Review Reports - Regulatory Roles of MYB Transcription Factors in Root Barrier Under Abiotic Stress

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Overall Assessment

This review article offers a comprehensive and thoroughly researched synthesis of the regulatory role of MYB transcription factors in the deposition of suberin and lignin in roots, especially in the context of abiotic stress conditions. The manuscript encompasses a substantial volume of contemporary literature and synthesizes findings from Arabidopsis, rice, tomato, and various other species, thereby elucidating both conserved and species-specific mechanisms.

The subject is pertinent and timely due to the increasing focus on root barrier plasticity and its effects on drought and salinity resistance. The manuscript is scientifically sound, but it needs to fix some problems with its structure, clarity, and figure quality before it can be published.

1. Length and Repetition

The manuscript is quite lengthy, and numerous sections reiterate concepts previously elucidated (e.g., hormonal crosstalk, ABA–MYB signaling, and overarching phenylpropanoid pathway descriptions). It would be easier to read if the presentation were shorter. Keeping the story focused will be easier if you cut down on the number of times you have to explain things.

2. Flow and organization of sections

The information is complete, but some parts get too dense because they have long lists of gene names, signaling pathways, or stress responses. It would be helpful for readers if some subsections ended with short summaries of the main point before moving on to the next topic.

3. Quality of Figures and Images

The numbers need to be better.

Some pictures are not very clear,

It's hard to read labels and short forms, and

Panels have too many parts for a review-type figure.

Since numbers are important in review articles, I suggest changing them to make them easier to read, with clearer layouts, higher resolution, and more readable fonts.

4. Comparisons between species

The manuscript often changes species. This is necessary, but the changes could be smoother. Short connecting sentences that explain why each comparison is important would help the reader understand the logic.

5. Style and Language in English

It would be helpful to lightly edit the manuscript to make it flow better. Some sentences are very long, and the words used aren't always the same. A language polish would make things clearer and look better overall.

Small Comments

The abstract could be made a little shorter without losing any important information.

Tables 1 and 2 are helpful, but they could be better if the formatting and terms were more consistent.

Some parts have long lists of references. If you can, it may be enough to cite a recent review.

When you first use an abbreviation, you should explain what it means (for example, ROL, CASP, or SGN3/CIF).

Overall Recommendation

This is a useful and well-documented review. The manuscript will be ready for publication in Plants after some changes, such as making the figures better, getting rid of extra information, and improving the writing.

Recommendation: Small Change

Author Response

Reviewer #1:

>. Length and Repetition

The manuscript is quite lengthy, and numerous sections reiterate concepts previously elucidated (e.g., hormonal crosstalk, ABA–MYB signalling, and overarching phenylpropanoid pathway descriptions). It would be easier to read if the presentation were shorter. Keeping the story focused will be easier if you cut down on the number of times you have to explain things.

>. Response

We appreciate the reviewer’s insightful comment regarding the length of the manuscript and the repetition of certain concepts in response; we have carefully revised the text to streamline the narrative and remove redundant explanations. Present only once in the introduction or the first relevant section. Improved readability while keeping the focus on the main story.

>. Flow and organization of sections

The information is complete, but some parts get too dense because they have long lists of gene names, signalling pathways, or stress responses. It would be helpful for readers if some subsections ended with short summaries of the main point before moving on to the next topic.

Thank you for this helpful suggestion. We agree that certain subsections contained dense information and could benefit from brief concluding summaries. In the revised manuscript, we have added concise summary sentences at the end of the key subsections.

>. Quality of Figures and Images

>. The numbers need to be better

We thank the reviewer for the constructive feedback on the quality and clarity of the figures. In response, we have thoroughly improved readability and presentation quality.

>. Some pictures are not very clear it's hard to read labels and short forms.

All figures have been changed to ensure that images and graphical elements appear clear

>. It's hard to read labels and short forms, and Panels have too many parts for a review-type figure.

Thank you for pointing this out. we have improved the clarity of the figures. Although the number of panels is necessary to present the full scope of the pathways discussed in the review, we have made several improvements to enhance clarity.

>. Numbers are important in review articles, I suggest changing them to make them easier to read, with clearer layouts, higher resolution, and more readable fonts.

Thank you for your thoughtful suggestion. We agree that figure clarity is essential. After careful evaluation, we believe that the figures were with standardized fonts and layout guidelines consistent with the journal’s format. Therefore, we have retained the current design. However, we appreciate the reviewer’s comment.

>. Comparisons between species

Thank you for this helpful comment. We agree that smoother transitions between species comparisons can improve clarity. We have reviewed the relevant sections and added short connecting sentences to clarify the rationale behind each species comparison and to guide the reader through the logical flow of the discussion.

>. Style and Language in English

Thank you for this valuable comment. We have carefully revised the manuscript to improve readability and flow. Long sentences have been shortened or restructured, and wording has been standardized to ensure consistency throughout the text. These edits enhance clarity while maintaining the scientific meaning.

>. The abstract could be made a little shorter without losing any important information.

Thank you for this helpful suggestion. We have revised the abstract by removing redundant wording and several sentences. The updated abstract is now more concise and easier to read.

>. Tables 1 and 2 are helpful, but they could be better if the formatting and terms were more consistent.

Thank you for pointing this out. We have reviewed Tables 1 and 2 and improved the consistency of formatting. Minor adjustments were made to ensure uniform presentation while keeping the original content intact. We believe these refinements enhance clarity and readability.

>. Some parts have long lists of references. If you can, it may be enough to cite a recent review.

Thank you for this suggestion. We agree that long lists of references can interrupt the flow of the text but in cases where specific original studies were essential to support detailed points, we retained those citations.

>. When you first use an abbreviation, you should explain what it means (for example, ROL, CASP, or SGN3/CIF).

Thank you for highlighting this issue. We have carefully reviewed the manuscript and ensured that all abbreviations, including ROL, CASP, and SGN3/CIF, are defined upon their first appearance in the text.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors,
Your manuscript presents complex information on plant adaptation to climate change stresses on plants. The study is based on a significant number of references, is well documented, but nevertheless it still needs some improvements. My recommendations in this regard are attached.

Best regard!

Comments for author File: Comments.pdf

Author Response

Reviewer #2

>. Use Plants Template (The writing model of personal data as well as the writing model of the manuscript).

Thank you for this suggestion. We have carefully reformatted the manuscript to align with the Plants journal template. All required sections, headings, and formatting now follow the journal’s guidelines.

>. Specify that this is a Review, not an article.

Thank you for this comment. The manuscript already clearly indicates in the abstract. We have also ensured that the journal’s submission system reflects the article type as a Review.

>. The abstract should represent the manuscript within the limit of 200 words.

Thank you for this comment. We have revised the abstract to better reflect the content of the manuscript and remain within the 200-word limit. Specifically, references to rice and tomato have been adjusted to match the level of detail presented in the main text, ensuring that the abstract accurately represents the scope and focus of the review.

>. I recommend replacing the term "climate-resilient agriculture" in the abstract with "climate-resilient crops."

Thank you for the suggestion. We have replaced “climate-resilient agriculture” with “climate-resilient crops” in the abstract to better reflect the focus of the manuscript.

>. At 55, after the word “inducible”, remove ".".

Thank you for pointing this out. The period after “inducible” has been removed as suggested.

>. Use Alignment for the entire manuscript.

Thank you for the suggestion. We have carefully adjusted the formatting so that alignment is consistent throughout the entire manuscript, in accordance with the journal’s guidelines.

>. You are using sentences that are too long. Reduce the text or insert more paragraphs.

Thank you for this valuable comment. We have revised the manuscript to shorten overly long sentences and add as paragraphs.

>. At 103 mention Table 1. This should be as close as possible to where it is first mentioned in the text. Move the table closer to the text.

Thank you for the suggestion. We have moved Table 1 closer to its first mention in the text at line 103 and ensured it is properly cited at that point.

>. According to the journal rules, subchapters are written in Italic and not Bold.

Thank you for pointing this out. We have revised all subchapter headings to italic in accordance with the journal’s formatting guidelines.

>.The number of tables and figures must be written according to the journal requirements.

Thank you for this comment. We have reviewed and revised all table and figure numbering to ensure they comply with the journal’s formatting guidelines.

>. In table 2, correct the species names (e.g. Cicer arietinum, Camellia sinensis, Vitis vinifera).

Thank you for pointing this out. We have corrected the species names in Table 2 to ensure they are accurate and follow proper scientific nomenclature.

>. What did you mean in line 316 ("both for annual plants and for some cereals"?) that there are no annual cereals? Rephrase.

Thank you for pointing this out. We have clarified the sentence in line 316. It now reads: to accurately convey that some cereals are annual species.

>. The sentence "Through improved lignin deposition in roots (by increasing H- and G-lignin biosynthesis), improved photosynthetic efficiency and reduced water loss in leaves compared to non-transgenic controls [77]." is not logical. Reformulated.

Thank you for noting this. We have rephrased the sentence for clarity.

>. On line 341 after "conditions" put "." or write in lowercase.

Thank you for pointing this out. We have corrected the text by adding a period after “conditions” as suggested.

>. Reformulated 381-382. The sentence is not logical

Thank you for pointing this out. We have revised the sentence in lines 381–382 for clarity.

>. In case 5.1, multiple sources need to be cited to confirm similarities or differences. One study is not enough. Develop the topic.

Thank you for this valuable comment. We have revised Section 5.1 to include references that support the reported similarities and differences.

>. In line 393 put "." before the reference.

Thank you for pointing this out. We have corrected the text so that a period now appears before the reference in line 393.

>. In line 514 put "." after "’’".

Thank you for pointing this out. We have added a period after the quotation marks in line 514 as suggested.

>. Arrange the data so that you cite the figure in order. Figure 3A should be cited first.

Thank you for this suggestion. We have revised the manuscript to ensure that all figures are cited sequentially, with Figure 3A cited first as recommended.

>. There is no in-text citation for Figure 3C. Cite all figures in the text.

Thank you for pointing this out. We have added an in-text citation for Figure 3C and ensured that all figures are properly cited throughout the manuscript.

>. Li et al. (line 575) should be cited with [].

Thank you for pointing this out. We have corrected the citation format to [Li et al.] as per journal guidelines.

>. “Conclusions” contain information that was not presented in the manuscript (apple). The conclusions should reflect only the results obtained in the study. Reformulate the paragraph.

Thank you for this comment. We have revised the Conclusions section to include only information and results that are presented in the manuscript.

Reviewer 3 Report

Comments and Suggestions for Authors

Ms. plants-4008531

Title: Regulatory Roles of MYB Transcription Factors in Root barrier under Abiotic Stress

The manuscript is a review devoted to the important problem of regulation of suberin and lignin biosynthesis in the endodermis and exodermis of plant roots, which play a key role of effective barriers to apoplastic water and ion movement. The authors focused on regulation of lignin and suberin biosynthesis by MYB transcription factors. The list of references consists of 117 publications on the topic, a part of them of recent years.

However, on the basis of the criteria provided to me in considering the current paper for publication, I feel that this paper is not acceptable for publication in an international level journal and, particularly, in Plants, as it stands. The manuscript contains important information on the topic, but this information is insufficient to fully cover the topic, it is presented illogically, the illustrative material requires significant revision. The text is not written in good English and contains multiple errors. The manuscript looks crude and sloppy. A more detailed analysis is provided in the comments to the authors. I would encourage the authors to consider submitting a new manuscript after extensive revision.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

The text is written in poor English and contains stylistic, grammatical and spelling errors.

Author Response

>. The authors should give a clear and concise definition of the endoderm and exoderm, their location in a plant and chemical composition.

We thank the reviewer for this valuable suggestion. However, due to requirements of the manuscript’s focus on MYB-mediated regulatory mechanisms rather than detailed anatomical descriptions, mainly we are focused on that part.

>. It would be appropriate to give brief information about the main pathways of regulation of lignin and suberin biosynthesis at the transcription level and what role MYB transcription factors play in this. Then, the authors should briefly describe MYBs, their diversity, structure, hierarchy, basic principles of functioning, regulation (hormones, ROS, etc.)

We sincerely thank the reviewer for this useful comment. To address the reviewer’s suggestion within the constraints of the manuscript, we have carefully revised the introduction to enhance clarity and ensure that the essential context regarding the role of MYB transcription factors in lignin and suberin biosynthesis is clearly stated. We have also added or updated comprehensive overviews of MYB diversity, structure, hierarchical organization, and their regulation by hormonal and stress signaling pathways (including ROS).

>.Table 1: What means “phenotypes” – mutant phenotypes?

Not a unified terminology: “Increased lignin”, “Increased lignin deposition”, “Enhanced suberization”, “Enhanced Lignin”

Thank you for pointing this out. “phenotypes” in Table 1 refer to mutant or transgenic phenotypes. In addition, we have changed terms such as “Enhanced lignin deposition” and “Enhanced suberization” according to suggestion.

>. Figure 1: Authors should provide an explanation of the diagram's construction and list all abbreviations in the legend. Not all TFs and enzymes shown in the diagram coincide with those in the text.

Thank you for this comment. We have added a detailed explanation in the figure legend All abbreviations used in Figure 1 are now listed in the legend for clarity. We have also reviewed the figure to ensure consistency with the text, although some TFs and enzymes were included to provide a more comprehensive overview of the regulatory network .

>. What principle was used to list the TFs in each block? It would be better to arrange them in ascending numerical order.

Thank you for the suggestion. In the revised manuscript, we have arranged all MYB TFs in each block in ascending numerical order.

>. Figure 2: “Hormonal signaling / ROS signaling / (ABA)” – ABA is a part of hormonal signaling. Obviously, the arrow between “Lignin and suberin accumulation” and “Enhances…” needs to be reversed.

Thank you for this helpful comment. We have revised Figure 2 to clarify that ABA is part of hormonal signaling and corrected the direction of the arrow between “Lignin and suberin accumulation” and “Enhances…” to accurately reflect the regulatory relationship.

>. Section 6 (Line 427): It would be better to give this section as a subsection of section 5 and write more clearly.

Thank you for the suggestion. We have moved Section 6 as a subsection of Section 5 and revised the text to improve clarity.

>. Line 397: “They, integrate both ABA-dependent and stress-induced signaling…” – Here the authors differentiate between ABA and stress signaling, although they wrote above (line 123): “Drought-induced accumulation of ABA represents a central signaling event in stress adaptation”.

Thank you for pointing this out. We acknowledge the apparent overlap between ABA-dependent and general stress signaling. In the manuscript, ABA is considered a key component of stress signaling, while other stress-induced pathways can act independently or in parallel. We believe the current explanation accurately reflects this distinction and provides a comprehensive view of the regulatory network.

>. Lines 425-429: This sentence repeats the sentence (line 255-258).

Thank you for pointing this out. We have reviewed these sentences and retained them as is to maintain context.

>. Line 476: “hydroxycinnamoyl-CoA esters that serve as substrates for both monolignol biosynthesis and the production of phenolic suberin constituents”. However, this important intermediate is absent in the diagram (figure 1).

Thank you for this comment. We have now added hydroxycinnamoyl-CoA to Figure 1 to accurately reflect its role as an intermediate in both monolignol and phenolic suberin biosynthesis.

>. Figure 3 is called as “Cross-talk between suberin and lignin biosynthesis”. However, diagrams of suberin biosynthesis (A) and lignin biosynthesis (B) are presented separately and not connected. Moreover, the diagrams A and B are not equivalent: (A) contains only 2 MYBs (MYB36 and MYB39), whereas B contains the full set of TFs. The authors should list all abbreviations used in the diagram in the figure legend or in the abbreviation list

Thank you for this suggestion. We have added a complete list of all abbreviations used in the diagram to the figure legend. We agree that the current title “Cross-talk between suberin and lignin biosynthesis” may indicate a fully integrated diagram, while the figure presents the two pathways separately. Due to figure layout constraints and the different evidence available for MYB regulators in each pathway, we were unable to combine the diagrams into a single interconnected scheme at this stage.

>. Line 63: “Suberin and lignin are the chief biopolymers building these barriers”

Thank you for pointing this out. We have retained this sentence as it accurately summarizes.

>. Line 70: “lignin offers structural reinforcement”

Thank you for this comment. We have revised the sentence to improve clarity

>. Line 71: “they form an apoplastic barricade”

Thank you for this comment. We have revised the sentence to improve readability.

>. Line 123: “Drought-induced accumulation of ABA represents a central signaling event in stress adaptation.” – overestimation

Thank you for this comment. We have revised the sentence.

>. Line 247: “Lignin is a complex polyphenolic polymer whose biosynthesis…”

Thank you for this comment. We have revised the sentence .

>. Line 212: “Root suberization is modulated by abiotic stress conditions and closely interacts with plant stress hormone signaling pathways”. etc.

Thank you for this comment. We have revised the sentence to improve clarity

>. Line192: “In Arabidopsis, the half size transporters ABCG2, ABCG6 and ABCG20 act to ensure proper suberin barrier formation in roots and seed coats triple mutants show increased permeability and defects in suberin lamellae [49].”

Thank you for the comment. We have revised and clarified this sentence to improve readability.

>. Lines 319-324: “For example, the OsNAC5 transcription factor activates the rice gene OsCCR10 (Cinnamoyl-CoA Reductase 10), thereby improving drought tolerance at the vegetative stage in overexpressing rice plants. Through enhanced root lignin deposition (via increased H- and G-lignin biosynthesis), improved photosynthetic efficiency, and reduced leaf water loss compared to non-transgenic controls [77].”

Thank you for the comment. We have revised and clarified this sentence to improve readability.

>. Lines 348-354: “StMYB168 activates phenylpropanoid pathway genes, while StMYB24 and StMYB144 regulate aliphatic suberin biosynthetic genes, together ensuring coordinated formation of the phenolic and aliphatic suberin domains. supplies monolignols whose polymerization is controlled by a NAC and MYB regulatory hierarchy; MYB15 [66], MYB36 [27] and lignin activators MYB58/MYB63 are central nodes that translate biotic and abiotic cues 353 into localized lignification [43]”. – too long

Thank you for this comment. We have changed the original long sentence into shorter, clearer sentences to improve readability while retaining all the scientific information.

>. Incorrect use of abbreviations (not consistently after the first introduction):

“MYB transcription factors (TFs)” (line 89, 119, 363, 585)

“Casparian strip (CS)” (line 403, 530), then “CS” (403-415, 510, 518, 534-543) and “Casparian strip” throughout the text.

Thank you for pointing this out. We have reviewed the manuscript and ensured that all abbreviations are defined upon first use and used consistently throughout the text (e.g., “MYB TFs” and “CS” for Casparian strip), improving clarity and uniformity.

>. Line 257: “AC elements” – clarify this

Thank you for the comment. We have clarified that “AC elements” refer to specific cis-regulatory motifs in the promoters of lignin biosynthetic genes.

>. Strange use of Latin names of species:

multiple times – tomato (Solanum lycopersicum) (line 368, 381, 446),

or not at first mention – wheat (Triticum aestivum) (line 453)

Thank you for this comment. We have reviewed and standardized the use of Latin species names throughout the manuscript.

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

Ms. plants-4008531

Title: Regulatory Roles of MYB Transcription Factors in Root barrier under Abiotic Stress

A review of version 2 of the manuscript, unfortunately, revealed that the authors had limited themselves to correcting some parts of the text, which had been cited in the first review as examples of fail text formatting, and making some cosmetic changes. As a positive example, Figure 1 has become better. However, overall, the presentation of the material hasn't improved. It remains not well thought out and structured enough. The authors again failed to provide a clear and concise general description of MYB transcription factors, their diversity and functions. Listing TFs by numbers does not provide an idea of their regulatory mechanisms. The text remains a rather chaotic list of interesting facts about lignification, suberization and the various ways in which they are regulated. Moreover, the manuscript still looks sloppy and contains many errors. Some examples of bad revision are provided in the comments to the authors. So, I still think that this article in its current form is unacceptable for publication in an international journal, in particular in Plants. The authors should completely revise their manuscript. It could be shorter, but it should provide a clear understanding of the subject matter and identify gaps in knowledge.

Comments

Examples of bad revision:

The authors have moved Table 1 from subsection 2.1 above. As a result, if you look at the first paragraph of section 2 (lines 94-105), in the middle of the story about hormonal regulation, a phrase is inserted: “Key MYB TFs involved in root suberin and lignin regulation are summarized in (Table 1).” (line 97). However, Table 1 does not contain any information about connection of MYB TFs with hormonal pathways. What this sentence and this table are doing here remains a mystery.

Table 1: “Enhanced lignin” – – about quantity, “Enhanced suberization” – about process

Reference to Figure 3C is given first (line 92), before Figure 1.

Line 97: The abbreviation “SHR” is given without an explanation, but the explanation is given later: “SHR pathway (a key endodermal regulator)” (line 84), “SHORT-ROOT (SHR)” (lines 137, 157, 535).

The same with “SCARECROW (SCR)” (lines 137, 157, 535)

“radial-oxygen-loss (ROL) barrier” (lines 96, 403).

“5.1. Tomato (Solanum lycopersicum)” (line 619), and then “tomato (S. lycopersicum)” (lines 361, 412), tomato (406, 408).

The authors corrected “CS” in the legend to Figure 3, but did not notice the repetition” …Interaction between Interaction between…”

I want to emphasize again that these are just examples of sloppy work with the text, and correcting these errors does not eliminate the need to rewrite the article so that it is clear, well-written, and easy to read.

Comments on the Quality of English Language

The text contains grammatical and spelling errors.

Author Response

Response to Reviewer Comments for Manuscript Number plants-4008531 “Title: Regulatory Roles of MYB Transcription Factors in Root barrier under Abiotic Stress”

We would like to sincerely thank the reviewer for the careful reading our manuscript and for providing constructive suggestions to improve the quality of the paper. We have carefully revised the manuscript according to the comments. Our point-by-point responses are as follows:

>. A review of version 2 of the manuscript, unfortunately, revealed that the authors had limited themselves to correcting some parts of the text, which had been cited in the first review as examples of fail text formatting, and making some cosmetic changes. As a positive example, Figure 1 has become better.

We sincerely thank Reviewer for their second evaluation of our manuscript. We apologize that our previous revisions did not effectively address the fundamental concerns regarding the manuscript's structure, clarity, and overall scholarly presentation. We accept the reviewer's evaluation that the manuscript remains lacks a clear, synthesizing narrative. We are grateful for the specific examples provided, which have helped us understand the required depth of revision. We now properly reorganized and rewrite the manuscript, not merely cosmetic edits and revised carefully.

>. The authors again failed to provide a clear and concise general description of MYB transcription factors, their diversity and functions.

We sincerely thank the reviewer feedback. We agree that this review lacking introduction to MYB, but now in our comprehensive revision, we have added a section introducing MYB TF Classification, diversification and general functions (Lines 61-80), supported by a new schematic (Figure 1). provide the missing clear, concise description and introduction to root endodermis and exodermis (Line 34-40)

>. The text remains a rather chaotic list of interesting facts about lignification, suberization and the various ways in which they are regulated. Moreover, the manuscript still looks sloppy and contains many errors.

We thank the reviewer for the comment. We have carefully revised the manuscript to improve its structure, clarity, and readability. Additionally, we have thoroughly checked errors in the manuscript. I hope these revisions have substantially improved the overall quality and readability of the manuscript.

Examples of bad revision:

>. The authors have moved Table 1 from subsection 2.1 above. As a result, if you look at the first paragraph of section 2 (lines 94-105), in the middle of the story about hormonal regulation, a phrase is inserted: “Key MYB TFs involved in root suberin and lignin regulation are summarized in (Table 1).” (line 97). However, Table 1 does not contain any information about connection of MYB TFs with hormonal pathways. What this sentence and this table are doing here remains a mystery.

We thank the reviewer for this observation, which highlighted a flaw. The reviewer is correct the previous placement of Table 1 within a paragraph discussing hormonal regulation was confusing, as the table did not relate to hormone signaling. In the revised manuscript, we have corrected this error as the out-of-place sentence (“Key MYB TFs involved in root suberin and lignin regulation are summarized in (Table 1)”) has been removed from the paragraph on hormonal regulation. Table 1 has been moved to a new, logical location at sub-section (2.2) line 132. Therefore, the structural confusion noted by the reviewer has been resolved.

>. Table 1: “Enhanced lignin” – – about quantity, “Enhanced suberization” – about process

We thank the reviewer for this valuable clarification. We agree that the term “enhanced lignin” refers to changes in lignin quantity, whereas “enhanced suberization” more accurately describes the activation or intensification of the suberin deposition process. To ensure conceptual consistency and precise terminology, we have revised Table 1 by replacing “enhanced lignin” with “enhanced lignification”, which appropriately reflects the biological process rather than solely lignin abundance.

>. Reference to Figure 3C is given first (line 92), before Figure 1.

We thank the reviewer for pointing this out. The order of figure citations has now been corrected to follow their sequential appearance in the manuscript. The reference to Figure 3C (which is now Figure 4C as we add Figure 1) has been relocated to a more appropriate position (line 434) and Figure 1 is now cited first in the text.

>. Line 97: The abbreviation “SHR” is given without an explanation, but the explanation is given later: “SHR pathway (a key endodermal regulator)” (line 84), “SHORT-ROOT (SHR)” (lines 137, 157, 535).

We thank the reviewer for noticing this. The abbreviation SHR is now defined at its first occurrence as SHORT-ROOT (SHR), and subsequent mentions throughout the manuscript consistently (line 400-401)

<. The same with “SCARECROW (SCR)” (lines 137, 157, 535)

We thank the reviewer for identifying this issue. The abbreviation SCARECROW (SCR) is now defined at its first occurrence in the manuscript, and all subsequent mentions consistently (line 400-401)

<. “Radial-oxygen-loss (ROL) barrier” (lines 96, 403).

We thank the reviewer for noting this. The term radial oxygen loss (ROL) barrier is now defined at its first occurrence in the manuscript, and subsequent mentions consistently use the abbreviation. (line 328)

<. “5.1. Tomato (Solanum lycopersicum)” (line 619), and then “tomato (S. lycopersicum)” (lines 361, 412), tomato (406, 408).

We thank the reviewer for pointing out this. We have revised the manuscript to ensure consistent nomenclature for tomato (Solanum lycopersicum), following with common name further.

<. The authors corrected “CS” in the legend to Figure 3, but did not notice the repetition” …Interaction between Interaction between…”

We apologize for this oversight and thank the reviewer for bringing it to our attention. The repeated phrase “Interaction between Interaction between” in the legend to Figure 3 has now been corrected, and the legend has been carefully revised to remove the duplication.

We sincerely thank the reviewer for their careful reading of the manuscript and for highlighting several issues. We have revised the manuscript in accordance with the suggestions, resulting in a shortened version with improved clarity and readability.

Round 3

Reviewer 3 Report

Comments and Suggestions for Authors

The third version of the manuscript is better than the previous ones. The authors considered my comments and made the necessary corrections: added a short introduction of MYB transcription factors, made the text more logical and clearer, identified some gaps in knowledge, and made a significant number of corrections. However, the text still contains many shortcomings that require revision. See the comments.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

The text contains grammatical errors.

Author Response

Response to Reviewer Comments for Manuscript Number plants-4008531 “Title: Regulatory Roles of MYB Transcription Factors in Root barrier under Abiotic Stress” All changes made are highlighted in yellow >. The third version of the manuscript is better than the previous ones. The authors considered my comments and made the necessary corrections: added a short introduction of MYB transcription factors, made the text more logical and clearer, identified some gaps in knowledge, and made a significant number of corrections. However, the text still contains many shortcomings that require revision. Thank you for your positive feedback on the improvements made in the revised manuscript and for your continued guidance in strengthening the work. We are pleased to hear that the new version is better and that our efforts to address your previous comments. We appreciate you taking the time to provide further detailed comments to help us improve the manuscript. We have carefully review each of your points and have make all necessary revisions to ensure the text meets the highest standards of clarity and accuracy. >. Figure 1: It looks like the view is missing information for 2R-MYB. Thank you for pointing out the missing information for 2R-MYB in Figure 1. We have now updated the figure to include the 2R-MYB data. >. In addition, in the figure or its legend the authors need to insert explanations for R1, R1/2… R2R3-MYB is described in the text but is missing from the figure. Thank you for your feedback. We have updated the figure legend to include clear explanations for the structural domains (R1, R1/2, etc.) and also added R2R3-MYB in Figure. >. 2.1 The subsection title doesn't reflect the content. It provides specific information about the regulation of hormonal processes. Therefore, the title should be changed. In addition, it would be better to swap 2.1 and 2.2. Thank you for your valuable suggestion. We agree and have made the following changes. The title of subsection 2.1 has been revised to better reflect its content regarding hormonal regulation. The order of subsections 2.1 and 2.2 has been swapped as recommended to improve the logical flow of the section. >. Line 101: the sentence repeats information given in line 93. Thank you for noticing. The repeated information in line 101 (line no. is changed which is now line 207 because of making changes) has been removed. >. Line 106: transcription factor -> TF Thank you for your suggestion. "Transcription factor" is now abbreviated as "TF" after its first use in each relevant section. >. 128-130: The authors write “lignification”, but the text above describes only suberization. Lignification is present in Table 1, which is located below. It should be changed. Thank you for your suggestion. The term “lignification” has been changed along with “suberization” >. Subsections 3.1 and 3.3 have different titles but contain similar information on suberization and lignification. It should be unified. An alternative is to combine 3.1 and 3.2. Thank you for this suggestion to improve the structure. we have combined subsections 3.1 and 3.2 into a single, cohesive section that now more logically and comprehensively presents the information on suberization and lignification. >. Lines 108-116: It would be better to move this part to section 3.2. Thank you for the suggestion. The text from lines 108-116 has been moved to improve the logical flow of the manuscript. >. Line 231: “hydraulic” should be added to “conductivity” Thank you for pointing this out. We have added "hydraulic" before "conductivity" on line 424, as suggested, to use the correct technical term. >. Lines 250-253: The sentence should be moved to the text about suberin. Thank you for your suggestion. The sentence from lines 250-253 has been moved to the section discussing suberin. >. Figure 3: MYBs in the figure do not fully correspond to the text. Moreover, coniferyl alcohol and sinapyl alcohol are mentioned only in the text on lignin (197). Authors should either describe this connection in the text or redo the figure. Thank you for this observation. We have updated the manuscript text to explicitly describe the connection between the MYBs in the figure and the monolignols (coniferyl and sinapyl alcohol), ensuring full alignment between the figure and the narrative line 275-278 >. Lines 289, 327, 333: The Latin name of the species has already been given in full Tomato (Solanum lycopersicum), 267), and after this either the shorted name “S. lycopersicum” without “tomato” or just “tomato” should be used further. The second option would be preferable, given that the authors use a trivial name for rice. Thank you for your valuable suggestion. We followed 2nd option used tomato further. >. 4.1 and 4.2: The material on Arabidopsis and the species in these subsections is presented in different orders. It's better to do it the same way: Arabidopsis – species or species – Arabidopsis. Thank you for this suggestion. We followed the sequence for Arabidopsis-species and make changes accordingly. >. Lines 362-364: imperfect sentence construction Thank you for your feedback. The sentence construction in lines 362-364 has been revised for improved clarity and grammatical correctness. (Line no. can be changed in revised version) all changes highlighted. >. Line 368: “Transcriptional control is essential in phenylpropanoid.” – phenylpropanoid metabolism Thank you for this correction. We have revised accordingly. >. Lines 375 and 376-377: The sentences repeat each other. Thank you for pointing out the repetition. The redundant sentence has been removed to improve conciseness. >. Line 396: CS Thank you for the correction. We make sure to change all CS. >. Line 462: “these promoters” – what promoters? Thank you for your suggestion. We have revised line 462 to explicitly specify the promoters being referred to, ensuring clarity for the reader. >. Lines 475-480: The sentence is long and unclear Thank you for your feedback. The long and unclear sentence in lines 475-480 has been revised for better clarity and readability. >. Line 483: “salt” was omitted Thank you for noticing. The word "salt" has been added as suggested. >. Figure 5: to all abbreviations are indicated in the legend. Thank you for your comment. We have updated the legend for Figure 5 to include all necessary abbreviations. >. Failed expressions: Line 65: “DNA binding protein–protein interactions” Thank you for your suggestion. Changed and highlighted. >. Line 68: “cell morphogenesis” Thank you for your suggestion. We have marked changes and highlighted. >. Line 103: “In various crops particularly in Arabidopsis” - Arabidopsis -a crop? Thank you for this important correction. You are right that Arabidopsis is a model plant, not a crop. We have revised line 103 to clarify the comparison. >. Line 111: “lateral root production” -> formation Thank you for the suggestion. We have changed "production" to "formation" as suggested. >. Line 208: 3.4 “Transcriptional Regulation of Lignin” Thank you for the suggestion. The title of subsection 3.3 has been updated to "Transcriptional Regulation of Lignin" for clarity. >. Line 253: “lignin activators” Line 405: “suberin activation” Thank you for your note. We have changed according to your suggestion. >. Line 484: “biosynthetic genes” Line 494: “…devoted to TFs for regulating genes at transcriptional level” Line 514: “By molecular, physiological, evolutionary, and applied perspectives” Line 496: It would be better to replace “living” with “flexible”. Thank you for these detailed corrections. We have made all changes accordingly. >. Grammar: Lines 122, 165, 223, 262, 289, 290, 293, 305, 311, 313, 324, 337, 347, 368, 369, 389, 405, 409, 436, 490, 510 Thank you for these detailed corrections. We have made all changes accordingly. And also checked grammar errors in whole Manuscript.