PONE-D-21-05714

Effect of exogenously applied alpha-tocopherol on vital agronomic, physiological and biochemical attributes of Lentil (Lens culinaris Medik.) under induced drought stress

PLOS ONE

Dear Dr. Sami Ullah,

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.

I agree with the reviewers that language editing is needed. Abstract need to be modified. Correction should be done in materials and methods according to reviewers. Quality of graphs need to improve. The discussion of the results must be done in scientific terms, explaining the meaning of the results. Conclusion should be improved.

Please submit your revised manuscript by May 06 2021 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:

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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.

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We look forward to receiving your revised manuscript.

Kind regards,

Basharat Ali, Ph.D

Academic Editor

PLOS ONE

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Whilst you may use any professional scientific editing service of your choice, PLOS has partnered with both American Journal Experts (AJE) and Editage to provide discounted services to PLOS authors. Both organizations have experience helping authors meet PLOS guidelines and can provide language editing, translation, manuscript formatting, and figure formatting to ensure your manuscript meets our submission guidelines. To take advantage of our partnership with AJE, visit the AJE website (http://learn.aje.com/plos/) for a 15% discount off AJE services. To take advantage of our partnership with Editage, visit the Editage website (www.editage.com) and enter referral code PLOSEDIT for a 15% discount off Editage services.  If the PLOS editorial team finds any language issues in text that either AJE or Editage has edited, the service provider will re-edit the text for free.

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• A clean copy of the edited manuscript (uploaded as the new *manuscript* file)

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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?

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: Partly

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

Reviewer #1: Yes

Reviewer #2: Yes

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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

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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: No

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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: The authors have presented the effects of drought induced stress by exogenous application of alpha-tocopherol on the agronomic and physiological attributes of lentil. The manuscript shows mechanistic approach and provides extensive results that could be useful in scientific community. However, this manuscript could not be considered for publication in its present form. Some comments are available for revision in order to improve the manuscript:

Major comments:

1. Highlights are missing. Add them substantially.

Minor comments:

Title should be revised. Either change the title by giving the punch line of your findings as a title or keep the same in this form “Effect of exogenously applied alpha-tocopherol on vital agronomic, physiological and biochemical attributes of Lentil (Lens culinaris Medik.) under drought induced stress”

Line 31: Revise it to “climatic conditions”

Line 32: “ill” must be replaced with “drastic”

Line 33: Revise “induced drought” to “drought induced”

Line 34: Mention the abbreviation of “Alpha α” at the first place (line 32) and then use it throughout.

Line 66: Add Comma after “findings”

Line 74: Insert Comma after “plants”

Line 79: Insert Comma after “regimes”

Line 83: Full stop after “Fabaceae”

Line 88: Change “induced drought stress” to “drought induced stress” in the whole manuscript.

Line 113: Revise “potassium available” to “available potassium”

Line 123: Please provide some detail about CVG in materials and methods.

Line 204: Seems like Alpha-tocopherol levels??

Line 205: Inset TAB at the start of the paragraph.

Line 211: Insert comma after “treatments”

Line 227: Add Full stop after “error”. Follow this for all the figures.

Line 239: Delete the word “group” after treatments. You have used this repeatedly. Please revise it carefully throughout as it is not making a good sentence structure.

Line 250: It is better to mention all the figure legends together after the references. Please revise it. Follow this for all the legends.

Line 262: Add comma after treatments.

Line 266: Delete the word “condition”

Line 338: Add Comma after “condition”

Line 338: “slow” must be replaced with “slows”

Line 342: Add comma after “study”

Line 343: Add comma after “contrary”

Line 350: Remove parenthesis after “in”

Line 354: Add Comma after “conditions”

Line 359: Add Comma after “study”

Line 361: Add Comma after “contrast”

Line 362: Add Full-stop after figure number.

Line 384: Add Comma after “condition”

Line 387: Add parentheses (100 mg/L)

Line 392: Add Comma after “condition”

Line 408: Add Comma after “results”

Line 421: Add “such as” after “enzymes”

Line 423: Revise it “Our results are”

Line 426: Add Parentheses (200 mg/L). You did this mistake repeatedly. Please revise it throughout the manuscript.

Line 429: Add Comma after “study”

1. It is better to polish the conclusion section. It is not up to the mark. Substantially revise this portion with justifications and logical statements.

2. For all the figures: you have missed the Alphabetical Letter within the figures. Please insert the letters to indicate the small figures within the main figures.

3. Secondly, all the small figures are not combined properly into one figure. Please revise it carefully.

Reviewer #2: The present study explored the possibility to using alpha-tocopherol as a mitigating agent against drought s tress on Lens culinaris. The manuscript provides an important set of data demonstrating the beneficial effects of alpha-tocopherol. The experiment is well conducted and data analyzed in an intelligible way. In its current state however, the manuscript is not suitable for publication. Major revision is needed. Substantial language editing is needed to reach standard for publication in this Journal. Long sentences should be avoided as much as possible. Besides, there are many inaccurate statements and/or incomplete information that need to be addressed throughout the manuscript.

Below are detailed comments:

The title of the study can be simplified… for ex. “Effect of exogenous of alpha-tocopherol on physio-biochemical attributes and agronomic performance of Lentil (Lens culinaris Medik.) under drought stress”

The introductive sentence of Abstract can be refined to highlight the link between drought stress and oxidative burst, and the potential mitigating effects of alpha-tocopherol. Further, the strategy used to induce drought stress (L33) can be mentioned in the Abstract.

The Abstract is too descriptive and doesn’t provide insights about specific mechanisms underlying the mitigating effects of alpha-tocopherol. A major overhaul is needed. Notably, it is important to highlight the interrelations of obtained data, and their relative relevance in influencing plant performance under drought stress.

An alternative word can be used for “Chaos” (L31). And throughout the manuscript, the term “environmental Chaos” could be avoided, as it seems quite subjective.

The Introduction section can be improved by underlining the regional biophysical context to highlight how severe is drought stress in Punjab. And the reason for choosing Lentil (Lens culinaris) as biological material in this study should be clearly stated.

There is also a need for more background information to justify selection of Alpha-tocopherol as alleviating agent against drought stress… Existing reports on Tocopherol changes under drought stress, or its other potential association with plant responses to stress could be useful. Information in L383-L387, L389-391 can also be explored for the purpose. This may smooth the formulation of the hypothesis of the study, which is not clear in current manuscript.

The “Methods” part needs to be rewritten in a more rigorous way, by providing necessary information required for possible replication of the study:

- L99-100: What the ratio 2:1 stands for, since physico-chemical analyses suggested that the soil is sandy loam. Please clarify this.

- What is the relative proportion of water and 70 % EtOH in the vehicule solution used for preparation of Alpha-tocopherol treatment?

- What was the basis for selecting the different levels of Tocopherol treatments (100, 200, 300 mg/L) and those of drought treatments (20 and 25d drought periods)

- Please rewrite statement for statistical analyses, and integrate the approach used for correlation analysis.

- Please accurately present Dubois methodology for measurement of soluble total content.

- It is apparent that the experiment was designed with two factors (tocopherol level, level of drought stress). However, authors appear to suggest three factors (L198). Please address this matter.

The quality of graphs is rather poor… Please improve their resolution. And for better readability, the legend of different figures can be modified as (within drought stress period): (i) Control, (ii) Drought + 0 mg/L tocopherol, (iii) Drought + 100 mg/L tocopherol, (iv) Drought + 200 mg/L tocopherol, (v) Drought + 300 mg/L tocopherol. Information about statistical inference, including the interaction of studied factors can also be integrated in legends.

It is important to indicate how strong is the coefficient of correlation (Table 2) rather than only mentioning that the correlation is significant. This way, results may be interpreted in a different way, notably drought effects on chlorophyll (a and b) content…

Besides, to gain more insights about the influence of studied factors, it would be interesting to analyze how studied parameters are related one with another, which would allow a deeper discussion of results.

Overall, the discussion didn’t help understanding the specific mode of action of Tocopherol in attenuating harmful effects of drought stress. Since drought stress is commonly related to stomatal closure, resulting in impaired Photosynthesis efficiency, it would have been interesting to clarify which parameter (s) in the photosynthesis apparatus was/were site(s) of Tocopherol beneficial action…

Following minor flaws could also be addressed:

- L303-306. This section is not at its appropriate place… Please check.

- L378: alternative expression can be found for “distinctly significant”.

- L344: “key metabolic activities”…What are they?

- For better readability, at the first occurrence in each section, please mention what different acronyms stand for.

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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: Yes: Shahbaz Atta Tung

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-21-05714R1

Effect of exogenous alpha-tocopherol on physio-biochemical attributes and agronomic performance of Lentil (Lens culinaris Medik.) under drought stress

PLOS ONE

Dear Dr. Sami,

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.

==============================

ACADEMIC EDITOR: Dear authors, plz have a look on the comments raised by reviewers. I agree with both reviewers that some minor changes are still required..

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Please submit your revised manuscript by Aug 20 2021 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: http://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,

Basharat Ali, Ph.D

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.

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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 #1: (No Response)

Reviewer #2: (No Response)

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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 #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: N/A

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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 #1: Yes

Reviewer #2: Yes

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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 #1: Yes

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 #1: The authors have made good efforts in revising the manuscript according to the comments. But at some points, I still feel that changes should be made. You should have to follow the below mentioned points before acceptance for publication.

1. Even you have revised your paper according to the suggestions but the newly added information is still the same in the Abstract and Conclusion. So, please be rational and make the information unique for both of these sections. They must not have similar sentences.

2. You have added the highlights and the information is good but please follow the standard rules for highlights. Make the bullet points and divide the information in these bullet points.

3. Please arrange the abbreviations in the footnotes of the first page to make them more obvious and precise.

Reviewer #2: The manuscript has been substantially improved, and authors have satisfactorily addressed previous comments, though some minor concerns persist.

For example, the sentence in paragraph L104-L111 is too long! It can be split in shorter sentences as follows: From [In plant (L104)] to [environment (L106)], from [Among (L104) to oxidative stress (109)], from [It is suggested (L109) to condition (L111)]. Please double-check the manuscript for similar flaws.

Further, I am not sure whether the style for presentation of “Highlights” (added section: L61-77) is conform to the Journal instructions…

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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 #1: Yes: Shahbaz Atta Tung

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.

Effect of exogenous alpha-tocopherol on physio-biochemical attributes and agronomic performance of Lentil (Lens culinaris Medik.) under drought stress

PONE-D-21-05714R2

Dear Dr. Sami Ullah,

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,

Basharat Ali, Ph.D

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: