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: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript “The knock-down of Death inducer-obliterator1 gene (DIDO1) would promote apoptosis and inhibit proliferation of leukemia induced Endothelial cells by inhibiting the expression of CDK6 and CCND1” by Cao et al. describes the effect of inhibition of DIDO1 on the proliferation of Leukemia induces endothelial cells. GeneChip analysis was performed to find out gene upregulation and downregulation. The effect of DIDO1 gene on cell proliferation and apoptosis has been analyzed. The study is interesting and can be useful in increasing survival of leukemia patients.

The manuscript needs significant improvement. It has to be improved for both English language and grammar as well as organization. It should be thorough and easily understandable. Even the results in the Abstract section loses its flow and does not provide the complete picture of the manuscript. The abbreviations should be introduced when they are used first time.
Experimental design: Is there any in vivo data showing that the knockdown of DIDO1 gene would either treat leukemia or increase survival? In vivo data will add a great value to the study.

Did the authors analyze different domains in DIDO1? For example, if the effect is same when NLS deleted DIDO1 is used. This will elaborate the mechanism of action of DIDO1.
Validity of the findings: Figures 1C and S2 are not clear. It seems that GFP is higher in shDIDO cells. It should mean that the cells proliferation is higher in these cells as compared to control. If it is so, it will contradict the entire results.

Did authors use an activator of CDK6 or CCND1 gene to check if there is any counter effect on DIDO1 action? It will confirm the mechanism of action of DIDO1.

All the GeneChip data can be seen in supplementary. Authors have written that 30 genes were analyzed by qPCR for the expression level determination. That data should be included.

Authors have discussed the inhibitory effect of DIDO1 on ERK/MAPK signaling pathway, however except the GeneChip analysis there is no other evidence provided. It will be great if authors can provide more qPCR/protein expression/phosphorylation data in control and shDIDO cells. Also, more can be discussed about this pathway how it can be beneficial in leukemia patients.

In schematic figure 4, it is not clear how or at which step DIDO1 is affecting ERK/MAPK signaling pathway. It shows general pathway, how does it relate to the current study?
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: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript needs substantial English language editing.

The background information for the molecule of interest, Dido1, is not sufficiently described.

Figures legends need to be more descriptive and axes need better definition
Ex: Fig 1G, Percentage can be revised to Percentage cell death

Raw data are shared.
Experimental design: The experimental system leading to the most critical data with Genechip is not described. It is not clear how the co-cultured cells were prepared for GeneChip analysis.
Validity of the findings: The importance of the molecule of interest Dido1 in the context of leaukemia-activated endothelial cells is unclear. The authors are suggested to discuss the potential role of Dido in endothelial cells.
Additional comments: In the manuscript by Cao et.al. the authors compare gene expression profiles of endothelial cells and leukemia-activated endothelial cells. They identified Dido gene expression to be downregulated in leukemia-educated HUVEC cells. They independently confirmed this using stable silencing of Dido1 in HUVEC cells and found that it resulted in cell death. They used publicly available data to identify that Dido may regulate cell cycle genes CCND1 and CDK6.
Listed below are the major concerns regarding this manuscript in its current form:
1.	The authors have not detailed the method for co-culture of HUVEC cells with K562. It is recommended to give a detailed description of the procedure used prior to gene expression profiling by Genechip.
Were the HUVEC cells separated from K562 for the GeneChip experiment?
2.	The authors do not describe clearly the cell type in which the genes were upregulated or down regulated.
3.	The authors do not sufficiently introduce the background information for Dido necessary to convince the audience of the importance of this protein in leukemia or endothelial biology. Previous results indicate that Dido is required for cancer cell proliferation (Neoplasma Vol.67, No.5, p.1074–1084, 2020) The authors are suggested to explain the relevance of Dido in the endothelial cells. The implications of this finding for cancer biology should be discussed.
Further, Dido1 has been described as a tumor suppressor gene. How does this current data align with previous literature on Dido1?
4.	With the knockdown of Dido 1 with shRNA, do the other splice variants (Dido2, Dido3) get affected?
5.	It is recommended that the authors revise the manuscript to check for
- typographical errors: such as Genichip (Ln 41),
- expand acronyms: such as MVD (Ln 82)
- usage of biological jargons: such as leukemic induced ECs.(Ln 40)
- sentences are fragmented and do not flow well. For instance: “In this study, we aimed to identify the genes function in the formation of leukemic induced ECs.”(Ln 39-40)
This current phrasing of sentences makes it hard to follow. It is suggested that the authors use professional English language editing services to improve the manuscript.


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

Input:
Title: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revised submission of the manuscript entitled “The knock-down of Death inducer-obliterator gene would promote apoptosis and inhibit proliferation of leukemia induced Endothelial cells by inhibiting the expression of CDK6 and CCND1”, concerning the effect of DIDO1 gene knockdown on leukemia induced Endothelial cells. Authors have answered all the comments/concerns point-wise and improved the manuscript accordingly.

English language must be improved further. At some places DIDO1 has mentioned, while at others DIDO. It should be consistent throughout the manuscript including figures.
Experimental design: no comments
Validity of the findings: Authors have adjusted contrast in figure 2C. However, contrast and other parameters should remain identical in control vs the experimental pictures. Is it possible to take new pictures if the authors have saved the slides?
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: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 2
Reviewer: 2

Output:
Basic reporting: The sentence construction still needs to be improved. The statements and ambiguious and sometime conflicting.

Additional reference are need to clarify basic concepts, please see additional comments.

The replacement of DIDO1 to DIDO has to be consistent in all of the Figures and legends. This change was made only to the text, as suggested in the review report.
Experimental design: Methods are not clearly described. Please see additional comments
Validity of the findings: No comment.
Additional comments: The authors have submitted an updated version of the manuscript addressing some of the points raised in the review report. The origin of cell lines has been described in the manuscript.
Listed below are some of the concerns that still remain:

1.	The authors use various terminologies regarding leukemia activated endothelial cells which make for a difficult read. At times the statements are even conflicting.
The authors use “leukemia induced endothelial cells” which they need to clarify. Do they refer to leukemia induced endothelial cell activation or differentiation?
In the title the authors mention “DIDO knockdown inhibits proliferation of leukemia induced endothelial cells”.
In the abstract the mention “formation of endothelial cells to leukemia” (Ln40)
Later in the text, the authors state “mechanism of how leukemia cell induces ECs formation”

The literature recognizes the cross talk between endothelial cells and leukemic cells and details two separate phenomenon:
1.	Leukemia mediated endothelial cell activation : Ref https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3613371/
2.	Endothelial to hematopoietic transition
https://pubmed.ncbi.nlm.nih.gov/27015586/

The authors need to clarify what they refer to in the above listed statements with suitable references.

3.	The authors described the cell used for Genechip, as requested. But it still lacks key details. The authors reported that the GeneChip analysis was performed with HUVEC cells vs HUVEC-K562 co-cultured cells. Did the authors wash away the K562 suspension cells before harvesting cells from the co-culture or use the entire co-culture set up for gene expression analysis ? Please specify.

4.	Similarly, for the proliferation assay reported in Figure S1, the authors would need to clarify the method used to ensure that only HUVEC cells were counted by Celigo and/or analyzed by MTT assay without the interference of K562 cells.

5.	The shRNA targeting DIDO gene may target all three isoforms so the authors agreed to change this to DIDO rather than DIDO1, but the data and legend in all of the Figures still reports DIDO1. Please correct this throughout the manuscript.

6.	In the final results section, the authors explore potential transcriptional regulators of the DIDO gene. However, the outstanding question from the manuscript is the mechanism by which Leukemic cell interaction may suppress the expression of DIDO in HUVEC cells. What do the authors speculate regarding this reported finding? It is suggested to include this in the discussion section.

Minor:
1.	Check abbreviations :
CEC should be changed to EC in Ln 70,72
LAML should be changed to AML in Ln 213, 215
2.	Check the statement “Human umbilical vein endothelial cells (HUVEC) are capable to differentiate into endothelial cells in vitro.” (Ln 181-182)


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: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 3
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No comments
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: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 3
Reviewer: 2

Output:
Basic reporting: No comment. Please see additional comments
Experimental design: No comment. Please see additional comments
Validity of the findings: No comment. Please see additional comments
Additional comments: In the revised manuscript, “In leukemia, knock-down of the death inducer-obliterator gene would inhibit the proliferation of endothelial cells by inhibiting the expression of CDK6 and CCND1”, the authors have addressed some of the concerns raised in the previous review report. While some changes in the rebuttal were satisfactory some other concerns remain. They acknowledged that some terminology in the manuscript was misleading to the readers and needed to be clarified. It appears they have corrected some of these instances however, the revised, track changed manuscript still contains significant instances of inaccuracies.
Listed below are concerns that still remain:
1.	The authors were asked to clarify the phrase “ Leukemia-induced endothelial cells”. They have addressed this in the rebuttal, however this statement still remains in the revised manuscript in the conclusion section of the abstract Ln 56-57 “The knock-down of DIDO will inhibit the formation of leukemia-induced endothelial cells”.
Another instance in Ln 92-93 In this study, we investigated the role of DIDO in the formation of leukemia-induced ECs.”

2.	There are instances of abbreviations that are not expanded OS for overall survival in the abstract Ln 50.

3.	Among the changes that the authors have made, they added this statement to the discussion section “In this study, we found that there are H3K4me3 and Pol II (POLR2A) signals near Exon1 and exon2, suggesting that there are isoforms with Exon1 and exon2 as transcription start sites.”
This statement is factually incorrect about the DIDO gene alternate transcripts generated by splicing. It is clearly defined in the literature that DIDO2 and DIDO3 isoforms generated with the inclusion of exons 6-8 and 15-16 respectively near the 3’ end of the gene, rather than exon 1 and 2 as transcription start sites like the authors suggest. (Ref: https://www.jci.org/articles/view/24177 , Dido gene expression alterations are implicated in the induction of hematological myeloid neoplasms, by Futterer et al in 2005)


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: IN LEUKEMIA, KNOCK-DOWN OF THE DEATH INDUCER-OBLITERATOR GENE WOULD INHIBIT THE PROLIFERATION OF ENDOTHELIAL CELLS BY INHIBITING THE EXPRESSION OF CDK6 AND CCND1
Review round: 4
Reviewer: 1

Output:
Basic reporting: no comment
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: B-BOX CONTAINING PROTEIN 1 FROM MALUS DOMESTICA (MDBBX1) IS INVOLVED IN THE ABIOTIC STRESS RESPONSE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The presented research describes the functional annotation of a BBX gene isolated from apple. Several experiments are performed to determine the gene-of-interest’s native expression in different tissue of apple seedlings, the protections it provided to transformed E. coli cells and the role it plays in transgenic plants overexpressing the GOI when exposed to abiotic stress. The presented data fall within the broad scope of general plant and molecular biology as required by PeerJ. The presented research conveys novel data regarding the function of an apple BBX TF gene.

The grammar used in the article is not good and the authors need to address all the linguistic aspects of the paper before resubmitting. I have indicated numerous mistakes and editorial issues on the attached pdf file.  In general, the authors should proofread and make the necessary corrections across the whole document. Also, the authors should decide on one tense and stick to that.

The abstract reflects the content of the paper but the grammar in some instances need to be improved by the authors. The introduction section covers relevant and recent information on the presented topic.

The Material and Method section is adequately described but numerous mistakes need to be corrected as indicated in the attached pdf. Give complete information regarding kits used.

Sections of the Discussion are repeating the Results too extensively and should be shorten.
Experimental design: The results are presented in a systematic and relevant manner. The statistical analysis is sound. However, a few issues the authors need to address:
The authors should remove the Discussion sections from the Results, or change it all together to a Results and Discussion section.
Fig 1A, the authors are working with one year old apple seedlings (as per fig legend), does such small plants really have fruits to analyse?
Unclear how tobacco plants were transformed for subcellular localization studies, this is not explained in the M&M, and details are missing. In the Results it seems like tobacco epidermal leave cells were transformed.  No controls are included for the subcellular localization results, it is likely the expression occur in the nucleus but without a control this cannot be concluded.
In Table 1, + and – positions for cis-elements are indicted but the authors said that the 1.5 kb upstream region of the gene was analyses, why then + and -?
Fig 2 – it would be better if the authors can standardise the y-axis of the graphs across treatments, at least in the graphs where the values are within the same range.
Why are the line graphs in Fig 3 in colour but not the line graphs in Fig 4?
Fig 4B graph – where is the stats for the Control treatment?
Why is the qPCR done with PEG but the seedlings germination/rooting tested with mannitol (Fig 6a,b) and seedling growth on PEG? Why the authors jumped between osmoticums are unclear.
Legend of Fig 7 do not correspond with the presented figures around ABA treatment.
Why did the authors only test one of the OE MbBBX1 transgenic lines for ABA-related gene expression (Fig 7)?
Figure 8 has a Fig 7 legend about ABA and images linked to ROS, this needs to be fixed. Similar confusion around legends for Fig 9.
Validity of the findings: Overall, experiments seem to be conducted in a scientifically sound manner and presented in a way that address the scientific aim of this paper. However, numerous mistakes and improvements need to be made to the paper before this data set can be considered for publication.
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: B-BOX CONTAINING PROTEIN 1 FROM MALUS DOMESTICA (MDBBX1) IS INVOLVED IN THE ABIOTIC STRESS RESPONSE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript entitled “B-box containing protein 1 from Malus domestica (MdBBX1) involved in abiotic stress response” submitted by Dai et al shows great effort in elucidating MdBBX1 might play an important role in the regulation of tolerance to abiotic stresses. However, the terms and content organization need to be rephrased to highlight importance of their work.
Experimental design: The experimental design is overall reasonable. Please state the evidence for the suitable choice of PEG and NaCl  concentrations.
Validity of the findings: MdBBX1 might play an important role inthe regulation oftolerance to abiotic stresses by reducing the
generation of ROS. The result in this study is informative, and the findings are novel.
Additional comments: 1.	In terms of the main issue existing in this manuscript, the language should be improved.
2.	Please carefully check the entire text and make sure the genes name should be in italic, but not the protein.
3. L24, remains should be remain.
4. L25-26, various abiotic stresses response should be stress responses.
5. L29, longer length should be rephrased.
6.L33, SOD, POD, AND APX should be summarized into antioxidant enzymes, noy peroxidases.
7. L104, what is the meaning of different time?
8.	Line 129, “Su et al., 2013” can be replaced with “Livak and Schmittgen 2001” https://doi.org/10.1006/meth.2001.1262
9.	Line 180-185, recently-published papers about POD, CAT, SOD and APX should be cited in the manuscript.
10.	In the methodology sections, there is no information about the statistical analysis. Please add a separate section of “statistical data analysis”.
11.	In figure 3, 4, and 7 legend, * should be indicate P< 0.05 usually, **, P < 0.01, ***, P < 0.001.
12.	Please check figure 7, 8, 9. The figure does not match the legend.


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: B-BOX CONTAINING PROTEIN 1 FROM MALUS DOMESTICA (MDBBX1) IS INVOLVED IN THE ABIOTIC STRESS RESPONSE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors improved the paper significantly. A few remaining gramma mistakes are indicated in the attached pdf.
Experimental design: A well constructed and executed set of experiments.
Validity of the findings: Valet data set.
Additional comments: Fig 1 – why not just use the Supplementary figure that show the DAPI control in the paper?
Fig 3B – The significant different indicator “b” at the control and Nacl treatment must be a mistake, its highly unlikely that the two samples do not differ significantly
Fig4B – The significant difference indicated (a) is highly unlikely.  Does the transgenic lines under control treatment really not differ from the NaCl treatment?
Why do the graphs in for example Fig 4B look different from the graphs in Fig 5. Use the same formatting across the whole paper
Fig 8B graphs – x-axis “Control” and y-axis “Content”– fix spelling mistakes.
Fix the use of in-text references, that the publication year is indicated in italics and sometimes not.


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: B-BOX CONTAINING PROTEIN 1 FROM MALUS DOMESTICA (MDBBX1) IS INVOLVED IN THE ABIOTIC STRESS RESPONSE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript can be accepted for publication.
Experimental design: I have no concerns about the experimental design.
Validity of the findings: The findings presented in this study is suitable for publication in PeerJ.
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: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: The hypothesis and addressed research gap should be clarified more precisely. Please find my comments for the Introduction section under “4. Additional comments”.

I identified some issues with the meta-analytical approach (e.g. imprecise reporting of data set building, not state of the art statistics, lack of addressing non-independence issue among effect sizes and lack of application of sensitivity analysis). Please find my comments for the methods section under “4. Additional comments”.
I have no expertise on structural equation modeling.
Validity of the findings: The data table comprising the effect size values was provided but the information about the included articles is missing. Please provide the reference list and indicate which article is associated to which data row.

The discussion has substance but is weak at some points. Please find my comments for the Discussion section under “4. Additional comments”. The conclusion section is just one sentence.
Additional comments: Title:

It is not clear from the title that this is a meta-analysis. Please add this term. You could also add it to the keywords. By this, you article can be easier found by a WoS search.

Introduction:

Line 67: I wonder how important AMF really are in agriculture. I guess they have their role in organic/ low input agriculture, but under conventional agricultural practices with high fertilizer input and tillage AMF play very likely only a minor part in improving crop production.

Line 75: What do you mean by “all kind of biotic and abiotic” constraints? This statement is very unspecific. It sounds like AMF can solve all plant related issues. That is not true. In the second half of this sentence you gave specific examples of what the AMF have proven to be capable off. This is a more precise and clear way to praise their beneficial symbiotic functions.

Line 86: This is again unspecific. Please make clear what exactly you mean and want to focus at in this work. AMF have many functions. You probably want to focus on functions related to crop productivity and nutritional quality?
What do you mean by "effect size of AMF"? The effect size represents the magnitude of a treatment effect compared to an associated control effect. Please rephrase and clarify.

Line 88: So there is no meta-analysis on AMF-mediated effects on crop production/growth etc. for agricultural systems under rainfed irrigation only? If this is true, then just state it here clearly. This is your identified research gap which you want to fill with your work. So make it clear so that the readers understand what is your new knowledge gained from this work you contribute to the science community.

Line 91: You do not mention in the methods that you also collected data on non-rainfed systems. How are these data incorporated in the following analyses? Were they just used in a sub-analysis or were they included in all analysis? Please clarify this very important point.

Methods:

Line 95. Excellent. Reporting guidelines are vital.

Line 96: Did you consider alternative terms to identify articles about this topic? Is there really only the term "rain*fed" used to describe water management without artificial irrigation? Especially in the context of applied science which affects humans (here in the form of food production), extra care has to be taken to make the meta-analysis as comprehensive as possible. What was your search term building strategy? Please have a look into this article which demonstrates with worked examples how to develop search strings following best practices. (Foo et al. 2021 https://doi.org/10.1111/2041-210X.13654)

Also, state more clearly what settings you chose for your search. You give search date and publication year range, but you do not say if you run a topic or title search, which database in WoS you used (WoS is the platform, not the database. CoreCollection is a database in WoS. So which did you use? Please specify.), if you preferred/ excluded any languages or citation indexes. Subscriptions with WoS vary widely across different research institutes and hence the available publication years, databases and citation indexes, could you please add your institution/university name in this section. Then it is transparent with which subscription setting you did your search.

Line 97: This is already a very small number of articles to begin with which makes the following meta-analysis quite challenging. Did you consider searching other possible sources for articles e.g. other databases or the reference lists of identified matching articles or even gray literature? Each article is important to increase statistical power and to correctly estimate AMF effects.
Also, please state how many articles were in the end matching your inclusion criteria. I know you give this information the PRISMA flow diagram, but also give this information here.

Line 98: I highly appreciate your effort to dual screen and cross-check during the screening process. This is done very rarely but a very important routine. Excellent.

Line 105: Your hypothesis focus only on biomass and yield, then why did you also collect data on proline and chlorophyll? And what do you mean by “nutrition”? Please clarify.

Line 116: You explain which effect size metric you chose and explain how to interpret it. But you do not mention the effect size variance at all. How did you calculate it? How did you deal with missing variance data? How did you deal with missing information on replication in experiments? (Which are needed to calculate the effect size variance.) By the way, how did you extract data from e.g. figures and graphs? Please add this important information to this section.

Line 119: Please add more information about the data you used for your effect sizes: e.g. biomass (What biomass data did you included, shoot, root, total?). Give concise information to allow readers not familiar with harvest index, proline and Co. to understand why these parameters are interesting and what the data you included in your data set looked like. How did you decide that different aspects of a metric can be combined (a hypothetical example: how to decide to include root and shoot biomass data in the biomass effect size but exclude total biomass data)?

Also, add information about your moderators (explanatory variable): e.g. N- and non-N-fixing crops. You mention these in your results section but never introduced them in the methods. The same for grain crops and non-grain crops in Fig. 2. Give concise rationales why these moderators or subsets are meaningful and interesting for your project and what is included in each group (e.g. which crop species are included in the grain and non-grain crops groups).

Line 121: How did you weighted the effect size, with the effect size variance data, I guess? Please add a statement.

Line 124: I recommend using rma.mv() because this allows inclusion of the data hierarchy and addressing the non-independence issue (see below). This is a mixed-effects meta-analysis model. In the random factor you can include the information of articles comprising specific effect sizes means (rows in your data table). Please, find this article on the topic by Song et al. 2020 https://doi.org/10.1002/ecy.3184. They explain very well the possible options to address this problem of non-independency of effect sizes and why it is so important to take care off it.

Ok, following your statement in line 103 “491 pairs of observations” where extracted from 17 articles. So, each article gave you multiple data pairs. This is absolutely normal in ecological MAs but you need to account for the resulting non-independence issue in your dataset or else your data analyses are flawed.

The non-independence issue. From your methods it is not clear to me if you have corrected for non-independence issues in your dataset. There are two typical sources: 1) multiple data from one study (which is definitely the case here) and 2) multiple studies from one author/lab (which is quite likely the case here). As mentioned above you can apply rma.mv()-models to account for hierarchical structure in your dataset, but it is still useful to “reduce” your dataset as far as possible. With this I mean, only have the absolute minimum of effect sizes (rows in the data table) per study. E.g. When potentially matching data for multiple harvests are presented you can decide to only use data from the last harvest. (You just have to report your decisions and give a rationale.) When data are given for different AM fungal species (actually I am surprised that you did not mention AMF genera/species as moderator variable) you could “merge” these data by applying a “within-article meta-analysis model”. You could even apply a phylogenetic model (Anderson 2015 https://doi.org/10.1111/nph.13693). Please check out this paper explaining this important issue (Song et al 2020 https://doi.org/10.1002/ecy.3184).
Concerning the dependency of articles published by the same author/lab, I recommend to you the paper by Moulin & Amaral (2020; doi: 10.1002/jrsm.1430). Correcting for this source of “bias” will improve the robustness of your model outcomes.
These methods to target the non-independence issue are extremely important especially for meta-analysis on small numbers of articles. You artificially increase your statistical power by including all ~ 500 single effect size values without properly merging them as far as possible and/or accounting for data structure via a mixed-effect MA model (rma.mv).

Line 124: Please, add the package name "metafor" and its citation.

I apologize if I missed it, but you seem not to apply any sensitivity analyses to test for biases or the robustness in general of your data. A very popular tool is the publication bias. With this you can evaluate how likely it is that you have missed a publication (either it was never published or you really physically missed it) and weather you over-or underestimate potentially the true effect with your data collection. Another option are the leave-one-out-approaches. Here, you test the individual impact of data points or whole studies, by excluding them from the analysis, re-run the analysis, include the data points or whole studies again, delete the next one, and so on and so on. By this you can identify data points or studies with disproportional impact on your results. There are many other techniques, but at least one should find its way into your manuscript to further improve its trustworthiness and robustness.
When using rma.mv() many diagnostic function from the metafor package are not applicable anymore. However, there is the Eggers Regression Test for publication bias test which can be applied as a simple sensitivity analysis.

Results:

Line 135: The increased crop yield for non-grain crops should be treated with caution. As you state some lines later, this effect bases on a limited number articles and trials. So, you should carefully phrase your conclusions based on data with such low statistical power.

Line 136: You plotted every single effect size. You already calculated them or else you could not run any analysis.

Line 152: I get confused here. Did you collect only data for wheat and chickpea? No. But it reads like it. You really need to make clear in the methods what plant species you included in your data base. And you should better add also a section in the methods where you explain the analyses you are doing in this project. In the results section the reader should have an idea what to expect but here new subset groups/moderators and subset analyses keep popping up. This makes it hard to follow you.
So you chose wheat and chickpea in this subset analysis because they are the most abundant plant species in your dataset? Please give a rational for your decision. The same applies later to the line 190.

You only focus on the AMF-mediated effect on crop growth/productivity/nutrition but neglect the performance of the AMF completely. E.g. do you have data on AM fungal root colonization? This could be used as a potential proxy of the intensity of the plant-host symbiosis and could explain some patterns you find. More colonization higher AM fungal mediated productivity effects? Crop species specific differences in percent AM fungal root colonization?

Discussion:

Line 246: The global scale part is not convincing when presenting data comprised from 17 articles. Better delete that term.

Line 251: “Glomalin” or glomalin-related soil proteins“ are a rather critical group of substances. As discussed in this article (Rosier CL, Hoye AT, Rillig MC. 2006. Glomalin-related soil protein: assessment of current detection and quantification tools. Soil Biology & Biochemistry 38: 2205-2211.) detection methods for glomalin are not as specific as had previously been assumed. Such I do not see that this group of compounds should be discussed here.

Line 253: Could you extract data on hyphal length. By this you could test this in your very own data set and check if it really can explain the beneficial impact of AMF in the rainfed systems.

Line 255: This comparison (whose effect size is bigger) is a bit odd. Your data base is way more limited than that of this Zhang meta-analysis, hence pointing on magnitude effect size percentages is not valid. In general, you need to be careful with overemphasizing your findings. Your meta-analysis is interesting and important but it also bases on “only” 17 articles and hence give us just a first glimpse on an area of application for AMF in rainfed agricultural systems but it does not give us percent values which are set in stone.

Line 256: Sorry, this argument is also odd. Yes, you include more crops but by this you also increase heterogeneity in your data set. It is difficult to compare the performance of AMF symbiosis with members of the Poaceae and Fabaceae. Poaceae crops can have very vast and fine root systems reducing the importance of the AM fungal extraradical hyphal network for nutrient uptake, while Fabaceae crops can form a tri-partite symbiosis with AMF and Rhizobia giving them the option under nutrient limited conditions to overcome P and N limitations. It is not an advantage or disadvantage of a synthesis to a have narrower or wider focus. So for me your argument is invalid.

Line 290: Yes, this information would be nice to have in the methods section when you explain why you include which plant parameters as effect sizes in your analyses.

Figures

The figures look a bit blurry. This is a resolution issue, I guess. Please upload your nice figures with a higher resolution. I like your clear figure layout and color scheme. Very aesthetic.
Please add also the number of articles contributing to each group.
Add the effect size to the y-axis. In each graph it is written “Effect size” but which one. This should be indicated clearly and should not require reading the figure legend. Please add something like e.g. response ratio of biomass or biomass effect size… Be creative.

Fig. 3: Add the 95% confidence intervals to your effect size means. Or how did you decide which effect size is negative, neutral or positive? Now I’m a bit confused. I do this by checking if the 95% CI is overlapping with the zero line (line of no effect). This applies to all the effect size range plots.
Fig. 6: Why did you abandon here your color scheme and colored everything in red. Stay consistent.

Could you combine some figures? E.g Fig. 3 and Fig. 2 would fit nicely together showing the distribution of negative to positive single effect sizes and overall model outcomes.

Figure legends.

Why did you present SE and not the 95% confidence intervals which are given by the metafor model outcomes?

Data table.

Please provide information on the articles you used to build your data table. Also indicate which article is associated to which data row.


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: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Abstract
- Underscore the scientific value-added to your paper in your abstract. Your abstract should clearly state the essence of the problem you are addressing, what you did and what you found and recommend. That will help a prospective reader of the abstract to decide if they wish to read the entire article.
- Lines 27-28: This sentence is not clear.
- Line 31: The sentence ‘Shoot biomass had a direct positive effect …’ should be deleted.
Introduction
- Line 43: Wikipedia is not a reliable source for reference in the article.
- Please add more information about climate change and increasing world temperature and drought area.
- the linkage between introduction paragraphs is missed.
- Lines 46-47: The sentence should be deleted.
- Lines 53-54: Please add another reasons for decreasing nutrient uptake under drought stress conditions; for example, low mobility, decreasing micro-organisms activity and etc.
- Please subject the manuscript to review made by English Native speaker.
- What is the innovation of this study? Please add in text. Many studies previously reported the effectiveness of AMF on the plant performance and productivity especially in drought stress conditions.
Materials and methods
- Lines 95-96: The authors used the paper that included two factors of AMF and Rain-fed or used simple mentioned factors effects?
- Line 130: Please add the method of standardized path coefficients calculating.
- The analysis method is not complete.
- The papers were used in this study was belonged to only one plant?
Results
- Please check again the increasing or decreasing percentage rate.
- Line 227: What is the SEM?
- Why the effect of the AMF in different climates (such as different between arid and semi-arid region) has not been studied.
Discussion
- In this section, the authors firstly reported the obtained results and after that noted the similar results of previous studies. The main reasons for increasing and decreasing of measured traits is missed. Actually, I don’t see any discussion in this section. This section should be deeply revised.
- What is the novelty of this study? The previously studies professionally reported the positive effects of AMF inoculation in plant performance. The novelty of this study is missed.
Experimental design: All comments are reported in the first section.
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: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript deals with broadly researched and quite a recurrent topic, but it is done in a transparent and easily understandable fashion. Quality of writing and presentation is generally high (with just a few syntax/vocabulary items to be improved). However, I was not able to locate a list of the specific studies included in the meta-analysis (including literature references), which should definitely be done - for example by including references to each ana every row in the raw data file (Excel table in the supplement).
Experimental design: This contribution is a meta-analysis aiming at providing a global picture on AM fungal inoculation effects in rainfed agriculture. Whereas this is a worthwhile and valuable and broadly interesting question/objective, this has previously been approached many times and there also are other reviews/meta-analyses available. I am afraid the approach taken (using "rain*fed" as a keyword) is much too restrictive and the search returns only a minutious fraction of the relevant science that has already been conducted and reported. This is because a large part of the world such as Europe and North America produces lots of food under rainfed agriculture - but does not call it so. My own search using keywords "arbuscul* AND mycorrh* AND field and inoc*" returned 1621 research articles (compared to 55 articles recovered by authors of the current manuscript using their search), and these should be all thoroughly screened to recover more or less the complete story, including such important crops like maize and potatoes, for example, which are currently not at all represented in the manuscript. Besides, meta-analyses like McGonigle (1988) collated results of 78 field inoculation studies and Hijri et al (2016) reported results of 231 field trials just with potatoes should be a great guide, but these are not even cited in the current manuscript.
Validity of the findings: Whereas the results are in line with previous observations and quite well presented, I consider this only a preliminary efforts, too early to be considered for publication in an international journal. This is because a large majority of previously published and relevant research has been missed by too restrictive/not quite relevant keyword search, effectively missing many previous studies from Europe and North America. Europe and North America is also missing from the list of regions presented in lines 41-43, where they should definitely be included (because lots of the research has previously been generated there and they are still large regions of the Earth).
To provide a balanced view, more critical literature on employing AM fungal inoculation (e.g., Ryan et al 2018) should also be included in the discussion, next to the overly positive opinions by Rillig, Thirkell and others. It should also be clearly stated that every soil (or nearly every soil) has an active indigenous AM fungal communities already present and inoculation effectively brings "invaders" to the soil. The inoculation is thus effectively always representing a competitive interaction between native and incoming genotypes/communities. There is hardly any soil on the planet devoid of AM fungi (unless previously sterilized/disinfected, which is really not a widespread practice as yet).
Specific comments:
line 52: there are many regions in the world (particularly in middle/northern Europe) where water is not limiting yields, and rain provides sufficient (or even excessive) water. So the global validity of the research from Mediterranean climate zones should be critically evaluated and the existence of regions where water is sufficient should be clearly acknowledged.
line 251: currently there is an intensive debate about what a glomalin or GRSP actually is and who produces it (several opinion papers recently published in high IF journals). The writing should be adapted to be less authoritative/definitive in this regards.
line 253: there is really very thin or no experimental evidence that AM fungi would transport water (in spite of widely spread myths). There are some good papers showing that drought tolerance of mycorrhizal plants is often higher than tolerance of nonmycorrhizal plants, though. However, this is probably not relevant to the fields where all mycotrophic plants (inoculated or not) are mycorrhizal, due to indigenous AM fungal communities (see above). Thus the writing should be very cautious/specific at this point.
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: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 1
Reviewer: 4

Output:
Basic reporting: The paper presents a meta-analysis of data collected in the literature in order to demonstrate that symbiotic associations with Arbuscular Mycorrhizal Fungi (AMF) are beneficial to crop productivity with particular emphasis on crop biomass production and grain yield in a variety of crops. In addition, the aim of the study was to show that such a beneficial effect is species-specific, notably if legumes and non-legumes are considered. Additional information is also provided on the physiological impact of the symbiosis on representative physiological traits related to Phosphorus (P) and nitrogen (N) metabolism.
The study is moderately original, since a significant number of studies presenting such kind of analysis have been published over the last decade, including impact on yield and mineral nutrition. The novelty of the present study should be more clearly emphasized with respect the studies cited by the authors and to the published literature that was not cited in the paper.
The introduction starts and is largely focused on rainfed agriculture which is not really the main aim of the study. The introduction should be primarily focussed on the importance of AMF with respect to agricultural practices and crop productivity.
In the Introduction a number of references are a bit old (e.g. 2008-2009); there are many more recent references on rainfed agriculture and on the importance of AMF on crop productivity.
Experimental design: One of the major problems in the presentation of the paper is that, although a database is presented as Supplementary Data, one would like to see in this database references to the papers used for the meta-analysis (1 to 133 e.g. in a different sheet). In the supplementary Data set, the units for the different measured traits must be indicated. In line, with this comment Figure 1 provides very limited information and is not really necessary for the understanding of the paper, just mention it in the text.
One would like also to see in the various figure legends a brief description of the crops used to perform the analysis together with a way to have access to the corresponding data in the dataset.
Validity of the findings: -	Unless I missed something, I do not see clearly the information we gain from the results presented in Figure 2 compared to that of Figure 1. Please explain a bit more in the Results section.
-	Line 209: what do you mean by uptake by seeds? An uptake of nutrient is performed by the roots and is generally expressed at the whole plant level.
-	In the Discussion there are a number of speculations related to water absorption when plants are colonized by AMF. However, nothing is presented on this aspect in the Results section.
-	I am not sure that the interpretation of the lower of Proline in AMF inoculated plants. If they are more resistant to a water stress, higher amounts of Proline could be expected. Please clarify by citing and discussing more extensively the literature in the field.
Additional comments: Minor comments:
-	Line 27, explain what is a crop functional group.
-	Line 28 and throughout the text: the term effect size is not very clear to me and thus not the most appropriate. See also line 36 and in the Supplementary Data set.
-	In the abstract, one would like to have a minimum of information on the crops used to performed the analysis.
-	Line 54: what means N resorption?
-	Line 57: management measures is not clear, please explain.
-	Line 78: It is drought or water stress, not arid stress.
-	Line 88: what is a normal grain production system?
-	Line 116: indicate which type of yield.
-	Line 227: Write Structural equation analysis at the beginning of the sentence.
-	In the legend of figure 2: grain crops and not grin crops.
-	In the Supplementary Data set it is Olive trees and not Olive tress.


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: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: yes
Experimental design: I have some minor questions concerning the methods. Please find my comments under "4. Additional comments"
Validity of the findings: yes
Additional comments: I thank the authors for the thorough revision. I just have a few comments left, please find these below.

I recognized that the English language of your text is a bit rough. As English is also not my native language, I would like to recommend to you to either ask an English native speaker or a language editing services to improve your text.

Introduction:
L 114: Maybe better write that no meta-analysis on that topic is available so far and that you going to fill this knowledge gap.
L 116: Could you maybe add a map with the study locations? In R there are several functions that allow you to plot decimal degree data for longitude and latitude on a world map. Most paper give location/GPS data for their field sites and in the worst case you could use the affiliation location of the first author. Then you could also highlight, for which rain-fed regions in the world no data could be found for your meta-analysis. These “data white spots” are a very important information when it comes to priorities future research (funds).

Methods:
As this point is not clear for me after reading the methods, I would like to ask if you only included field studies. I imagine that rain-fed systems are “outside” in the field but you never made a statement about it. Or did I overlook it.
If it really is only field studies, you should state it because the biological relevance and generalizability of your analysis outcomes would drastically increase. The majority of the crops you comprised in your analysis are cultured in outdoor agricultural systems and not under controlled greenhouse conditions.

L 173: I’m not convinced by the use of the fixed-effects meta-analysis model. I agree that merging the data from the same year into one effect size, is the right step to reduce the non-independence issue. But by using the fixed effects model you assume that the data for these different observations share the same true effect. This is unlikely, because these observations differ in some way from each other and because of that they were presented as individual data. Imagine in year 1 biomass data is given for sandy soil and clayey soil. These two observations have inherent heterogeneity because they differ in texture (and everything else that gets affected by texture). So, you have to account for this variability and this is done by random-effects models even on the level of the same article. This will give you higher values for your effect size variance, but this approach is more robust.

L 175: Please add here where this data table can be found (e.g. supplementary data of this article…).

L 203: Please add the publication bias outcomes to your supplementary data, thus people can see that there is no case of publication bias evident. You show the plot in your response on my comments from the last round.

Results:
L 299: There is a point missing: “12.7% (CI=-011-0.37)” should be “12.7% (CI=-0.11-0.37)”

Discussion:
L363-365: I cannot follow your thoughts here. First you write that AMF effects on yield in (conventional) agriculture is often overstated. I agree on that point. In especially conventional agriculture AMF play almost no role, due to high fertilizer input and soil disturbance (e.g. tillage). These management practices tend to reduce community richness and functionality leaving only “pioneer-type” AMF species in the soils. On the other hand, in an organic system, with well aggregated soil and active soil biota community, AMF can indeed provide valuable service to soil and crops.
So your second sentence about tillage, seems a bit detached from the prior sentence. Maybe you could rephrase these liens to make your statement clearer.

Others:
I like your protocol on the search string development (in your response to reviewer comments). Maybe you want to consider to add it to your supplementaries. People rarely show how they came up with their search terms. This adds a new layer of transparency and reproducibility to your work.


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

Input:
Title: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The article is well written and (formally) well presented. Comments made previously by a number of reviewers have been addressed, though the critical weaknesses of the meta-analysis (search keywords) remain unchanged, resulting in the dataset being thin and not covering the entire range of available literature.
Experimental design: Search keywords did not cover the field sufficiently. Whereas authors only identified a handful of papers quoting "rainfed", my previous ciriticism was that there is lot more literature which actually address rainfed agriculture in the broadest definition of the term, which were not covered because the authors of those original papers did not use the word "rainfed". The authors of the presented meta-analysis now clearly write that (in their definition) rainfed agriculture occupies 80% of worlds cropland (beginning of the abstract) - but these 80% of the worlds croplands have not been representatively covered in the meta-analysis. The focus of the study is thus not clear, although the authors give, in their response, some other arguments such as in their understanding the rainfed agriculture is "naturally drought-prone croplans". Such definition is not given in the article, though, nor it is generally understandable/acceptable. So I dearly miss the specific focus of this meta-analysis, and the delimitation from other rainfed croplands which have not been covered only because they did not write "rainfed" in their texts. This is not sufficiently justified. Hundreds of inoculation trials from non-irrigated agriculture have been published and these ought to be covered shall the ambition of the presented paper be "representative". Besides, the claims presented in the text that synthesis of results from rainfed conditions has not been made so far is incorrect for reasons that I brought up previously and now again.
Validity of the findings: Given the thin dataset which is not corresponding to available knowledge, the validity remains quite limited - although the conclusions are in line with expectations. Unless the few papers which were included have a common denominator AND could clearly be distinguished from the other inoculation trials done under rainfed conditions elsewhere, but not calling it so, I still consider this manuscript a preliminary effort to what its aims declare and what should in fact be done. Just an idea - maybe include all literature on AMF inoculation under field conditions, and filter out those that were done under "irrigated" conditions - most of the remainder will correspond to what we all understand under "rainfed" conditions. I am sure there will be many more papers remaining in the pot.
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: ARBUSCULAR MYCORRHIZAL FUNGI INCREASE CROP YIELDS BY IMPROVING BIOMASS UNDER RAINFED CONDITION: A META-ANALYSIS
Review round: 2
Reviewer: 3

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: I am satisfied with both the answers to my comments and the relevant modifications to the manuscript which has been greatly improved.


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

Input:
Title: A REVISION OF PLECTANOCOTYLE (MONOGENEA, PLECTANOCOTYLIDAE), WITH MOLECULAR BARCODING OF THREE SPECIES AND THE DESCRIPTION OF A NEW SPECIES FROM THE STREAKED GURNARD CHELIDONICHTHYS LASTOVIZA OFF ALGERIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: I believe it is journal policy to include the name of the newly described species in the title. Also, since the article presents much more than a single species description, with a lot of insights in the systematics of Plectanocotyle and the Plectanocotylidae, I would suggest revising the title.

The Introduction session for the moment does not contain a lot of background information (e.g., to have a clearer idea of the previous work on this genus, it would be useful to read already at this point from which host species and which geographical area the two known species have been described). I wonder whether the overview of plectanocotylid species, and the nomenclatural history of Plectanocotyle, would not make more sense as part of the introduction, at least insofar as this is the result of a literature scan rather than of the empirical work contributed in this study. The novel discoveries thanks to the morphological and molecular work in this study can then more easily be put in perspective in subsequent sections of the manuscript.

The text refers a number of times to Adiyodi et al. (2017), but this reference is not in the reference list.

While the English text is generally fine, some formulations strike me as a bit strange, such as (L. 55): “processed for molecules”, (L. 83-84): “using Maximum Likelihood method based on the Tamura 3-parameter”, (L. 99-100) “by DNA barcoding approach”, (L. 100): “present study fish species”, (L. 124): “clamps that measures”, (L. 168): “worthy to mention”

Fig. 1: Can the families be indicated on this figure?

Fig. 2: Since this paper presents an overview of the genus and family, and hence will be very useful to beginners working on Plectanocotyle, I would suggest that the different structures used in the description (posterior and anterior jaw; sclerites a-b-c-d-e; median and lateral hamulus; sclerite types in the male genital apparatus…), or at least those used in the differential diagnosis, be indicated at least once on the figure. This will improve usability to people with no previous experience on this group of monogeneans (like me!).

Table 4: both “Euzetplectanocotyle” and “Euzeplectanocotyle” are used in this manuscript.
Experimental design: L. 42-45: It would be useful to state here that both fish species are gurnards. How were the fish caught and sacrificed?

L. 83-84: Which criterion was used for model selection? What was the proportion of invariable sites?

L. 85-86: Why use a different model in distance calculation and in tree inference? There are certainly reasons thinkable (e.g. comparability with other studies, or with datasets where no model correction was used), but if no such arguments are provided, why not (also) at least provide the distances under the same model as used in tree building?

L. 106: could you explain the discrepancy in length between the amplified fragment for the specimens in the present study and the alignment used for the phylogenetic reconstruction? It is a difference of almost 150 bp, is this because of lack of overlap (other primers) or length differences with published sequences?
Validity of the findings: L. 117-118: It is mentioned that the interspecific distances of over 10% support the species status of the monogeneans under study. Could you comment a bit on this: how does this compare to interspecific divergence over the COI gene in other congeneric (polyopisthocotylean) monogeneans? I know that the amount of existing sequence data for this is not enormous, but I think it is necessary to offer some comparison to have some idea of the cut-offs that the authors use (or propose).

L. 137: The text states 15 as the maximal number of testes, but I think I see 16 of them on Fig. 2?

L. 179-193: This overview of the nomenclatural history of the genus is extremely useful, but I don’t understand everything, could the authors please clarify a bit?
- both 1950 and 1850 are mentioned for Diesing’s work;
- what is the type species of this genus? Is it P. elliptica, but what is the status of this species now? It is only mentioned that the species was never found again and that representatives of this genus are not expected on moronid hosts.
- if I understand it well, P. lorenzii and P. caudata were synonymised with other congeners then? With which ones?

The abstract mentions that the authors “confirm that Plectanocotyloides Euzet & Suriano, 1974 is a junior synonym of Triglicola Mamaev & Parukhin, 1972” but this is not discussed in the text?
Additional comments: I really enjoyed reading this paper. The methodology is sound and thorough, the illustrations are of high quality, and the authors are convincing in arguing why the newly described species indeed represents a distinct species. Importantly, the authors frame the description of this new species in a broader overview of the diversity of the genus and family it belongs to. This overview is both concise and complete, and therefore I think this manuscript will become a handy future reference for scholars working on these marine monogeneans. I commend this approach to a taxonomic study, and this manuscript definitely merits publication. My suggestions only concern the presentation of the paper, where I believe that mainly (1) the methods of the genetic analyses need some more explanation and justification and (2) the taxonomic description and nomenclatural history could use some clarification to be better useable by neophytes.


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 REVISION OF PLECTANOCOTYLE (MONOGENEA, PLECTANOCOTYLIDAE), WITH MOLECULAR BARCODING OF THREE SPECIES AND THE DESCRIPTION OF A NEW SPECIES FROM THE STREAKED GURNARD CHELIDONICHTHYS LASTOVIZA OFF ALGERIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Some literatures cited in the introduction (line 39) are not relevant with the MS. I don’t know if it is a kind of self citation!
Experimental design: Authors applied rigorous methods to ensure traceability of host and parasite with adequate barcoding but for Plectanocotyle species I suggest to use anterior half (and not the posterior half) for molecular analysis and the posterior half mounted as voucher slide for morphological assessment. The most differential characters are clamps and testes which are located in the posterior half of the body.
Also, I strongly recommend to deposit an hologenophore or a paragenophore (sensu Pleijel et al., 2008). This would have made possible to verify the sequences deposited by Jovelin and Justine (2001) as Octoplectanocotyla obscura and synonymised  by the author as Plectanocotyle major.
The reference used for molecular analysis is Ayadi et al. 2017 (and not Adiyodi et al. (2017) (line 62 and line70). I Think that is a typological error
Validity of the findings: The new species Plectanocotyle lastovizae is adequatlilly described. The second species found on the same host Cheilodichthys lastoviza attributed by the authors to Plectanocotyle gurnardi without any comment and discussion. The type host for P. gurnardi is  Cheilodichthys gurnadus, one of the authors in a last papers have sequenced partial 28 S from P. gurnardi of Trigla gurnardus (and unfortunately not COI!) it is strongly suggested to have the COI sequence from the type host for comparison
Additional comments: This is an interesting manuscript in a number of respects. This paper is the first attempt to bring order in the genus Plectanocotyle using morphological and molecular analysis. Until now distinction between species is based essentially on the measurement of the clamps which certainly lead to many confusions in the literature (e.g. Llewellyyn, 1941). The table given for the species of Plectanocotylidae and their host can be particularly useful if all synonyms for parasites and host can be added.

Specific comments
-	The valid name of the streaked gurnard (as given by worms and Fishbase) is Chelidonichthys lastoviza and not Trigloporus lastoviza. This must be checked throughout the text.
-	L 55 : for Plectanocotyle species I suggest to use anterior half (and not the posterior half) for molecular analysis and the posterior half mounted as voucher slide for morphological assessment. The most differential characters are clamps and testes which are located in the posterior half of the body.
-	L62 and 70: the reference is Ayadi et al. (2017) and not Adiyodi et al. (2017)
-	L80: the reference Olson & Littlewood (2002) or Jovelin & Justine (2001) who made molecular phylogeny is more appropriate here. Phylogeny of Boeger & Kritsky is based on anatomical and ultrastructural characters’
-	L 124: the size of the clamps seems to be very variable (90-180). Way only 11 clamps were measured? Is the size of anterior, median and posterior clamps the same? In specimens examined from C. lastoviza in my collection anterior clamps seem to be  smaller .
-	L125 & 126 the sentence “we use the nomenclature of Llewellyn (1956)” should be moved in the paragraph Nomenclature
-	L130 According to Llewellyn (1956) nomenclature used for (e) is the “slender rib-like sclerite in the inner walls of the clamps “and not the posterior most sclerites of the clamps as used by the authors.
-	L137. 16 testes is given in the figure of Holotype (Fig 2 A) and (11-15) in the description. this should be verified!
-	L143. Add figure for egg if possible
-	L153 For comparative material authors should easily examine Paratypes of Plectanocotyle major who is deposited in the MNHN
-	L 157 lastoviza with “e”
-	L160 The type host of P. gurnardi is  Cheilodichthys gurnadus. If they are no type material deposited authors should compare with available bibliography or with specimens of P. gurnardi from Cheilodichthys gurnadus
-	Table 3 : measurement is based on carmine stained specimens or Berlese mounted specimens. Standard deviation is not given for all measurement. Prevalence can be added in table 3 for all the parasites
-	Table 4 : change Euzeplectanocotyle by Euzetplactanocotyle
-	Table 4 : change Peristedionella by Peristedionelia
-	Table 4: Another species Octolabea turchinii Euzet & Trilles, 1960 is palced by Euzet & Trilles among the Plectanocotylidae. Yamaguti (1963) establishes new family Octolabeidae Yamaguti, 1963 but Mamaev & Parakhin (1972) replace this species among the Plectanocotylidae. A discussion is needed here.
-	Line  314 : change ?? by 6


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 REVISION OF PLECTANOCOTYLE (MONOGENEA, PLECTANOCOTYLIDAE), WITH MOLECULAR BARCODING OF THREE SPECIES AND THE DESCRIPTION OF A NEW SPECIES FROM THE STREAKED GURNARD CHELIDONICHTHYS LASTOVIZA OFF ALGERIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: L. 34, end of the “Results” section of the Abstract: I prefer not to use the word “significant” instead of in cases of a formal statistical test.

L. 276-278: this comparison of your results with earlier work on pairwise genetic distance in polyopisthocotylean monogeneans is highly interesting and a strong element in support of your conclusion – but please let the merit of this approach show by mentioning the (approximate?) distances Bouguerche et al. reported intraspecifically and interspecifically.

L. 288-293: at this point it is not clear to the reader what the nomenclatural relationships are between Plectanocotyloides obscurum, Octoplectanocotyla obscura and Triglicola obscura. It is not stated explicitly how the authors assess these names until Table 4 at the end of the text. Please clarify or announce that this will be clarified later on in the paper.

L. 323: minor typo: “issueD”

Figures 1 and 2: please mention in the caption what the scale bar represents.
Experimental design: L. 84-86: why is the voucher slide based on the anterior part? Does this mean the haptor is not necessary for species-level identification with plectanocotylids? It would be good to clarify this, as practices differ strongly between monogenean taxa here. I am wondering because the differential diagnosis, in the Discussion session, contains a lot of information on the haptor, and lines 279-281 seem to suggest that the MCO is not very informative for species-level identification in Plectanocotyle. Which diagnostic feature is hence visible on a voucher slide of the front part of the worm? The testes seem too posterior for that, unless really only the haptor is removed for genetic work?

L. 125: the word “clean” here suggests that something was wrong in the previous alignment matrix. I don’t think that is necessarily the case – incomplete sequences may be a result of low-quality starting material or some hick-up somewhere in the amplification/sequencing process, of course, but can also simply result from e.g. the use of primer sets that amplify regions that only partially overlap. And certainly indels can simply signal meaningful differences between sequences. Did the author consider these indels as problematic or untrustworthy so that they removed these columns in the first matrix? Or did they remove them because of the methodological challenges this poses for downstream analysis? (Though the model-based phylogenetic inference used here could probably handle that in large part anyways.)

L. 165-166: but I thought these indels were removed? Did that not improve the statistical support? (That being said, I also do not find it problematic nor alarming that just over 300 bp of COI sequences do not yield good support values for deeper nodes. This in no way harms answering the questions of this study, I am merely enquiring about how the authors handled indels.)
Validity of the findings: L. 235-243: I appreciate the caution of the authors with respect to characterising specimens when insufficient data is available from the type locality and type host of the species they may belong to. In this case, it is careful, and probably even commendable, to use “Plectanocotyle cf. gurnardi”. However, I do think that doubt should be specified in such cases: e.g. are there morphological differences, of which it is at this point impossible to evaluate whether they represent interspecific or intraspecific variation? Is the original description insufficiently detailed? Or is it because type material is lacking (in which case this should be explicitly stated)? I think the authors should at least make a comparison of what is published about the morphology of P. gurnardi, compare it with their observations they made on the specimens for this study, and assess whether there is reason not to assign them to P. gurnardi. When no potential contradictions arise with the original description, I personally would assign the specimens (supposing they fall within the diagnosis of the described species) to the formally described species. To play the devil’s advocate: why not follow the saying “if it looks like a duck, and quacks like a duck, it’s a duck”? Perhaps that makes me a lumper :-) and I in no way challenge the authors’ decision here; I just would like to see a little bit more justification.

L. 261: on L. 248, the authors mention a type specimen from a representative of Plectanocotyle, P. major, with these spines being 195 µm long. Even though this is only a single specimen, and even though there may be multiple environmental or artefactual reasons for some sort of outlying measurement result, it does make me question whether it is safe to use the length of MCO spines as a highly informative diagnostic trait to distinguish between members of these two genera.
Additional comments: I was already supportive of this MS in its first version, and I was delighted to see that the authors continued this interesting work. Thanks for the opportunity to re-referee this work. I am satisfied with the way the authors incorporated my suggestions, and with the other changes to the MS. However, in view of the study becoming more elaborate now, there are some additional minor things where I think some additional justification or clarification would be needed to render the text palatable for non-Plectanocotyle-specialists. I will detail these below. Actually, I think these are mainly cases where the authors undersell their work or are too cautious; these remarks do not question the soundness of the work. Only under “validity of the findings” I will mention two instances where I would not reach the same conclusion as the authors did based on the current information; I am not necessarily disagreeing so perhaps some sentences in the MS detailing how the authors reached this conclusion would make everything clear.


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

Input:
Title: A REVISION OF PLECTANOCOTYLE (MONOGENEA, PLECTANOCOTYLIDAE), WITH MOLECULAR BARCODING OF THREE SPECIES AND THE DESCRIPTION OF A NEW SPECIES FROM THE STREAKED GURNARD CHELIDONICHTHYS LASTOVIZA OFF ALGERIA
Review round: 2
Reviewer: 2

Output:
Basic reporting: Compared with the last version, the manuscript is  improved but need some revision before publication.
For me, non-native english speakers, English is clear and unambiguous overall, literature is appropriately referenced apart from those in the last paragraph of the introduction (line 64-67) that have absolutely nothing to do with the subject at hand!
Figures and tables are relevant to the content of the article, appropriately described but legend need to be improved:
Figure 1: The  phylogenetic tree  presented is not  of all the members of the Plectanocotyle but based on some sequences from this study and those of Jovelin & Justine (2001). for this figure the legend should be: ML phylogenetic tree constructed based on COI sequences of Plactanocotylidae from this study and the others from Jovelin & Justine (2001).
In the text (line 157) authors Comment NJ tree but in figure they give ML Tree?
Figure 2: It would be more appropriate to say: Molecular phylogenetic analysis of selected sequences of the Plectanocotylidae.
il the end of line 6 of the legend...P. major (90 ML; 88 NJ) and not (80 ML; 88 NJ)
Figure 3: legend: for the male copulatory organ, authors use interchangeably "spine" and "sclerite". I think it's more appropriate to use sclerite. For D change lapped by terminal lapped
for P. cf. gurnardi, It would be advisable to give unfolded claps as for P. major and P. lastovizae
Table 1: the table include 12 new sequences and not 8. I suggest to add sequences for fish with not retrieved monogeneans in the table.
I think there's been some confusion in table 1 the ascension number MG761759 in given for for the same host Br 29 and Monogenea Br 23 MO1!!! please check
Table 2: genetic distances between COI distances of Plectanocotylidae and not of Monogeneans
Table 3: Measurements of Plectocotyle spp. instead of species of Plectanocotyle
In this Table, I suggest to add measurement of Triglicola from Algeria since the authors studied them
Line 33 add congeneric after other
Experimental design: As I mentioned in my last review, posterior half is not appropriate to be in voucher slide for Plectonocotyle spp. because measurements overlap for what remains of the anatomy making traceability difficult
Validity of the findings: line 165-166. Can authors give some references to support this ascertainment
line 187: between Plectanocotyle spp. interspecific distance is ranging between 7.8 and 11.8 % and not 13.8% ( please correct this also in the abstract)
Line 197: the sentence we use the nomenclature of... will be transferred in the M & M
Line 207: in table 3 authors give length and wide for the pharynx but in the description they give diameter!
Line 210: Testes 13 and not 12 as given in table 3
in the taxonomic summary, I suggest to add: synonym:  Plectanocotyle sp. of Jovelin and Justine (2001)
line 243: can authors give some others arguments why they suspect that P. cf. gurnardi could be a distinct species?
Line 256 sclerite instead of spine
line 261: Plectanocotyle species: 90-162 instead of 125
line 266: size of the lateral hamuli instead of sclerites
line 281: 162 instead of 125
line 301: Plectanocotyle gurnardi is also recorded from Chelidonichthys  cuculus by Llewellyn (1941)
line 327: Lewellyn instead of Llewelyn
le list of
Additional comments: General remarks: please give, in the text, the accepted host name  followed in parentheses by the original name used by the authors.


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: IDENTIFICATION OF KEY GENES AND BIOLOGICAL PATHWAYS IN CHINESE LUNG CANCER POPULATION USING BIOINFORMATICS ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article is trying to identify hub genes in Chinese lung cancer.
Experimental design: The overall design of this study is quite general.
Validity of the findings: The report of the methods needs some improvement. The validity of the results is reliable.
Additional comments: This article is trying to identify hub genes in Chinese lung cancer. The overall design of this study is quite general. The report of the methods needs some improvement. The validity of the results is reliable. The findings and conclusions were well supported.
 Thus, I suggest accepting this paper. My major concerns are that the sample size is small for lung cancer research. I suggest the author add more GEO dataset which contains samples from the Chinese population
1.	Line 97, please specify which R package was used.
2.	Line 99, log2, or log10 fold change?


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: IDENTIFICATION OF KEY GENES AND BIOLOGICAL PATHWAYS IN CHINESE LUNG CANCER POPULATION USING BIOINFORMATICS ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: not good
Experimental design: not good
Validity of the findings: not good
Additional comments: Author performed reanalyzed work
 GEO accession no GSE136043 was previously analysed, work done and published.
1. The roles and mechanisms of the circular RNA circ_104640 in early-stage lung adenocarcinoma: a potential diagnostic and therapeutic target (PMID: 33569440 PMCID: PMC7867959 DOI: 10.21037/atm-20-8019)
2. Combined Metabolomics with Transcriptomics Reveals Important Serum Biomarkers Correlated with Lung Cancer Proliferation through a Calcium Signaling Pathway (PMID: 34056907 DOI: 10.1021/acs.jproteome.0c01019)

GEO accession no GSE130779  was previously analysed, work done and published.
1. Identification of key genes in lung adenocarcinoma based on a competing endogenous RNA network (PMID: 33281971 PMCID: PMC7709547 DOI: 10.3892/ol.2020.12322)
2. The relationship between LncRNAs and lung adenocarcinoma as well as their ceRNA network (PMID: 33896828 DOI: 10.3233/CBM-203078)

GEO accession no GSE118370  was previously analysed, work done and published.
1. Identification of therapeutic targets and mechanisms of tumorigenesis in non-small cell lung cancer using multiple-microarray analysis (PMID: 33126319 PMCID: PMC7598833 DOI: 10.1097/MD.0000000000022815)
2. LINC00476 Suppresses the Progression of Non-Small Cell Lung Cancer by Inducing the Ubiquitination of SETDB1 (PMID: 33370431 DOI: 10.1667/RADE-20-00105.1)

GEO accession no GSE85841  was previously analysed, work done and published.
1. Prognostic value of BIRC5 in lung adenocarcinoma lacking EGFR, KRAS, and ALK mutations by integrated bioinformatics analysis (PMID: 31093306 PMCID: PMC6481100 DOI: 10.1155/2019/5451290)
2. Identification of Hub Genes Associated with Lung Adenocarcinoma Based on Bioinformatics Analysis (PMID: 31698633 DOI: 10.3934/mbe.2019384)

Re analysed work is not acceptable Meanwhile, Author  not provided  Differential gene expression (DEGs) table with probe id, logFC, pValue, adj.P.Val, t value and Gene Name, which are more fundamental and basic in this work. No evedance in this work without this DEGstable.
Author not performed construction and analysis of  target genes - TF regulatory network and target genes - miRNA regulatory network
Author also not performed validation of hub gene ((FABP4, LPL, PDK4, and PPARGC1A ) by  ROC analysis and  RT-PCR.

 No originality and  no novelty in this work.

I strongly recommend rejection


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: IDENTIFICATION OF KEY GENES AND BIOLOGICAL PATHWAYS IN CHINESE LUNG CANCER POPULATION USING BIOINFORMATICS ANALYSIS
Review round: 1
Reviewer: 3

Output:
Basic reporting: There still have grammatical errors and typos in this manuscript. For examples:
- ... in progression of lung canxer ...
- ... we validated he expression levels ...
- The results of GO annotation contains three parts ...
- Some parts miss punctuation.
- ...
Therefore, the authors are suggested to re-check and revise carefully to meet the quality for publication.

Quality of figures should be improved.
Experimental design: The authors merged different data without considering batch effect removal.

Source codes should be provided for replicating the methods.

A critical concern is the use of a very small sample size in this study. This number of data cannot ensure a consistent/reliable result.

GO database or analysis has been used in previous studies i.e., https://doi.org/10.1016/j.neucom.2019.09.070 and https://doi.org/10.1016/j.csbj.2019.09.005. Therefore, the authors are suggested to refer to more works in this description.

Multivariate analysis on hub genes should be performed.

ROC analysis should be performed also.
Validity of the findings: More results/discussions on the differences between Chinese and other populations should be conducted.

There should have validation data.

The authors raised some limitations, however, they did not show how to avoid the limitations in further studies.

How to define the cut-off values when selecting the top GO terms (Table 2)?
Additional comments: No comment.


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

Input:
Title: IDENTIFICATION OF KEY GENES AND BIOLOGICAL PATHWAYS IN CHINESE LUNG CANCER POPULATION USING BIOINFORMATICS ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No new comments
Experimental design: No new comments
Validity of the findings: No new comments
Additional comments: No new 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: IDENTIFICATION OF KEY GENES AND BIOLOGICAL PATHWAYS IN CHINESE LUNG CANCER POPULATION USING BIOINFORMATICS ANALYSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Thanks for addressing my previous comments. However, some comments were not addressed well and there must have a room for improvement, including:
- RT-PCR should be conducted to validate the results as well as hub genes.
- My previous comment #5 were not answered.
- The meaning of immunohistochemistry images in Fig. 7 should be explained.
- What are positive and negative classes in ROC curves?
- How to define the cut-off values when selecting the top GO terms (Table 2)? Why top 5 is selected as optimal ones?


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: IDENTIFICATION OF KEY GENES AND BIOLOGICAL PATHWAYS IN CHINESE LUNG CANCER POPULATION USING BIOINFORMATICS ANALYSIS
Review round: 3
Reviewer: 1

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


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

Input:
Title: THE EFFECT OF 2,4-DICHLOROPHENOXYACETIC ACID ON THE PRODUCTION OF OAT (AVENA SATIVA L.) DOUBLED HAPLOID LINES THROUGH WIDE HYBRIDIZATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. English language of the manuscript could be improved. To provide some examples from the text where comprehension may be difficult – lines 167-168, line 183, line 190. I suggest using professional editing service to polish the language.

2. The introduction provides sufficient background to show previous research and to explain the aim of the presented study. The literature is relevant for the field of study. My only suggestion is adding some information (if known) about the possible explanations of the fact that oat is recalcitrant to haploidization.

3. The tables and figures are for relevant. Figures are of high quality. Tables are well designed and allow easily to find relevant results. All figures and tables are properly referred in the text. The captions of tables 2-4 and the captions of figures 4-6 should be completed. In tables 2-4 it should be explained which values are numbers and which are percentages. Moreover, in table 6 it is not clear what Authors would like to show by calculating “seeds/DH lines” (should be number of DH lines). Is it better to obtain more seeds from less DH lines or less seeds from more DH lines and what these values show? This value could be similar for genotypes were low number of seeds were produced by one line and high number of seeds were produced by multiple lines (e.g. DC11003 and DC11027).  Moreover, maybe it would be useful to somehow marked “the best” and “the worst” genotypes in tables 2-4. Figures 4-6 the caption should be complemented i.e. the description how the percentages were calculated (and what they really represent) should be added. Could Authors explain why the sum of percentage for 50 mg/L 2,4-D and for 100 mg/L 2,4-D is always 100% (for all genotypes and for all traits)? What really represent the values on figures 4-6? It would be interesting to see how many haploid plants on MS media, successfully acclimatized haploid plants and plant that survive the colchicine treatment there is for a particular genotype in comparison to the number of haploid embryos obtained for a particular genotype. Please use the same colours for 2,4-D concentrations on all charts.

4. Raw data are supplied however some more detailed description should be added.
Experimental design: 1. The research is well planned, and the materials and methods are carefully described. The level of details is sufficient to easily repeat the presented experiments. The only information that should be added to materials and methods section are as follows: (i) when (how long after transfer the haploid lines from MS to soil) was the procedure of chromosome doubling performed, (ii) when (how long after the chromosome doubling procedure) was the ploidy level measured, (iii) when the seeds were obtained.
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: THE EFFECT OF 2,4-DICHLOROPHENOXYACETIC ACID ON THE PRODUCTION OF OAT (AVENA SATIVA L.) DOUBLED HAPLOID LINES THROUGH WIDE HYBRIDIZATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper investigates the post-pollination application of different 2,4-D concentrations in the efficiency of oat double haploid production by wide hybridization with maize. The expression of this paper is clear and unambiguous. Literature references and research background were sufficiently provided.
I read this paper with interest. The flow of the experiment seems thorough and the data provided is sufficient, I believe that this paper can be published, but improvements are necessary.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Below I provide recommendations for improvements.
Line 51-53, a comma should be added before in which.
Line 53-55, a comma should be added before DH technologies
Repeated expression in the caption of figure 2 “(A) control 2n, (B) haploid 1n and (C) doubled haploid 2n.”
I recommend combining your raw data in table 2, 3, 4 to some figures.


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

Input:
Title: THE EFFECT OF 2,4-DICHLOROPHENOXYACETIC ACID ON THE PRODUCTION OF OAT (AVENA SATIVA L.) DOUBLED HAPLOID LINES THROUGH WIDE HYBRIDIZATION
Review round: 1
Reviewer: 3

Output:
Basic reporting: The work of Juzoń et al. reveal a new aspect of DH production. The manuscript showed interesting results, however, there are some points for discussion and revision.

Specific comments:
Your introduction needs more detail. I suggest that you improve the description at 2,4-D and auxin to provide more justification for your study (specifically, you should describe the difference between 2,4-D and auxin or IAA et al.). Also introduce the mechanism and advantage of 2,4-D in DH production.

2,4-D is one of the auxin analogues. In your experiment, there was no exogenous auxin treatment. Please consider to use this term more carefully.

More details should to provided in the method of pollination. The time, the temperature and so on.
Experimental design: The authors reported the tested genotypes produced in total 287 haploid plants after 100 mg/L treatment and 27 haploid plants after 50 mg/L 2,4-D. So how about higher concentration 2,4-D treatment, like150mg/L?

Figures 4-6 should be more standardized.
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: THE EFFECT OF 2,4-DICHLOROPHENOXYACETIC ACID ON THE PRODUCTION OF OAT (AVENA SATIVA L.) DOUBLED HAPLOID LINES THROUGH WIDE HYBRIDIZATION
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: My all previous comments has been taken into consideration. Thank you.


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

Input:
Title: THE EFFECT OF 2,4-DICHLOROPHENOXYACETIC ACID ON THE PRODUCTION OF OAT (AVENA SATIVA L.) DOUBLED HAPLOID LINES THROUGH WIDE HYBRIDIZATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: This paper is written with clear and unambiguous, professional English throughout, provided sufficient literature references,  field background/context,  shared the professional article structure, figures, tables, as well as relevant results to hypotheses.
Experimental design: The experiment was designed within the Aims and Scope of the journal,  well defined, relevant & meaningful research question, and sufficient detail & information to replicate.
Validity of the findings: I do not think the raw material is necessary for the paper as described in Tables 2 and 3. These raw data could be provided as supplementary materials.
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: DIMENSIONAL CHANGES OF COMMERCIAL AND NOVEL POLYVINYL SILOXANE IMPRESSION MATERIALS FOLLOWING SODIUM HYPOCHLORITE DISINFECTION
Review round: 1
Reviewer: 1

Output:
Basic reporting: Please expand more on the introduction of why it is important to conduct this study and in particular, why is dimensional changes important and what implication can it be lead to if it is not achieved.

Statement of the problem and justification of study need to be enhanced in introduction section. Why use 1% sodium hypoclorite in this study (and what other solutions is used commonly)
Experimental design: ok
Validity of the findings: ok
Additional comments: number of whole sample size should be included and written clearly
Line 108 n=10 is refering to which group?
Line 123, to add statistical significance

Why do you choose 30 mins to check for dimensional stability - add reference or justify
Line 145- Remove add discussion here
Line 147- correct quoting of referencing