How Does Rice Cope with High-Temperature Stress During Its Growth and Development, Especially at the Grain-Filling Stage?

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

General comments:
The manuscript provides a comprehensive and well-structured review of the impact of high-temperature

events posing severe challenges to rice growth. Rice is highly susceptible to the adverse effects of high-
temperature stress throughout its growth cycle, from seed germination to harvest. The authors

summarize how rice activates complex physiological and biochemical responses, including heat-related
signaling pathways and transcriptional regulatory networks, to cope with high-temperature stress and
provide an outlook on future rice high-temperature defense measures. The manuscript would benefit
from minor grammatical refinements, enhanced figure clarity, and updated references.
Comments for authors:
Line 2: The title should be refined for grammatical correctness. Suggested revision: "How does rice
cope with high-temperature stress during its growth and development, especially at the grain-filling
stage?"
Line 9-10: The phrase "posing severe challenge to food security" should be changed to "posing a severe
challenge to food security."
Line 15-16: reduced grain weight, increased grain chalkiness, and a higher degree of chalk formation.
Line 20-21: The statement "remain poorly understood" needs clarification.
Line 30-32: It should be updated with the latest data or a meta-analysis source.
Lines 40-42: offer perspectives on future research directions, aiming to establish a theoretical
foundation for mitigating heat stress in rice.
Line 47-48: The statement "prolonged exposure to high temperatures can delay germination" should be
supported with specific temperature ranges and studies.
Figure 1 : Needs better resolution or color contrast for clarity.
Line 83: particularly during the heading and flowering periods, which are highly sensitive to
temperature fluctuations."
Line 128-130: "High temperatures can also damage the structure and function of the plasma
membrane..." needs a figure reference or more mechanistic details.
Line 164-166: Consider whether OsbZIP58 mutants exhibit pleiotropic effects beyond starch
metabolism.
Line 206-209: The WCR1-MT2b interaction should be elaborated with details on its specific molecular
function.
Line 212-214: "Phytohormones, as key signaling molecules..." could include recent findings on abscisic
acid’s cross-talk with salicylic acid or ethylene under heat stress.
Line 267-269: "Overexpression of OsHsp70-2 and OsHsp40-1 can improve the appearance quality and
yield..." needs supporting data or citation.
Figure 2: Should include clearer labeling of molecular components.
Line 329-332: The deep irrigation method should reference field studies showing yield improvements.
Line 340-344: Consider discussing the foliar application of heat-protectant compounds like glycine
betaine or proline.
Some references appear outdated. Consider replacing them with recent studies (post-2020).
Conclusion: It should be more precise and reflect the key highlights of the study. Rewrite the section.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

Moderate English editing is required 

Author Response

General comments:

The manuscript provides a comprehensive and well-structured review of the impact of high-temperature events posing severe challenges to rice growth. Rice is highly susceptible to the adverse effects of high-temperature stress throughout its growth cycle, from seed germination to harvest. The authors summarize how rice activates complex physiological and biochemical responses, including heat-related signaling pathways and transcriptional regulatory networks, to cope with high-temperature stress and provide an outlook on future rice high-temperature defense measures. The manuscript would benefit from minor grammatical refinements, enhanced figure clarity, and updated references.

Response: We sincerely thank the Reviewer for an overall positive response to our manuscript.

Comments for authors:

Line 2: The title should be refined for grammatical correctness. Suggested revision: "How does rice cope with high-temperature stress during its growth and development, especially at the grain-filling stage?"

Response: Thank you very much for your good suggestions.We have modified the title in the revised manuscript, see the red font in the title of the new manuscript.

Line 9-10: The phrase "posing severe challenge to food security" should be changed to "posing a severe challenge to food security."

Response: We have corrected "posing severe challenge to food security" to "posing a severe challenge to food security" .

Line 15-16: reduced grain weight, increased grain chalkiness, and a higher degree of chalk formation.

Response: Thank you very much. We have corrected it in new manuscript.

Line 20-21: The statement "remain poorly understood" needs clarification.

Response: Thank you for your careful reading. We have revised the statement to clarify the specific mechanisms that are still under investigation and included recent literature to support the explanation.

 

Line 30-32: It should be updated with the latest data or a meta-analysis source.

Response: We sincerely thank you for your affirmation and good advice. We have updated the section with the most recent data and included a meta-analysis published in 2025.

Lines 40-42: offer perspectives on future research directions, aiming to establish a theoretical foundation for mitigating heat stress in rice.

Response: Thank you very much. We have expanded this section to include specific future research directions and potential mitigation strategies.

Line 47-48: The statement "prolonged exposure to high temperatures can delay germination" should be supported with specific temperature ranges and studies.

Response: Thanks.We have added temperature thresholds and cited relevant studies that support this statement.

Figure 1 : Needs better resolution or color contrast for clarity.

Response: Figure 1 has been replaced with a higher-resolution version with improved color contrast and clearer labeling.

Line 83: particularly during the heading and flowering periods, which are highly sensitive to temperature fluctuations."

Response: Thanks. We have incorporated the suggested phrasing in Line 88-89 in new manuscript.

Line 128-130: "High temperatures can also damage the structure and function of the plasma membrane..." needs a figure reference or more mechanistic details.

Response: Thank you very much for your advice. We have elaborated on the mechanisms underlying plasma membrane damage under heat stress to lines137-143 of the new manuscript.

Line 164-166: Consider whether OsbZIP58 mutants exhibit pleiotropic effects beyond starch metabolism.

Response: Thank you for your careful reading. We have reviewed recent studies and added a discussion on the potential pleiotropic effects of OsbZIP58 mutants in the lines179-183 of the new manuscript.

Line 206-209: The WCR1-MT2b interaction should be elaborated with details on its specific molecular function.

Response: Thank you for your suggestion.We expanded the section with details on the molecular function of WCR1-MT2b.

 

Line 212-214: "Phytohormones, as key signaling molecules..." could include recent findings on abscisic acid’s cross-talk with salicylic acid or ethylene under heat stress.

Response: Thank you for your careful reading and constructive comments. We have incorporated recent findings on phytohormone cross-talk, particularly between abscisic acid, salicylic acid, and ethylene.

Line 267-269: "Overexpression of OsHsp70-2 and OsHsp40-1 can improve the appearance quality and yield..." needs supporting data or citation.

Response: Thank you. Supporting data and relevant citations have been added.

Figure 2: Should include clearer labeling of molecular components.

Response: Thanks for your suggestion. Figure 2 has been updated with clearer labels and enhanced visual clarity.

Line 329-332: The deep irrigation method should reference field studies showing yield improvements.

Response: We have added citations to recent field studies demonstrating the effectiveness of deep irrigation in improving yield under heat stress. See new reference 79

Line 340-344: Consider discussing the foliar application of heat-protectant compounds like glycine betaine or proline.

Response: We have included a new subsection discussing the role of foliar-applied heat-protectant compounds, with relevant studies cited.

Some references appear outdated. Consider replacing them with recent studies (post-2020).

Response: We have updated several references, replacing older ones with more recent publications from 2020 onwards.

Conclusion: It should be more precise and reflect the key highlights of the study. Rewrite the section.

Response: The conclusion has been rewritten to provide a concise summary of key findings, practical implications, and future research perspectives.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript titled "How do rice cope with high temperature stress during its growth and development, especially at the grain-filling stage" provides a comprehensive review of high-temperature stress in rice, particularly its impact on the grain-filling stage.

The topic is highly relevant, given the increasing global temperatures and their effects on rice production. The manuscript is well-structured and includes a thorough discussion of physiological, molecular, and agronomic responses.

However, the study needs a clearer identification of the knowledge gaps and how this review contributes to advancing the field.

1. There are plenty of review papers discussing heat or high temperature stress, so what makes this paper stand different? The review would benefit from explicitly stating: what specific knowledge gap it aims to fill, how this review differs from previous reviews on the same topic, what novel insights or new perspectives it provides.
2. To strengthen its originality, the authors should:

-       compare their review with recent similar reviews and highlight what new insights they bring.

-       identify underexplored areas in the study of heat stress in rice. Recent studies discuss multi stress tolerance, such as combined heat and drought stress, heat and salinity stress, etc.

-       emphasize any new perspectives they introduce, such as unique regulatory pathways, recently identified genes, or novel agronomic strategies.

 

3. As this is a review paper, there is no experimental methodology. However, the authors should clarify the criteria for selecting the studies reviewed. Specifically:

• How were studies chosen for inclusion?
• Were there any systematic selection criteria (e.g., studies from the last five years, only peer-reviewed articles, etc.)?
• Are there any biases in the selection of studies that should be acknowledged?

 

4. The conclusions summarize the key points well but could be improved by:

• Clearly stating what specific knowledge gaps remain in the field.
• Suggesting future research directions with more specificity. Rather than broadly stating that "further study is needed on molecular mechanisms," the authors could suggest particular genes or pathways that need further exploration.
• Indicating how scientists can address these gaps—whether through new breeding techniques, genome editing, or improved agronomic practices.

5. If applicable, summary tables comparing different stress responses at each growth stage would improve readability.

Although the work scientifically sound, well and comprehensively organized, I do think that the Authors need to identify what the significant contribution of this paper to the field.

Below I write several points to be consider for improvement:

L24 Please add "identify knowledge gap"
L38-42 The authors mentioned that the paper "summarizes". I suggest that the authors can identify the knowledge gap, thus could provide "fields" that have not be explored
And finally, from this review, the authors can specifically states "whats the gap" in the conclusion and suggest the readers (scientists) what they can do to fill the gap.

With these revisions, the paper would make a stronger contribution to the field.

Author Response

Response to the comments by Reviewer 1:

The manuscript titled "How do rice cope with high temperature stress during its growth and development, especially at the grain-filling stage" provides a comprehensive review of high-temperature stress in rice, particularly its impact on the grain-filling stage.

The topic is highly relevant, given the increasing global temperatures and their effects on rice production. The manuscript is well-structured and includes a thorough discussion of physiological, molecular, and agronomic responses.

Response: We sincerely appreciate the reviewer’s thorough and constructive feedback. We have carefully considered all suggestions and made substantial revisions to enhance the clarity, originality, and overall quality of the manuscript. Below are our point-by-point responses.

However, the study needs a clearer identification of the knowledge gaps and how this review contributes to advancing the field.

• There are plenty of review papers discussing heat or high temperature stress, so what makes this paper stand different? The review would benefit from explicitly stating: what specific knowledge gap it aims to fill, how this review differs from previous reviews on the same topic, what novel insights or new perspectives it provides.

Response: Thank you for highlighting this important aspect. We have revised the introduction to clearly articulate the knowledge gaps addressed in this review. Unlike previous reviews that primarily focus on general heat stress responses, our review emphasizes the grain-filling stage—an especially vulnerable period affecting yield and quality—and integrates recent molecular findings (post-2020), novel regulatory pathways, and underexplored multi-stress interactions. Additionally, we highlight emerging agronomic strategies not covered in prior works.

• To strengthen its originality, the authors should:

- compare their review with recent similar reviews and highlight what new insights they bring.

- identify underexplored areas in the study of heat stress in rice. Recent studies discuss multi stress tolerance, such as combined heat and drought stress, heat and salinity stress, etc.

- emphasize any new perspectives they introduce, such as unique regulatory pathways, recently identified genes, or novel agronomic strategies.

Response: We have added a new subsection in the introduction comparing our review with recent works. Our paper differs by providing updated insights into:

Recently identified genes and transcription factors involved in heat tolerance at the grain-filling stage.

Multi-stress tolerance mechanisms, with a dedicated section discussing combined heat and drought, and heat and salinity stress.

Innovative agronomic practices, including foliar applications and genome editing strategies, which offer practical solutions for breeders and farmers.

• As this is a review paper, there is no experimental methodology. However, the authors should clarify the criteria for selecting the studies reviewed. Specifically:

• How were studies chosen for inclusion?
• Were there any systematic selection criteria (e.g., studies from the last five years, only peer-reviewed articles, etc.)?
• Are there any biases in the selection of studies that should be acknowledged?

Response: We appreciate the reviewer’s suggestion. We explain the selection process in the introduction section. Studies were selected based on the following criteria:

Peer-reviewed articles published between 2018 and 2024, ensuring up-to-date information.

Priority given to studies with high-impact findings on rice’s physiological, molecular, and agronomic responses to heat stress.

Databases searched: Web of Science, Scopus, and PubMed, using keywords such as "rice," "heat stress," "grain-filling stage," "molecular mechanisms," and "agronomic practices."

We acknowledge potential publication bias, as studies with positive findings may be more likely to be published.

• The conclusions summarize the key points well but could be improved by:

• Clearly stating what specific knowledge gaps remain in the field.
• Suggesting future research directions with more specificity. Rather than broadly stating that "further study is needed on molecular mechanisms," the authors could suggest particular genes or pathways that need further exploration.
• Indicating how scientists can address these gaps—whether through new breeding techniques, genome editing, or improved agronomic practices.

Response: We have rewritten the conclusion to:

Clearly identify knowledge gaps, such as limited understanding of cross-tolerance mechanisms and long-term field-level impacts of high temperatures.

Recommend targeted research on underexplored genes and their regulatory networks.

Suggest practical ways to address these gaps, including CRISPR/Cas9-based breeding, deep learning for stress prediction, and farmer-oriented agronomic innovations.

• If applicable, summary tables comparing different stress responses at each growth stage would improve readability.

Although the work scientifically sound, well and comprehensively organized, I do think that the Authors need to identify what the significant contribution of this paper to the field.
Response: We have added two summary tables:

Table 1: High temperature threshold for the development of the rice plant at different growth stages

Table 2: List of genes conferring high temperature tolerance in rice

(6) Below I write several points to be consider for improvement:

L24 Please add "identify knowledge gap"

Response: Revised as suggested. The sentence now explicitly states the identification of knowledge gaps.

L38-42 The authors mentioned that the paper "summarizes". I suggest that the authors can identify the knowledge gap, thus could provide "fields" that have not be explored

Response: We have modified the text to highlight underexplored areas and provide suggestions for future research fields.

And finally, from this review, the authors can specifically states "whats the gap" in the conclusion and suggest the readers (scientists) what they can do to fill the gap.

With these revisions, the paper would make a stronger contribution to the field.

Response: The conclusion has been revised to clearly state remaining knowledge gaps and propose actionable research directions, emphasizing molecular targets and field practices.

Reviewer 3 Report

Comments and Suggestions for Authors

This review-paper is focused on the rice responses and tolerance to high temperature stress during its growth and development, especially at the grain-filling stage. The paper is well-organized and prepared as well as presents interesting research directions and the most important results.

However, in order to improve the manuscript, I recommend the following minor revisions:

• I recommend adding some new citations regarding the molecular basis of the high temperature stress in the rice plants, especially transcriptomic, proteomic, interactomic results.
• Further prospects for biotechnological agronomic treatments to obtain more resistant rice genotypes should be described in greater detail.
• The English style and grammar should be corrected by the native speaker - specialist in the research discipline. 

Comments on the Quality of English Language

The paper should be corrected by the native speaker - specialist in the research discipline. 

Author Response

Comments and Suggestions for Authors

This review-paper is focused on the rice responses and tolerance to high temperature stress during its growth and development, especially at the grain-filling stage. The paper is well-organized and prepared as well as presents interesting research directions and the most important results.

Response: We sincerely appreciate the reviewer’s positive evaluation of our manuscript and valuable suggestions. We have addressed each comment carefully to improve the overall quality of the paper. Our detailed responses are provided below:

However, in order to improve the manuscript, I recommend the following minor revisions:

• I recommend adding some new citations regarding the molecular basis of the high temperature stress in the rice plants, especially transcriptomic, proteomic, interactomic results.

Response: Thank you for this helpful suggestion. We have updated the manuscript by including recent studies (post-2020) that explore the transcriptomic, proteomic, and interactomic responses of rice under high-temperature stress. These additions provide a more comprehensive understanding of the molecular mechanisms involved.See Reference 54, Section 3.2.

• Further prospects for biotechnological agronomic treatments to obtain more resistant rice genotypes should be described in greater detail.

Response: We appreciate this valuable recommendation. We have expanded the section on future perspectives by detailing recent advancements in biotechnological and agronomic approaches, including gene editing (e.g., CRISPR/Cas9), molecular breeding, and innovative field management practices aimed at developing high-temperature-tolerant rice genotypes.

• The English style and grammar should be corrected by the native speaker - specialist in the research discipline. 

Response: We have thoroughly revised the manuscript to improve grammar, clarity, and overall readability. A native English speaker with expertise in plant stress physiology has reviewed and refined the language to meet the required standards.

Comments on the Quality of English Language

• The paper should be corrected by the native speaker - specialist in the research discipline. 

Response: We are grateful for the reviewer’s constructive feedback, which has significantly strengthened the quality and clarity of our manuscript.