Dear Dr. Moonmoon,

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Ramegowda Venkategowda, PhD

Academic Editor

PLOS ONE

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7. Please ensure that you refer to Figure 19 and 20 in your text as, if accepted, production will need this reference to link the reader to the figure.

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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: Yes

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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: No

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: No

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Reviewer #1: Dear authors,

In this work, it has been studied the effects of exogenous application of different compounds on strawberries during field study. A detailed analysis of photosynthetic parameters is provided, and evidence that the treatments contribute for reducing the leaf temperature differential. The research is relevant in the context of alleviating heat stress, which is expected to increase due to climate change.

However, there are some major points that should be addressed. Certain aspects of the methodological setup require further clarification. I also provide a list of minor points for consideration. I see potential in the data acquired by the authors, and I hope my comments help to improve their work.

Kind regards

Major points

• English writing requires deep revision.

• Please provide the precise identification (variety) of the tested genotypes, as research reproducibility is fundamental for its acceptance. This information is also very relevant for future studies as, for instance, aiming at understanding why G1 is tolerant, while G2 and G3 are susceptible to heat stress.

• I think that is necessary a better description of how the G and P lines are related (sections 2.3 and 2.6). I suggest the authors to create a schematic representation.

• In the context of the comment above, it was also not clear for me which variety was used for the treatments. For instance, in Figures 6-18, three varieties are represented on panels b, d and f (P1-3). Do they correspond somehow with the genotype used for the treatments test (panels a, c, and e)?

• Please provide the full details of soil fertilization (concentration and manufacturer, amount and source of the manure, etc), fungicide and pesticide (manufacturer, concentrations).

• Please also mention at which time of the day, and at which day of the month the measurements were made. This is important for understanding why some of the observed effects did not appear on the first month of the analysis.

• The manuscript would benefit from a deeper bibliography input. That goes from the decision for the selection of the compounds for treatments to the analysis of the results. Statements in the Results should be supported by a source reference, even if the information is taken for granted. The impact of heat stress on the photosynthetic apparatus has been greatly studied in other species, often in combination with biotic or chemical stimulants. The deeper bibliographic review would also help to improve the Discussion, which has only 5 cited references.

Minor aspects

• Regarding the experimental setup (point 2.2), the authors mention that the treatments were performed during constant intervals over three months and repeated thrice, between October-November 2023 to April-May 2024. Next (point 2.7), I understand that an initial set was prepared and from this set, the three experiment rounds were started in parallel. However, this point is elusive and I do suggest to clarify it. Was there a time window between the start of each experimental round? If not, I would consider these not as three independent experiments, but rather as one larger experiment with a good number of replicates (Figure 3). While this design can be valuable for statistical analysis, especially in a field experiment, I would still recommend repeating the study to confirm the results.

• Scientific names should be italicizes (e.g. Fragaria, Rosaceae)

• Some abbreviations are lacking their full description next to its first mention (e.g. RCBD, PGR; RH);

• Please provide the reference for the RCBD model.

• I suggest to move the description of treatments that are now in the section 2.7 to section 2.8, so it would be next to the full description of the treatment application. Also, please provide the manufacturer of each reagent used. I would also suggest to clearly specify the temperature at which the reagents were stored;

• All Figures would benefit from better legends. Please introduce more details, including full description of what they are representing, description of the error bars, statistical test applied, number of individuals tested per condition (n), etc.

• Please also provide the version of each software and R package used.

• I suggest that each treatment be described by both its full name and abbreviation (T1–T5) when presenting the results. This would make the reading easier. Also, please add the description of each treatment o the figures’ legend.

• I suggest to increase the font size of Figure 19 for easier reading. Also, although not really critical, blue and red are usually used for denoting negative and positive values/correlation, so I suggest to swap them to avoid any misinterpretation. I understand that data across all genotypes and treatments were combined and used to plot the Pearson’s correlation. This information should be clear. Furthermore, I do suggest to review the data analysis and interpretation, as for instance the observation that “SPAD increases ambient temperature”, which is unlikely. Still, I wonder whether it might be more informative to represent the correlations of the tested parameters separately for each genotype and treatment. This way one would be able to evaluate their inputs independently.

Reviewer #2: Comments to the author (PONE-D-25-14863)

The manuscript titled “Mitigation of heat stress using hormones during summer season for survivability of strawberry plants” is an interesting study. The study by Salwa et al is mainly focusing on the use of five exogenous substances (Molasses, Melatonin, Kaolin, CaCl₂, and Abscisic Acid) to mitigate heat stress in three strawberry genotypes under tropical summer conditions which involves physiological and fluorescence measurements across different months. To be honest, the study is interesting and I truly enjoy the reading of manuscript. Overall, the research is valuable and relevant, particularly in the context of climate resilience in horticultural crops. However, some extensive improvements are needed to enhance the scientific clarity, rigor, and writing quality. As per my observation, in the current form, it lacks hypothesis-driven design clarity, and some methodological and interpretational issues need attention. Some suggestions are as follow:

It is extremely embarrassing to review the paper without line numbers to point-out the specific corrections. But I have following suggestions to consider:

I would strongly suggest to change the title as: Exogenous Application of Hormones and Chemicals to Mitigate Heat Stress in Strawberry Under Tropical Summer Conditions.

Overall, the abstract should be carefully revised as it lacks clarity and scientific soundness. Terms like “some sort of capability” are vague and has incomplete phrasing. Please provide better summary of the most effective treatments and genotype response needed.

In the introduction section, it needs better framing of research gaps. For example, what is unknown about hormone-based mitigation strategies in strawberry under field-like summer stress particularly in Bangladesh. Further, inconsistent tenses and awkward phrasing are present throughout, e.g. “It is establishing climatic change...” is an unclear statement. Further, “…mandatorily has to endure high heat” is overly informal. In Introduction, it doesn’t clearly state the hypothesis and research question as well as key gaps.

Material and methods are the most important section and it should have clear citations/protocol used in the present study. There are some key parts to improve. In this section, genotypes are inconsistently labeled, as sometimes G1/G2/G3, elsewhere P1/P2/P3 there must be uniformity throughout the manuscript. Please use scientific or cultivar names of strawberry genotypes if available and specify the concentration basis for each treatment (e.g., ppm or % w/v, and how these concentrations were finalized/selected. Please provide justification why these 5 treatments were chosen or provide appropriate literature citations/physiological rationales.

It is unclear that how many number of replicates per treatment/per genotype were used. Was the fluorometer standardized/calibrated before use? Any internal checks? Please elaborate the statistical models in more depth. Was random effect for replication included and interaction terms analyzed?

In the results section, many figures show overlapping trends please consider consolidating. Leaf surface temperature being higher in treatment than control contradicts expectation. Explanation is speculative; check methodology or measurement error. Graph captions needed improvements as some are mislabeled (e.g., gH shown but figure caption mentions ϕNPQ Fig. 18).

Please provide numerical values with statistical summaries (e.g., mean ± SE) in text for major parameters.

In statistical analysis part, please clarify what variables were considered random vs fixed in LMM. Further, Pearson correlation matrix (Fig. 19) is dense but informative and interpret more in text.

Overall discussion is descriptive, not mechanistic. Please include some information on the roles of ABA in stomatal regulation under heat, Melatonin as antioxidant/protective compound, Kaolin and molasses as physical shields against radiation, and also genotype differences are needed to be discussed. Please provide recent literature and add more context from global strawberry stress physiology.

Conclusion Needs to state clear practical recommendations, e.g., which combination(s) are best for growers. It must be concise to the point. Also, clarify that this is a first step, and in future long-term, field-based, and reproductive/yield trait studies are needed. add these as future directions.

Reviewer #3: Comments for Author

� This manuscript “mitigation of heat stress using hormones during summer season for survivability of strawberry plants” is very interesting in which Author focused on effect of heat stress on strawberry and its alleviation through Hormones.

� In abstract Author did not mention the levels or concentrations of hormones used to alleviate heat stress.

� Authors required to write novelty statement about their research

� Introduction is insufficient, Author should compare their research with studies of previous researchers

� Hypothesis of study is not clear in introduction section

� In material and methods section too much headings are used by Authors, it is required to write under minimum headings. Particularly second and third heading of the material and methods.

� Why Author used RCBD design? Whereas experiment was conducted under pots.

� Results are written well by authors but still revision is required to improve results. Author should add significance level or p values in results section.

� Graphical presentation also required to improve. Authors need to write in correct manners on axis of graphs.

� Pearson’s correlation diagram is not clear. It is advised to add clear diagram

� In discussion section Author no need to indicate treatments as T1, T2.. etc Moreover, Author needs to write discussion in well mechanized form.

� Conclusion section needs improvement and required to write limitation and future prospects of their study.

� References should be balanced in reference list as well as in manuscript text.

Reviewer #4: In the present manuscript, the authors evaluated the effect of different PGRs sprays on three different strawberry genotypes to observe the tolerance response of plants from the onset of tropical summer to peak summer. The experimental approach is well justified and well described; however, the results are not adequately presented. In the result sections 3.2, 3.3 (number repeated in this section) and 3.4, there is no indication of which phenotype the data for the different treatments are (Figures, a, c, & e) or from which treatment the data in the phenotype graphs are (Figures, b, d & f). All of the above does not allow for understanding the interpretation of the results or their discussion.

Other minor concerns are:

- All abbreviations must be defined in the abstract and when they are first used in the introduction or materials and methods sections (RCBD, RH, SPAD, PGRs…).

- P1, P2 & P3 must be G1, G2 & G3 in section 2.6 of MM

- In the result sections 3.2, 3.3 (number repeated in this section) and 3.4, there is no indication of which phenotype the data for the different treatments are (Figures, a, c, & e) or from which treatment the data in the phenotype graphs are (Figures, b, d & f).

- In the MM section, three phenotypes are indicated: G1 (heat tolerant), G2, and G3 (heat susceptible). However, all the corresponding graphics are labeled with the P1, P2, and P3 varieties. Please clarify. These labels are repeated in all the results sections and in the discussion or conclusion sections.

Finally, I recommend that a native English speaker revise the entire manuscript…

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Reviewer #1: Yes:  Gustavo Turqueto Duarte

Reviewer #2: No

Reviewer #3: No

Reviewer #4: No

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Attachments

Attachment

Submitted filename: Plos one.docx

Dear Dr. Moonmoon,

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 Aug 28 2025 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 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 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,

Ramegowda Venkategowda, PhD

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:

Dear Authors,

Kindly sees the comments from a reviewer especially Reviwer 4. There is a serious issue on labeling the figures and given proper legends or methodological explanation. Kindly correct the same.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #3: All comments have been addressed

Reviewer #4: (No Response)

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

Reviewer #3: Yes

Reviewer #4: Partly

**********

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

Reviewer #3: Yes

Reviewer #4: Yes

**********

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

The PLOS Data policy

Reviewer #3: Yes

Reviewer #4: Yes

**********

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

Reviewer #3: Yes

Reviewer #4: Yes

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Reviewer #3: Author has addressed all the comments properly with reasons. Therefore my decision is accepted for further publication process

Reviewer #4: The authors have greatly improved the manuscript's language but some revisión is still needed. However, my concerns remain regarding the presentation of results as explained in the first review. The graphs in Figures 6 through 18 (the number in Figure 17 is repeated) represent the different parameters recorded (Ambient temperature on the leaf surface, Leaf internal temperature, Leaf thickness, SPAD, etc., for each treatment (T0-T5)). The primary concern is that authors do not specify whether the data in the graphs in sections a, c, and e of each figure correspond to all plants or a specific variety. Furthermore, the graphs in sections b, d, and f of each figure, which refer to the three varieties used, do not specify which treatment (T0-T5) the data are from. The authors should present the data collected for each Strawberry variety and each treatment used to allow for better interpretation, comparison, and discussion of the results.

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Reviewer #3: No

Reviewer #4: Yes:  Homero Reyes de la Cruz

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Attachments

Attachment

Submitted filename: accept.docx

Exogenous application of hormones and chemicals to mitigate heat stress in Strawberry under tropical summer conditions

PONE-D-25-14863R2

Dear Dr. Moonmoon,

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,

Ramegowda Venkategowda, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: