PONE-D-23-39597The small iron-deficiency-induced protein OLIVIA and its relation to the bHLH transcription factor POPEYEPLOS ONE

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Dear Authors,Please check the Reviewers' comments carefully and revise your work accordingly.Thank you very much for submitting an interesting work to PLOS ONE.Best regards,Nguyen 

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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?

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

Reviewer #1: I Don't Know

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: Yes

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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: Figures:

Comment 1.

In Figure 4D and E, proOLV::GUS activity was induced by -Fe treatment in root differentiation zone and hair. Did the authors test the transcript level of OLV by qPCR at same developmental stage to show that induction is relevant? Seems the expression of OLV is not increased by -Fe in WT (see Fig S5 C).

Comment 2.

In Figure 4F and G shows, proPYE::GUS mainly detected in root tissue only. Why proPYE::GUS activity did not induce in leaves (or shoot) although PYE expression was highly induced both root and shoot by -Fe in Fig 5C (in WT). How do you explain?

Comment 3.

Different letters need improving in Fig5 and Fig6 diagrams. For instance, same letter indicated for all samples in Figure 6F. Seems there are some significant differences between treatment. Similar other errors: Fig5C “ab”, Fig5C “abc”, Fig6B “ab”, Fig6C “ab”.

Comment 4.

Y axis needs improving in Fig 5A and Fig 6A. Please double check.

Comment 5.

Authors used two independent OLV overexpression lines (OX7 and OX11) in all experiments. Expression of OLV was not increased in OX11 (see Fig S5 C and D).

Methods:

Comment 6.

What type of pye-1 mutant was used. Describe it in Plant material section.

Comment 7.

Please, complete the description of protocol. What kind of secondary antibody was used.

Typo errors:

Typographical error in “rot” instead of root (Page9)

Typographical error in “bHLHIVc” (Page5)

Typographical error in “wasused” (Page15)

Typographical error in Fig S4 legend “Subcelllar”.

Reviewer #2: Article review

1. EVALUATION OF THE PAPER MANUSCRIPT

Title of the Manuscript:

"The small iron-deficiency-induced protein OLIVIA and its relation to the bHLH

transcription factor POPEYE"

Manuscript number: PONE-D-23-39597

The article PONE-D-23-39597 has comprehensively demonstrated that Fe deficiency-induced bHLH transcription factor (POPEYE) can bind to the small protein OLIVIA (OLV), which is induced by Fe scarcity. The article PONE-D-23-39597 is slightly suited to the PloS One as per its aims and scope. In general, the manuscript is written in good logical style with a suitable introduction, interesting and reasonable Results and Discussion sections. Therefore, the manuscript can be conditionally accepted. However, to improve the quality of the article, some minor points should be noted:

- Could the authors re-analyze RNA-Seq datasets to see how the OLV gene is expressed in different organs/tissues during growth and development? If so, please write several sentences describing the expression level of the OLV gene.

- The authors should include more information about the OLV in the conclusion section. Also, if possible, please simplify and combine the perspectives into a single paragraph.

- All abbreviations should be applied after the first occurrence, such as transcription factor (pages 5 and 11).

- List of references should be exactly followed the PloS One’s style.

- A minor issue is that the reviewer is unable to access the link to jmicrovision. If possible, offer a new link.

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Reviewer #1: No

Reviewer #2: Yes: Ha Duc Chu

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PONE-D-23-39597R1The small iron-deficiency-induced protein OLIVIA and its relation to the bHLH transcription factor POPEYEPLOS ONE

Dear Dr. Bauer,

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 Apr 25 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,

Nguyen Hoai Nguyen

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.

Additional Editor Comments:

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I also agree with the Reviewer 1 regarding the usage of biomass in this case. Please amend your writing in the manuscript according to the obtained data (e.g., plant size...). 

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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: All comments have been addressed

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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: Yes

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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

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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: Comment1:

In page 5, you mentioned “OLV also affects biomass”. Indeed, olv mutant showed smaller phenotype compared to WT (supported by rosette size data), but you do not have fresh weight value. Please, think on that, or just remove “biomass”.

Typo errors:

Typographical error in “BHLH” (Page5). Usually, we use bHLH.

Typographical error in “Sesanum indicum” (Page 7).

Typographical errors in FigS2. Please, double check the name of genus: “Actinida chinensis “, “Sesamum idicum”, “Viits vinifera”.

Typographical errors in FigS3. Please, double check “Sesamun indicum “, “Gingko biloba”.

Typographical error in FigS4 “Subcelllar”.

Reviewer #2: All comments have been addressed. The reviewer believes that this manuscript version meets the requirements for publication in PloS ONE.

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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: Shapulatov Umidjon

Reviewer #2: No

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The small iron-deficiency-induced protein OLIVIA and its relation to the bHLH transcription factor POPEYE

PONE-D-23-39597R2

Dear Dr. Bauer,

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,

Nguyen Hoai Nguyen

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: