Dear Dr. Kammer,

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.

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Mayank Anand Gururani

Academic Editor

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?

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

Reviewer #1: Adaptation of plants to environmental conditions, in particular to altitude ranges, is important for understanding the adaptation of photosynthesis processes. The results confirm the regulation of stomata conduction in response to changes in the amount of CO2. It is possible to make a forecast of changes in the conductivity of stomata with an increase in the level of CO2 in the atmosphere. However, the question arises whether all plants will react equally to the CO2 content. There may be a need for research on other plants.

Reviewer #2: I found this manuscript interesting to read and fairly well-written. However, there are some issues that the authors need to address before the manuscript could be accepted for publication. Detailed comments are given below.

Abstract

L19: “Understanding the stomatal responses …” instead of “Comprehension of the stomatal …”.

L21: remove “, for example”.

LL22-25: not very clear to read. Please revise to improve clarity as to what treatments were used and how the treatments were applied (under controlled lab experiments).

L27: “increasing Co2 levels ..” instead of the current.

L32: “2,970 m a.s.l” instead of “2’970 m a.s.l”.

L36: local environmental conditions like what? Give examples.

Introduction

L45: “into the atmosphere” instead of “in the atmosphere”.

LL50-51: the reference should be placed at the end of the sentence, not the in the middle.

L51: better to use Co2 levels throughout the manuscript instead of [Co2].

LL51-52: continuously increasing atmospheric Co2 due to what?

L54: remove “, for example”.

L60: “periods of drought” instead of “drought periods”.

LL63-66: the terminology adopted here should be referenced.

LL66-67: needs a reference.

LL69-71: needs a reference.

L74: field or lab experiments?

L75: “an inverse relationship with Co2 levels” instead of the current.

LL77-79: were those findings documented under lab or field experiments, or both?

LL80-81: “the present-day concentrations” instead of the current.

L82: comma after “[26]”.

LL90-92: responses of what is being discussed here?

LL107-111: “ .. we tested plants originating from populations that have grown for prolonged …” instead of the current.

LL107-111: the language here needs be streamlined and the sentences need to be simplified a bit.

L114: please add” to minimize the confounding effects of those environmental factors in the experiment” at the end of the sentence.

LL117-119: the terminology adopted here should be referenced.

L123: “grown” instead of cultivated.

Materials and Methods

L140: “have been growing” instead of “have been living”.

LL148-152: references are needed.

LL155-157: unclear to me if seeds were collected from plants and grown in growth chambers for experiments or if seeds were obtained from the Col-0 wild type, please clarify which seeds were used in the experiments. Also, why seeds of the Col-0 wild type were used?

L158: “could be” instead of “can be”.

LL186-188: but those were lab conditions not field conditions, as the growth chambers were placed in a lab. How would the fluctuation of temperature and humidity under lab conditions be similar to their fluctuation under field conditions?

L189: “grown” instead of “cultivated”.

LL192-194: why the plants in the first group were grown for 28 days and for 35 days in the second group? Explain to the reader.

L195: “to prevent any damage to the root system” instead of the current.

L199: “Measurements of Plant Traits” instead of the current.

L200: “For measuring stomatal aperture …” instead of the current.

L206: “For measuring stomatal frequency …” instead of the current.

LL209-217: relevant references should be cited here.

L229: “after the end of the experiment.” instead of the current.

Statistical Analysis

L234: “as the main effects” instead of “as explanatory variables”.

L235: why chamber ID was used as a random effect?

L236: those are “liner models” not regression analysis. Please make sure you use the correct terminology throughout the manuscript.

L247: “as main effects” instead of “as explanatory variables”.

LL249-250: is there a refence to support this approach?

L253: “as main effects” instead of “as explanatory variables”.

LL257-258: “as main effects” instead of “as explanatory variables”.

LL263-264: I did not see any co-variate analysis in the manuscript. So how could that be? Was there bias in the experiment? If yes, this is not the right way to reduce bias in the experiments. Please explain.

LL266-273: seems to align with the policy of the Journal, yet I do not like to see AI tools being used for generating or composing texts in scientific papers.

Results

L278: why is the stats table placed in the appendix? They should be a core part of the manuscript.

LL292-298: As I could see in Figure 2, the responses to temperature and pCo2 were also analyzed but not mentioned here. Why is that?

L321: why is the stats table placed in the appendix?

L335: “grown” instead of “cultivated”.

L340: why is the stats table placed in the appendix?

LL357-358: which figures show this trend?

L362: why is the stats table placed in the appendix?

L387: why are the figures placed in the appendix?

L389: why is the stats table placed in the appendix?

L395: why is the stats table placed in the appendix?

Tables

Table 1: “n=1,200” instead of “1’200”.

L303: “main effects”.

Why all degrees of freedom are 1 even in interactions?

Table 2: “n=1,926” instead of “1’926”.

Why all degrees of freedom are 1 even in interactions?

Table 3: Why all degrees of freedom are 1 even in interactions?

Discussion

L495: “diffusion” instead of “diffusivity”.

L552: “grown” instead of “cultivated”.

When the authors say, “plant grew under natural conditions,” I am not sure what they mean by that. Plants were grown in growth chambers, so how does that relate to natural conditions?

LL560-568: so we have leaves collected from natural sites and leaves collected from plants grown in growth chambers, which ones were included in the analysis? The authors did not describe their methods well to clarify this, which makes the Discussion confusing for the reader. The authors need to address this and improve the clarity of their Methods and Discussion.

Conclusions

OK.

Figures

Figure 1: The plant species under study should be specified in a better way, the reader does not need to figure out which is which on the figure. Also, the figures should show any significant differences between/within treatments, or lack thereof.

Figure 2: The plant species under study should be specified in a better way. The figures should show any significant differences between/within treatments, or lack thereof. L308: : “n=1,200” instead of “1’200”.

Figure 3: The plant species under study should be specified in a better way. Also, the figures should show any significant differences between/within treatments, or lack thereof.

Figure 4: The plant species under study should be specified in a better way. Also, the figures should show any significant differences between/within treatments, or lack thereof.

Figure 5: The plant species under study should be specified in a better way. Also, the figures should show any significant differences between/within treatments, or lack thereof.

Figure 6: The plant species under study should be specified in a better way. Also, the figures should show any significant differences between/within treatments, or lack thereof.

Figure 7: The plant species under study should be specified in a better way. Also, the figures should show any significant differences between/within treatments, or lack thereof.

Figure 8: The plant species under study should be specified in a better way. Also, the figures should show any significant differences between/within treatments, or lack thereof.

**********

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

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Reviewer #1: Yes: Plant physiologist Dr. Oksana BelousPlant physiologist Dr. Oksana BelousPlant physiologist Dr. Oksana BelousPlant physiologist Dr. Oksana Belous

Reviewer #2: No

**********

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Attachments

Attachment

Submitted filename: The review report for the manuscript, Aug. 2025.docx

Dear Dr. Kammer,

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 12 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.. 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.. 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.. 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 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 . 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 . 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 . 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,

Raffaella Balestrini

Academic Editor

PLOS One

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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:

The manuscript has been improved with respect to the original version. Thanks. If possible, I suggest to try to reply to the additional comment by the reviewer to increase the reading.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #2: All comments have been addressed

**********

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

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.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??>

Reviewer #2: Yes

**********

Reviewer #2: I found the revised version of the manuscript improved, well-written, and well-structured. I thank the authors for addressing my comments and concerns on the previous version. I have a few minor suggestions as below. Other than that, the manuscript is in a good shape.

Abstract

Should be one paragraph, not separated into multiple paragraphs.

Introduction

L55: “to variations in Co2 levels” instead of “to Co2 levels variations”.

Materials and Methods

L205: “nutrient solution” instead of “nutrition solution”.

Statistical Analysis

Good.

Results

Good.

Tables

Good.

Discussion

Good.

Conclusions

Good.

Figures

I still think that statistical differences between treatments and/or species or lack thereof should be shown on the figures using labels or letters. I understand that overlapping and non-overlapping error bar tell the story, but it is always better to have the figures labeled appropriately so they are very straightforward and easy to understand by the reader without referring to the text. I think the Editor will agree with me on this.

**********

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

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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 For information about this choice, including consent withdrawal, please see our For information about this choice, including consent withdrawal, please see our For information about this choice, including consent withdrawal, please see our Privacy Policy..-->

Reviewer #2: No

**********

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Stomatal responses of differently CO2-acclimated plants to natural and experimental CO2 gradients

PONE-D-25-38428R2

Dear Dr. Kammer,

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: