PONE-D-22-33646Mycorrhizal fungi reduce the photosystem damage caused by drought stress on Paris polyphylla var. yunnanensisPLOS ONE

Dear Dr. Huang,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Mar 24 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Mayank Gururani

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at 

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and 

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. 

When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

3. Thank you for stating the following financial disclosure: 

"Liu Tao conceived, designed the experiments and wrote this paper. Huangcan analyzed the data. Zi Shuhui, Xi Congfang and Li Xiaoxian Preparing plant materials All authors read and approved the final manuscript."

At this time, please address the following queries:

a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution. 

b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.”

c) If any authors received a salary from any of your funders, please state which authors and which funders.

d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.”

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

4. Please include a copy of Table 2 which you refer to in your text on page 6.

5. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 

6. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

[Note: HTML markup is below. Please do not edit.]

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: Partly

Reviewer #2: Yes

********** 

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

Reviewer #1: No

Reviewer #2: Yes

********** 

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: No

Reviewer #2: Yes

********** 

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: No

Reviewer #2: No

********** 

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: This manuscript investigated the protective mechanism of AMF on plant photosynthetic systems by inoculating Paris polyphylla var. yunnanensis (P.py) with two mycorrhizal fungi (AMF) under drought stress (DS). The authors have underlined that the DS decreased the RWC and increased the OKJIP curve of the plants, and the authors claim that AMF inoculation could reverse it. This paper also reports the photosystem damage caused by the DS and shows how AMF helped to reduce this damage. The study appears to be sound, but the language is unclear in the results, and the discussion needs more relevant references to support the findings.

Comments:

• The statistical tests and significance values of different comparisons should be clarified for tables/figures to ensure that readers understand exactly what the researchers studied.

• The results need to be described well, e.g., “and the decrease in the rate of increase in Fo after inoculation with AMF can indicate a decrease in its damage.” The authors should clarify the results section to avoid confusion.

• The authors can also report the difference between the protective mechanism of Glomus eburneum (G.e) and Paraglomus occultum (P.o) in the results section and support these findings with relevant literature in the following discussion.

• In the first paragraph of the discussion, the authors try to highlight the effects of drought stress on the symbiosis of AMF and P.py; I suggest citing more studies to show how the findings of this paper relate to previous research on symbiosis.

• 2nd paragraph of discussion: Please add more references to support the claim that “AMF inoculation of plants significantly enhanced plant photosynthetic rate under drought.” These findings need more explanation (with references to specific studies) in the context of underlying molecular processes that show how AMF rescues P.py photosynthesis under drought stress.

Minor changes/Typos:

• Page 1: Please add more supportive references in addition to reference [7]

• Page 1: please add more literature in addition to reference [8] to establish the link between drought stress and photosynthesis

• Page 2, last paragraph of introduction: Change “P.py” to “Paris polyphylla var. yunnanensis(P.py)” in the first sentence

• Page 2, last paragraph of introduction: Build up the importance of explicitly using P.py for studying DS responses in AMF and non-AMF plants.

• Page 2, last paragraph of introduction: The author states that “While shade plants should be more sensitive to changes in photosynthesis in the face of drought stress, most current studies have focused on the interaction between drought and photosynthesis, as well as the interaction between drought and AMF, and there are few studies on the interaction between the three.” Please cite relevant studies that explored these interactions.

• Material and Methods, Page 2: Italicize “Paraglomus occultum (P.o)”

• Material and Methods, Page 2: Please provide references to relevant articles/websites for protocols and software discussed in materials and methods

• Material and Methods, Page 2: “resu4=lts”

• Material and Methods, Page 2: Inconsistent styles used, e.g., “determined by the method of [17]” and “method of Sartory et al. [19]”, it needs to be consistent throughout the manuscript

• Material and Methods, Page 2: inconsistent usage “O-J-I-P” or “OJIP.”

• Table & Figures: Please add detailed captions for multiple-part figures and tables

• Table & Figures: The treatment labels used in Table 1 are different “WX”, “WYL”,”DX”, and”DYL” and no description is provided

• Table & Figures: Table 2 is missing

Reviewer #2: Using Paris polyphylla and arbuscular mycorrhiza fungi (AMF), the authors examined AMF colonization rate, relative water content, chlorophyll content and photosynthesis under drought stress. The experiments were designed well and executed efficiently. Except for typos, the data is presented coherently, and the MS is well written. For researchers engaged in plant stress research, I believe the MS is valuable. It is important that the authors address the following issues, however:

1. In Abstract, Capitalize the beginning of the sentence like Chlorophyll a fluroscence curve measuresments…, DS also caused a descrease…

2. Explain YI, YII, ETRI and ETRII in abstract.

3. I found some inconsistencies, such as arbuscular mycorrhiza fungi (AMF) is written as AMF plant and AM plant throughout the MS.

4. Kindly type in bold the sub-headings and Capitalize the first letter such as Drought treatment.

5. Typo error in the drought treatment, the germination of the D P.py

6. In drought treatment, how much water did you use for normal wet condition and drought condition. Did you withdraw water completely in drought condition?

7. Include the microscopic images of colonization.

8. Typo error in Relative water content such as oil to soil.

9. In determination of chlorophyll content repeat of the sentence the filter paper was placed on a funnel, moistened with ethanol.

10. In data analysis, typo error in results were analysed

11. In table 1, explain what is WX, WYL, DX, DYL.

12. Where is table 2? Most of the table numbers are incorrectly written.

13. Expand photo, cond and ci in Table 3.

14. In results, effects of drought stress on quantum yields of PSI and PSII, inoculation of Eleutherococcus balsamifera is given. This part was not seen in materials and methods.

15. A proofreading should be done for typos.

********** 

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

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

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PONE-D-22-33646R1Mycorrhizal fungi reduce the photosystem damage caused by drought stress on Paris polyphylla var. yunnanensisPLOS ONE

Dear Dr. Huang,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Oct 14 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Arun Kumar Shanker

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments:

The authors are requested to revise according to reviewer 2's comments

[Note: HTML markup is below. Please do not edit.]

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 #2: All comments have been addressed

**********

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 #2: Yes

**********

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

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 #2: Yes

**********

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 #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 #2: Using Paris polyphylla and arbuscular mycorrhiza fungi (AMF), the authors examined AMF colonization rate, relative water content, chlorophyll content and photosynthesis under drought stress. The experiments were designed well and executed efficiently. The methods and results are well justified towards the primary goal of this study. This study could be interesting to the Plant cell, Tissue and Organ Culture audience. A couple of minor things need to be solved before it is published.

1. In last sentence it is written that it is necessary to study the relationship between Dianthus, AMF and photosynthesis. The term Dianthus is not used anywhere. Is it Dianthus or P.Py.?

2. In materials and methods, how P.py seedlings were planted without 1 Kg of soil mixture. Sentence error?

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

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 #2: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachments

Attachment

Submitted filename: Review.docx

Mycorrhizal fungi reduce the photosystem damage caused by drought stress on Paris polyphylla var. yunnanensis

PONE-D-22-33646R2

Dear Dr. Huang

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.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Arun Kumar Shanker

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The revision can be accepted as the authors have incorporated the suggestions of the reviewers.

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 #2: All comments have been addressed

**********

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 #2: Yes

**********

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

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 #2: Yes

**********

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 #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 #2: Using Paris polyphylla and arbuscular mycorrhiza fungi (AMF), the authors examined AMF colonization rate, relative water content, chlorophyll content and photosynthesis under drought stress. The authors have underlined that the DS decreased the RWC and increased the OKJIP curve of the plants, and found that AMF inoculation could reverse it. This paper also reports the damage to the photosystem that the DS caused and demonstrates how the AMF assisted in mitigating the damage.The experiments were designed well and executed efficiently. The methods and results are well justified towards the primary goal of this study. This study could be interesting to the PLOS ONE.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

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 #2: No

**********