PONE-D-24-15543Site climate more than soil properties and topography shape the natural arbuscular mycorrhizal symbiosis in maize and spore density within rainfed maize (Zea mays L.) cropland in the Eastern DR CongoPLOS ONE

Dear Dr. Byamungu,

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 Sep 05 2024 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,

Marcela Pagano, Ph.D, M.D.

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. In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why.

3. Please provide a complete Data Availability Statement in the submission form, ensuring you include all necessary access information or a reason for why you are unable to make your data freely accessible. If your research concerns only data provided within your submission, please write "All data are in the manuscript and/or supporting information files" as your Data Availability Statement.

4. We note that [Figure 1 and Supporting Information Fig 1] in your submission contain [map/satellite] images which may be copyrighted. All PLOS content is published under the Creative Commons Attribution License (CC BY 4.0), which means that the manuscript, images, and Supporting Information files will be freely available online, and any third party is permitted to access, download, copy, distribute, and use these materials in any way, even commercially, with proper attribution. For these reasons, we cannot publish previously copyrighted maps or satellite images created using proprietary data, such as Google software (Google Maps, Street View, and Earth). For more information, see our copyright guidelines: http://journals.plos.org/plosone/s/licenses-and-copyright.

We require you to either (1) present written permission from the copyright holder to publish these figures specifically under the CC BY 4.0 license, or (2) remove the figures from your submission:

a. You may seek permission from the original copyright holder of Figure 1 and Supporting Information Fig 1 to publish the content specifically under the CC BY 4.0 license.  

We recommend that you contact the original copyright holder with the Content Permission Form (http://journals.plos.org/plosone/s/file?id=7c09/content-permission-form.pdf) and the following text:

“I request permission for the open-access journal PLOS ONE to publish XXX under the Creative Commons Attribution License (CCAL) CC BY 4.0 (http://creativecommons.org/licenses/by/4.0/). Please be aware that this license allows unrestricted use and distribution, even commercially, by third parties. Please reply and provide explicit written permission to publish XXX under a CC BY license and complete the attached form.”

Please upload the completed Content Permission Form or other proof of granted permissions as an "Other" file with your submission.

In the figure caption of the copyrighted figure, please include the following text: “Reprinted from [ref] under a CC BY license, with permission from [name of publisher], original copyright [original copyright year].”

b. If you are unable to obtain permission from the original copyright holder to publish these figures under the CC BY 4.0 license or if the copyright holder’s requirements are incompatible with the CC BY 4.0 license, please either i) remove the figure or ii) supply a replacement figure that complies with the CC BY 4.0 license. Please check copyright information on all replacement figures and update the figure caption with source information. If applicable, please specify in the figure caption text when a figure is similar but not identical to the original image and is therefore for illustrative purposes only.

The following resources for replacing copyrighted map figures may be helpful:

USGS National Map Viewer (public domain): http://viewer.nationalmap.gov/viewer/

The Gateway to Astronaut Photography of Earth (public domain): http://eol.jsc.nasa.gov/sseop/clickmap/

Maps at the CIA (public domain): https://www.cia.gov/library/publications/the-world-factbook/index.html and https://www.cia.gov/library/publications/cia-maps-publications/index.html

NASA Earth Observatory (public domain): http://earthobservatory.nasa.gov/

Landsat: http://landsat.visibleearth.nasa.gov/

USGS EROS (Earth Resources Observatory and Science (EROS) Center) (public domain): http://eros.usgs.gov/#

Natural Earth (public domain): http://www.naturalearthdata.com/

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.

Additional Editor Comments:

The manuscript was improved; however minor details are needed:

--Figure 3: Mycorrhizal Status of maize (Zea mays L) in the three selected territory

--Replace by in the three selected territories or better, use <sites>

[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: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

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: Yes

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: Yes

Reviewer #2: Yes

**********

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 study investigated the impacts of site climate, soil properties, and topography on the natural arbuscular mycorrhizal symbiosis in maize and spore density within rainfed maize (Zea mays L.) cropland in the Eastern DR Congo. The data were harvested from the 32 sites across three territories. The study employs a well-thought-out experimental design, encompassing site climate variables, soil physicochemical properties, topography, and vegetation variables. This design aids in analyzing the influence of different environmental factors on maize MC and the density of AMF spores and provides reliable data to support the study's conclusions. The article provides a detailed interpretation of the experimental results, highlighting the positive role of site climate factors on arbuscular mycorrhizal fungi colonization and spore density of maize rhizosphere soil. The explanation of the results is clear and well-structured, making it easy for readers to comprehend the study's findings. The authors mention the potential value of the research results in practical field applications. This connection to real-world applications is helpful in translating research findings into actionable recommendations for agricultural production in rainfed maize cropland systems. It is clear that the authors are not concerned about the latest publications in this field in the sections of Introduction and Discussion. The presentations are not sufficient to clarify the key issue and background in terms of AMF colonization. The latest publications might be considered as Rehman et al., 2022, Plant Soil, 1–17; Khan et al., 2024, Science of The Total Environment, 917, 170417, and Abrar et al., 2024, Plant Physiology and Biochemistry, 213, 108839.

In summary, this manuscript has several positive attributes, but minor improvements could further enhance the research.

1) Lines 27: Units of spores density should be uniform.

2) Lines 38: Zea mays should be italicized.

3) Lines 115-117: Reference should be added.

4) Line 118: Replace “bbelow” with “below”.

5) Line 134: A comma should be added after specifically.

6) Line 166: How did plants were uprooted?

7) Lines 168-169: How did rhizosphere soil was collected?

8) Lines 169-170: Arbuscular Mycorrhizal Fungi should be written in lowercase.

9) The equation for the total mycorrhization percentage should be mentioned.

10) Lines 179 and 192: The magnification unit should be standardized.

11) Reference should be added for pH measurement.

12) Reference should be added for exchangeable cations measurement.

13) The instruments and chemicals should be mentioned which were used to determine soil analysis.

14) Line 211: Extra full-stop should be deleted.

15) Lines 38: Full-stop should be deleted after algorithms.

16) In headings 2.7.1, 2.7.2, and 2.7.3, names and versions of software used for analysis should be mentioned.

17) The headings format should be uniform throughout the manuscript.

18) p should be italicized throughout the manuscript.

19) Line 278: Anova should be capitalized and followed throughout the manuscript.

20) Lines 285-287: It should be mentioned that these sites belong to which territory.

21) Line 289: Spore density unit should be corrected (100 g-1 soil) and followed throughout the manuscript.

22) Lines 365-366: Arbuscular Mycorrhizal Fungi should be written in lowercase and you have already mentioned the abbreviation earlier, so you have to use an abbreviation instead of using the full name. Follow it throughout the manuscript.

23) Lines 432-435: What are the favorable conditions that promote spore germination?

24) Line 443: Carbon (C) and phosphorus (P) abbreviations should be added at the start and no need to repeat it later.

25) References should be updated by adding journal names, italicizing the scientific names, and keeping the text in a similar pattern such as sentence form.

26) Figure 1 quality should be enhanced.

27) Figure 3, p should be italicized in the figure and caption, Anova should be capitalized, and p = < should be written in a standardized form and followed throughout the manuscript.

28) Figure 3, What do highland and lowland stand for? The author did not mention it in the whole manuscript.

29) Table 2, Zea mays L should be written as Zea mays L.

Reviewer #2: Dear,

The paper “Site climate more than soil properties and topography shape the natural arbuscular mycorrhizal symbiosis in maize and spore density within rainfed maize (Zea mays L.) cropland in the Eastern DR Congo” brings interesting data and relevant analyzes for understanding some edapho-climatic variables on some mycorrhizal parameters. Here are some observations about the text:

Summary

- It is well written, but it is important to add the study hypothesis.

Introduction

- Extremely long, with paragraphs with poorly connected construction;

- It is important to add the hypothesis at the end of the introduction text.

Methodology

- Inform whether the Trypan Blue, used to color the roots, was prepared in lactoglycerol or lactophenol;

- Inform the concentration of sucrose used to extract the glomerospores;

- It is important to build a heat map between the variables studied;

- Add algorithm analysis (K-means), to check which variables are most relevant in the study;

- It is important to add data on the Most Probable Number of AMF infective propagules, production of Soil Proteins Related to Glomalin (PSRG) and spore viability.

Discussion

- Needs adjustments, as there are excessive comparisons with other works. It is important to explain the observed behaviors, especially based on whether or not the hypothesis is confirmed;

- There is a need to further explore the effects of C and P on the mycorrhizal parameters evaluated;

- The discussion on sporulation needs to consider what is already well established for fungal reproduction, such as the buffering effect of the substrate on the production of glomerospores.

Conclusion

- They must be direct and written with the verb in the present tense; are written as a mix of results and final considerations.

Figures

- Figures 1 and 7 – improve resolution;

- Figures 2 and 3 – supplementary material.

Tables

- Delete table 3;

- Tables 5 and 6 – supplementary material

References

- It is important to add more up-to-date references on the topic.

After making the aforementioned adjustments, the paper can be considered for publication

Sincerely,

**********

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.</sites>

Attachments

Attachment

Submitted filename: Evaluation Report.docx

Site climate more than soil properties and topography shape the natural arbuscular mycorrhizal symbiosis in maize and spore density within rainfed maize (Zea mays L.) cropland in the Eastern DR Congo

PONE-D-24-15543R1

Dear Dr. Adrien Ndeko Byamungu,

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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, 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,

Marcela Pagano, Ph.D, M.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Manuscript improved accordingly to suggestions

Reviewers' comments: