PONE-D-22-12165Many foliar endophytic fungi of Quercus gambelii are capable of psychrotolerant saprotrophic growthPLOS ONE

Dear Dr. Koide,

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PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: No

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The paper is well written, well organized, and absolutely clear about how the authors conducted the research. The study could be of certain interest to the target of Plos One.

Nevertheless, the overall quality of the manuscript needs to be slightly improved before publishing in Plos One, since minor issues are present.

I suggest the authors to better explain the second hypothesis in the material and method section since just a few words were spent about the goal of this second part of the paper.

Moreover, it is important to improve the clarity and quality of the tables provided in the text deriving from ANOVA analyses; it is not specified the number of ways (one way or two way) as well as the meaning of the acronyms.

Furthermore, It would be useful to provide more details on the result of the statistical analysis reported in the tables within the text. Which variance gave greater significance? Between groups or within groups? This is important since It seems that these parameters differ a lot in terms of explained variance.

Minor issues

The figures contains a lot of data, but are not properly legible. Is it possible to rotate the figures? I understand that could negatively affect the figure quality, but It surely could improve from a legibility point of view.

Reviewer #2: I think the authors satisfactorily replied to the reviewer 1 which raise some issues that go beyond the aim of the work. Moreover, some comments are inappropriate (e.g., since mid-2000s direct PCR is successfully applied to fungal material without any problem about the sequence quality). The aim of the work is clear and the biological hypotheses which the work is based on are appropriate and fully met. Experimental planning and procedures are detailed and they support appropriately what the authors intended to demonstrate although some parts of the M&M section can be improved (see the below comments). However, it is not clear because you selected as “warm” temperature 17 °C that in many parts of the world is not so “warm”. I think this has to do with the climate where Q. gambelii grow. In this case the choice of this temperature level should be justified and an in-depth description of the study site should be reported (e.g., min and max temperatures of coldest and warmest months). The results about strain identification (ln 114-120) should be moved at the beginning of the result section and the phylogenetic tree generated to identify the unknown isolates should be shown. This part is a result and not a methodology and, in addition, this shift would compensate the gap of length between M&M and Result sections.

Accession numbers and site of collection of each isolate should be added in the supplementary Table 1

Specific comments

Ln 40, a brief description of the differences between psychrotolerant, psychrotrophic and psychrophilic should be provided to ensure that the reader understand why you chose the first term.

Ln 85, delete “2000”

Ln 88, how many sub-samples (disks) from each leaf?

Ln 92, “very few” is too subjective, “less than xx” would be better

Ln 92-96, I suppose the plates were inspected every day to isolate strains before R selected fungi (e.g. Penicillium, Trichoderma,…) form and release conidia

Ln 103, “ITS is considered a universal barcode for fungi” delete this sentence because superfluous

Ln 116, 119,…, “sp.” must not be italicized; What is “Ophiognomonia 1”? perhaps “Ophiognomonia sp.”?

Ln 132, for a total of 640 plates, is it correct?

Ln 133-136, it is not clear which is the size of the plates. I do not think that 200 ml of medium was added to each plate. Moreover the amount of each component of the medium should be reported as weight (g, mg,…) per L.

Ln 137, was the leaf litter autoclaved?

Ln 143, how did you separate mycelium biomass from residues of litter?

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Reviewer #1: No

Reviewer #2: No

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Many foliar endophytic fungi of Quercus gambelii are capable of psychrotolerant saprotrophic growth

PONE-D-22-12165R1

Dear Dr. Koide,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Raffaella Balestrini

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: After a careful and considered review of the content of this paper by the authors, the article was now found to be available for publication.

Reviewer #2: The revised version of the manuscript can be now published on PlosOne. All comments have been satisfied appropiately.

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Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

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