Dear Dr. Beyene,

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 19 2026 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.

• A 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'.
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We look forward to receiving your revised manuscript.

Kind regards,

Debasis Mitra

Academic Editor

PLOS One

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2. When completing the data availability statement of the submission form, you indicated that you will make your data available on acceptance. We strongly recommend all authors decide on a data sharing plan before acceptance, as the process can be lengthy and hold up publication timelines. Please note that, though access restrictions are acceptable now, your entire data will need to be made freely accessible if your manuscript is accepted for publication. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If you are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process.

3. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

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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?

Reviewer #1: Partly

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: No

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: No

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

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: No

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Reviewer #1: General Assessment: The manuscript investigates AMF spore density and diversity in two Ethiopian Afromontane forests. The topic is relevant to ecology, biodiversity, and restoration, and the study contributes useful data from a region where such research is limited. However, the manuscript requires major revision before it is suitable for publication. Key issues include English language quality, methodological clarity, consistency in taxonomy, statistical rigor, and redundancy in presentation.

Major Comments:

1. Language and Grammar: There are numerous grammatical and typographical errors starting from the title through to the conclusion (check lines 1, 9-31, 43-45). I recommend that the manuscript undergo professional English editing.

2. Methods and Reproducibility: The Materials and Methods section lacks clarity on sample replication, randomization, and statistical assumptions. For example:

Line 138-141: “Five to ten dominant plant species were considered from each plot” is vague. Specify the exact number and criteria for selection.

Line 153–156: Clarify how composite samples were created and how many replicates per species per forest were analyzed.

Add soil classification for both sites (lines 116–135).

3. Statistical Analysis: The authors reported ANOVA and Tukey’s HSD but did not justify statistical assumptions (normality, homogeneity of variance) or provide measures of variation (SD/SE).

4. Results Presentation: The results are clear but mostly descriptive without statistical backing (lines 254 – 261). Explanations about potential causes (e.g., soil type, altitude, disturbance) appear in the Results (lines 266–268) and should be moved to the Discussion.

5. Discussion and Synthesis: The discussion repeats results excessively and lists numerous studies without deeper analysis (Lines 323–338 and 369–396). Authors should synthesize findings into clear messages: (a) what environmental factors drive differences between the two forests? (b) how AMF-host specificity is expressed, and (c) what are the implications for restoration practices?.

6. Conclusion: The conclusion currently reads more like a summary of the results. It should be revised to emphasize the novelty of the findings, acknowledge study limitations, and outline their practical applications.

7. References: Several references are outdated (e.g., Schenck 1990; Gerdemann & Nicolson 1963; Walkley & Black 1934) and should be updated with more recent studies. In addition, reference formatting should be standardized by italicizing species names, and consolidating redundant citations (e.g., Birhane et al. [6,7,23] cited multiple times for similar points).

Minor Comments:

1. Abstract (lines 9–31): Correct multiple typos, spacing issues, and species name inconsistencies.

2. Introduction: Literature review is too long and repetitive. Condense into a focused summary.

3. Figures/Tables: Ensure captions and units are clear. For example: “AMF Spore Density" ( 100 g⁻¹ soil).”

4. Terminology: Use consistent species spelling throughout (e.g., Cupressus lusitanica not Cupressu lucitanica; Olea europaea not Olea europea).

Reviewer #2: In order to be considered for publication, this manuscript's research paper requires some modifications and enhancements. Comments for revision are as follows:

General Assessment

The manuscript entitled “Arbuscular mycorrhizal fungi spore density and species diversity varied within plant species and between dry Afromontane forests in northwestern Ethiopia” addresses an important and underexplored topic related to soil microbial diversity and forest ecosystem restoration. The study is well designed, data are clearly presented, and the findings contribute useful baseline information for dry Afromontane forests in Ethiopia. Overall, the manuscript is suitable for publication after moderate revisions as outlined below.

Major and Minor Comments are imposed to do corrections at author level

1. Title and Language Clarity

The title is informative but contains minor grammatical issues (for example, “pants species” should be corrected to “plant species”). Overall language is understandable, but moderate English editing is required to improve grammar, sentence structure, and consistency throughout the manuscript.

2. Abstract

The abstract clearly summarizes the objectives, methods, and key findings. Minor corrections are suggested to improve readability and correct spacing issues, especially around scientific names.

3. Introduction

The introduction is comprehensive and well referenced. However, the research gap and novelty could be highlighted more explicitly in the final paragraph to strengthen the justification of the study.

4. Study Design and Sampling

The sampling strategy and site description are clearly explained and appropriate for the objectives. A brief justification for the number of plots and sampling period would further strengthen methodological rigor.

5. Soil Physicochemical Analysis

Standard and accepted methods were used. It is suggested to briefly discuss how soil parameters may have influenced AMF spore density differences between the two forests.

6. AMF Identification and Enumeration

The use of wet sieving, decantation, and morphological identification is appropriate. However, limitations of morphological identification without molecular confirmation should be briefly acknowledged in the discussion.

7. Statistical Analysis

The statistical methods used (ANOVA and Tukey’s test) are appropriate. Please ensure uniform reporting of significance levels (for example, p < 0.05) throughout the text and figure captions.

8. Results Presentation

Results are clearly presented with appropriate tables and figures. Minor formatting issues exist in tables (alignment and consistency in species names) and should be corrected.

9. Discussion

The discussion is detailed and well linked with previous studies. Some sections are repetitive; concise restructuring would improve clarity and focus. Practical implications for forest restoration using AMF could be emphasized further.

10. Conclusion and Implications

The conclusions are supported by the results. Adding a short paragraph on future research directions, such as molecular AMF diversity and inoculation trials, would enhance the manuscript’s impact.

Final Recommendation

Recommendation: Accept with moderate revisions.

The manuscript presents scientifically sound data and meaningful ecological insights. The required revisions are mainly related to language polishing, minor methodological clarifications, and formatting improvements, and do not affect the core scientific merit of the study. After addressing the above comments, the manuscript will be suitable for publication.

Reviewer #3: The manuscript “Arbuscular mycorrhizal fungi spore density and species diversity varied within pants species and between dry Afromontane forests in northwestern Ethiopia” provides an interesting study, which is timely and relevant to the current scenario. However, few points need to be addressed before considering it suitable for acceptance to this esteemed journal. These are-

1. Page 9, line no. 75-76….”In their study these authors revealed a strong negative correlation between AMF spore density and an altitude.”….please rewrite the sentence.

2. Please check the spacing in between words, also in between two sentences throughout the manuscript. At several position, spacing are missing.

3. Page 9…line no. 80-84…”These AMF inoculations can improve seedling establishment and success rate due to their role in nutrient and water absorption and translocation, help the seedlings share growth limiting plant nutrients via common mycorrhizal 82 3 83 networks[15], help seedlings to tolerate abiotic stresses (drought, salinity, acidity) and biotic stresses (pathogens, competitions for nutrients and water)[10,15].”… please rewrite the sentence.

4. Page 10, line no. 85-97………no need to give those previous study in details, give the research gap and novelty of this study.

5. Introduction section needs to be shorten and focused to improve the clarity of this study.

6. It would be better to give the study location through location map.

7. Also, give the plant species authentication numbers in table 1.

8. Page 15….”Enumeration of AMF spores from each subsample was carried out under stereomicroscope at x40 magnification according to the methods and procedures described in the International Culture Collection of Vesicular-Arbuscular Mycorrhizal Fungi (INVAM), (http://invam.wu.edu/)” give full stop at the end of the sentence.

9. Page 16, line no. 207…remove the underline from the full stop.

10. 4.3 and 4.4 sections lacks critical discussion.

11. Conclusion section is very weak. Please improve

12. Include the limitation and future perspective of this study.

13. Figure 1 A and 1B…..Y axis title should be shorten, and X axis plant species name should be made uniform for both.

14. Please shorten the gap of x axis in between two forests bar.

Reviewer #4: The manuscript explores the ‘Arbuscular mycorrhizal fungi spore density and species diversity varied within pants species and between dry Afromontane forests in northwestern Ethiopia’. However, there are several critical comments and suggestions that can be made to improve the clarity and effectiveness of the manuscript.

1. The specific research questions are not explicitly stated in question form. While the objectives imply what the study aims to address, the manuscript would benefit from stating them directly.

2. The manuscript does not clearly state a hypothesis. A well-defined hypothesis would strengthen the scientific rigor of the study and help guide the reader's expectations.

3. The research gap is mentioned but should be sharpened to explicitly state what is new compared to previous AMF studies in Ethiopian Afromontane forests.

4. The sampling design needs clearer justification, including rationale for plot number, plot size, and selection of dominant plant species.

5. The pooling of soil samples should be justified, and its implications for variability and statistical power should be discussed.

6. The statistical analysis section should specify the experimental unit, assumptions checked for ANOVA, and how normality and homogeneity of variance were addressed.

7. Methods for AMF identification should clarify how uncertain morphotypes (identified only to genus level) were handled in analyses.

8. The presentation of species richness and diversity indices could be strengthened by including additional diversity metrics (e.g., Shannon or Simpson indices) or by justifying their exclusion.

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what does this mean?). If published, this will include your full peer review and any attached files.

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: Yes:  Dr. Kartik Chandra Guchhait, Post-doctoral Fellow, IIT Kharagpur, WB, India

Reviewer #4: Yes:  Dr. DASARI SREEKANTH

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Attachments

Attachment

Submitted filename: plos one.docx

Attachment

Submitted filename: Comment.pdf

Dear Dr. Beyene,

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.

• A 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.

As the corresponding author, your ORCID iD is verified in the submission system and will appear in the published article. PLOS supports the use of ORCID, and we encourage all coauthors to register for an ORCID iD and use it as well. Please encourage your coauthors to verify their ORCID iD within the submission system before final acceptance, as unverified ORCID iDs will not appear in the published article. Only  the individual author can complete the verification step; PLOS staff cannot  verify ORCID iDs on behalf of authors.

We look forward to receiving your revised manuscript.

Kind regards,

Erika Kothe

Academic Editor

PLOS One

Journal Requirements:

If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

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 (if provided):

The review indicates a small change to be considered.

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

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #3: All comments have been addressed

**********

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

Reviewer #3: Yes

**********

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

Reviewer #3: N/A

**********

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

The PLOS Data policy

Reviewer #3: Yes

**********

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

Reviewer #3: Yes

**********

Reviewer #3: Authors have addressed most of the comments satisfactorily. However, line no. 112-116 should be written in a better way to meet the standard of this journal.

**********

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 #3: Yes:  Dr. Kartik Chandra Guchhait

**********

[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.]

To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures

You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation.

NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications.

Arbuscular mycorrhizal fungi spore density and species diversity varied within plants species and between dry Afromontane forests in northwestern Ethiopia

PONE-D-25-33914R2

Dear Dr. Beyene,

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.

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Kind regards,

Erika Kothe

Academic Editor

PLOS One

Additional Editor Comments (optional):

Reviewers' comments: