etc.
- Detection and annotation of broaden metabolic groups from the nontargeted metabolomics will be beneficial.
- QC samples used for nontargeted metabolomics should be clearly defined.
- Information on how the system suitability is assessed is missing.
Validity of the findings: Validation study is recommended as number of sample used in this study is too low.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NONTARGETED AND TARGETED METABOLOMICS APPROACHES REVEAL THE KEY AMINO ACID ALTERATIONS INVOLVED IN MULTIPLE MYELOMA
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript by Yue et al.  uses nontargeted and targeted metabolomics approaches to investigate possible metabolic differences between 20 newly diagnoses MM patients and 20 normal controls. They explored the differences in amino acid metabolites between the two groups and claim that their findings will provide” new insight for elucidating the possible mechanism of MM and exploring promising biomarkers or therapeutic targets for this disease”.
Based on previous studies in this area, these results are not new because many authors have already showed amino acid alterations in MM patients (references already cited in introduction) when compared to normal controls.  An important issue currently is if these alterations are related to the tumor or are in born errors that predispose to MM development. The reference Biochemical phenotyping of multiple myeloma patients at diagnosis reveals a disorder of mitochondrial complexes I and II and a Hartnup-like disturbance as underlying conditions, also influencing different stages of the disease. da Silva IDCG, de Castro Levatti EV, Pedroso AP, Marchioni DML, Carioca AAF, Colleoni GWB.Sci Rep. 2020 Dec 14;10(1):21836. doi: 10.1038/s41598-020-75862-4. was missed and can be useful to improve this discussion.

Worldwide, MM diagnosis is based in laboratorial criteria. Therefore, it is hard to believe that metabolomic profile could be used as a more efficient biomarker than the currently used.
The authors should also develop the argument of how their findings could be therapeutic targets for this incurable disease.
Minor points:
Summary: Please review the entire text to include the term "amino acid metabolites" rather than just amino acids when referring to choline, creatine and creatinine.
Incorrect: “ROC curve analysis showed the high diagnostic accuracy of key amino acids, including choline, creatinine, tryptophan, and valine. “
Correct: “Key differential amino acid metabolites, such as choline, creatinine, tryptophan, and valine, may discriminate MM patients from NC. “
Correct: “The changes in amino acid metabolites may provide some clues for elucidating the pathogenesis of MM.”
Introduction: “and there are approximately 21,000 new diagnoses each year”.  Please specify where. China? US? World?
“Although there have been some achievements in MM remission, several reports have demonstrated that the real success of MM treatment depends on its early diagnosis.” Please review this concept. Genetic-molecular background associated to clinical staging (R-ISI) are currently considered the gold standard to define prognosis of MM patients.
“The metabolic changes in MMC cells include the generation, accumulation and inhibition of metabolites as well as metabolic shifts”. Please correct MMC to MM.
Figure 1:  Please change for “20 normal controls”.
Raw data was shared.
Professional English was used.
Experimental design: This manuscript by Yue et al.  uses nontargeted and targeted metabolomics approaches to investigate possible metabolic differences between 20 newly diagnoses MM patients and 20 normal controls. They explored the differences in amino acid metabolites between the two groups and claim that their findings will provide” new insight for elucidating the possible mechanism of MM and exploring promising biomarkers or therapeutic targets for this disease”.
Based on previous studies in this area, these results are not new because many authors have already showed amino acid alterations in MM patients (references already cited in introduction) when compared to normal controls.
Validity of the findings: This manuscript by Yue et al.  uses nontargeted and targeted metabolomics approaches to investigate possible metabolic differences between 20 newly diagnoses MM patients and 20 normal controls. They explored the differences in amino acid metabolites between the two groups and claim that their findings will provide” new insight for elucidating the possible mechanism of MM and exploring promising biomarkers or therapeutic targets for this disease”.
Based on previous studies in this area, these results are not new because many authors have already showed amino acid alterations in MM patients (references already cited in introduction) when compared to normal controls.
Worldwide, MM diagnosis is based in laboratorial criteria. Therefore, it is hard to believe that metabolomic profile could be used as a more efficient biomarker than the currently used.
The authors should also develop the argument of how their findings could be therapeutic targets for this incurable disease.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NONTARGETED AND TARGETED METABOLOMICS APPROACHES REVEAL THE KEY AMINO ACID ALTERATIONS INVOLVED IN MULTIPLE MYELOMA
Review round: 2
Reviewer: 1

Output:
Basic reporting: In the manuscript the authors reported their observation on regulation of some amino acids in multiple myeloma patients in comparation to that in normal controls. The manuscript is clearly written with proper professional English. The literature references are appropriately referenced in this manuscript.
Experimental design: In the manuscript there is no data about confounders, e.g., medications taken by the individuals, smoking habits, etc. Confounders can influence the metabolic trend. It must be reported and incorporated in the data analysis.

In materials and methods part the authors didn't mention about type of QC samples applied. The labelled IS should be described more detail about the mass ranges and retention times.
Validity of the findings: There are already some published data on metabolomics of multiple myeloma, including data on amino acids, e.g., Chanukuppa et al, 2019; Gonsalves et al, 2020. I suggest not to restrict the observation on amino acids instead broaden the metabolic classes to find novel biomarkers.
Additional comments: In the manuscript the authors reported their observation on regulation of some amino acids in multiple myeloma patients in comparation to that in normal controls. However, there are already some published data on metabolomics of multiple myeloma, including data on amino acids, e.g., Chanukuppa et al, 2019; Gonsalves et al, 2020. I suggest not to restrict the observation on amino acids instead broaden the metabolic classes to find novel biomarkers.

In the manuscript there is no data about confounders, e.g., medications taken by the individuals, smoking habits, etc. Confounders can influence the metabolic trend. It must be reported and incorporated in the data analysis.

In materials and methods part the authors didn't mention about type of QC samples applied. Please elaborate this in the manuscript. The labelled IS should be described more detail about the mass ranges and retention times. For the metabolic annotation for nontargeted metabolomics, did you use retention time or retention index?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NONTARGETED AND TARGETED METABOLOMICS APPROACHES REVEAL THE KEY AMINO ACID ALTERATIONS INVOLVED IN MULTIPLE MYELOMA
Review round: 3
Reviewer: 1

Output:
Basic reporting: The manuscript is written in proper professional English. The introduction is well written with sufficient field background and references.
Experimental design: Research question is meaningful. Methods is decently explained, however, I miss the information about biochemical annotation on nontargeted metabolomics part. Also, the authors didn't mention confidence level of the annotation.
Please provide these information.
Validity of the findings: The authors have responded to my concern in the first round of reviewing process.
Additional comments: N/A


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A NEW GOLDEN SPECIES OF DIASPORUS (ANURA: ELEUTHERODACTYLIDAE) FROM SOUTHWESTERN COLOMBIA, WITH EVALUATION OF THE PHYLOGENETIC SIGNIFICANCE OF MORPHOLOGICAL CHARACTERS IN DIASPORUS
Review round: 1
Reviewer: 1

Output:
Basic reporting: I think the article is pertinent, since the contribution of this new species is valid and is a contribution to the diversity of this group. In general, the article is well written. But the phylogenetic analyses are poor, they require a detailed review, in particular the alignments on which they are made. I also believe that it is very important that the results of Maximum likelihood are explicitly put in the document and that the results of the phylogenetic part are not only based on the results of the Parsimony analyzes, since all previous works for this group report results. Based on ML and to be clearly comparable, it is necessary that they be exposed, which is not clearly the case in the article.
Experimental design: The methods are not complete, data on the protocol used for the RAG1 nuclear gene are lacking.

Information is also lacking to clarify the number of individuals included in the analysis and the length of the sequences analyzed.

Why the authors performed alignments with default values of the programs if the nature of each of the genes is different, they should not be the same costs in the alignments for protein-coding genes as the ribosome-associated genes (16S)
the cost of insertion and extension of gaps must be different, this will affect the alignments and therefore the results of the phylogenetic inferences
Validity of the findings: The taxonomic results are valid, but the phylogenetic approach is poor and requires that the alignments and therefore the analyses and results of the phylogenetic inferences be reviewed.
Additional comments: line 95 Add to the line a more integrative approach, to include molecular and morphological data is not enough, also is required behavior, acoustics, and maybe biochemical signaling to understand this group of frogs.

Line 233, The authors should also include the most recent phylogenetic hypotheses about the positions of this group of frogs and which are based on massive data from the position of Diasporus tales Hutter 2017, Barrientos 2021. This may give further support to the relationships with respect to their group brother (Eleutherodactylus) which are quite clear.

Line 235 and fig1 Authors should show ML results or summarize them in the parsimony tree that authors show in figure 1 or show both trees


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A NEW GOLDEN SPECIES OF DIASPORUS (ANURA: ELEUTHERODACTYLIDAE) FROM SOUTHWESTERN COLOMBIA, WITH EVALUATION OF THE PHYLOGENETIC SIGNIFICANCE OF MORPHOLOGICAL CHARACTERS IN DIASPORUS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: The authors gives relevant nformation for the understanding of the genus, and only minor revision is required to be published.

The authors must add figures of a hand and a foot to see details of characters (fringes, and webbing) that they have stated are important for the recognition of the species.
Other comments are in the revised manuscript
Additional comments: Other comments are embedded into the revised manuscript


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A NEW GOLDEN SPECIES OF DIASPORUS (ANURA: ELEUTHERODACTYLIDAE) FROM SOUTHWESTERN COLOMBIA, WITH EVALUATION OF THE PHYLOGENETIC SIGNIFICANCE OF MORPHOLOGICAL CHARACTERS IN DIASPORUS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The text is well-written and good to understand. However, it would certainly be good to check the text once again. There are some minor grammar errors. However, the authors use clear, unambiguous, and technically correct language. Literature references are sufficient.
The Introduction should include all the relevant work on Diasporus. I noticed that e.g., the works of Arias et al. 2019 and Chavez et al. 2009 are not mentioned in the introduction. The work of Garcia-Rodriguez et al. 2016 (Neotropical Biodiversity Volume 2, 2016 - Issue 1) is not mentioned at all.
A weak point in this description is the lack of any bioacoustics data or analysis. Diasporus calls are easy to record and the analysis is cheap and easy. Call descriptions have been proven a powerful tool for species identification, especially in species groups that lack good morphological characteristics. Describing a new species of Diasporus without presenting any bioacoustic data, call description, and a comparison with its congeners is not appropriate any more.
The general structure of the text is good.
Experimental design: I generally welcome this study and I agree with the authors that there is a lack of work in this genus from South America, while there has been considerable progress in Central America. I also don´t doubt that the species presented by the authors is new to science.
I, however, have some issues with the approach the authors use to delimit the new species.
First, I think that sample size of five appears a little low. Diasporus species are usually abundant where they occur. Especially coloration may be very variable in Diasporus species. This is one of the main characteristics (two chrome orange spots (=glandlike structures) on sacral region) the authors use to distinguish the new species and I see the small sample size as a weak point here.
An even bigger weak point in this description is the lack of any bioacoustics data or analysis. Diasporus calls are easy to record and the analysis is cheap and easy. Call descriptions have been proven a powerful tool for species identification, especially in species groups that lack good morphological characteristics. Describing a new species of Diasporus without presenting any bioacoustic data, call description, and a comparison with its congeners is not appropriate any more.
Validity of the findings: I think describing the species as new is justified. The genetic distance to the closest relatives is large enough and there are some morphological characters that support that idea. However, including a bioacoustics analysis would provide an additional line of evidence, especially when comparing with calls of Diasporus gularis and the other yellowish species.
Further, I would be careful with putting too much emphasis on the idea of a D. diastema clade that includes only bright yellow species. First, not all species are bright yellow and second the clade is not supported (<50% jackknife support) in the phylogenetic analysis.
Additional comments: I have made some other, mostly minor, comments directly in the pdf.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A NEW GOLDEN SPECIES OF DIASPORUS (ANURA: ELEUTHERODACTYLIDAE) FROM SOUTHWESTERN COLOMBIA, WITH EVALUATION OF THE PHYLOGENETIC SIGNIFICANCE OF MORPHOLOGICAL CHARACTERS IN DIASPORUS
Review round: 2
Reviewer: 1

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: I have read the rebuttal letter and the manuscript, and I do not have additional comments on it.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A NEW GOLDEN SPECIES OF DIASPORUS (ANURA: ELEUTHERODACTYLIDAE) FROM SOUTHWESTERN COLOMBIA, WITH EVALUATION OF THE PHYLOGENETIC SIGNIFICANCE OF MORPHOLOGICAL CHARACTERS IN DIASPORUS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Dear authors,

I am happy with most of your reply and welcome, the additional work you have spent to improve your manuscript.
I am still in a way disappointed that there will be no information on the male advertisement call of this species. I have never doubted that you discovered a new species and I think and have always thought your two lines of evidence are enough for species delimitation. I also don´t think that call data are more important than the other lines of evidence and I never said that. However, for a complete description of a new species that emits a sound, reporting on that sound is very important, as it is a very important aspect in the life of the animal and a significant information to identify it in the future. Imagine you would describe a frog on molecular and bioacoustic evidence only, but would not give a description on how it looks like. Yes, of cause you can do your species description without bioacoustics and I intentionally did not ask for an extensive bioacoustic analysis (although for many Diasporus species there is excellent bioacoustic material available), but asked for any bioacoustic data like a simple recording you made with a cell phone, likewise the photos you took. Simple parameters like the call length or the frequency range would have been interesting for researchers working on that genus. To keep it short, I accept that you don´t have call data available at the moment and hope anyone will describe Diasporus lynchi´s call someday in the future.

Comments in the PDF
Abstract: Since the new species is not the only yellow species in the genus, this sentence is kind of redundant.
Reply: We respectfully disagree with this suggestion. We think that it is not redundant; on the contrary, it is very informative in the context of the results, discussion and conclusions of our work.
Reply to the reply: No, you got me wrong here! My comment was unclear, sorry my fault. My comment was a pure linguistic thing. You wrote “The new species, along with Diasporus citrinobapheus, D. gularis, and D. tigrillo are the only species in this genus known to exhibit a yellowish coloration in life.” First, it must read “the new species is” not "are"! And, what I meant with my comment is basically the word “only”. It would be like saying “Bananas, along with cherries, apples, pears, and peaches, are the only sweet fruits produced by plants”. You know what I mean? I would just reword the sentence saying something like: “The new species exhibits a yellowish coloration in life, a character that it shares with other species in the genus like Diasporus citrinobapheus, D. gularis, and D. tigrillo.” Or something in that way.

Apart from that I am completely fine with the manuscript.

Thanks for choosing me as a reviewer!
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EVALUATING THE THERAPEUTIC EFFECT OF TUMOR TREATING FIELDS (TTFIELDS) BY MONITORING THE IMPEDANCE ACROSS TTFIELDS ELECTRODE ARRAYS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article is written clearly, with the introduction providing motivation for the problem addressed. TTF is a therapeutic modality that gaining wider interest, and having a real-time reporting/monitoring system would be beneficial. The literature and background/context for the proposed solution of EIT is well motivated. The article structure, figures and tables as presented are acceptable.
Experimental design: The authors present an original result where the use a head phantom to test whether electrode configurations used for TTF can detect a tumor mass via impedance measurements. The results are in align with the hypothesis and demonstrates the feasibility of the approach.
Validity of the findings: The only perceivable weakness of the topic is that it reports on the feasibility of impedance measurements using TTF electrode arrays, that would be further strengthened if actual treatment vs. response were to be measured. Perhaps these experiments could be performed in a petri dish setting.

Further, the use of a potato, which is a heterogenous mass, in an otherwise homogeneous background is expected to provide distinct results as seen here. More nuanced studies or measurements where the inclusion has much less dramatic conductivity differences in comparison to the background would be of interest.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EVALUATING THE THERAPEUTIC EFFECT OF TUMOR TREATING FIELDS (TTFIELDS) BY MONITORING THE IMPEDANCE ACROSS TTFIELDS ELECTRODE ARRAYS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is a fairly basic paper evaluating the potential of applying electrical impedance using tumor treating field electrodes as a means of assessing tumor response to therapy. In other words, one set of electrodes could serve as both a means for the treating the tumor and measuring the effect of treatment. The paper uses a fairly basic model of a skull, a potatoes of different sizes to mimic the tumor and a gel to mimic brain tissue. The demonstrate that impedance changes can be detected and these mirror tumor size. A sensitivity analysis is also completed.

The language is clear, the figures, sufficient, references look complete.

But the article is incredibly sparse and many details are omitted (see below).

Table 1 does not need to be a table.
Experimental design: The experimental design is very straightforward, but the model is a very weak approximation of human anatomy and the tumor behavior. GBMS are not simply solitary masses but rather are often deeply infiltrating tumors, with scattered neoplastic cells extending out far from any observable mass on MRI. Moreover, the brain has ventricles which can take up a substantial volume of the cranial vault. It is entirely unclear how the non-homogeneities in normal brain would impact this model as well. Thus the model is really quite unrealistic, yet there is absolutely no discussion as to these huge limitations.

Moreover there is often considerable edema associated with these tumors...again another limitation that this model does not include.

We are told very little about the impedance methods used except the frequency (200 kHz), the rationale of which is not provided. It is also unclear which electrodes were used for the impedance measurements, and were these 4-electrode measurements (I think so)? And if so, which are the voltage and which are the current electrodes.

What impedance system was used?

Only the impedance magnitude was measured. Why? They mention simulations showed that phase was less sensitive, but why not provide that information too?
Validity of the findings: The validity is limited as noted above based on the very simplistic nature of this model.

There is no inclusion of any kind of skin layer which could also impact the findings.

The conclusions are vastly overstated the limitations of this very simplistic model are not included.
Additional comments: See comments regarding limitations of the study design above.

Overall, the results decided unsurprising and I think this adds relatively little to the literature, although the basic idea of using electrical impedance along with TTFs is good. I really which the authors had spent more time constructing a more realistic model than that presented here. The article is also short on many details. While the results here could be useful for further developmental work, they could also be highly misleading.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EVALUATING THE THERAPEUTIC EFFECT OF TUMOR TREATING FIELDS (TTFIELDS) BY MONITORING THE IMPEDANCE ACROSS TTFIELDS ELECTRODE ARRAYS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Appropriate as presented.
Experimental design: Acceptable.
Validity of the findings: Acceptable.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GENOME-WIDE IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF CDPK GENE FAMILY REVEAL THEIR INVOLVEMENT IN RESPONSE TO DROUGHT STRESS IN GOSSYPIUM BARBADENSE
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript present clear and unambiguous conclusions. I commend the authors for their extensive data set, however, the authors should provide the raw transcriptomics data in the public database (i.e. NCBI), so that the original data are available to the public.
Experimental design: The methods are thorough.
Validity of the findings: The manuscript presents the results which are well presented and appropriately described. Conclusions are appropriate, and supported by the data.
Additional comments: Shi et al presented a genome-wide identification and characterization of 84 CDPK genes in G. barbadense. In this study, the authors used bioinformatics to identify the members of CDPK gene family and predict the structure, physiological and biochemical features of the protein. Further, the authors analyzed the evolution and classification, and the expression of the CDPK genes. Finally, the authors demonstrated that these CDPKs are associated with the drought.

The manuscript presents the results which are well presented and appropriately described. The methods are thorough. Conclusions are appropriate, and supported by the data. The manuscript is overall sound, and below I have highlighted a few minor concerns.

1.With regarding to data availability: the authors should provide the raw transcriptomics data in the public database (i.e. NCBI), so that the original data are available to the public.

2. Line 29: it should be CDPKs for the short name Calcium dependent protein kinases. The author should check other places in the manuscript for the correction.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GENOME-WIDE IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF CDPK GENE FAMILY REVEAL THEIR INVOLVEMENT IN RESPONSE TO DROUGHT STRESS IN GOSSYPIUM BARBADENSE
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Calcium dependent protein kinases (CDPKs) are a class of important calcium signal sensing response proteins, which play an important regulatory role in response to abiotic stress. Shi et al identified the CDPK family genes in sea-island cotton genome and performed a series of bioinformatics analysis, and curried out qRT-PCR for some genes under different stresses and further selected some genes for VIGS analysis of drought tolerance. These findings provide a solid foundation for the development of future molecular mechanism in sea-island cotton. This study is very carefully done and the paper is organized very well, except a few minor mistakes.
Minor
1.	Some English spellings, such as line 59, 67-68,89,165,231,246,249,303,341,379.
2.	Please cite this paper and compare with your result in the manuscript. Liu W, Li W, He Q, Daud MK, Chen J, Zhu S. Genome-wide survey and expression analysis of calcium-dependent protein kinase in Gossypium raimondii. PLoS One. 2014 Jun 2;9(6):e98189
3.	Please provide the detailed methods for RWC, RE,MDA, PROLINE, SOLUBLE SUGAR AND WATER LOSS RATEE in the material and methods section.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GENOME-WIDE IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF CDPK GENE FAMILY REVEAL THEIR INVOLVEMENT IN RESPONSE TO DROUGHT STRESS IN GOSSYPIUM BARBADENSE
Review round: 1
Reviewer: 3

Output:
Basic reporting: ..
Experimental design: .
Validity of the findings: .
Additional comments: This manuscript conduct the CDPK gene family characteristics analysis and also analyze the potential function of some of them respond to drought stress. The manuscript was well written and the story is appealing. However, there are several questions need to be clarified before the publication.
1)	The figures are not clear enough, I wonder this is because the figure was inserted into the manuscript, so please check the figure quality
2)	Can you add the relationship analysis about the subcellular location and the response to drought stress of some CDPK gene in the DISCUSSION Part.

Minor questions:
1)	Some minor mistakes need to be noted: such as the figure 7 legend, t-test, the first t should be italic; P value, the P should be italic.
2)	Line 38 sea should be Sea
3)	Line 72-73 should be rewritten
4)	N-myristoylation should be N, the N should be italic
5)	Line 267 CDPK should be italic


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GENOME-WIDE IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF CDPK GENE FAMILY REVEAL THEIR INVOLVEMENT IN RESPONSE TO DROUGHT STRESS IN GOSSYPIUM BARBADENSE
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have answered all my concerns, I suggest accept it in the present form.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GENOME-WIDE IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF CDPK GENE FAMILY REVEAL THEIR INVOLVEMENT IN RESPONSE TO DROUGHT STRESS IN GOSSYPIUM BARBADENSE
Review round: 3
Reviewer: 2

Output:
Basic reporting: Clear and unambiguous, professional English used throughout.

Literature references, sufficient field background/context provided.

Professional article structure, figures, tables. Raw data shared.

Self-contained with relevant results to hypotheses.
Experimental design: Clear and unambiguous, professional English used throughout.

Literature references, sufficient field background/context provided.

Professional article structure, figures, tables. Raw data shared.

Self-contained with relevant results to hypotheses.
Validity of the findings: Impact and novelty not assessed. Meaningful replication encouraged where rationale & benefit to literature is clearly stated.

All underlying data have been provided; they are robust, statistically sound, & controlled.

Conclusions are well stated, linked to original research question & limited to supporting results.
Additional comments: No additional comments