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: PREDICTING MIGRATION ROUTES FOR THREE SPECIES OF MIGRATORY BATS USING SPECIES DISTRIBUTION MODELS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This study uses quantitative modeling and occurrence data for several migratory bat species in North America to derive models and maps of species distributions and possible seasonal migration patterns for these species on a continental scale. The occurrence dataset was collected using Global Biodiversity Information Facility (GBIF) data.  Once species distribution models (SDMs) were developed, several approaches to modeling seasonal migratory patterns (circuit theory, 95th percentile suitability, and least cost path analyses) were used to develop possible seasonal migration patterns on the North American continent. The manuscript is well-written with an introduction to the study and some natural history characteristics of these species.

My comments on the manuscript focus both on the quantitative methods and the level of circumspection in the analysis. I am not as well-read as I would like to be on the methods used to develop the migratory pattern models (circuit theory, least cost analysis, etc.), so I can’t offer much serious feedback regarding the modeling done using these methods. I do feel more comfortable in the species distribution modeling domain and so provide some suggestions for this piece of the analysis. Based on the details presented I suspect that the quantitative analysis is done reasonably well throughout. However, I don’t think the quantitative analysis is as well-explained as is could be, and the rationale for the approaches taken in the analysis are not always quite clear to me. I would encourage the authors to address the following methodological questions more thoroughly so other researchers and managers in this and other domains will more clearly understand the rationale and perhaps use this work as a framework to consider in their own areas of research. However, if the handling editor does not feel that these issues need to be addressed, I do not necessarily think they need to be addressed prior to publication, though some more detail (as described below) would likely help some readers, including myself.

Throughout the paper: consider defining “migratory corridor” early in the introduction so that readers know what you mean by this term, and then consider returning to this topic in detail in the discussion. I’m not convinced that the migratory tree bat species you analyze use migratory corridors per se. It is possible they do, but it seems to me that a reasonable hypothesis would be that they tend to more or less diffuse from winter to summer grounds and then back to winter grounds along broad geographic fronts. For example, hoary bats, being relatively solitary, may very well migrate from the southeastern United States where some populations spend winter into the interior of the continent where they spend the warmer summer months, and then back to the southeast in the fall. I think it would be reasonable to hypothesis that this general movement pattern may very well happen in a way that is more like gas particles moving from higher to lower pressure, or a liquid moving across a surface from higher to lower ground. If this is the case then describing the phenomena as a migratory corridor would probably not be an accurate description of the phenomena. Consider being clear about what you mean by the term corridor, and also consider explicitly stating that this is one possible hypothesis about migratory movement, but not the only possible pattern. This detail would add some length to the manuscript, but I believe it would be worth discussing in some detail.
Likewise, I believe your analysis assumes that these species follow gradients of habitat suitability and that the species are never really allowed to “decouple” from contact with the habitat suitability surfaces that shape the migratory patterns you are deriving. Some readers (myself included) will wonder whether these species, being long-distance migrants that often fly long distances at high altitudes above the ground, do indeed regularly decouple of the influence of these habitat suitability surfaces, and will thus wonder how this potential phenomenon is dealt with in your analysis. Some readers may also wonder if these species may regularly pass through and over lower quality habitat in order to move quickly between high quality winter and summer habitat. It would certainly seem reasonable to me to conclude that some if not all of the individuals of these species regularly decouple from the surface habitat suitability of a given pixel in your modeling template, and likewise regularly pass through lower quality habitat in transit. It is not clear to me how you have considered and resolved these questions, and I would encourage you to consider discussing these questions in some detail, especially in the discussion.

Thanks for the opportunity to take an early look at this manuscript. I enjoyed reading it and look forward to seeing it in print.
Experimental design: Generally I think the study design is very good and represents a novel and creative approach to analyzing seasonal migratory patterns of these species.

Line 113. Rationale for using only records from last 50 years. I don’t find this argument to be very convincing. For example, there are numerous records that could be used for specimens collected before about 1970. Many of these records can be definitively identified to species and many of these records can be reliably identified to their described occurrence location. See Cryan (2003) for a discussion of these details; Cryan compiled a comprehensive North American data set of the three species you analyze going back to the late 1800’s and in considered an authoritative sources.

Line 119. Consider adding roughly how many km 1 degree of latitude and longitude is equivalent to. I believe this is about 111 km or so. It seems to me that this would substantially thin the data, possibly in a way that causes the resulting models to not resolve relative habitat suitability very well. If implementing this course thinning procedure I would be particularly concerned that this would be the case in mountainous areas and in areas with substantial topographic relief. I would encourage you to discuss this in a bit more detail in this part of the methods. My opinion is that it’s likely that you have over-thinned the occurrence data.
Validity of the findings: My perspective is that the analysis is very likely done reasonably well, and thus the results are valid given the data and approaches used.

If you are not doing this already I would encourage you to post your data and code on a publically-accessible repository.

Consider comparing and contrasting your results and conclusions with those of others who have analyzed the migratory patterns of these species in North America and beyond. You very briefly mention the results of other workers but I think the manuscript would be much improved if you made sure to compare and contrast your results and conclusions with earlier workers in more detail. For example, the pathway analysis map in Figure 4 strikes me as a gross oversimplification of the seasonal migratory patterns that are occurring in reality. What have earlier workers said about these patters? Do your results confirm or contradict these ideas, and why?
Additional comments: Line 58 and throughout the paper, “latitudinal migrant”. My sense is that these bats aren’t really latitudinal migrants, per se, but rather tend to move from high quality winter to high quality summer habitat in ways that sometimes follows latitudinal gradients. So for some individuals and populations these patterns may follow latitudinal gradients, but not always. For example, some prior researchers have proposed that hoary bats move from areas of the North American continent that are closer to oceans and other large water bodies in winter to the interior of the continent in summer. This does not really follow latitudinal gradients. The same may be true along mountain fronts like the Rocky Mountains. Consider being clear about what you mean by latitudinal migrant and consider whether this is the term you want to use here and elsewhere in the paper.

Figs 2 & 3 could be sharper. I have a very hard time interpreting what’s being presented in these figures in the pdf. I think this is just a result of the lower resolutions of the figures in the pdf version offered for review, and it looks like the supplementary figures are reasonably sharp. But these are important figures in the overall argument of the paper and readers will want to see them clearly.

Fig 2. The color ramps for LABO and LACI extend from 0-0.6, but from 0-0.5 for LANO. Consider standardizing the color ramps so they are the same for all three maps. I would suggest scaling them all from 0-1.0. I also find the color ramp colors difficult to view. For example, summer and fall for LACI is very difficult for me to visualize and interpret.

Line 14. Consider removing “accurate” from the “describe accurate seasonally resolved distributions” phrase. This seems to imply that prior efforts are inaccurate.

Lines 17 & 16. Consider saying how many locations for each of the three species; and indicate which decades.

Line 93. Consider describing what the WorldClim data is. This will be helpful for readers who don’t know this lingo.

Methods and throughout. Consider checking to make sure that you have provided a good reasonably-recent citation associated with each SDM algorithm you use. You provide good citations for Maxent, but not for some of the other SDM approach. Some readers, including myself, will want to see high quality citations for these approaches in case they want to follow up and get more details about the approach.
Consider adding a figure to the manuscript that shows the locations of the occurrence locations used in the analysis, for example by showing a panel of maps, one for each species, that shows the locations of the points. Some readers, including myself, will want to see this. This would also allow readers to consider any sampling bias that may be associated with the locations, for example by looking for clusters of points and/or large gaps in occurrence records, etc.

Line 123. Consider saying approximately how many km are in 2.5 minutes of resolution (I believe about 4.5 km).

Lines 125-6. Consider adding the units used for each variable considered. For example, temperature (°C), winds


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: PREDICTING MIGRATION ROUTES FOR THREE SPECIES OF MIGRATORY BATS USING SPECIES DISTRIBUTION MODELS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors present an interesting underlying pair of hypotheses (that seasonal migration habitats and pathways of certain bats can be predictively modeled from presence-only occurrence records), but in my judgement fall a bit short in a few areas of basic reporting.  First, although things start out well and the writing is passably professional, things come a bit unwound in the latter half of the manuscript, where the writing begins to include contractions, grammatical errors, and generally begins to wander in terms of thematic consistency and focus.  Hypotheses that are either explicitly stated or alluded to during the earlier parts of the manuscript are not consistently revisited in the context of the findings, whereas new ones appear in the latter parts of the discussion that surprised me.  An example of where consistency checks could improve the professional feel of the writing include statements in the abstract saying there are "...many species of migratory bats...'' followed later by "...only 12 of 500 Vespertilionid bats undertake long-distance migration...".  Overall, I think the depth and breadth of the background literature is somewhat shallow but probably sufficient, although several key concepts presented in the manuscript do not cite relevant primary literature (e.g., prior occurrence-record and stable-isotope evidence of longitudinal components to migration in these species).  In general, I got the impression that the authors were under-describing the prior effort to hypothesize seasonal distributions of the hoary bat using these same SDM methods.  I was not convinced the SDM elements of the study described here were unique enough to downplay those similarities (and spend text comparing results and addressing previously highlighted problems/biases inherent in that type of analysis) to the prior effort.   Other things that rubbed me the wrong way while reading through this manuscript were things like use of the term 'latitudinal migratory bats' when that is not common use or necessarily accurate, as well as potential false dichotomy of migrating bats following linear features or not.  The writing and logic style of the manuscript in its current form leave open opportunities for improvement.
Experimental design: If I read between the lines and ignore the somewhat opportunistically scattered hypotheses that surface later in the text, I think the real strength of this study is the novel use of what I interpreted as diffusion models to try and improve understanding of where migrating bats might concentrate in landscapes between seasonal occurrences.  This is a really neat idea and it makes sense that given the best information at hand (temporally variable and spatially explicit occurrence records), the next logical step in trying to infer the seasonal movements of these elusive migratory animals is to look at transitions between known and hypothesized known locations.  The earlier work to model seasonal species distributions of these tree bats using museum occurrence records, then stable isotopes, and then SDM methods all took understanding from a single, static species distribution map to what is essentially a seasonal and now monthly time series of species distribution maps.  Extending the analogy, the prior work focused on the data points along the time series, and now the current study begins addressing what might happen between those points.  In my opinion, this manuscript could be much improved by focusing less on the tangential and tenuous underlying justifications of the work (e.g., that yellow warblers are observed more often because they are active during the day...a rabbit hole I will avoid here), claiming less novelty with the SDM ensemble modeling, and honing the manuscript to support the assumptions, methodological details, and findings of the transition model elements.  You can tell from my loose use of terms that I'm not sure whether to refer to the intra-month modeling as diffusion, transition, flow, or something else, but further making that part pop out with terminology consistent with the scientific literature would really help.  Put another way, future scientists wanting to replicate this work can go to other studies to get details of the SDM methods used, but will not have other leads in the bat literature for repeating the diffusion modeling to study changes in density of migrants across landscapes at various spatial and temporal scales.
Validity of the findings: The authors' use of diffusion modeling methods to generate spatially and temporally explicit hypotheses of variation in density of migrating tree bats was the strongest element of this work that deserves further fleshing out and exploration.  It was good to see additional SDM modeling using methods very similar to a prior effort, but the claimed increase in resolution brought with improving the sampling period (from seasons to months) and spatial resolution of environmental covariates (down to 2.5km grid cells for climate info) was not presented with comparatively convincing evidence to support the claim of implied improvement.  That seems incidental to the overall impact if focus were shifted to the month-to-month transition analysis.  More information exists in the literature presenting alternative migration patterns to the simple latitudinal model implied in this manuscript as the prevalent standout (e.g., multiple stable isotope studies of all three species of tree bat)---inclusion of those alternatives in the context of the diffusion model findings could also provide new and unique context.
Additional comments: Despite my reservations about the way the information is currently presented.  The underlying findings of the diffusion modeling to explore and model density of migrants are exciting and promising.


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: PREDICTING MIGRATION ROUTES FOR THREE SPECIES OF MIGRATORY BATS USING SPECIES DISTRIBUTION MODELS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The reporting in the revised manuscript is clear and thorough.
Experimental design: The experimental design is appropriate for this analysis and a creative approach to the problem.
Validity of the findings: I believe the results are valid given the data and approaches used.
Additional comments: Nice work on this paper and keep up the creative modeling and analysis.


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: EVALUATION POTENTIAL OF PGPR TO PROTECT TOMATO AGAINST FUSARIUM WILT AND PROMOTE PLANT GROWTH
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript by Rizwana begum Syed Nabi et al. evaluated the ability of plant growth-promoting rhizobacterium (PGPR) Bacillus aryabhattai strain SRB02 to alleviate the effects of tomato wilt disease caused by Fusarium oxysporum f. sp. lycopersici (strain KACC40032) and promote plant growth, which is very beneficial to protect crops from vascular wilt, one of the most important fungal diseases of crops.

Major concerns:
1. Lack of the research data on the mechanism of bacteria inhibiting fungi;
2. Figure 1 has poor clarity and the name of the bacteria and fungus in the plate was not indicated;
3. Figs 4 and 5, the meaning of the significant difference mark are not stated in the legend. The difference between tolerant or susceptible plants should be marked with different signs;
4. Table 1 and Table 2, Table names and Tables are mismatch; data with significant differences between different treatments should be marked with symbols;
5. Line 88 and 279, pointed out that one of the mechanisms of PGPR to promote plant growth is that PGPR synthesizes ethylene, which is incorrect. Instead, PGPR degrades the ACC synthesized by plants through ACC deaminase, thereby reducing the plant ethylene synthesis and the inhibition of ethylene on plants.

Minor concerns:
Line 325, 343, 346, 355, 371, 383, 387, 404, 418, 433, the reference format is incorrect.


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: EVALUATION POTENTIAL OF PGPR TO PROTECT TOMATO AGAINST FUSARIUM WILT AND PROMOTE PLANT GROWTH
Review round: 1
Reviewer: 2

Output:
Basic reporting: The research work described the ability of plant growth-promoting rhizobacterium Bacillus aryabhattai strain SRB02 to control tomato wilt disease caused by Fusarium oxysporum f. sp. Lycopersici. The paper presents an interesting research topic and good ideas, but the manuscript is not acceptable for publication in its present form.  The research work contains insufficient methodological detail. The plant growth-promoting activity and bio-control of fungal disease of tomato by B. aryabhattai were reported (see comments)., thus it should be indicated and cited. Most of references are outdated and should be replaced by recent reports. Tables and figures should be worked out and statistical analyses should be added.
Experimental design: The research work contains insufficient methodological detail. I have made some comments that should be improved and more detailed descriptions should be provided. Research questions should be also worked out because PGP traits of used bacteria were not described in detail.
Validity of the findings: no comment
Additional comments: Comments
Introduction section: Please reduce outdated citations and replace them with recent reports. The introduction part should focus on the biological control of plant pathogens by PGPR and, mechanisms of action.
Line 23: “to alleviate the effects of tomato wilt disease caused” to alleviate or to control?
Line 96-98: add citations.
Line 111: “there is a lack of information about the growth-promoting activity of B. aryabhattai”. There are some reports on the effect of this bacteria on plant growth of tomato, and biological control, please add references. https://pubmed.ncbi.nlm.nih.gov/31216607/
https://link.springer.com/article/10.1007/s40858-019-00283-2

Line 132:” tomato seeds of four Korean cultivars” include cultivar names and describe each cultivar characterization.
Line 134: Horticultural soil, describe soil physical and chemical characteristics and origin
Line 136: big pots, the volume?
Line 142: 14 days growth, is the plant growth duration not so short to show symptoms of the disease?
Line 146: biotic stress? Please describe
Line 153: why plants were grown in high soil humidity? Plant growth conditions should be described in detail.
Line 155: “. to determine the response of plants to infection in the presence or absence of SRB02”, dry weigh or chlorophyll content does not relate to plant-pathogen interaction, other parameters should be discussed to support the response of the plant to fungal infection.
Line 164:” was determined using relevant standards.” Describe in detail
Line 199: Response of tomato cultivars against pathogen?
Figure 1, please add as supplementary material

Figure 2. This figure is not clear, clarify control and infected plant, and also describe means of tolerant, moderately tolerant and susceptible varieties. How these parameters were determined?
Figure 6 demonstrate general mechanisms used by PGPR to control plant disease and stimulate plant growth, but not findings of this study, thus should be omitted.
Table 2 has some format problems, statistical analyses are missing
Table 1. Statistical analyses are missing, add significant differences among treatments


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: NGSCLOUD2: OPTIMIZED BIOINFORMATIC ANALYSIS USING AMAZON WEB SERVICES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Language throughout is clear, professional, and unambiguous, though I highlight some typographical errors in "General comments".

Intro and background insufficiently show context for the work. For example, much more could be done to elaborate on "Next Generation Sequencing (NGS) technologies and the algorithms and applications employed". Overall the Introduction feels very rushed.

The results and discussion appear to be largely methods, in my view. I would move all the content of results and discussion to methods, and then provide more results and discussion as relevant.

Much could be done to improve accessibility of this framework by providing more information for the reader about the details of AWS cloud infrastructure. For example, on line 92 "architecture of instances and volumes" will be entirely opaque to the average reader. What is a "volume"? What is an "instance"? What are "m4, c4 and r4 instance types"? Try to clarify technical jargon for the non-specialist.

The figures are very complicated, do not have legends, and are not described sufficiently in the main text, so it's difficult to understand their meaning.

Raw data is supplied in the SRA, which I verified.
Experimental design: The correct operability of the pipeline is referenced in the materials and methods, yet no evaluation of this is presented in the results. I would like to see more elaboration on the methods for this evaluation and presentation of the results.

Typically a new method will involve some validation with simulated data to ensure correct results. I would recommend this here.
Validity of the findings: Results and discussion are largely methods, so I can't evaluate validity of the findings.
Additional comments: The authors present NGSCloud2, a bioinformatic system for interfacing with Amazon Web Services (AWS) to facilitate running next-generation sequencing analysis workflows on flexible AWS cloud computing infrastructure. It is clear that the authors have done a significant amount of work to provide this resource to the community, and I applaud their effort. The documentation is extensive, which is wonderful, but the install process may be beyond the reach of non-specialized users, so I would recommend trying to ease this burden. As it was not part of the manuscript, I did not consider the state of the documentation or the install process during my review, I only highlight this here for the benefit of the authors.

L53: I would not call AWS a "public cloud service" as this is a private service which the user must pay for.
L60: M3, C3, and R3 will not mean anything to the average reader. Can you make it more clear what the difference is between these?
L65: Delete "NGSCloud2" here as it's redundant.
L75: As the methods for these bioinformatic pipelines haven't been described yet, it's a little confusing to be referred to the Results and Discussion on this line.
L93: "Figure 1" <- Add space

Table 1
"Read alignment" <- Bowtie2
"Transcriptome filtering" <- CD-HIT-EST

Table S1
I know supplementary materials are not supposed to be reviewed, but this appears to be a pricing list for AWS cloud compute usage. I don't think this is a result of the work of the authors, and I also think this will become rapidly outdated, so I would suggest removing.


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: NGSCLOUD2: OPTIMIZED BIOINFORMATIC ANALYSIS USING AMAZON WEB SERVICES
Review round: 1
Reviewer: 2

Output:
Basic reporting: This work presents next-gen sequencing analysis tool based on Amazon Web Services (AWS) platform. The work is clearly presented. English is Ok.
Raw data (SRA) are shared. The manuscript has sufficient (but rather short) background for cloud computing and existing tool. The introduction could be extended.
Some abbreviation might be not understandable for broad readers’ audience (“m4, c4 and r4 instance...”)
The work meets PeerJ standards, overall.
However, reader can see just presentation of the tool, but not the novel results. It might be appropriate for bioinformatics paper. I believe PeerJ is the journal for Life sciences research rather than just software tool description.
PeerJ Computer Journal seems more appropriate for such publication (Just suggestion to consider)
Experimental design: The work describes the pipeline, but not biological results. I expect more biological applications rather than just software updates in the Results. Presentation of the Methods section is quite detailed.
Validity of the findings: This work presents NGSCloud2, the  bioinformatics tool for cloud computing. As the software it is quite useful, presents novel solution and diversity current platforms in sequencing analysis. Thus, such work is valid for science community and should be published.
However, the findings/results are not given in classical form. IT makes this paper interesting for the specialists, but not a reader with general background.
Computer jargons and unclear technical presentation including figures should be fixed.

Amazon Web Services named as "public cloud service" though it is commercial project. Please state it clearly. Supplementary Table S1 with the pricing looks not relevant for science publication. Need either remove or update it.
Please comment on the abbreviations M3, C3, and R3.
And explain CD-HIT-EST.
Additional comments: Please avoid technical details to make this work close to broad reader audience. The Results section should be reformatted.
The figures' legends could be more informative.


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: NGSCLOUD2: OPTIMIZED BIOINFORMATIC ANALYSIS USING AMAZON WEB SERVICES
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Fernando et al provide an updated toolkit to analyze RNA-seq data in Amazon cloud. The toolkit adopts several most popular packages for RNA-seq analysis and the author shows a distinct flowchart and structure on Amazon platform. The online resource and service faciliate the personal bioinformatic analysis and teamwork in different loci. I agree the authors provide a solution of simple and convenient protocols for RNA-seq data and the manual is quite readability for a beginner on Amazon cloud.
However my concern is about the usage or experience for the user. Currently, we have too many choices for the tools selection (https://en.wikipedia.org/wiki/List_of_RNA-Seq_bioinformatics_tools) and hardly find a best pipeline for all types of data(Genome Biology.2016,17:13). Some tools have limited maintenance and are out-of-date in scientific community (eg. "cufflinks" for DEG calling). There are still some popular tools for DEGs calling (Bioinformatics.2019,35(18):3372–3377 ). I suggest author to list the activity status for database (eg: https://bigd.big.ac.cn/databasecommons/) and the latest version info to provide additional information for the user.  Another suggestion is to add the steps of "enlarge the volume in EBS" in the manual book.


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: NGSCLOUD2: OPTIMIZED BIOINFORMATIC ANALYSIS USING AMAZON WEB SERVICES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: In my view, the section "Comparison with other cloud platforms for bioinformatics" isn't really a "result". I would move this to the discussion.
Additional comments: The authors have satisfactorily addressed all my initial comments and suggestions. The added text describing NGS and AWS in the introduction are very helpful. My main remaining concern is the redundancy between the results and the discussion. For example, the first paragraph of the discussion
largely revisits the "Comparison with other cloud platforms for bioinformatics", and the last two paragraphs of the discussion largely recapitulate the results section "Validation of NGScloud2 operability". I suggest reorganizing the results and the discussion to reduce the redundance.

Line 264: "functional"


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: NGSCLOUD2: OPTIMIZED BIOINFORMATIC ANALYSIS USING AMAZON WEB SERVICES
Review round: 2
Reviewer: 2

Output:
Basic reporting: Thank you for the manuscript update. It is interesting material from the point of view of a user.
Experimental design: Experimental design of the application fits to the journal scope.
Validity of the findings: Novel software tool is presented, useful for wide range of classical NGS applications in transcriptomics. The results are based on the data, well reproduced. All the tests and resulting data are provided in archive, stored in Github.
Additional comments: Due to extensive text update there are some logic duplication (repeating phrases in the discussion part of the text (advantages of NGScloud2). It gives no new information. Recommend remove it or rephrase.

There are some technical comments to be fixed before publication.
RNA-seq name usually is published with a dash. But the authors use RNAseq throughout the text, except of Table 1. I recommend at least write first in the text “RNA-sequencing (RNAseq)” to keep consistency.
I have no more critical remarks. Endorse publication of this manuscript.
Line 29: de novo - could be in Italic
Line 81: “several applications” - please mention several, not only Galaxy. Or remove word ‘several’.
I’d recommend mention iPlant, miCloud (doi: 10.1089/cmb.2018.0218), DolphinNext (doi: 10.1186/s12864-020-6714-x) to name a few.
Line 100: GiB - it is GiB through the text, more common is GB abbreviation. May keep as is, however
Line 111-112: Mora-Márquez, Vázquez-Poletti & López de Heredia U, 2018
The citation could be by first author name only, with ‘et al.’
Line 131 - repeat of the full abbreviation from line 121 -
Restriction site Associated DNA sequencing (RADseq)
- may just use ‘RADseq’ second time
Line 132: and functional annotation -
More common is ‘gene functional annotation’
Line 142: ‘n virtual machines’ - need mark ‘n’ in Italic.
Maybe specify here maximal (tested) number of n machines. Or remove it, since number n is not used anymore in the text.
Line 180: JCR -
Please add the link (https://www.scimagojr.com), or full abbreviation for JCR
‘Web-of-Science’ is written with dash, or as ‘WoS’. Might give here reference or link to the database used.
Line 212: (Anders, Pyl & Huber, 2015) - citation could be by first author name et al.
Line 231: psedudogenome - typo in ‘pseudo’
Line 240: TOA - give the abbreviation in full (it is given in the Table only)
Line 241: (PLAZA) - the database could be with the link or reference
Line 242: NCBI's RefSeq
May write NCBI without ‘s’
Line 251: Gene Ontology -
There are different ontology databases, please give the link (http://geneontology.org/ or preferably direct to the data source)
Line 265: Illumina TM -
I think it is quite standard, may write without (TM), or at least mention only once, not need repeat in line 267.
Line 292: PE -
Please comment as (pair-end) since there are two many abbreviations in the text, it is better avoid extra abbreviations.
Same for line
298: SE
- write single end, minimize the use of abbreviations
Line 305: compute oriented
- compute-oriented
Lines 307-308: ‘because’ is repeated, try to rephrase
‘TOA.db’ maybe write as TOA database? This name is not used as .db in the text
Line 322: ‘in a flexible way’ - what means flexible? Could give more detail here since it is discussed as the advantage of the tool.
Maybe ‘flexible’ in terms of user time? Or optimal in some other sense?
Lines 346-350: --
I believe it is redundant text repeat, already mentioned in the text section before.
Rephrase or just remove it for clarity.
Line 417: ‘Table 1’ - it is not paper title, check the reference
There is some improper reference formatting for line  454 too.
Figure 5 -
A plot right to ‘Summarize & plot’ box - it is too small, not clear what is it? Assume it is just illustration of a plot?

Overall all the remarks are really minor for authors’ discretion to fix.
I will not check it the text again.


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: ANTIFUNGAL PROPERTIES OF VOLATILE ORGANIC COMPOUNDS PRODUCED BY DALDINIA ESCHSCHOLTZII MFLUCC 19-0493 ISOLATED FROM BARLERIA PRIONITIS LEAVES AGAINST COLLETOTRICHUM ACUTATUM AND ITS POST-HARVEST INFECTIONS ON STRAWBERRY FRUITS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Self-contained with relevant results to hypotheses.
Experimental design: Original primary research within Aims and Scope of the journal.
Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap
Validity of the findings: All underlying data have been provided; they are robust, statistically sound, & controlled.
Additional comments: This article aimed to isolate endophytic fungi from Barleria prionitis plants grown in Thailand and to investigate the antifungal properties of the volatile compounds produced by these endophytic fungi against Colletotrichum acutatum.

The article is straightforward. Although the article is not innovative, it contains original and interesting information. The authors may consider removing the term “room temperature” throughout the manuscript. The term may misguide readers. Besides, this article would be improved if the authors clarify or revise the following:

Line 59. Italicize “C. acutatum”.
Line 104. Leave a space between “al.,” and “2008”.
Line 181. Revise to “… CD-AX, Japan)”
Line 273. Revise to “(HP 6890, Agilent Technologies)”.
Lines 350-351. Revise to “varying degrees (Fig. 1).” And remove “The results of the in vitro assay are shown in Fig. 1.”
Line 353. Revise to “(80.79%, p<0.05).”
Line 361. Revise to “incubation period (Figures not shown).”.
Line 398. Revise to “respectively (Figures not shown).”.
Line 403. Revise to “ treated with elemicin than the positive control sample”.
Line 428. Revise to “According to previous studies (Singh et al., 2005; Amoo et al., 2011), B. prionitis …”.
Lines 465-468. However, results shown in Figure 3 were not impressive. Elemicin demonstrated minimal effect against C. acutatum”.
Line 478. Revise to “… this plateau for additional 3 days.”

Table 1. The table should be self-explanatory. Inhibition, severity, and incidence percentages of what? observed from the in vivo experiments. Clarify how the authors obtained these data, referring to Figure 3. The strawberries treated with elemicin shown in Figure 3 seem to have less inhibition than 70.87%.

Figures 1, 2, 3, 4, and 6. Italicize fungus names in the titles.

Revise Figure 3 title to “In vivo antifungal activity of the volatile compounds produced by the Daldinia sp. MFLUCC19-0493 against C. acutatum infections on day 7 after incubation at 27°C.”


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: ANTIFUNGAL PROPERTIES OF VOLATILE ORGANIC COMPOUNDS PRODUCED BY DALDINIA ESCHSCHOLTZII MFLUCC 19-0493 ISOLATED FROM BARLERIA PRIONITIS LEAVES AGAINST COLLETOTRICHUM ACUTATUM AND ITS POST-HARVEST INFECTIONS ON STRAWBERRY FRUITS
Review round: 1
Reviewer: 2

Output:
Basic reporting: There were missing references that appeared in the text and are not in the reference list: (Sahu 1984) and (Guo et al 2018).
The discussion is too long because there is a lot of redundancy from the Results section.
Table 3 is unnecessary - the data is presented in Fig 4.
Figures 2 and 5 should be removed to supplemented materials.
Experimental design: The molecular identification of the endophytic fungus was performed using only one set of primers. The authors should validate their identification with at least 3 sets of primers. Therefore, the authors should sequence at least two more genes in order to confirm their results.
Validity of the findings: No comment
Additional comments: General comments:
Since the volatiles that were examined in this study are organic molecules, please change to VOCs (volatile organic compounds) instead of volatile compounds throughout the manuscript.
In the third paragraph of the introduction, the authors give examples of bioactive endophytic fungi. Since this study is about the endophytic fungus Daldinia, the authors should elaborate on this fungus and especially on its bioactivity described in the literature. See articles by Marc Stadler for example.
Molecular identification of the endophyte BP11 should be done with at least 3 different sets of primers. Therefore, the authors should complete the identification with 2 more sets of primers.
It is known that Daldinia produces volatiles with a pronounced, sweet and fruity odor [Stadler M, Læssøe T, Fournier J, Decock C, Schmieschek B, Tichy HV, et al. A polyphasic taxonomy of Daldinia (Xylariaceae). Stud Mycol. 2014;77: 1–143. pmid:24790283]. Did the author noticed a sweet smell from isolate BP11, and if so, please indicate it in the results, since this smell is also a feature of Daldinia.
From the screening of the bioactive endophytes, did the authors identified the second-best bioactive endophyte (BP24)?
The concentration units used in the manuscript are not clear. What do you mean by “µL/mL headspace”? Please clarify and change throughout the manuscript.
Did you check the viability of the pathogen in the in vitro experiment with the VOCs? This data is missing.
Did the authors examine the bioactivity of isolate BP11 against other phytopathogenic fungi (in in vitro assays)? In my opinion, a screen of the activity of this isolate against various phytopathogenic fungi in vitro may strengthen the importance of this isolate. For example, volatiles from D. cf. concentrica fully inhibited the growth of 12 phytopathogenic fungi (Use of the Endophytic Fungus Daldinia cf. concentrica and Its Volatiles as Bio-Control Agents
Liarzi O, Bar E, Lewinsohn E, Ezra D (2016) Use of the Endophytic Fungus Daldinia cf. concentrica and Its Volatiles as Bio-Control Agents. PLOS ONE 11(12): e0168242. https://doi.org/10.1371/journal.pone.0168242).
Did the authors try to prepare mixtures comprising different ratios of the most abundant VOCs? Perhaps a synthetic mixture might be more potent to protect the strawberry fruits from C. acutatum. If so, it will be easier to use the synthetic compound or mixture instead of the organism itself because it will not be necessary to maintain life-supporting conditions.
Can the author compare the concentration of elemicin used in the study and the concentration of this compound emitted by the BP11 isolate?
The discussion section is much too long and the parts describing the results of this study should be shortened.

Specific comments:
Line 42 - write "their volatile" insead of "the". Delete "by these endophtyic fungi"
Line 43 - delete "that usually causes" and write "the causal agent of"
Line 45 - write "a" instead of "the"
Line 46 - "in vitro" should be in Italic. Please correct throughout the manuscript.
Line 48 – write 80.3% - one number after the decimal point is enough. Please correct throughout the manuscript.
Line 48 - "in vivo" should be in Italic. Please correct throughout the manuscript.
Line 61 – the molecular identification of the BP11 endophyte should be described after the description that this endophyte possesses the highest activity – line 48.
Line 61 – instead of “based on the results of PCR reaction assay” write “molecular identification of BP11 endophyte revealed…”
Line 70 – change degrees to Celsius, since all the other temperatures in the manuscript are in Celsius.
Line 93 – delete “which are readily accessible”
Line 102 – please add reference to reinforce your statement.
Line 109 – I disagree with the statement that biofumigation does not require a sophisticated system. This is because you need maintain life-supporting conditions for the bioagent. The author should rephrase this sentence.
Line 119 – the reference for (Sahu, 1984) is missing.
Line 144 – didn’t the authors added antibiotic such as tetracycline or chloramphenicol to prevent bacterial growth during the isolation procedure?
Line 147 – delete “to prevent microbial contamination”.
Line 164 – it is not clear which primers did you use for the molecular identification of C. acutatum? Did you sequence the whole genome? Please clarify.
Line 188 – change to “viability” test
Lines 196 & 303 – delete the sentence. It belongs to the Results section and not to the Materials and Methods section.
Line 210 – “a 7-days old”
Line 210 – the endophyte culture plate in this test was without its cover. Please clarify.
Lines 224 & 299 – this is the same formula as in line 184. Therefore, please refer to the formula in line 184 and delete the formula in lines 224 & 299.
Line 231 – “as follows” and not “as followed”.
Line 238 – change “exposing to” instead of “treating with”.
Line 254 – why 30oC and not 27oC as the temperature used in the experiments?
Line 292 – please clarify how you calculated the estimated concentrations. You can calculate the moles of each compound used in the experiment (see for example Liarzi O, Bucki P, Braun Miyara S, Ezra D (2016) Bioactive Volatiles from an Endophytic Daldinia cf. concentrica Isolate Affect the Viability of the Plant Parasitic Nematode Meloidogyne javanica. PLOS ONE 11(12): e0168437.)
Line 305 – fashion? Perhaps manner is better word.
Line 315 – replace “above” instead of “previously”.
Line 348 – the reference to Fig. S1 should be moved to line 350, because the figure demonstrates only the bioactive isolates.
Line 350 – delete the word “but”.
Line 358 – this section should be combined with the previous paragraph and not as a separate section. Also, this section is written in a passive mode instead of active mode. Please rephrase.
Line 366 – please elaborate on each treatment. For example, the difference between the control and negative control treatments.
Line 368 – add (Fig. 3 C versus D) after “isolate BP11”
Lines 370-372 – delete, it is a repetition of the data presented in Table 1.
Lines 384-386 – write “The in vitro antifungal activity of the six most abundant VOCs against C. acutatum are summarized in Table 3.
Line 402 – change to Fig. 3E.
Line 405 – add “and similar to those obtained with the endophyte BP11”.
Line 413 – please clarify exactly what is each control.
Line 416 – please add “these results suggest that Both BP11 and elemicin are not phytotoxic to strawberry fruits”.
Lines 429-431 – please rephrase your hypothesis. This plant may serve as a candidate for isolation of endophytes that might be active against phytopathogens. I disagree that the bioactivity is only against C. acutatum.
Lines 432-444 – please refer to the suitable figures and tables in the text.
Lines 445-455 - compare the VOCs emitted by Daldinia isolate BP11 to published VOCs of Daldinia. For example: (Liarzi O, Bar E, Lewinsohn E, Ezra D (2016) Use of the Endophytic Fungus Daldinia cf. concentrica and Its Volatiles as Bio-Control Agents. PLOS ONE 11(12): e0168242. https://doi.org/10.1371/journal.pone.0168242) (Pažoutová S, Follert S, Bitzer J, Keck M, Surup F, Šrůtka P. et al. A new endophytic insect-associated Daldinia species, recognised from a comparison of secondary metabolite profiles and molecular phylogeny. Fung Divers. 2013;60: 107–123.)
Line 474 – the reference to Guo et al 2018 is missing.
Line 476 – why there is loss in the concentration of the synthetic VOCs? Possible reasons might be degradation of oxidation.
Line 515 – it is not clear if the farmers should use the endophyte itself or elemicin. Please clarify. The author should add disclaimers about the use of the fungus, for example, the need maintain life-supporting conditions.

Figs 2 & 5 – should be presented as supplemented materials.
Fig. 3 – write isolate BP11 instead of Daldinia sp. MFLUCC19-0493, since the results of the molecular identification is described only later in the manuscript. Also, the figure legend is not clear. Please elaborate and clarify.
Fig. 4 – the results demonstrated in the figure are redundant to Table 3. I would suggest deleting Table 3 or presenting it as a supplemented material.


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: ANTIFUNGAL PROPERTIES OF VOLATILE ORGANIC COMPOUNDS PRODUCED BY DALDINIA ESCHSCHOLTZII MFLUCC 19-0493 ISOLATED FROM BARLERIA PRIONITIS LEAVES AGAINST COLLETOTRICHUM ACUTATUM AND ITS POST-HARVEST INFECTIONS ON STRAWBERRY FRUITS
Review round: 2
Reviewer: 1

Output:
Basic reporting: There is still a problem with the reference of Guo et al 2018. In the reference list, there is no such reference. Perhaps, the authors mean 'Gao et al 2018' and not 'Guo et al 2018', which was added to the revised manuscript.
Experimental design: The authors molecularly identified the phytopathogen C. acutatum using ITS primers. However, there is no indication in the text if indeed the pathogen was confirmed as C. acutatum (not % of identity, coverage, etc.).
Also, despite my previous suggestion to verify the molecular identification of the endophyte with two other sets of primes, the authors deposited only the ITS sequence of the endophyte in the GenBank database. The authors identified the endophyte using a phylogenetic tree. I would recommend verifying this identification by sequencing two more sets of primers and to deposit the resulting sequences in the GenBank, as the authors did for the ITS primers. This would strengthen the validity of the endophyte of interest.
Moreover, please add the accession numbers of the sequences used for the construction of the phylogenetic tree.
Validity of the findings: No comment
Additional comments: Line 114 - the organism 'Streptomyces globisporus' is a bacterium and not a fungus and therefore may not serve as an example for endophytic fungi.
Line 228 - replace 'in' to 'and'.
Line 359 - delete the '.' after (Fig. S1).
Line 361 - there is a space missing between '3.3' and 'and'.
Lines 373-374 - include in the parentheses the corresponding figures.
Line 468 - check the reference. Apparently, it should be 'Gao' and not 'Guo'.
Table 3 and Figure 2 - elaborate on the treatments: emphasis the differences between the controls.

I would also recommend deleting the first paragraph of the Discussion in order to shorten this section.


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

Input:
Title: ANTIFUNGAL PROPERTIES OF VOLATILE ORGANIC COMPOUNDS PRODUCED BY DALDINIA ESCHSCHOLTZII MFLUCC 19-0493 ISOLATED FROM BARLERIA PRIONITIS LEAVES AGAINST COLLETOTRICHUM ACUTATUM AND ITS POST-HARVEST INFECTIONS ON STRAWBERRY FRUITS
Review round: 3
Reviewer: 1

Output:
Basic reporting: This is my second review of the manuscript. The authors fulfilled all of my comments and changed the manuscript accordingly.
However, there are some minor comments:
Line 172 - instead of "A gene region ITS" change to "The internal transcribed spacer (ITS)".
Line 172 - please add references to the ITS4 and ITS5 primer sequences.
Line 175 - replace "a" instead of "the".
Line 175 - write "performed" instead of "confirmed"
Line 176 - after the word "strawberries" please add "to confirm pathogen identification".
Line 329 - add "s" to the word "belong"
Line 427 - add "s" to the word "number"
Line 457 - there is a missing space between "19-0493" and "is"
Experimental design: The authors sequenced more genes for the molecular identification of the endophyte and also confirmed the pathogen using a pathogenicity test. In addition, the authors submitted the genetic sequences to the GenBank and listed their accession numbers. However, one of the accession numbers - MW495050 was not found in the Nucleotide database (due to today, 1st of February, 2021), whereas the two other accession numbers were valid. The authors should check why this accession number is missing and solve this issue.
Validity of the findings: No comment
Additional comments: Dear author
Thank you for revising your manuscript thoroughly. Yet, there is a missing accession number - the sequence for the RPB2 is not found in the Nucleotide database. The two other accession numbers - for the ITS and LSU sequences are valid and can be found in the Nucleotide database. Please check this issue. In addition, a few very minor changes in the manuscript are listed above. In my opinion, the manuscript is much better now and should be published once all accession number problem will be resolved.


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: RECOMBINANT FUSION PROTEIN BY LYSOZYME AND ANTIBACTERIAL PEPTIDE ENHANCES ISCHEMIC WOUND HEALING VIA ANGIOGENESIS AND REDUCTION OF INFLAMMATION IN DIABETIC DB/DB MICE
Review round: 1
Reviewer: 1

Output:
Basic reporting: All is OK except some images and figures which are not convincing
Experimental design: The high technical standard is not reached in some experiments like western blots
Validity of the findings: see my general comments to the author
Additional comments: The article “Recombinant fusion protein by lysozyme and antibacterial peptide enhances ischemic wound healing via angiogenesis and reduction of inflammation in diabetic db/db mice » is a well written paper in a good english with the aim to answer the question if the fusion protein « lysozyme and antibacterial peptide » could enchance the wound healing in diabetic mices via angiogenesis and reduction of inflammation. I have two main critics :
1-	1- The experiments were carried out with appropriate techniques and the results are convincing except for a few images and western blots like those in fig. 4, 6B and 7B; yet the statistical analyzes seem convincing.
2-	2- The second remark concerns the use only of diabetic mice in this work whereas the authors should have compared diabetic mice to non-diabetics, even if they would have been forced to reduce their investigations either on angiogenesis, or on reducing inflammation.

Other minor remarks:
Line 66- 67	preventing infection from pathogenic bacteria and viruses within wounds and fractures in trauma patients (Callewaert&Michiels 2010).>> This reference didn’t speak about Virus!

Line 112 	NONISCH+PROT and ISCH+PROT mice were injected with fusion protein (0.1 mL, 50 μg/mL) >> You didn’t show or verify if the fusion is still active?; What about the purification of this enzyme and why did you use only one concentration ?
Line 123 	Wound samples were homogenized in cold PBS s and centrifuged at 10000rpm for 20min at 4°C.>>please remove the “s”
Lines 152-153 	2 mmol/L phenyl methylsulfonyl fluoride and protease inhibitors (10 μg/L leupeptin and aprotinin).>>  the phenyl methylsulfonyl fluoride (PMSF) is also a protease inhibitor!

Line 168 	infiltrationt and increase of the number >>please remove the “t” at the end of infiltration
Line 188 The fusion protein also elevated the levels of PDGF and VEGF in serum (P<0.05) (Figs. 3A and 3B)>> in the legend, it is said p<0.01
Line 198 	enhanced expression of PDGF and CD34 in ischemic adductor muscle compared to the ISCH group (Figs. 4C and 4D) >> in the legend, it is said the inverse: (C) Representative images of adductor muscle sections stained for CD34 (x400). (D) Representative images of adductor muscle sections stained for PDGF (x400)


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: RECOMBINANT FUSION PROTEIN BY LYSOZYME AND ANTIBACTERIAL PEPTIDE ENHANCES ISCHEMIC WOUND HEALING VIA ANGIOGENESIS AND REDUCTION OF INFLAMMATION IN DIABETIC DB/DB MICE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Manuscript Number: 53150v1
Manuscript title: Recombinant fusion protein by lysozyme and antibacterial peptide enhances ischemic wound healing via angiogenesis and reduction of inflammation in diabetic db/db mice

This manuscript written by Wei Li et.al, to demonstrated the effect of recombinant fusion protein on wound healing in diabetic conditions. They have conducted a number of experiments and collected much valuable data for such a paper. The paper was written very well and the results are interpreted well in the aspect of diabetic wound healing. This manuscript is suitable for publication in this journal after minor revision of the manuscript based on the following queries.

1. The manuscript has issues with grammatical mistakes and should be corrected before publication.
2. Add the detailed methodology of fusion construction in materials and methods section. Whether you construct this fusion protein used in this manuscript? Or you obtained this fusion protein from any commercial manufactures?, please add this details.
3. again add some details about how many days you treated the animal with fusion protein, please exactly mention treatment period in materials and methods section.
4. How the authors picked the concentrations of fusion proteins in their experiments – Justify.
5. add the wound morphology image of control and all treated animal groups with wound area closure calculations.
Experimental design: 1. Add the detailed methodology of fusion construction in materials and methods section. Whether you construct this fusion protein used in this manuscript? Or you obtained this fusion protein from any commercial manufactures?, please add this details.

2. again add some details about how many days you treated the animal with fusion protein, please exactly mention treatment period in materials and methods section.
Validity of the findings: No comment
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: RECOMBINANT FUSION PROTEIN BY LYSOZYME AND ANTIBACTERIAL PEPTIDE ENHANCES ISCHEMIC WOUND HEALING VIA ANGIOGENESIS AND REDUCTION OF INFLAMMATION IN DIABETIC DB/DB MICE
Review round: 3
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I found that the authors have answered all my questions and remarks; I thank them for their efforts


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: OPTIMIZING SPELEOLOGICAL MONITORING EFFORTS: INSIGHTS FROM LONG-TERM DATA FOR TROPICAL IRON CAVES
Review round: 1
Reviewer: 1

Output:
Basic reporting: I found this to be a very well written paper, and I didn't have my usually minor comments. With one exception, discussed below, the literature review is sufficient.
Iron ore caves are very interesting both from a biological and conservation point of view, and the authors do a good job conveying this.
Experimental design: I am not an expert on statistical modelling, but the authors persuaded me that they used appropriate tests and procedures.
Validity of the findings: The relatively low explanatory power (line 250) is disappointing, but I thought that perhaps one of the reasons is that much of the habitat is not caves per se but rather than MSS in the canga itself.  Most iron ore caves, at least in the Iron Triangle where I have worked, are very small, and many don't even have a dark zone.  Perhaps sampling in caves is not sampling the bulk of the species or populations. I am not saying that this appeal to the MSS invalidates the study but somehwere it is surely  worth a mention.
Additional comments: I found that there was a fresh approach to monitoring and i think it is a significant contribution to the literature.


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: OPTIMIZING SPELEOLOGICAL MONITORING EFFORTS: INSIGHTS FROM LONG-TERM DATA FOR TROPICAL IRON CAVES
Review round: 1
Reviewer: 2

Output:
Basic reporting: I did not complete the review of this paper. The paper seemed rather underdeveloped. There were also organizational issues, as well as several key references were overlooked. As such, I recommend a major revision. Once these issues are addressed, I'd be more than happy to review the paper again.
Experimental design: Major revision requested. Experimental design is not clearly written. They vacillate between stating they used existing data and collected field data. I wasn't sure which.
Validity of the findings: This section was surprisingly underdeveloped. They need to present their results using the same structure provided for their methods. In other words, they use apply the same subsections in the results there were presented in the methods. And thus fully develop their results using that approach.
Additional comments: Please find general comments as sticky notes on the PDF.


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

Input:
Title: OPTIMIZING SPELEOLOGICAL MONITORING EFFORTS: INSIGHTS FROM LONG-TERM DATA FOR TROPICAL IRON CAVES
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No Comment
Additional comments: My comments were addressed.


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: RELATIONSHIP BETWEEN EMOTIONAL INTELLIGENCE AND EMPATHY TOWARDS HUMANS AND ANIMALS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: The authors provide meaningful replication of previous findings on the relationship between human empathy and animal empathy and emotional intelligence and empathy. That said, I do have a concern that the authors do not control for current or childhood pet ownership within their analyses. Paul (2000) mentions that animal empathy was significantly different between owners and non-owners, which could fundamentally change the findings if it is controlled for in the current analyses. Without that control, the paper may contribute inaccurate findings to the literature. I should note that I'm not concerned with the current lack of findings between EI and animal empathy (and if they still lack significance when controlling for ownership, that would also be noteworthy), but that the findings could be wrong. At minimum there should be a discussion of this point in the limitation section, as it is a primary finding of the seminal work on animal empathy and a paper that is cited.
Additional comments: Overall an interesting paper but I would strongly encourage reporting if past/current ownership was assessed. If so, test it. If not, be sure to mention it as a limitation of the study to provide a clear path for future research.


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: RELATIONSHIP BETWEEN EMOTIONAL INTELLIGENCE AND EMPATHY TOWARDS HUMANS AND ANIMALS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Thank you for the opportunity to review this interesting paper. It is well-written and the Introduction provides a very sound review of the literature on EI and empathy. The presentation of the article, including its tables, is professional; and the raw data and ethics statements have been shared. There is a major problem with the paper, however, that makes it difficult to evaluate in regard to its contribution to the scientific literature. Specifically, I needed much more information about the construct of 'empathy for animals' before I could assess the logic of the hypotheses. What does the construct mean, and how has it previously been measured? Is it about compassion for animals and if so, all of them or just a few species? Is it about cognitive empathy, involving respect for animals and a moral imperative not to cause them unnecessary harm? Is it about feeling a close, empathic relationship with animals that means one tends to treat them more like humans? These are just speculations, because no information about the scale used to measure the construct was provided in the Method section, making it impossible to understand the nature of the associations reported in the Results. My suggestion for improvement (because I think the topic is interesting) is for the authors to re-conceptualize their study with a very clear focus on the construct of empathy for animals - make this the central feature of the study and describe in detail how the construct is measured. This will make the interpretation and discussion of the Results much more meaningful.
Experimental design: No further comment
Validity of the findings: No further comment
Additional comments: I commend the authors on putting together such a well-written, professionally presented paper and encourage them to submit a new paper with a strong conceptual and methodological focus on empathy for animals and its implications.


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: RELATIONSHIP BETWEEN EMOTIONAL INTELLIGENCE AND EMPATHY TOWARDS HUMANS AND ANIMALS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I appreciate the author(s) efforts to strengthen their work in response to my comments. That said, I think displaying the findings by ownership may be beneficial for readers to better understand those findings, esp. since that seems to be a major finding (i.e., that direct interaction is needed to empathize with animals).
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: RELATIONSHIP BETWEEN EMOTIONAL INTELLIGENCE AND EMPATHY TOWARDS HUMANS AND ANIMALS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Thank you for the opportunity to review this revised version of the paper, and please accept my apologies for lateness. Overall, I believe the authors have done a very good job in revising their paper in line with previous comments and recommendations. The coverage of the relevant literature is both wider and deeper, and the rationale for the study is more compelling than previously. In particular, the authors have clarified their construct of 'empathy for animals' and I appreciated the addition of scale items to demonstrate how the construct was measured. The quality of English used throughout the paper is good, with just a few words that are a little out of place: e.g., line 154, "objects" should probably be "entities"; and pg. 188, 'stimulated' should probably be 'simulated'. Line 194 also needs attention: specifically, an individual observing an animal in a state of suffering MAY (not will) experience distress.." This may not apply to a hunter, for example, who may experience happiness at shooting an animal. Finally, the first paragraph of the Results also needs some attention - it's always good to see effect sizes reported but this section is a bit hard to follow.
Experimental design: The Methodology is fine and. as noted above, it was good to see some example items from the Animal Empathy Scale. The sample size is a particular strength. My only remaining caveat concerns the rationale, expressed just before the Methods section, for operationalizing '"degree of proximity that the person has with animals" in terms of pet ownership only. My thoughts throughout the paper go to farmers and others who have working relationships - sometimes very close - with animals that are not pets (including race horses; experimental animals in laboratories such as mice or even chimps), and that may be hurt or even slaughtered at some point.  Many farmers claim to feel enormous empathy with their stock and do everything in their power to give them "happy" lives; others regard them as units of production and have no emotions towards them at all. I think the "pet" relationship is too narrow and doesn't tap into the wide range of potential relationships that people may have with animals. I realize that this can't be remedied in the current paper, but I believe it is a limitation worth mentioning in the Discussion, especially in the context of future research.
Validity of the findings: One point: in response to the editor's query about why the author(s) did not use a (more economical) regression analysis, they claim that they didn't do this because they were not predicting causality between the constructs analysed. However, regression is a correlational model and does not imply causality - the use of the word "predictor" in regression models often trips people up! I still think it would be worthwhile - and cleaner - to take a regression approach.
Additional comments: Overall, a great topic that I hope you will pursue further.


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

Input:
Title: A QUANTITATIVE ASSESSMENT OF SITE-LEVEL FACTORS IN INFLUENCING CHUKAR (ALECTORIS CHUKAR) INTRODUCTION OUTCOMES
Review round: 3
Reviewer: 1

Output:
Basic reporting: From an Invasion Biology perspective it is interesting that this paper promotes the use of modelling for release of a globally recognised invasive species. I think the manuscript would benefit from a brief species description and invasive status, particularly in North America where this species is promoted. Additionally including the use of a model to predict potential invasive range could broaden the scope of this MS and its impact. In a global context (of interest), introductions in South Africa have also failed, yet a population persists on Robben Island just off the coast. Regardless I think a brief framing of the context of this species (even just for North America) would improve the MS.
Experimental design: Authors have addressed concerns raised by previous reviewers.
Validity of the findings: Findings are clear and well presented. Some clarity on the ebird map is needed. Is this reflecting the authors refined data set or the whole ebird data base? It seems some countries are not represented where known presence of this species is.
Additional comments: Some text in the methods section can be moved to the discussion and there are a few typing errors which can be corrected (in particular spacing between brackets and commas in the methods section lines 216-222). See also lines 86, 89, and 133 for typing errors.


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: MORPHOLOGICAL GROUNDS FOR THE OBLIGATE AERIAL RESPIRATION OF AN AQUATIC SNAIL: FUNCTIONAL AND EVOLUTIONARY PERSPECTIVES
Review round: 1
Reviewer: 1

Output:
Basic reporting: I am very pleased about the present contribution. Descriptions, preservation of tissues and quality of photos are all of high quality.

Accordingly, all following comments and recommendations are made in the sense to make an already very good paper excellent.
Experimental design: The experimental design, light and electron (SEM, TEM) microscopy, appears adequate to the questions to be studied.
Validity of the findings: The paper continues, widens and summarizes the previous studies by the authors on the same species, most of them being published in PeerJ.
Additional comments: General:
Term „vessel“: I am aware that (unfortunately) this term is regularly used for all molluscan blood tubes. However, a true „vessel“ ist clearly defined by the presence of an endothelium, a condition which  - as outlined (line 340ff) - simply is not the case in ampullariids (contrary to e.g., cephalopods or vertebrates). In particular concerning gas transport the difference is crucial, because in case of true vessels an additional cell-layer (the endothelium) needs to be overcome. Accordingly, I recommend to use the widely used term „sinus“ instead of „vessel“ to express the difference (see also Ponder et al. 2019: Biology and Evolution of the Mollusca. Vol. 1: pp. 295-296, CRC Press).

Details:
Line 55: Replace Helicidae by Stylommatophora. You may mention that in case of terrestrial slugs epidermal gas exchange is a major part of respiration.
Line 66: You should add here the detailed anatomical and histological descriptions by Starmühlner (1969: Malacologia 8: 1-434; available via BHL) on Pila cecillei (p.119ff) from Madagascar.
Fig. 1: You may mention that the sections are shown in frontal view. Add bm to the legend.
Fig. 6: Text is somewhat confusing and may cause misunderstandings (see above about „vessels“): Blood chambers are in principle intercellular spaces (micro-sinuses) between the epithelial cells of the lung, the latter do not form an endothelium, however.


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: MORPHOLOGICAL GROUNDS FOR THE OBLIGATE AERIAL RESPIRATION OF AN AQUATIC SNAIL: FUNCTIONAL AND EVOLUTIONARY PERSPECTIVES
Review round: 1
Reviewer: 2

Output:
Basic reporting: English: Recommend active voice
Literature: Appears comprehensive - could better address what elements contribute int he paper
Figures - professional
Hypotheses - none really present - could be better structured around questions

See additional comments below
Experimental design: Research aims could be more clear
Could better identify how this work expands past work and how it fills in an important knowledge gap
Methods seem comprehension but not in a position to evaluate

See below for more comments
Validity of the findings: Novelty could be more clear.
Data images archived.
Conclusions well done - rest of paper could be more succinct.
Additional comments: Morphological grounds for the obligate aerial respiration of an aquatic snail: Functional and evolutionary perspectives
Summary: This paper describes the structure of the lung for Pomacea canaliculata, a species of concern given its ability to tolerate a wide range of environments.  The anatomical descriptions appear thorough and the figures professionally rendered.
The MS would benefit from:
1.	A better description of how this work advances what has previously been done in the field
2.	A broader context for why/how a detailed description of this anatomy and functional morphology adds to our understanding of the ecology and evolution of apple snails.
3.	A set of clear objectives or questions that the investigation answered
a.	Could complement the “Conclusions” found at the end
4.	A synthesis of what this work found in terms of expected, new or different from past studies (i.e. dividing into the main morphological features)
5.	Partitioned use of references to refer to specific information or simply cite them chronologically
6.	Comparisons of lungs between Pomacea canaliculata and Pomacea maculata – not sure how one can describe as “most developed” without more comparative work. Pomacea maculata lays much larger clutches out of water and may spend more time air breathing than P. canaliculata.
7.	Grammatically – chose active voice and avoid use of semicolons and colons. Simplify sentence structure.
Note to Editor: While I do work on apple snails, my experience and strengths do not lie in physiology, morphology or histology.
Abstract:
•	Would start with a bigger picture than the focus on a single organism such as the importance of adaptations for amphibiousness
•	Little clarity – In this study, we offer supporting evidence (?) versus complimentary?
•	Some examples where the language could be improved include increased use of active voice:
o	Example: A pavement epithelium lines the lung cavity … instead of “is lined by”

Introduction:
•	Could combine sentences – Higher oxygen concentrations and diffusion rates of air, coupled with its lower viscosity, make oxygen update by an animal easier in air than water. Not surprisingly, air-breathing has…
•	58 – “with later development of a lung”
•	66 – “only a few studies examine”
•	69 – how does this work advance the work of Rodriguez
•	77 – this piece of information seems out of place
Methods – outside area of expertise – cannot comment on appropriateness
•	Some examples where the language could be improved include increased use of active voice:
o	Methods: preferred use of “we”  - We obtained samples and stained them….
•	83 – why males? And how many?
Results –
•	145 – 147 – can the authors support this difference in muscle layer thickness quantitatively?
•	Figure 2 includes 3 ventral views and only one dorsal view. It would be better if balanced or show an informative diagram for C/D (same view) that appears between A/B (mirror image).
•	I would make a table of the anatomical parts and their function
•	223-226 – does not warrant its own paragraph
•	227 – 229 – sounds more like a discussion item
•	It seems Figure 10 would be better placed earlier in the manuscript as an overview and then have boxes or arrows that point to the more specific SEM photos
•	Figure 11 should be drawn to scale to increase its usefulness

Discussion –
•	292 – usually cite references in chronological order
•	297 – “their lungs” – what is the reference here? Apple snail lungs appear homologous to the pallial cavity of other gastropods?  “In these gastropods” – which ones? Hard to discern the meaning behind these sentences.
•	303 – 308 – reads as Results (possibly redundant)
•	311 – 313 – what’s the mechanism for this vessel only appearing in some Pomacea species? What do those species have in common? Do they belong to the same clade? Could expand Discussion of their lower dependence on water with more examples.
•	323 – comma after “through it,” but it also…
•	335 – form and not forms
•	336 – “Our results agree” – worth being specific on which results
•	346 – 349 – Good information but complex sentences makes it a little hard to decipher
•	354 – do the authors mean “phospholipidic”
•	380 – I would disagree that the knowledge of phylogenetic relationships in Ampullariidae only depends on molecular characteristics.  I
•	382 – This section on comparative information provides some interesting insights and should be developed further and more clearly connected to the results of this study.
•	394 – end the sentence instead of using a colon. Follow with “For instance,”
•	The Discussion overall seems partially redundant and overly extensive
•	The subtitles inform the reader as to the subject but not its importance
•	498 – cool that you found something new – the rhogocytes should be given more emphasis – “These cells had not been reported before in the Ampullariidae.”


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: MORPHOLOGICAL GROUNDS FOR THE OBLIGATE AERIAL RESPIRATION OF AN AQUATIC SNAIL: FUNCTIONAL AND EVOLUTIONARY PERSPECTIVES
Review round: 1
Reviewer: 3

Output:
Basic reporting: Rodriguez et al. present a follow up study of the respiratory system of an ampullariid species. Ampullariid snails are easily accessible as they are abundant invading pests and aquarium pets. For this reason their very well investigated also regarding anatomical details such as the respiratory system.
This study provides an extensive description of details of anatomical and histological details surrounding the lung. These results appear valuable as they contain details (at the TEM-level - type of small lacunar system at the base of the epithelium) that may have been overlooked or confused in previous literature.
However, the theoretical treatment of the results (Discussion) is extremely poor. The topics (e.g. comparison with vertebrates, function of rhogozytes, how is the tissue tightened together?) are speculative and/or meaningless. It seems there is a shortage of ideas accompanying the findings, which may due to oversplitting their results (compare Rodriguez et. al, 2019).
Experimental design: The morphological examination has been well done with adequate methodology.
Validity of the findings: Mediocre.
Additional comments: In summary I CLEARLY vote against the publication of this MS UNLESS the theoretical part is FULLY new prepared using different approaches.

For this I recommend to remain within the systematic vicinity of ampullariids and related gastropods (functionally related - pulmonates). A review of previous data of amullariids would be helpful/desirable (tables on data/results). Detailed reasons why previous/other results should be dismissed would be interesting. How does a pulmonate (basommatophoran, sylommatophoran) lung surface compare in detail? Diagrammatic comparative figures could be beneficial. Functional comparisons to other lung breathing gastropods (e.g. lung always air filled or sometimes water filled?) in general could be exploited.

Please find attached the PDF MS with specific comments of mine.


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: MORPHOLOGICAL GROUNDS FOR THE OBLIGATE AERIAL RESPIRATION OF AN AQUATIC SNAIL: FUNCTIONAL AND EVOLUTIONARY PERSPECTIVES
Review round: 2
Reviewer: 1

Output:
Basic reporting: This is a valuable contribution to our understanding of the structure and function of the ampullariid lung serving as a model of terrestralization in gastropods. It continues a high-quality series of papers on the overall subject.

There are only certain minor points which should be considered (see below): .
Experimental design: I am pleased about the various methodologies applied and about the high quality of the LM, SEM and TEM images.
Validity of the findings: The paper provides further progress in our understanding of the model system Pomacea.
Additional comments: This is a valuable contribution to our understanding of the structure and function of the ampullariid lung serving as a model of terrestralization in gastropods. It continues your high-quality series of papers on the overall subject.

I am pleased about the various methodologies applied and about the high quality of the LM, SEM and TEM images.

There are only certain minor points which should be considered: .

Line 55: no – the other way around: originally gastropods (molluscs) had a mantle cavity, and the inner mantle epithelium has been always used for respiration too, the gill (better: ctenidia) are basically used for water ventilation rather than for respiration, in particular in small animals. In case of air-breathing the gills (ctenidia) are no longer necessary and are usually reduced or lost.

Line 278: Better to cite the new textbook of Ponder & Lindberg 2019: Biology and Evolution of the Mollusca. Vol. 1. CRC Press, Taylor & Francis Group, Boca Raton FL, 864 pp

Line 392: hematopoietic

Wells RGM (1980): add „The Institute of Biology´s Studies in Biology 127, ISBN 10: 9780713128062, 71 pp"

Fig. 7ff: the labeled “hemocytes” (hcy) may be specified as amoebocytes with their typical pseudopodia.

Table 1: Haszprunar (1996) instead of “(1995)”


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: MORPHOLOGICAL GROUNDS FOR THE OBLIGATE AERIAL RESPIRATION OF AN AQUATIC SNAIL: FUNCTIONAL AND EVOLUTIONARY PERSPECTIVES
Review round: 2
Reviewer: 2

Output:
Basic reporting: The MS is sufficiently improved. The discussion appears to be completely reworked, it is clear and interesting and does not contain unfounded consideration anymore in this version.
It is generally well written now.
Experimental design: Well done.
Validity of the findings: Interesting in the field of invertebrate biology.
Additional comments: I placed a few comments and suggestions in the PDF MS.


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: MEIOGYNE OLIGOCARPA (ANNONACEAE), A NEW SPECIES FROM YUNNAN, CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article is well written and the English is of good scientific quality. The literature and references are good. I would however add an introduction to the Annonaceae family with some references as the start of the introduction.
The results are relevant and well presented. the hypothesis of a new species is very well supported by different types of data (morphological, anatomical, and molecular to a certain extent).
Experimental design: The article is well designed with a solid experimental design. The authors use different methods and approaches to reach their conclusions.
The methods are well described.
I would have liked to have some more details on where the rest of the genetic data comes from (the markers of the rest of the species, and outgroups).
Validity of the findings: Overall this MS provides sufficient data to support the description of the new species. All data point towards this conclusion. The phylogenetic methods are well explained and robust and sufficient for the conclusions.
The conclusions and discussion are well explained. the key to the two species of Meiogyne is very useful.
Additional comments: This article presents the description of a new species of the genus Meiogyne (Annonaceae). I am not a specialist of the genus or the region, but the data provided are convincing.
This species was mainly described for a few collections growing in the Xishuangbanna Tropical Botanical Garden, which is quite interesting, and unusual for Annonaceae. The authors provide substantial evidence for the status of new species, from morphology, to anatomy and molecular phylogenetics.
I have no major comments, and only minor ones. The methods are appropriate and well done. The English is very good, the figures really nice. The short key to the two species of Meiogyne in China is parallel and precise. Overall a very good and complete manuscript. Congratulations to the authors.
Minor:
Line 35: maybe a short intro to the Annonaceae family?
Line 117: where did the rest of the sampling come from? Could the authors cite a publication? Idem for the outgroups?
Line 209: when and where was this species transferred? Is this transfer official? Please provide some details.
Line 300: “of” secondary veins
Line 305: “inflorescences”, with an s.
308: flowers per se cannot be leathery, do you mean the petals?


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: MEIOGYNE OLIGOCARPA (ANNONACEAE), A NEW SPECIES FROM YUNNAN, CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: 'no comment'
Experimental design: 'no comment'
Validity of the findings: 'no comment'
Additional comments: The manuscript expands the number of Chinese species of Meiogyne from one to two. It's a straightforward thing to do, but the authors have been conscientious in their approach: the phylogenetic placement of the new species within the genus Meiogyne is convincingly demonstrated, the species is accurately described and nicely illustrated.

However, there are some things that I would like to see changed or added before this manuscript would be suitable for publication:

1)	Introduction: the introduction should be improved and it is necessary to present hypotheses about the phylogenetic position of the new species, about the anatomy of its petals and about its biogeographic origin.

a)  one of the authors' objectives is to infer the function of the longitudinally grooved or verrucose base surface at the inner petals, however, they do not mention anything relevant in the text, beyond it is a characteristic present in all species of the genus Meiogyne. They cited some references but did not mention anything about it. These modifications in the petals are also present in other genera of Miliuseae. Specifically in the genus Sapranthus, a lineage relatively close to Meiogyne (An ancestral character?). Schatz in his doctoral thesis (1987) suggests that those "topographical" modifications in the petals are associated with sites where flower visitors bite (eat? = reward?).
b)	The second species described provides the necessary material to hypothesize about the origin and evolution of the genus Meiogyne in China. For example, it could be expected that both species are sisters and that they originated in the same geographical region where their evolution is related to the same factors; or that according to their phylogenetic position the invasion of China by the ancestral lineage happened during a certain period of time and from a certain geographical point. I'd like to see something similar (or other hypotheses) postulated in the introduction.
The results of this manuscript suggest that they are independent invasions and in different periods of time, something very interesting, right?
A previous study (where the first author participated) could be very useful to address the above: see Thomas et al. 2012 where they analyzed the biogeographic history of the genus Meiogyne but including fewer species than those presented here.

2) Materials and methods
a)	It is necessary to include dating analysis and reconstruction of ancestral areas (related to the previous comment).
b)	Phylogenetic trees (MP, ML, Bayesian) must be presented separately, even if not in the main text.
c) It would be very interesting for the authors to construct a dendrogram of morphological similarity with Meiogyne species (although this is not mandatory from my point of view). If the topology of the dendrogram is similar to that of the phylogenetic tree, this data could be used to include it in the phylogenetic reconstruction and assess if it improves the resolution. What do you think about it?

Results & Discussion
a) Results and discussion must be separated. There are some notes after the taxonomic treatment section (eg "Meiogyne species in China") that are part of the discussion and therefore the paragraphs should be rearranged.
b) The comparison of the new species with its relatives (this could also be derived from the dendrogram) should be in a table for easier tracking. Meiogyne hainanensis should be included in this table.
c)	There are three accessions of M. cylindrocarpa with a different phylogenetic position, why not present them as new 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: MEIOGYNE OLIGOCARPA (ANNONACEAE), A NEW SPECIES FROM YUNNAN, CHINA
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is a standard manuscript reporting a new species, with detailed description and phylogenetic corroboration. The new species status is warranted by the integrative evidence provided. The anatomical finding is valuable for the understanding of petal evolution in Annonaceae.

Nevertheless, there is a room for the improvement of the manuscript. My comments are embedded in the PDF file of the manuscript, most of them minors. I would like to see a further discussion on the corrugated structures. In the PDF file I have explained this in details.