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: AN INTEGRATIVE MONOGRAPH OF CAREX SECTION SCHOENOXIPHIUM (CYPERACEAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: This is a very well done treatment of a group for which there was a lot of confusing information. It is concise, considering the large amount of information presented, and well written.

The organization is appropriate I spot checked bibliographic citations and found all correct.

The drawing are very nice, as are the photos and other illustrations. It is a beautifully illustrated work.

It is comprehensive and detailed, and laid out in a logical, parallel manner. I had no substantive comments placed in the text.
Experimental design: no comments
Validity of the findings: I think the findings are solid and this will set the standard for all future work.

My  notes do include one item that must be corrected The one new species published herein, Schoenoxiphium gordon-grayii, has a problem that needs to be fixed.. Article 60.8. ex (a) of the International Code of Nomenclature treats the “y” in Gray as a vowel, therefore substantival epithets should be “grayi” (male) and for Kath Gordon-Gray it should be  Schoenoxiphium gordon-grayae. This is a small but important matter.

On a more minor note, there are 6 names in Schoenoxiphium in IPNI not accounted for in this treatment. There can be little doubt that these are not relevant for this monograph, but there is an opportunity here for completeness and also to prevent any confusion, so these name could be disposed of  briefly in a few sentences, if the authors choose.

Schoenoxiphium	clarkeanum Kuk. Bull. Herb. Boissier Ser. II. iv. 49.
Schoenoxiphium	fragile C.B. Clarke Bull. Misc. Inform. Kew, Addit. Ser. 8: 67. 1908
Schoenoxiphium	hissaricum Pissjauk. Bot. Mater. Gerb. Bot. Inst. Komarova Akad. Nauk S.S.S.R. 12: 72. 1950
Schoenoxiphium	kobresioideum Kuk. Bull. Jard. Bot. Buitenzorg ser. III, xvi. 312 (1940).
Schoenoxiphium	kuekenthalianum	N.A. Ivanova Bot. Zhurn. S.S.S.R. 24: 501, in obs. 1939
Schoenoxiphium	laxum N.A. Ivanova Bot. Zhurn. S.S.S.R. 24: 501, in obs. 1939
Additional comments: none


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: TRP-574-LEU MUTATION IN WILD MUSTARD (SINAPIS ARVENSIS L.) AS A RESULT OF ALS INHIBITING HERBICIDE APPLICATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors report Trp-574-Leu MUTATION IN WILD MUSTARD (Sinapis arvensis L.) AS A RESULT OF ALS INHIBITING HERBICIDE APPLICATIONS in Turkey. However this is not the first time this resistance has been reported. Thus the manuscript has no novelty for PeerJ and should be rejected
Experimental design: The experiments are well designed
Validity of the findings: The results are valid and obtain from a well performed experiment with many samples
Additional comments: Reporting for ALS resistance due to Trp574 mutation has been reported before for Turkey


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: TRP-574-LEU MUTATION IN WILD MUSTARD (SINAPIS ARVENSIS L.) AS A RESULT OF ALS INHIBITING HERBICIDE APPLICATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Figures are not appropriated. The authors should use a better map image (Fig 2), give the legends (Fig 3) and remove the software layout (Fig 4 and 5).
The English must be strongly reviewed specially in M&M section.
Experimental design: The investigatin was not rigourous due to other positions at ALS gene those can harbor target-site mutations. I my point of view, the authors sould investigate all ALS sequence to conclude about the remaining resistant populations.
Validity of the findings: As far as my concern, resistance factor must be calculate based on I50 values. 50% control or 50% mass reduction.
Additional comments: Congrats for your job.
However, there are some concerns about the methods and discussion.
- Why do not the authours investigated other target site mutations over the ALS gene?
- Why do you prefer showing ED90 instead of ED50. We know that ED50 is the best parameter to estimate the levels of resistance.
- The discussion must be improved. If you did not find Trp574Leu in the remaining resistant wild mustard populations, what would be the causes of resistance on those? I suggest you implement two or three more paragraphs in the discussion section talking about what it should change on weed control praticies at wheat fields in Turkey. Is there any other resistance mechanism evolved in wild mustard against ALS herbicides? If no, how about discuss around related species.


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: TRP-574-LEU MUTATION IN WILD MUSTARD (SINAPIS ARVENSIS L.) AS A RESULT OF ALS INHIBITING HERBICIDE APPLICATIONS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Sinapis arvensis L. is a worst weed in wheat fields in Turkey. This study detected the level of resistance and investigate the TSR to tribenuron-methyl in Sinapis arvensis. These results are helpful for understanding the resistance status and the mechanism of Sinapis arvensis. to ALS inhibitors. However, there are also some shortcomings. The organization need to be improved some sentences about herbicide resistance description were also not rigorous. Therefore, I'm afraid that the authors should pay much attention to carefully improve the manuscript before it can be considered for publishing in Peer J.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The major concerns are as following:
1. The detailed information on the methods used for collecting the seeds is needed. Did the seeds bulk？
2. How many copies of ALS gene in Sinapis arvensis.
3. How about the ALS enzyme activity, suggest to supplement the In vitro ALS activity assay.
4. L151 "Unite Taq DNA Polymerase" is a high-fidelity thermostable DNA polymerase ?
5. The discussion section need to be significantly improved. In this section, based on the obtained results, the authors should compare the similarity and difference between this current research and other reports，and discuss the possible reasons, then make conclusion and pointed out the novelty and importance of this research.

Some of the spelling and/or grammar errors in the manuscript：
1. L25: Change "Tribenuron methyl" to "tribenuron-methyl".
2. L31: Italics for Latin names.
3. L48-49: Change to "75.6 billion dollars ", "2.5-2.7 billion dollars ", "5 billion dollars ".
4. L64: "As wheat is a hoe plant weed control" ?
5. L77-79: Check to the tense.
6. L95: Change to "Ala 122".
7. L96-97: Delete the sentence " se Ala-122……...(Powles and Yu, 2010; Vencill et.al., 2012) ".
8. L106: Change "have" to "had".
9. L113 "pre"?
10. L115: suggest change "gr da-1" to g a.i. ha-1".
11. L119: Change to "Do -responses to tribenuron-methyl"
12. L120-L121: Change to "1, 2, 4, 8 and 16-fold".
13. L129-130: "in %"?
14. L137: "%90"?
15. L150-151: Change to "0.5mM", "3.5mM", "0.9mM"."0.5 Unite Taq DNA Polymerase"?
16. L152: Change "sek" to "sec". Change "cycle" to "cycles".
17. L158-160: Check to the tense and italics for Latin names.

The English writing is somewhat poor. These are some tense and grammar errors here and there. I just correct some of these errors. The authors should ask for help from native English speaker to polish the manuscript to improve the quality and readability.


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: TRP-574-LEU MUTATION IN WILD MUSTARD (SINAPIS ARVENSIS L.) AS A RESULT OF ALS INHIBITING HERBICIDE APPLICATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No coments
Experimental design: No coments
Validity of the findings: No coments
Additional comments: Dear authors,
Congrats to improve your manuscript. The discussion section was really revised and well corrected.
Figures are now appropriated.
I have a few suggestions before make the manuscript accept.
- Change the description of ED50: I think welcome call it: Dose to 50% of mass reduction.
- I keep do not understanding "endurance or durability coefficient" in the Table 1. I believe it might be the "resistance factor" or "resistance index". If so, please specify in the Table which wild mustard population was your Susceptible standard

Once the authors make these minor revisions, the article can be considered welcome to publication!
Congratulations for your work and your phD project.

Sincerely,


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: TRP-574-LEU MUTATION IN WILD MUSTARD (SINAPIS ARVENSIS L.) AS A RESULT OF ALS INHIBITING HERBICIDE APPLICATIONS
Review round: 2
Reviewer: 2

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: GROUP B STREPTOCOCCUS AND THE VAGINAL MICROBIOME AMONG PREGNANT WOMEN: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article is well-written and clear. The background is sufficient to understand the data presented and tables and figures are clear. Supplemental tables are helpful and could even be included in the main text.
Experimental design: Experimental design is clearly laid out and appropriate. The research question is meaningful and highlights the lack of data on GBS and other potential pathogens in the vaginal microbiota as well as the problem with variability between microbiome studies. This paper brings together many relevant studies and presents a collection of data that will be useful to the GBS and vaginal microbiota scientific communities.
Validity of the findings: The rationale for why the study was done is clear and well-reasoned. This review will benefit the literature by bringing together disparate microbiota studies to examine together. Statistics and conclusions are sound and well described.
Additional comments: Overall this paper was insightful and I believe it will be a helpful addition to the scientific literature on this topic. Striving for less variability in metagenomics approaches as well as increased access to raw data will benefit the field.

A minor issue to address in editing would be to check for the capitalization and italics issues when referring to Lactobacillus as a genus.

Overall interesting and well-written 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: GROUP B STREPTOCOCCUS AND THE VAGINAL MICROBIOME AMONG PREGNANT WOMEN: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well-written and figures and tables are appropriately formatted.
Experimental design: Meta-analysis study design is appropriate and cohesive.
Validity of the findings: Conclusions are valid and study strengths and limitations are adequately addressed.
Additional comments: Lim et al present a new meta-analysis on the prevalence of Group B Streptococcus reporting in vaginal microbiome studies in pregnancy. GBS is an important pathogen in pregnancy and the neonatal period, however, the inconsistencies in reporting GBS status, or GBS taxonomic identification, in studies examining the role of the vaginal microbiome in pregnancy outcomes may miss the true rate of GBS-associated adverse events. The authors perform a comprehensive assessment of the current literature, using the widely accepted PRISMA guidelines. They identified 45 studies, and further stratified based on GBS reporting. They found no differences between sample sizes, methods, or data analyses. A limitation to restricting the inclusion criteria to pregnancy only is that the breadth of non-pregnant vaginal microbiome studies is missed, however, the authors provide adequate rationale for limiting this meta-analysis to pregnancy studies only. Overall, the manuscript is well-written, and the study design and rationale are excellent. My specific comments below are minor.

Minor comments:
Tables S1 and S2: How many of the studies deposited raw sequencing data? It would be useful to readers to include a column indicating studies that have publicly deposited sequencing data (and the accession numbers if applicable).
Line 76: Add a specific reference for the CDC here, or consider a recent meta-analysis of GBS colonization rates (PMID: 29117327).
Line 188: The wording of this sentence is awkward and could be interpreted that Streptococcus and Lactobacillus would be combined at the genus level. Consider re-wording/separating “as well as when studies only speciate lactobacillus” to a separate sentence.


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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English language should be improved to ensure that an international audience can clearly understand your text.

Some examples where the language could be improved include line 254 – the current phrasing makes comprehension difficult (the term "Insignificantly").
Experimental design: In methods the authors should give more details about independent variables (sleeping disorder, mental health problems), on how they code and measure these variables
Validity of the findings: Findings are correct. Please, add the limitations of your study
Additional comments: Please, revise my comments. Furthermore, I suggest to improve for clarity and grammar the presentation of the work.


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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: a.	Throughout the manuscript, English used is ambiguous and not clear. Also the manuscript has many typing errors

•	Eg: 650 returned back the completed surveys 118 giving a response rate of 72%
•	Depression and anxiety are common and cited problems that generally encounter individuals and more 188 specifically frontline healthcare workers
•	COVID-19: Coronavirus disease 201; HCWs: Health care works;
•	This The authors declare no conflict of interest

b.	Referencing in the literature is not as per the standards of the journal

a.	According to [11], lack of social support and communication, maladaptive coping, and lack of training on 94 how to deal with such pandemic were identified as risk factors for developing psychological problems 95 such as depression, anxiety, insomnia and distress especially to those healthcare workers who interact 96 directly with suspected or confirmed COVID-19 patients [11]
b.	According to the study by [16], show a more satisfactory fit to the data than the results produced by 90 the (confirmatory factor analysis)
c.	Overall study design, methodology and results have been poorly presented
Raw data supplied
Experimental design: 1.Title:  The Impact of COVID-19 Pandemic on the Level of Depression among Health Care Workers: Cross-sectional Study
The title of the study doesn’t give information about the site of the study and is misleading as in a cross sectional study no cause effect relation can be established. So, COVID-19 can’t be causally attributed to depression in frontline workers but only associated with it.
2. Aim and research question
The aim of the study as given in the abstract is different from that in the methodology. Identification of interventional strategies for depression in front line workers is only seen in the abstract. No attempt was made to design these. Till date many studies have been carried out to assess the psychological impact of COVID-19 on health workers, what is the need of the current study?? This is not clearly evident from introduction.
3. Inclusion criteria
The authors didn’t specify in their inclusion criteria about the exposure of included frontline workers to COVID-19, neither was the duration of working hours, specialty included in their assessment.
No objective measures were used for the assessment of mental disorder and sleeping disorder, how was the assessment done??
4. Results and discussion
a. Methodology shows the number of respondents to be 650 but table-1 shows socio-demographic characteristics none of which account to 650. Same has been observed in the text also.
Eg: Almost three quarters of the 152 respondents 450 (70.4%) were non-Saudis while 189 (26%) were Saudis
b. Findings contrary to the previous studies- Our results show that those who did not suffer from sleeping disorder perceived more depression
c. Authors show new results in discussion section, the assessment of which wasn’t shown anywhere in the methodology/results section
Eg: In addition to the stress and anxiety associated with working at hospitals with COVID-19 patients, non-Saudi encounter more stressors such as not being able to visit their families or to return to their home countries, the fear and worries of getting infected, facing death alone, or having a family member die while they are away
Validity of the findings: Impact and novelty not assessed.
Inconclusive results portrayed as positive.
Eg In our study, there was an association between having previous mental health problems and depression, although it is not significant, it is very close to significant 222 association (p= 0.052)

Conclusions are just mere rewriting of the results
Additional comments: Overall English needs to be improved. Authors need to stick to their aim and present their results and discussion accordingly.


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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 3

Output:
Basic reporting: This is a significant cross-sectional study reporting the prevalence of depression in Saudi Arabia healthcare workers. Nonetheless I have a number of concerns that I think must be considered and responded to, in order to strengthen the manuscript for potential publication. Please see below:
Comment 1: Needs a thorough English check.
Comment 2: Abstract - The methods need to be more informative. Please include where and when this study was conducted.
Comment 3: Background - the authors have pointed to the conducted studies in China, please, provide the results of the related systematic review and meta-analysis studies.
Comment 4: Background - Please give some examples of related studies, in Saudi Arabia and their results. Also, specify the differences between them and your study.
Comment 5: Background – Some sentences are hard to understand, please clarify:
•	Working in hospitals in general was associated with clinically significant depressive symptoms whether the workplace is in COVID-19 associated department or other departments.
•	According to the study by [16], show a more satisfactory fit to the data than the results produced by the (confirmatory factor analysis) CFA of the original version indicating that the newly constructed version can allow for a more accurate estimation of burnout levels.
Experimental design: Comment 6: Aim of the study – I think it is better to add a paragraph at the end of the Background section to describe the purpose and importance of the research.
Comment 7: Please insert a section titled “Methods” after the Background section.
Comment 8: Study design – Please specify where and when the study was conducted.
Validity of the findings: Comment 9: Needs a limitations section to address possible weaknesses before the Discussion section.
Comment 10: Discussion – Also, as suggested in Comment 3, compare the results with the other studies in Saudi Arabia. Also, it could be interesting to compare with the studies in other Arabian countries.
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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: 1) There are still some minor typos (e.g. in introduction:
"Over the last decade, mental  56 health problems increased by 13% worldwide WHO, [1]. "
"The most common mental health conditions are 57 depression and anxiety which cost approximately US $ 1 trillion yearly from the global economy see 58 WHO, Mental Health, [2]."
"Therefore, studies in this filed are needed.."
 2) Some sentenced in introduction need to be referenced.
"The spread of COVID-19 pandemic caused huge stress on general population, however, the case was worse for healthcare workers due to their close interaction with COVID-19 patients". "Frontline healthcare workers (HCWs) are dealing with suspected and positive cases of COVID-19 patients on daily basis which puts them at high risk of infection and death"  (you can cite: Chirico F, Magnavita N. Covid-19 infection in Italy: An occupational injury. S Afr Med J. 2020 May 8;110(6):12944. Doi: 10.7196/SAMJ.2020.v110i6.14855."
"Such exposure has a substantial impact on their mental health" (you may cite: Chirico F, Nucera G, Magnavita N. Protecting the mental health of healthcare workers during the COVID-19 emergency. BJ Psych International. 2020. 1-6.  Doi: 10.1192/bji.2020.39"
Experimental design: Aim of the research is clear. Methods are rigorous and are well described
Validity of the findings: All data sound statisticall robust
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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have revised their manuscript to address most of my original points, which I believe has improved the manuscript. However, some of the changes have not satisfactorily solved the issue. Please see below:
Comment 1: Abstract - The authors inserted some information in this section but did not mention about the date and location conducted the study.
Comment 2: Background – There are a lot of studies in different countries around the world that evaluate the mental health problems of medical staff during the COVID-19 outbreak, I expect the authors to refer to some countries using the review studies, not just China.
Comment 3: Background – the authors mentioned to some studies in Saudi Arabia. I think it values to insert their results. Also, specify the differences between them and your study and explain the necessity of your study, please.
Comment 4: Aim of the study – I suggest delete this section.
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: THE IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 3
Reviewer: 1

Output:
Basic reporting: English language could be improved
There are typographical mistakes (e.g. filed instead of field in the introduction),
"The soci-demographic profile" in methods
As suggested, background/context on mental health in HCWs should be improved (Cite:Chirico F, Nucera G. Tribute to healthcare operators threatened by COVID-19 pandemic. J Health Soc Sci. 2020;5(2):165-168. 10.19204/2020/trbt1. Chirico F, Nucera G, Magnavita N. Protecting the mental health of healthcare workers during the COVID-19 emergency. BJ Psych International. 2020. 1-6.  Doi: 10.1192/bji.2020.39. Pappa S, Ntella V, Giannakas T, Giannakoulis VG, Papoutsi E, Katsaounou P. Prevalence of depression, anxiety, and insomnia among healthcare workers during the COVID-19 pandemic: A systematic review and meta-analysis. Brain Behav Immun. 2020 Aug;88:901-907. doi: 10.1016/j.bbi.2020.05.026. Epub 2020 May 8. Erratum in: Brain Behav Immun. 2021 Feb;92:247.)
Experimental design: The authors have addressed all my previous comments
Validity of the findings: The authors have addressed all my previous comments
Additional comments: Please, address some minor suggestions in introduction and revise the english language that may be improved.


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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 3
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors have revised their manuscript in light of my previous comments and they have improved it to a level of satisfaction.


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 IMPACT OF COVID-19 PANDEMIC ON THE LEVEL OF DEPRESSION AMONG HEALTH CARE WORKERS: CROSS-SECTIONAL STUDY
Review round: 4
Reviewer: 1

Output:
Basic reporting: The authors have addressed all my comments
Experimental design: The authors have addressed all my comments
Validity of the findings: The authors have addressed all my comments
Additional comments: Congratulations!
it's a good work!


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: MULTI-SCALE ANALYSIS OF THE CHARACTERISTICS OF THE CHANGING LANDSCAPE OF THE TYPICAL MOUNTAINOUS REGION OF SOUTHWEST CHINA OVER THE PAST 40 YEARS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper deals with an important research question in a large mountainous area with unique ecological and geographical characteristics in southwest China, using data from almost 40 years (1980 – 2018). After reviewing the paper, I think it can be published in this journal after some minor/intermediate revisions. I have provided detailed comments in the annotated PDF document.
Experimental design: The experimental design is suitable for the research objective.
Validity of the findings: Multi-level spatial and temporal variations of landscape patterns are revealed in the study area. Reasons or possible reasons behind the variations are also explained. The findings are important contributions to relevant literature in the study area, and can be used as baseline datasets for future studies.
Additional comments: The paper involves a large amount of datasets covering Yunnan Province in southwest China, employs solid methods, and presents significant results with high-quality figures. The paper is publishable after minor/intermediate revisions, mainly in the writing style and some grammatical errors. I have suggested changes in the annotated PDF document, and I’d like to suggest that the authors check the following points throughout the paper:
(1)	Line 36, the reference for Turner is missing.
(2)	Change the old name “Nu Chiang” to “Nu Jiang” or “Nu River”, and use a consistent name for the river in the paper.
(3)	Pay attention to uppercase and lowercase in river names. For example, the following words are used interchangeably in the paper: “Nu Chiang”, “Nu River”, and “Nu river”. Consistent names should be used.
(4)	Check if “river basin” should be used in some paragraphs, instead of “river”. For example, when discussing fragmentation in Lines 193-200, “river basin” should be used, not “river”. A river as a linear feature does not have fragmentation, while river basins can be fragmented.
(5)	The landscape indices are listed in Table 1. It is suggested to put the full names of the indices in the third column of the table to improve the readability of the paper. For example, for the “NP” index, the full name “Number of patches” should be added to the third column. The title of the third column in Table 1 can be “Description”.


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: MULTI-SCALE ANALYSIS OF THE CHARACTERISTICS OF THE CHANGING LANDSCAPE OF THE TYPICAL MOUNTAINOUS REGION OF SOUTHWEST CHINA OVER THE PAST 40 YEARS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Thank you for the opportunity to review the manuscript “Multi-scale analysis of the characteristics of the changing landscape of the typical mountainous region of southwest China over the past 40 years”. Generally, the academic contribution and innovation of this study are not clear to me. My comments and questions are as follows:

1.Please clearly explain the innovation and contribution of this study.
2.The draft is very long, some parts are not necessary, e.g. Line 103-112, it’s not necessary to describe the secondary categories and this can be included in the 2.2 Data and sources. There are many cases that the long draft is not closely related to the study. Fig.2 didn’t show the secondary classes.
3.The experiments are easy and the data is products. What’s the original data that you produced?
4.Line 97-101 is not precise. The used dataset need to give out direct websites. Projection is not needed.
5.LUCC data is the most base of the analysis of this study, what’s the accuracy of this dataset in Yunnan Province? Please adding some materials about the data accuracy and methods. There are many LUCC datasets, why use this one?
6. Adding a map to show the six river basins.
7.Why used nine indicators in this study? What are their differences, advantages, or disadvantages? How to understand the results, other than just showing the numbers?
8.Line 185-365: The results of the nine indexes are very long. Can the authors summarize the results and give out some main points? Rather than introduce nine indicators one by one.
9.The conclusion needs to be improved.
10.The LUCC in Fig.2 is strange (construction land is too small). Adding the year to the figures and the legends are not clear. The four sub-figures can be put together as one figure.
11.The titles of the figures should be above the figures. The titles can be improved.
12.Table 1 needs citation.


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: MULTI-SCALE ANALYSIS OF THE CHARACTERISTICS OF THE CHANGING LANDSCAPE OF THE TYPICAL MOUNTAINOUS REGION OF SOUTHWEST CHINA OVER THE PAST 40 YEARS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The aim of this manuscript is to analyze temporal and spatial changes of landscape patterns at county, watershed, and provincial levels over past 40 years. It is interesting to address the landscape changing at multi-scale in Yunnan Province, China, one of the most biodiversity richness regions in the world. However, this work needs substantial improvement in several related critical matters. Moreover, the sections of introduction, methodology, results, are needed to be clarity and language editing.
Experimental design: 1) The title, “the characteristics of the changing landscape”, what are the characteristics? And what is specific landscape? Forest? Agricultural? Urban? River? Mountain? and so on. Actually, this study is more focus on the land cover changes of Yunnan Province, China at different scales. Thus, the title can be change to “Spatial and temporal changes of land-cover at multi-scale in mountainous region of Southwest China over the past 40 years”

2) In section 1 introduction and literature review, in this section, it is lack of literature review of LUCC across multi-scale. Moreover, the research questions should be well defined. Why the land cover changes should be analyzed at county, watershed, and provincial levels? What is the research rationale? Why 40 years? Again, I think that landscape pattern changes more focus on specific landscape not general landscape.

3) line 43-45, “an important center of biodiversity within the Southwest part of China, represent ecological security barriers”. What is “center of biodiversity”, and why it represent ecological security barriers? The ecological security barriers for what? “The pattern of the landscape within Yunnan Province has not been improved” What is “the landscape has not been improved”? what’s this mean? “

4) line46-51, literature review listed previous studies have addressed the LUCC of the Lancang watershed, Nu Jiang watershed, Red River watershed. However, what are the main findings of these studies? Why the Jinsha, Pearl, and Irrawaddy River basins have not been studies and why do you want to study LUCC for those watershed?

5) line 53-55, these two sentences should be re-written. LUCC is the first time be used in this paper, the full name should be listed.

6) Line 53-65, in this paragraph, authors always mentioned the GIS spatial analysis was used in this study. What specific spatial analysis was used in this study?

7) Line 53-65, in this paragraph, authors always mentioned the GIS spatial analysis was used in this study. What specific spatial analysis was used in this study?

8) Line 92-101, in this paragraph, “the LUCC data for Yunnan Province from 1980 to 2018”, it should be land cover or land use maps, is not LUCC (land cover and land use change). Moreover, the methodology of how to make those land cover or land use maps and the quality (accuracy assessment) of those maps should be provided.

9) Line 102, landscape types, as mentioned above, landscape, land cover, land use, landscape change, LUCC, those concepts are confused. Authors should clarify those concepts, particularly what is landscape types.

10) Line 124, spatial correlation analysis, why? For what?.
Validity of the findings: no comments
Additional comments: Summary, It is interesting to address the landscape changing at multi-scale in Yunnan Province, China, one of the most biodiversity richness regions in the world. However, this work needs be improved, and the language need to be edited by native speaking people.


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: MULTI-SCALE ANALYSIS OF THE CHARACTERISTICS OF THE CHANGING LANDSCAPE OF THE TYPICAL MOUNTAINOUS REGION OF SOUTHWEST CHINA OVER THE PAST 40 YEARS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments.
Experimental design: No comments.
Validity of the findings: No comments.
Additional comments: Line 235, remove "Analysis of".
Line 251, use "The changing trend in PAFRAC varies ...." or "The changes in PAFRAC vary ..."
Line 259, insert a space before "stable".
Line 276, replace "bansins" with "basins".
Line 279, replace "biasin" with "basin".


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: MULTI-SCALE ANALYSIS OF THE CHARACTERISTICS OF THE CHANGING LANDSCAPE OF THE TYPICAL MOUNTAINOUS REGION OF SOUTHWEST CHINA OVER THE PAST 40 YEARS
Review round: 2
Reviewer: 2

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: Thank you for the opportunity to review the manuscript “Multi-scale analysis of the characteristics of the changing landscape of the typical mountainous region of southwest China over the past 40 years”. Generally, the manuscript has been improved after the reversion, especially the language and expression. My comments and questions are as follows:

1.	In the Rebuttal Letter, the authors wrote “There are many LUCC products in China, such as GLOBE 30 by Gong Peng from Tsinghua University”, which is not true. GLOBE 30 is developed by Dr. Jun Chen from National Geomatic Center for China. The reasons such as “It is the Institute of Geography of the Chinese Academy of Sciences. The technical strength of UCAS is at top level in China.” is not very convincing. The authors could have a deeper understanding of the LUCC products themselves.

2.	Line 58-66 is verbose. The actual contents of this research are not complicated and it’s fine to emphasis the significance. But the similar expression has been repeated many times, including the manuscript and Rebuttal Letter.

3.	Section 2.3 is not necessary, because Section 2.2 has some description. Or the authors can include some contents to Section 2.2.

4.	The English needs improved and the grammar of line 140 is not right. Line 144 “more descriptions could be also found in the user’s guide of FRAGSTATS”. Did the authors pick the nine indexes from the FRAGSTATS?

5.	It’s a big problem that there is no actual discussion in the Section “Results and discussion”. Generally, it’s only about the results of the nine indexes. How to understand the numbers? What about the comparison between this study and other studies? What’s the policy implication? What are the limitations and future perspective? A lot can be discussion.

6.	The conclusion is too long and some are discussion, not inclusion.

7.	In Fig.2 the LUCC changes of the four figures are not obvious. Please add some contents about the accuracy of this data in Yunnan Province and the detailed areas of the main LUCC types. Also, there are some strange lines in the figure (e.g. triangle in the snow and lines in the high coverage grass)


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 MULTI METAL-RESISTANT BACTERIUM CUPRIAVIDUS METALLIDURANS CH34 AFFECTS GROWTH AND METAL MOBILIZATION IN ARABIDOPSIS THALIANA PLANTS EXPOSED TO COPPER
Review round: 1
Reviewer: 1

Output:
Basic reporting: •	The objective of the manuscript is rather clear.
•	English require verification (check by a native speaker)
•	Intro & background should be further developed, at the moment too general.
•	Literature well referenced & relevant, however more literature review for the discussion section is necessary.
•	Structure conforms to PeerJ standards: Results and discussion should be separately following the journal’s requirements.
•	Figures are definitely too small and need some improvement. Table are missing important information (see my specific comments).
•	Raw data supplied, however, metadata not provided (description of columns’ names is needed)
Experimental design: •	Original primary research within Scope of the journal.
•	Research question well defined, relevant & meaningful.
•	It is not clearly stated how the research fills an identified knowledge gap.
•	Investigation performed to a rather low but sufficient technical standard.
•	Some methods described with insufficient detail & information to replicate (see my specific comments).
Validity of the findings: •	Moderate impact and novelty.
•	Inconclusive results.
•	All underlying data have been provided, however not always presented in a clear way.
•	Data are robust, statistically sound, & controlled.
•	Conclusions are not well stated, rather vague and general. Not enough linked to original results
Additional comments: •	Authors should highlight the importance, novelty, and significance of this work.
•	All the experiments were conducted in growth media and hydroponic cultures. Did the authors consider using more environmentally relevant matrix such as soil? That could significantly change the uptake and translocation of metals and certainly, it would give a better idea about the potential use of CH34 strain in bioremediations. It should be mentioned in the text…
•	The plant root exudes (PRE) analysis is barely mentioned in the discussion and results are not presented in any form (figure/table). Whether authors further elaborate on this result or completely remove it. At the moment I don’t feel it contributes much to the discussion and interpretation of results.
•	Generally, many sentences are written in a clumsy way. I strongly recommend giving the text for a revision to a native speaker.


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 MULTI METAL-RESISTANT BACTERIUM CUPRIAVIDUS METALLIDURANS CH34 AFFECTS GROWTH AND METAL MOBILIZATION IN ARABIDOPSIS THALIANA PLANTS EXPOSED TO COPPER
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors should thoroughly check the style and grammar of the complete manuscript (including the abstract.
Some examples:  L21: “but it excess”?; L31: Capital of sentence; L54-60: very long sentence; L67: chromid instead of chromide; L75; L222: “live strain CH34 cells”, remove “strain”; L286: “The effects on metal levels reported here have”; L236: “because it provoked”; L305: “located on”.; L320: Cd2+
Experimental design: - Why are the results with heat-killed cells not shown?
- Experiments with colonization (Table 2): agar plates without plants were used as control but the data are expressed as CFU/mg FW. Furthermore, this shows a 4-log decrease (initial inoculum is 10^8 CFU/ml), which is not discussed.
- The data on changes in PRE composition are not shown in detail.
- It would be interesting to also add impact of Cu on metal accumulation (% increase or decrease) without CH34 in Table 4.
- qPCR: It is unclear if gluconate was included in both the plant and control conditions, which could have a great impact on expression levels. It would be interesting to use the delta delta ct method and compare expression in plants with or without Cu. In addition, was primer efficiency tested?
- L297-298: is there information available to justify the correlation between 50 µM in agar and 25 µM in hydroponics?
- L305 and further: for the copC on the chromid, please refer to copC2
- L309 and futher: copK was clearly expressed 60 min after inoculation, but only in the presence of Cu, right? (Figure 2); copF was not changed: but significantly different at 30 and 180 min (plants vs plants with Cu)? Please revise this section.
- L332: impact on phaC1 expression: Is there any difference in phosphate content of the medium in the experiments with and without plants? If so, this could have an impact on phaC1 expression and should be discussed.
Validity of the findings: - The differences between beneficial and detrimental effects based on Cu concentration should be reported and discussed much more nuanced in both the abstract and body of the text (results and discussion) as the impact on the different parameters. For instance, L202-205, however there is no effect on FW and DW at 25 µM. L208: 10.3% increase of rosette area at 0 µM, but not significant?; The cutoff at 50 µM (positive/negative effects) is not that clear cut.
- What is the evidence for the presumably new PGPR traits in CH34?
- The expression data concerning Fe and siderophores metabolism should also be discussed in light of the metal accumulation experiments, indicating increased Fe uptake (167 µg/DW without and 484 µg/DW with Cu) that could impact Fe availability for CH34.
Additional comments: The authors provide an interesting study on the impact of strain CH34 in the presence of absence of Cu on Arabidopsis. Although they show interesting findings, the should present them with more details and nuances.


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 MULTI METAL-RESISTANT BACTERIUM CUPRIAVIDUS METALLIDURANS CH34 AFFECTS GROWTH AND METAL MOBILIZATION IN ARABIDOPSIS THALIANA PLANTS EXPOSED TO COPPER
Review round: 1
Reviewer: 3

Output:
Basic reporting: The English language could be improved.
Some examples where the language could be improved or shown below:

in general: try to be consistent in the spelling of Cu: sometimes Cu, sometimes Cu(II) and sometimes Cu+2

Line 21: it excess -> in excess, it
Line 22: interacts -> interact
Line 25: on the growth of
Line 26: gene
Line 32: reveal the complexity
Line 33: on the use of
Line 51: are the inhibition
Line 52: capture of ->  capturing
Line 54: metal -> metals
Line 54-59: very long sentence. Can be split in two or three sentences to make it more readable
Line 63: in the soil
Line 67: chromide -> chromid
Line 68: comprises
Line 83: In this work,
Line 121: add space between roots & 10 mM
Line 126: the sterile conditions? Sterility of the conditions?
Line 131: To obtain PRE,
Line 135: as for the
Line 137: one -> medium
Line 161: mL-1 cells? Cells/ml?
Line 196: on the Cu concentration
Line 198: to test the effects
Line 208: remove a
Line 236: because this concentration
Line 256-line 261: very long sentence. Can you rephrase to make it more readable?
Line 286: the effects on metals levels? -> the effect of metal levels?
Line 293: the effects
Line 304: shows the relative
Line 307: remove gene
Line 309: remove gene
Line 310: remove gene
Line 320: the Fur protein

The introduction gives a comprehensive overview of but could be improved by a more detailed description of the copper resistance mechanisms in strain CH34 as this is important in one of the results sections.

In general, the figures and tables are readable, but
Figure 2 legend: do you mean plants + CH34? All conditions are the result of a comparison Cu-induced or without Cu? To make a better comparison, it can help to put the Y-axes at the same scale so it is immediately clear that there is a big difference is relative expression levels of the different genes (e.g. almost no expression of copF)
Figure 3 is not readable. Can it be adapted to a larger format?
Experimental design: The research question from this paper is clear: the authors want to investigate whether CH34 can be beneficial for copper tolerance of Arabidopsis thaliana.

The experiment how rhizosphere and plant colonization was tested is not really clear to me. How was the agar treated? also rinsed with MgSO4?

I have a specific question regarding the gene expression: why did you use a different medium for the samples without plants? How will you discriminate for the differences due to the different medium?
Validity of the findings: Some general remarks:
As you mention in the discussion: CH34 is not able to metabolize sucrose. This you also see in the cell counts of the non-rhizosphere conditions, which remain 104 CFU/ml. Here, you also see that copper itself does not have negative impact on the growth or survival of CH34. It shows that CH34 needs the rhizosphere or plant environment to grow. As plants excrete more organic compounds due to the copper treatment, CH34 has maybe more carbon available to grow and that can explain the difference? Did you check if CH34 can metabolize the exudates? This could be elaborated more in the text. Maybe it is rather a matter of available energy sources to grow instead of a change in the microenvironment?
You did a sterility check but results are not shown? It is good (even in supplementary data) to prove that your samples are still sterile.
Regarding the effect on CH34 on the growth of A. thaliana: you mention that CH34 resulted in positive effects such an increase in root growth: indeed for the conditions without copper, it is visible. Also at 25 µM it is still visible, however, at 50 µm, you see a decrease compared to the conditions with 25 µM and this drop is higher than the drop that you see in similar conditions without CH34. Also, for the secondary roots: without copper, the levels with CH34 are significantly lower than without CH34. With the addition of 25 µM copper, the number does not change compared to the conditions without copper but afterwards, you a decrease. Same as for most other conditions where you already see a significant effect of CH34 in conditions without copper (and in most cases, except for root growth, the effect is negative). In conditions with 50 µM of copper, the effect clearly disappeared as the decrease from 25 µM to 50 µM is much more than that in conditions without CH34. So except for root growth, it rather has a negative impact on plant growth?  Showing results with heat-killed cells would valuable for the reader.

Some smaller remarks:
Line 182: general remark but the expression of the 16S rRNA gene has been shown not be stable and expression levels can vary depending on the conditions. One of the uvrD genes are much better housekeeping genes for CH34.
Line 230-231: any idea why you don’t see an increase in secondary roots?
Line 256 -257: Cu-concentration dependent mode? You see similar effects without CH34?
Line 283: CH34 promote plant growth? Only root length?
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 MULTI METAL-RESISTANT BACTERIUM CUPRIAVIDUS METALLIDURANS CH34 AFFECTS GROWTH AND METAL MOBILIZATION IN ARABIDOPSIS THALIANA PLANTS EXPOSED TO COPPER
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The quality of the manuscript significantly improved compared to the previous version. Although, I believe that authors could further improve the style of the manuscript and better expose their findings. My main criticism is about the Discussion section that doesn’t seem to refer enough to the authors' results but rather to the previous study. It feels that results are not sufficiently discussed in a way that can be easily interpreted. I suggest incorporating authors' observations into the text in a better, more clear way and discuss them in the context of other studies and potential implications/applications.

Line 307-309 – why condensing all the results in one sentence. It makes the sentence really loaded. Please split. This will also better highlight the results.

Fig.1 Please provide the label of the x-axis in panels B-F --> Cu concentration [µM]


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 MULTI METAL-RESISTANT BACTERIUM CUPRIAVIDUS METALLIDURANS CH34 AFFECTS GROWTH AND METAL MOBILIZATION IN ARABIDOPSIS THALIANA PLANTS EXPOSED TO COPPER
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors addressed (almost) all my comments. Nevertheless, the authors do not use the delta delta Ct method as indicated in their rebuttal (which would still be interesting to show, e.g. plant Cu vs plant; plant Cu vs Cu). They use delta Ct (difference in Ct values for GOI and HK). Delta delta Ct uses Ct values for GOI and HK for both the treated and control samples.


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 MULTI METAL-RESISTANT BACTERIUM CUPRIAVIDUS METALLIDURANS CH34 AFFECTS GROWTH AND METAL MOBILIZATION IN ARABIDOPSIS THALIANA PLANTS EXPOSED TO COPPER
Review round: 2
Reviewer: 3

Output:
Basic reporting: English grammar is much approved compared to the first version.
Experimental design: no comments
Validity of the findings: * The same medium was used in all experiments therefore gluconate was always present. This is clearly stated in lines 163-165.
However, Line 110-112: Square Petri dishes were prepared with half-strength Murashige-Skoog medium (MS1/2) (Murashige & Skoog, 1962), 0.8% agar, and 1.5% sucrose; were either inoculated or not inoculated with strain CH34.
This medium is with sucrose instead of gluconate so CH34 will not grow. And this is for most conditions throughout the paper. How can it optimally deal with the copper concentrations and influence plant growth? Results can be biased as it is able to grow on root exudates but in the non-rizosphere environment it has no carbon source to grow. This can have an important effect on the results.

Line 210: the 10,3 % is not significantly different than without CH34 inoculation so the number of rosetta’s does not increase with CH34. The fact that the difference is significant at 24 µM is because in the absence of CH34, the number of rosetta’s decreases, while with CH34, it remains the same as without Cu. That should be better explained in the text. Similarly for the secondary roots.
Additional comments: Line 33: Cop -> cop
Line 74: across
Line 75: remove over 20 different as at line 70 at least 24 metal resistance… is stated
Line 82: it is not clear what is meant by “the mobility of trace metals that are relevant to plant nutrition in this bacterial model”. Please rephrase
Line 89: A. thaliana is not in bold
Line 105: A. thaliana is not in bold
Line 277: Please change copSRABC in copD2C2B2A2R2S2 with the 2 in subscript as this is the official name and order of the cluster
Line 227: Remove “the sake of”
Line 364: may be explained by
Line 369: rephrase this sentence. Strains do not encode organic compounds. The encode genes to degrade organic compounds


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: DZNEP, AN INHIBITOR OF THE HISTONE METHYLTRANSFERASE EZH2, SUPPRESSES HEPATIC FIBROSIS THROUGH REGULATING MIR-199A-5P/SOCS7 PATHWAY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Hepatic fibrosis is reversible damage to liver tissue and leads to the formation of Collagen I extracellular matrix. The study of molecular mechanisms by therapeutic drugs is novel ideas to understand the process of disease progression and improvement. Authors in this manuscript understand the molecular mechanism of a drug molecule DZNep in the prevention of hepatic fibrosis and propose to prevent liver cell injury and repair hepatic cells from damage in detailed observation. This study would help to understand the model of the DZNep axis in liver damage prevention and improve hepatic fibrosis by DZNep/miR-199a-5p/SOCS7 pathway. In addition, the authors propose that DZNep could act as a therapeutic agent to treat patients with reversible liver damage.
Experimental design: The authors applied qualitative and quantitative experimental designs to show the indispensable features of the DZNep axis. Further, they also applied in vitro and in vivo models to show the molecular basis of DZNep inhibition of liver damage. Moreover, the results in this manuscript are well presented according to their data. Quantitative analysis of results by authors displayed that DZNep is critical in regulating miR-199a-5p levels and in turn regulating SOCS7 levels.
Validity of the findings: Further, they presented several lines of evidence to show the DZNep effect on the SOCS7 pathway, like TGF-B1 levels, SMA levels, etc,. Based on the analysis I would recommend this study for PeerJ with the following improvements in the manuscript.
Additional comments: Major comments:
1.	Figure 3A methodology and analysis are not clearly written. I suggest the authors to update methods for this section. Please also indicate the labels 1,2,3 in figure 3A.
2.	Introduction can be improved by highlighting the role of SOCS7 in inflammation and how do SOCS7 levels abrogate liver fibrosis.
3.	Authors may mention miR-199a-5p have complementary base pairing to SOCS7 3`UTR and they also indicate Figure 5A show mutated 3` UTR of SOCS7.
4.	It would be very interesting if authors were able to show morphological differences DZNep treated or untreated HSCs or the effect of miR-199a-5p or SOCS7 levels on liver fibrosis phenotypes.
5.	In the discussion maybe authors should propose a few best future directions to pursue in the DZNep field of liver fibrosis instead of claiming limitation of their study
Minor comments:
1.	Add “Forward” for SOCS7 at line 128 in methods.
2.	Line 219 miR-199a-5p should be “SOCS7”
3.	Line 244 authors should mention where does H3K37me3 occurs in COX-2 gene, here statements seem to be incomplete.
4.     Line 262 “Previous studies identified” is duplicated, please remove
5.	In some figure legends (4E and F), a number of replicates were not mentioned, please indicate them in the next update.
6.	In figures label DZNep (capitalization) appropriately to match the text in manuscript.
7.	Please correct errors in the list of abbreviations:
FBS- fetal bovine serum
K-lysine not X
MMP9 repeated twice,


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: DZNEP, AN INHIBITOR OF THE HISTONE METHYLTRANSFERASE EZH2, SUPPRESSES HEPATIC FIBROSIS THROUGH REGULATING MIR-199A-5P/SOCS7 PATHWAY
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1)	The English language should be improved in order to increase the readability of the manuscript. Ideally a scientist with high English proficiency should proofread the manuscript. Examples of sentences that require attention include but are not limited to
-	Line 56: liver fibrosis can be regressed
-	Line 58: Recent studies verified the effect
-	Line 104: There were no survived rats
-	Line 111: control (siNC) were structured by Thermo Fischer
-	Line 243-244: EZH2 catalyzes H3K27me3 in cyclooxygenase-2
-	Line 269: The pulmonary miR-199-5p level is in idiopathic form patients and regulates
-	Line 278: miR-199-5p inhibition, observably inhibited
Throughout the manuscript “additional DZNep treatment” (as well as “additional treatment with SOCS7-siRNA” and similar statements) should be replaced by “DZNep treatment”.

2)	The introduction shows the context and contains relevant references. However, this part could be better structured. Lines 72-73 state the aim of the study, then lines 74-79 present general information on miRNAs and lines 80-85 summarize the findings of the study. They authors may want to increase quality of their manuscript by improving their narrative.

3)	Structure appears to conform to PeerJ standards.

4)	While overall the Figures are appropriate and informative the labelling of Figures and data description should be improved.
-	E.g. in Figure 1 (nor in the other figures) “NC” is not defined in the figure legends. It may be clearer to use “Control”. “Model” is not informative. Could be replaced by “CCl4” and “DZNep” could be replaced by “CCl4+DZNep” to enable readers to immediately understand the data.
-	Figure 1: it is difficult to appreciate the subcellular location of the immunostaining in Figure 1: cytoplasmic or nuclear? What do the arrowheads point at? This data should be more clearly described in the results section and discussed. The most abundant cells in the liver are hepatocytes and likely the cells with positive EZH2 staining in the shown images are hepatocytes. However, the rest of the study is based on LX2 cells (fibroblasts) since fibroblasts play a major role in fibrosis. This is confusing and needs clarification. Additional immunostaining in cell culture experiments could be performed to assess the subcellular localization of EZH2 in LX2 cells in vitro.
-	Molecular weights of proteins detected by Western blot should be indicated next to the images.
-	Figure 3: the legends states “mRNA” expression for miRNA but per definition miRNA are not mRNA… this should be corrected in the manuscript wherever appropriate.
-	Figure 5G should be replaced by a higher quality Western blot as it is one “representative experiments picked from” several technical replicates.
-	Figure 5 is not easy to understand as in contrast to the other figures the effect of TGF-b1 is not represented as compared to control but TGF-b1 is represented as the control. Maybe the authors can find a way to both show that TGF-b1 increases in vitro markers of fibrosis and the effect of siSOCS2 in the presence and absence of DZNep. In Figure 2 the decrease in collagen I expression observed in the presence of DZNep is approximately 50% while in Figure 5 it appears to be only approximately 25%. The “rescue” of collagen I expression by siSOCS7 does not appear very robust, although an asterisk indicates statistical significance (please also see the comment on statistical analysis). Can this effect be reproduced when the initial decrease in collagen I is stronger?
Experimental design: 1)	The research question has been defined but could be presented in a more efficient way (see comment on the introduction).

2)	The study design and statistical analysis need attention and appear not to meet the expected standards.
-	The “Statistical analysis section” states that results are presented as means+/-SD. It should also include how many times each experiment has been performed. The figure legends indicate that representative images were picked from 3 separate experiments (technical replicates). Technical replicates are not independent experiments (biological replicates). Experiments are expected to have been performed e.g. 3 times on independent dates/with different cells using technical replicates within each of the experiments. Pooled data should show the data from the different experiments, not only technical replicates from one experiment.
-	Furthermore, it states that statistical analysis was done using Student’s t test or ANOVA but there is no reference to the Normality test that should have been performed prior to the use of parametric tests. In case of a non normal distribution of data (which is commonly the case for small number of samples/data), non parametric tests should be used instead.

3) The methods are described in a global way and lack details in order to enable reproduction of data. Furthermore, the rational for some experiments should be indicated.
- 	E.g. lines 111-115 the amount of miRNA mimics/inhibitors and the timing of transfection relative of cell plating and readout of the experiment should be clearly stated.
- 	Lines 131-and following: the amount of protein used for Western blot experiments should be clearly indicated.
-	Lines 139-143: how much vector was transfected? At what time following plating of the cells? For dual luciferase reporter assay LX2 cells were used, i.e. cells that express miR-199-5p. Why did the authors choose this cell line to perform the reported assay? Usually, a cell line that does not express the miRNA of interest (e.g. as assessed by PCR) is chosen in order to avoid background inhibition of the reporter by the endogenous miRNA.
Validity of the findings: 1)	As indicated above, experimental design/data analysis/representation need attention.
- 	While appropriate controls (control siRNA/miRNA/inhibitors) appear to have been used in most of the experiments, the data cannot be seen as robust and statistically sound at this stage (also see comments on statistical analysis).
-	In Figure 2, the authors did not include a control of DZNep in control cells. What is the effect of DZNep in cells not treated with TGF-b1?

2)	Several conclusions appear to be overstated.
-	They authors should more carefully phrase their conclusions in order to limit their conclusions to the shown results. E.g. an effect on collagen I and a-SMA shown in vitro with LX2 cells cannot be interpreted as an effect on fibrosis as fibrosis relates to pathological changes seen in an organ (as stated by the authors in their introduction). Therefore, throughout the manuscript the authors should replace “hepatic fibrosis” (or equivalent term) when referring to in vitro data by more appropriate description of their data.
-	Since the authors have access to an animal model of hepatic fibrosis, why did they not also assess the role of miRNAs in vivo in rat livers? It should be clearly stated why they solely chose LX2 cells for their mechanistical study. Is miR-199-5p conserved between humans and rats? What is miR-199-5p/SOCS7 expression in rat liver following DZNEp treatment and control condition?
-	Line 220 description of Fig 5A: “partially” should be added to read “DZEp treatment partially inhibited TGF-b1-induced downregulation of SOCS7”
-	Furthermore, the authors base all their conclusions related to EZH2 on DZNep, an inhibitor that globally inhibits histone methylation that is known not to be selective for EZH2. Indeed, while it was the first EZH2 inhibitor that has been developed, several other more selective EZH2 inhibitors have been developed since 2012 and it would be more appropriate to use one of those EZH2 inhibitors, e.g. an inhibitor in clinical development. Given the absent selectivity of DZNep, it is difficult to conclude about a direct role of EZH2 in TGF-b1-induced increase of collagen I/a-SMA. These data should be confirmed using a more selective EZH2 inhibitor, siEZH2 and/or EZH2 rescue experiments.
Additional comments: This study deals with an interesting topic: the molecular mechanisms of hepatic fibrosis and the potential of EZH2 inhibitors for treatment of liver fibrosis. While this is a very interesting field of research, the present study suffers several flaws and could be improved by addressing the comments detailed above.


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: DZNEP, AN INHIBITOR OF THE HISTONE METHYLTRANSFERASE EZH2, SUPPRESSES HEPATIC FIBROSIS THROUGH REGULATING MIR-199A-5P/SOCS7 PATHWAY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The English language and narrative have been improved thereby increasing the readability of the manuscript.

The title is an overstatement since the in vitro data have not been linked to the mouse model and could be toned down to better reflect the actual data.

The labelling of Figures and data description has been improved. Nevertheless, there are mistakes in the molecular weight description of the Western blots that should be corrected (in Figure 5B SOCS7 is labelled 63 kDa and in Figure 5G 85 kDa).
Experimental design: Statistical analysis section has been updated to include all tests that were used. It also states that means+/- SD were calculated on several independent experiments. Nevertheless, it is still not clear whether Western blot quantifications have been performed on independent experiments or solely on technical replicates from one experiment as the figure legends indicate that “representative images were picked from 3 separate experiments (technical replicates)”. This should be clarified.

The methods section has been updated and now includes more details in order to enable reproduction of the experiments. However, it does not include the description of immunofluorescence analysis that were added to Figure 1 nor the new data that are presented in the new Supporting Figure 1. This should be added.

The authors should include a legend for the new Supporting Figure 1. Furthermore ‘TGFb1” should be added to panels A and B of this Figure (as done in Figure 5) to clearly indicate that the experiment was done in the presence of TGFb).
Validity of the findings: Using DZNep in control cells would enable to assess the role of EZH2 in basal expression of the genes studied in addition to TGF-beta induced gene expression.

The authors indicate that miR-199-5p is conserved between humans and rats. It would thus have been easy to assess miR-199-5p expression in rat livers following DZNEp treatment and control condition to extend their finding in LX2 cells to the rat model shown in Figure 1 that does by itself not provide new findings as compared to previously published data and is not directly related to the rest of the figures.

Since the authors agree that it would be interesting to confirm their data obtained with DZNep using a more selective EZH2 inhibitor, siEZH2 and/or EZH2 rescue experiments, it would have been interesting to provide at least a few additional data.

Last sentence (“indicate that DZNep may be applied as a therapeutic target for fibrosis”) should be rephrased. According to the authors, DZNep would be used as a therapeutic strategy (no target…).
Additional comments: The English language, narrative, and data presentation have been improved in the revised manuscript.
Nevertheless, there is still some information lacking, particularly regarding the newly added data.
The manuscript could have been further strengthened by addressing the previous comments that the authors chose not to address.


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: OXIDATIVE STRESS INDUCES CELL DEATH PARTIALLY BY DECREASING BOTH MRNA AND PROTEIN LEVELS OF NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE IN DIFFERENTIATED PC12 CELLS
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this study, Zhou et al showed that Nampt plays significant roles in both the NAD+ synthesis and survival of the cells under basal conditions, their study indicated that oxidative stress can decrease both the mRNA and protein levels of Nampt, which indicated a novel mechanism underlying the NAD+ deficiency in aging and multiple neurological disorders. Overall, these findings suggesting a new mechanism underlying oxidative cell death.

There are a few concerns that the authors should address, which are detailed below.

1.	In this study, the author used differentiated PC12 cell as a cell model, however, the cell culture condition seems not a differentiation condition, please clarify. Does your differentiated PC12 cell have neurite outgrowth?
2.	Fig 1 B and C legend, “PC12 cells were treated with 2, 5, 10, 20, and 50 nM FK866 for 24 h rs.” please remove 2, 5, since your figure did not include these two concentrations.
3.	Does author have Supplementary figure legend?
4.	Treatment of the cells with 0.1 or 0.3 mM H2O2 led to significant increases in the Nampt mRNA level of the cells at 12 hrs, in contrast, H2O2 led to significant decreases in the Nampt mRNA level of the cells at 20 hrs after the H2O2 treatment. Does author have any explanation?


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: OXIDATIVE STRESS INDUCES CELL DEATH PARTIALLY BY DECREASING BOTH MRNA AND PROTEIN LEVELS OF NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE IN DIFFERENTIATED PC12 CELLS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is an interesting work that intends to elucidate how oxidative stress regulates the expression and protein levels of the enzyme NAMPT, involved in the salvage pathway of NAD synthesis.
The abstract needs improvements, the objective of the work is not clearly expressed, also the results.
The introduction has the elements necessary to understand the work, however the summarizing of main results do not follow the sequence of the results.
Experimental design: The methods are adequate to respond the objective of the work. However, some interpretation of the data seems to be mistaken.
For example, in figure 1A the authors show that FK866 induces a decrease in the NAD levels, but the effects are similar between all the concentrations tested. However, there is an increase in cell death concentration-depended, How the authors explain these results? It's possible that the increase in cell death is independed of reduction in NAD levels?

The make the data more robust, may include the data of cell death in response to H2O2 with the NAMPT siRNA

In figure 2, the authors show a reduction in the expression and protein levesls of NAMPT induced by oxidative stress, but what is the impact of this reduction in NAD levels?

In figure 3, the authors included a inductor of NAMPT activity, but what is the effect in NAD levels?, also the authors should include the flow citometry assay for this experiment.

Figure 4, its show the effct over ATP levels, however the reduction could be related to the reduction in cell viability induced by H202. It´s important to define what do you want to respond with this experiment.
Validity of the findings: According to the description in the text of the figures, the experiments are pooled from three independents experiments, with an n=6-8. However, to be exact the n =3 with replicates, thus to perform the analysis you must calculate the average of your replicates and perform the statistical analysis with an n=3.
Additional comments: The manuscript presents interesting data regarding the regulation of NAMPT by oxidative stress. This result is the most interesting of the work, however, the authors do not profundity in the mechanisms associate with this regulation. It´s important to unveil if the oxidative stress induces degradation of NAMPT or a reduction in the traduction of the protein. Also, the use of a precursor of NAD+ such as nicotinamide riboside would improve the quality of the work.


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: OXIDATIVE STRESS INDUCES CELL DEATH PARTIALLY BY DECREASING BOTH MRNA AND PROTEIN LEVELS OF NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE IN DIFFERENTIATED PC12 CELLS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Satisficed with  the author's answer.
Experimental design: experimental design  well
Validity of the findings: Nampt plays important roles in oxidative stress-produced decreases in NAD+ levels and cell survival. The  findings indicated the roles of Nampt in maintaining NAD+ levels and cell survival under both basal and oxidative stress conditions.
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: OXIDATIVE STRESS INDUCES CELL DEATH PARTIALLY BY DECREASING BOTH MRNA AND PROTEIN LEVELS OF NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE IN DIFFERENTIATED PC12 CELLS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors respond to all my queries satisfactorily
Experimental design: The authors respond to all my queries satisfactorily
Validity of the findings: The authors respond to all my queries satisfactorily
Additional comments: The authors respond to all my queries satisfactorily


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 RELATIONSHIP BETWEEN PLOD1 EXPRESSION LEVEL AND GLIOMA PROGNOSIS INVESTIGATED USING PUBLIC DATABASES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
Experimental design: The experimental design is original and the research is within the scope of the journal.. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical.
Validity of the findings: All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.
Additional comments: This paper investigates the function of PLOD1 gene expression and mutation in glioma pathology and prognosis using CGGA, TCGA, GEO and CCLE datasets. The authors identified PLOD1 as a potential oncogene in glioma by comparing the expression level in glioma samples and normal controls. The authors demonstrate that PLOD1 is functionally related to ECM signaling pathways utilizing the GSEA method.

Editorial Criteria
BASIC REPORTING
The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
EXPERIMENTAL DESIGN
The experimental design is original and the research is within the scope of the journal.. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical.
VALIDITY OF THE FINDINGS
All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.

Overall, I think this paper is novel and will be of interest to the community of glioma genetics. The statistical part is valid and makes sense. The authors make it comprehensive by integrating analysis of multiple sources including CGGA, GEO, TCGA and CCLE. The main strengths of this paper is that it addresses an interesting and timely question, finds a novel solution based on a carefully selected set of rules and filters. As such this article represents an excellent and elegant bioinformatics study which will almost certainly influence our thinking about the function of PLOD1 in glioma. Some of the weaknesses are the lack of in vitro or in vivo validation experiments. In general, the work is convincing except some major and minor comments below:

Major Comments:

I’m wondering if there are any ongoing clinical trials focusing on PLOD1 in GBM or glioma patients? It will be very strong evidence for the significance of the current study if so.

I’m just wondering if GSE4290 is the only GEO dataset with glioma expression data? Given the sample size for GEO datasets are relatively small, I strongly recommend the authors to incorporate more data into their finding datasets. The authors may also check if there are some consortiums focusing on glioma.

To assess PLOD1 expression in different cancers, the authors used the CCLE data, which is a collection of cancer cell lines. I would recommend exploring the NCI's Genomic Data Commons (GDC) to see if you can find some evidence in the GBM and LGG patient samples in TCGA. Please focus on the “mutation” tab and “oncoGrid” tab.
(https://portal.gdc.cancer.gov/exploration?filters=%7B%22op%22%3A%22and%22%2C%22content%22%3A%5B%7B%22op%22%3A%22in%22%2C%22content%22%3A%7B%22field%22%3A%22genes.is_cancer_gene_census%22%2C%22value%22%3A%5B%22true%22%5D%7D%7D%5D%7D)

Minor Comments:
It is great that a session of abbreviations was there to list all the abbreviations for the database names. I would recommend to also include abbreviations like GBM, LGG, etc in that list.

All the gene names should be italic for all the gene names.

Please add the sample size of both glioma samples and controls for TCGA and GEO datasets in the Data collection and download part, although they were labelled in the flowchart. And for TCGA dataset, how many samples are LGG and how many for GBM? It’s better to make it clear in the Data collection as well.

Line 284: please include the lack of in vitro validation experiment as one of the limitations.

Figure2: the labels are difficult to read for Figure 2D, 2E, 2F. They were too small and not clear.


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 RELATIONSHIP BETWEEN PLOD1 EXPRESSION LEVEL AND GLIOMA PROGNOSIS INVESTIGATED USING PUBLIC DATABASES
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The paper addresses an important issue of relationship between PLOD1 expression level and glioma prognosis. It is well written, interesting and suggests a potential future therapeutic strategy for cancer patients. I think this article should be published.


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: DNA BARCODING OF AUSTRIAN SNOW SCORPIONFLIES (MECOPTERA, BOREIDAE) REVEALS POTENTIAL CRYPTIC DIVERSITY IN BOREUS WESTWOODI
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is clearly written, the used of language is good and easy to understand. The background is well described and the current knowledge is well summarized. The gap in knowledge and the aim of the study is well identified. The figures are of high quality. I would just like to see an additional figure, a map displaying the distribution of genetic variants. The data is all publicly available (after acceptance).
Experimental design: The data presented is new. The data is suitable to answer the posed questions. The aim of the study is meaningful, as the authors study a widely unexplored taxon with a unique and interesting ecology.
The analyses are performed well and represent the state-of-the art. Some of the methods could be described in a bit more detail. Similarly, the results would benefit from some additional details, such as general parameters of genetic diversity in the dataset.
Validity of the findings: The findings are solid and represent important new data for an unexplored taxon. All interpretation is sound and includes not too much speculation. Potential weaknesses are well identified and addressed.
Additional comments: The manuscript is a nice descriptive study that provides important data for an unexplored taxon.

I have just few additional minor suggestions (besides the additional figure above).

Line 35: I would avoid the verb proved, but would rather say suggested.
Line 104: Name of the used kit spelled wrong.
Line 214: Maybe provide the ABGD graph to display the barcode gap.

Discussion
The pattern observed is interesting and very similar to what is seen in sedentary scorpions and darkling beetles in the Atlas, where it is aside from philopatry attributed to the mountainous terrain. May this be an explanation here as well?


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: DNA BARCODING OF AUSTRIAN SNOW SCORPIONFLIES (MECOPTERA, BOREIDAE) REVEALS POTENTIAL CRYPTIC DIVERSITY IN BOREUS WESTWOODI
Review round: 1
Reviewer: 2

Output:
Basic reporting: Boreidae is an interesting group that the adults normally reach sexual maturity in the Winter, and copulate on the snow, so that the studies on this group are quite limited. This manuscript provides a species delimitation analysis in Boreus westwoodi using DNA barcoding techniques. In general, the paper is well written and can be published in PeeJ.
Experimental design: The authors used the popular DNA Barcoding to resolve the species problem of Austria Boreus. The experimental desighn is normally right.
Validity of the findings: The findings are interesting, and the conclusions are robust.
Additional comments: Boreidae is an interesting group that the adults normally reach sexual maturity in the Winter, and copulate on the snow, so that the studies on this group are quite limited. This manuscript provides a species delimitation analysis in Boreus westwoodi using DNA barcoding techniques. In general, the paper is well written and can be published in PeeJ. However, there are some major concerns that have to be addressed before publication:
Introduction:
Line 44: change "(Insecta: Boreidae)" to "(Mecoptera: Boreidae)"
Line 83: please insert a comma "," before "the present study"
Line 87: change  “iv) to test"  to "iv) testing...".
Materials and Methods:
Line 103: "For DNA analyses...." should be in a separate paragraph.
Line 106: "the primer combinations" to " the primer set"
Line 108: What algorithm did the authors use to align sequences in MEGA? Clustalx or Muscle? The authors should give more details.
Line 120: Does "10 Mio" mean "10 million generation"? If so, please correct it.
Line 119-122: Why did the authors use the HKY model and a strict molecular clock (clock rate=1.0)?
Line 136-137: change "as implemented in PopART v.1.7...."  to "Phylogeographic relationships were visulized using PopART"
Results:
Line 159: "six distinct clusters and one singleton" is not very clear, please mark them in the tree.
Line 163: "As we only had two B. hyemalis,...." , the sample size is too low to make a persuasive conclusion. I suggest to increase the sample size of B. hyemalis from different localities.
Line 165: "Interspecific distances" should be calculated with more species of Boreus added. It seems insufficient to sample only two species, B. westwoodi and B. hyemalis.
Line 167: How did the authors reconstruct the NJ tree? The authors should clarify it in the Materials and Methods. Besides, "NJ tree" and "ML tree" should be in full name because the authors have never mentioned them before.
Line 171: "for conspecifity" or "for conspecificity"?
Line 173-174:  change "phylogeographic substructuring" to ""phylogeographic structure", and "sharing" to "shared".
Line 180: "the B. westwoodi clade most closely related to B. hyemalis" is not right. According to the topology of the phylogenetic tree in Fig. 2, B. westoodi is apparently a paraphyletic grade, not a monophyletic clade.
Line 181-182: change "different to" to "different from", and "haplotype sharing" to "the shared haplotype".
Discussion:
Line 237: The authors claimed "we found a remarkable geographic structure", but we can’t see it clearly in the network. If the authors want to clarify geographic structure, please give details about the haplotype diversity (Hd), nuleotide diversity (Nh), genetic differentiation (Fst) using DNAsp, Arlequin, and SAMOVA, etc.
Line 243: How did the authors translate the pairwise distances into divergence times? Did the authors estimate the divergence time based on COI? If so, please show it in the supplementary materials.
L249: change "large distance”to "long distance"


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: DNA BARCODING OF AUSTRIAN SNOW SCORPIONFLIES (MECOPTERA, BOREIDAE) REVEALS POTENTIAL CRYPTIC DIVERSITY IN BOREUS WESTWOODI
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1. The authors incorrectly quoted the Figure 1 (e) from Kreithner (2001). According to the original reference, only 33 and 34 are represented TAs of Boreus hyemalis, and 35-39 were those of taxonomically uncertain forms (35-38: Form I, 39: Form II).

2. The legend of Figure 1 seems to have errors or conflicts. There are two legends for C-D, "schematic drawings and images of TA and GS" and "Digital microscopy images of TA". In addition, labels for each figure are (A), (B) ... in legends but a), b) ... in figures. Arrows shown TA should be added into the microscopy images (C) and (D). Please revise the captions.

3. In Introduction, the authors do not propose some basic information, such as the number of described species in Europe, and a species list known from the Central Europe. In addition, the current situation of elucidation of Austrian Boreus fauna is not fully introduced, so readers cannot clearly understand which record of which species is new information for Austrian fauna. Please consider to add these information.

4. The reference format is inconsistent. For example, full titles and acronyms of journal titles are mixed; only two references have DOI, and one is represented by DOI itself but another is quoted as URL. A thorough review is needed.

5. In Introduction, about the relationship of Boreus hyemalis and B. westwoodi, there is another opinion that two species are conspecific (Sauri, 2003, Entomologische Nachrichten und Berichte 8; I do not check the original resource but some websites such as https://www.insekten-sachsen.de/Pages/TaxonomyBrowser.aspx?id=223691 follow this opinion). I recommend the authors to insert a short comment about this in the historical review.
Experimental design: 6. In Table 1, the authors represented the GS shapes by the numbers "41" or "45", which is the figure number in Kreithner (2001), quoting Kreithner's drawing of GS in Figure 1 (G) and (H). But, they do not provide any mention on specific differences in shapes which distinguish from one another; they only wrote as "distinctly different form" (Line 152). The detailed differences should be described so as to replicate the analysis.

7. A threshold value to show the branch support value (>0.95?) and a scale for branch length are not provided in Figure 2. In addition, Is the branch support value not Bayesian posterior probability but maximum likelihood bootstrap for ML tree?

8. I do not understand the reason why the authors chose six characters (TA, EA, HA, caput, antenna and front wing bristles: Line 97-100). Were the former three characters used as important diagnoses in previous taxonomic papers? And why they added latter three characters? Please add some information about the strategy on selection of morphological characters.

9. The authors identified the females based on the geographic location (Line 100-102) but they do not mention the validity. Why the authors did not adopt the ovipositor as a diagnosis of female written in Kreithner (2001)? Why they regard the location as a key for identification? In other words, are the Boreus species show exclusive allopatric distribution pattern or separated emergence season? I recommend the authors to add short comment about this issue.

10. For phylogenetic analysis, they do not provide any information to judge the validity of Boreus borealis as outgroup (Line 111-113). What kind of species is this, and how does this species relate to the target Austrian populations? I recommend the authors to add short comment about this issue.

11. About the records of Boreus hyemalis, one male and one female were recorded in manuscript (Line 142-144), but two males are shown in supplementary table (Panorpa294, 295). Please check which is correct.
Validity of the findings: 12. Related to comment 1, in results the authors said "a distinction between B. westwoodi anvil shapes and B. 150 hyemalis anvil shapes like Kreithner (2001) deduced (Fig. 1e, 25-32 for B. westwoodi; 33-39 for 151 B. hyemalis), was not possible" but this statement is due to misinterpreation (33-34: B. hyemalis, 35-38: Form I, 39: Form II). Comparing Figure 1 (E) and 1 (F), TAs of some Austrian individuals seem to be more similar to those of Form I or II than those of B. westwoodi (e.g. f6 and e38). How the authors regard that TAs of all Austrian specimens except B. hyemalis as that of B. westwoodi? Anyway, discussion on the comparison of Kreithner's forms should be added.

13. Related to comment 8, I recommend the authors to clearly state, 1) characters which have been regarded as good species diagnoses in previous papers, 2) characters from which the authors found unrecognized large variation. This helps readers to understand new findings of this research.

14. Please note if any BIN clusters distinguish from remaining ones morphologically.

15. An addition of a map showing geographic distribution of BIN clusters is welcome to understand the complicated situation. Please note if there are any differences of habitat preferences between species and/or BIN clusters.
Additional comments: In summary, the cryptic diversity of this attractive and wingless insect group which the authors reported is an interesting phenomenon, and this is included in the scope of PeerJ journal. On the other hand, there remains issues and errors on the manuscript, and significant revision is required.


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: DNA BARCODING OF AUSTRIAN SNOW SCORPIONFLIES (MECOPTERA, BOREIDAE) REVEALS POTENTIAL CRYPTIC DIVERSITY IN BOREUS WESTWOODI
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: No comments.
Validity of the findings: No comments.
Additional comments: The revised manuscript has been much improved. However, the reviewer has still the following concerns, and should be happy to recommend it to be published after a minor revision.
1.	The insect mutation rate of mtCOI is commonly considered as 2.3% per Ma (i.e., 0.0115 substitutions/site per lineage). We recommend the authors estimate the divergence time based on Brower (1994). Please see the references Brower (1994) "Rapid morphological radiation and convergence among races of the butterfly Heliconius erato inferred from patterns of mitochondrial DNA evolution" (PNAS, 91: 6491-6495) and Hu et al. (2018) “Evolutionary history of the scorpionfly Dicerapanorpa magna (Mecoptera, Panorpidae)” (Zoologica Scripta, 48: 93-105);
2.	A Yule model may be more suitable for the BEAST analysis than a birth-death model;
3.	Line 55, please insert a comma “,” between Hagen 1866;
4.	Line 58, please add a space prior to “Field”;
5.	Line 73, please change “to” into “with”. In this context, “with” should be used in combination with “compared”;
6.	References: the authors used different format for different references as for colon “:” or comma “,” after the issue number of journals;
7.	Penny (1977) is repeated between Lines 442 and 448, please delete Lines 442-443.


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: DNA BARCODING OF AUSTRIAN SNOW SCORPIONFLIES (MECOPTERA, BOREIDAE) REVEALS POTENTIAL CRYPTIC DIVERSITY IN BOREUS WESTWOODI
Review round: 2
Reviewer: 2

Output:
Basic reporting: 1. About the authors' response on my previous comment 3 contain some interesting information, that is, Kreithner (2001) did not find B. hyemalis in Austria, probably because B. hyemalis is not a common species, and it is not surprising that some previous records of B. westwoodi from Austria are misidentifications of B. hyemalis. I suggest the authors to add the information to discussion (around Line 242). The new information may give readers chances to re-check their previous Boreus records and specimens (even though in the next paragraph they mention the possibility of existence of more species in Austria).

2. The revised legend for Figure 1 (d) should be "pointed TA of B. hyemalis (d) from Austria (indicated by ...)".

3. In reference part, please check the format of DOIs of Huemer et al. (2019) and Raupach et al. (2016). They are not represented by URL.
Experimental design: No more comments.
Validity of the findings: No more comments.
Additional comments: I think the manuscript is revised well and only I mentioned a few minor points as above.