The Molecular Biology Analysis for the Growing and Development of Hydrangea macrophylla ‘Endless Summer’ under Different Light and Temperature Conditions
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsRow 41:five categories of hydrangea? species? which is the difference between Hydrangea areborescense and H. arborescens? H. panniculata?
you need to make the species clearer!!
Row 95: Why did you choose to carry out this study on H. m. Endless summer?.Endless Summer is a brand of several cold-hardy reblooming hydrangeas.
These cultivars have a peculiar plant physiology, maybe it was better if you chose a cultivar with a more habitual behavior
Row 116:Can you explain better and in more detail the 4 treatments you have adopted? WL01, WL02 and WL03 are at T0 ? WL04?
Row 272:Where is the Table A2?
Row 328: where is the table A3?
The introduction can be improved by trying to better explain the physiology of the plant and the response of Hydrangea to light. It is necessary to explain the particular behavior towards the flowering and light of Endless summer
The discussion is too long, it is scattered. It is necessary to reorganize it trying to make more punctual references to the hydrangea
Author Response
Dear reviewer:
Thank you very much for your hard work to review my manuscript. We have carefully revised our manuscript in response to your feedback and provided detailed responses to each comment. The following are our responses to the reviewer's comments.
Q1：
Row 41:five categories of hydrangea? species? which is the difference between Hydrangea areborescense and H. arborescens? H. panniculata?
R1：
We apologize for the oversight in our manuscript regarding the classification of Hydrangea species. We have now clarified the species names to ensure accuracy.
We appreciate the opportunity to further enhance the quality of our contribution to the scientific literature.
Q2：
Row 95: Why did you choose to carry out this study on H. m. Endless summer?.Endless Summer is a brand of several cold-hardy reblooming hydrangeas.
These cultivars have a peculiar plant physiology, maybe it was better if you chose a cultivar with a more habitual behavior
R2：
The decision to focus our study on H. macrophylla ‘Endless Summer’ was grounded in its widespread cultivation and significant usage in the Beijing region. This cultivar boasts a robust cold-hardiness, rendering it particularly well-suited to the northern climatic conditions prevalent in China. However, despite its prevalent cultivation, it exhibits suboptimal performance during the summer months. Our preliminary observations indicate that under semi-shaded environments during summer, these cultivars display favorable growth characteristics. Thus, by dissecting the underlying mechanisms behind this phenomenon, we seek to leverage this knowledge to improve their performance during the summer months, thereby refining the cultivar to its fullest potential. We aim to lay a foundation for enhancing its overall environmental adaptability through genetic breeding methods.
In summary, our choice of studying H. macrophylla ‘Endless Summer’ is not merely a reflection of its widespread cultivation or cold-hardiness, but rather a strategic approach aimed at addressing its existing limitations and enhancing its overall suitability through targeted genetic interventions. We trust that our rationale aligns with the objectives of our research and provides a compelling justification for our choice of cultivar.
Q3：
Row 116:Can you explain better and in more detail the 4 treatments you have adopted? WL01, WL02 and WL03 are at T0 ? WL04?
R3：
We sincerely apologize for any lack of clarity in our description of the treatments. Allow us to provide a comprehensive explanation of the four treatments adopted in our study.
The experimental treatments were designed to simulate varying degrees of shading conditions to investigate their effects on plant growth and physiological responses. We established three distinct microclimate conditions, each representing different levels of shading, namely WL01, WL02, and WL03. Additionally, we introduced a transitional treatment, WL04, to examine the recovery response of plants transferred from full sunlight to partial shade.
WL01: This treatment represents full shading conditions, with the highest level of shade. We defined WL01 as the "full shade" condition, where the plants experienced minimal direct sunlight exposure. Light intensity was recorded at 351 μmol∙m-2·s-1, and the average leaf surface temperature was 34.5°C.
WL02: In contrast to WL01, WL02 represents partial shading conditions, termed as the "half shade" condition. Here, plants received moderate sunlight exposure, with a recorded light intensity of 751 μmol∙m-2·s-1 and an average leaf surface temperature of 36.3°C.
WL03: This treatment denotes full sunlight exposure, representing the "full light" condition. Plants under WL03 experienced the highest levels of light intensity, recorded at 1503 μmol∙m-2·s-1, accompanied by an average leaf surface temperature of 41.7°C.
WL04: Unlike the other treatments, WL04 involves a transition from full sunlight (WL03) to partial shade (WL02). This transition period allows us to observe the recovery response of plants following exposure to high light intensity. Samples for WL04 were collected after one week of acclimatization to WL02 conditions, with environmental parameters matching those of WL02.
In summary, our experimental design encompassed a spectrum of shading conditions, ranging from full shade to full sunlight, with a transitional phase to investigate plant recovery.
We hope this detailed explanation clarifies the treatments adopted in our study. Please feel free to reach out if further clarification is required.
Q4-5：
Row 272:Where is the Table A2?
Row 328: where is the table A3?
R4-5：
In response to your inquiries, we would like to clarify that the tables referred to as A2 and A3 are indeed included in the supplementary information section of the manuscript. We have renumbered these tables as Table S2 and Table S3, respectively, to align with common conventions for supplementary materials.
The decision to include these tables in the supplementary information section was made to streamline the main body of the manuscript and to maintain focus on the primary findings and analyses. Placing them in the supplementary section allows readers to access supplementary details without cluttering the main text.
Q6：
The introduction can be improved by trying to better explain the physiology of the plant and the response of Hydrangea to light. It is necessary to explain the particular behavior towards the flowering and light of Endless summer.
R6：
In response to your concerns, we would like to clarify the limitations of our study. The experimental materials utilized in our research were planted two years prior to this manuscript. While we acknowledge the importance of incorporating physiological indicators, such as photosynthetic parameters, into our analysis, we encountered challenges in obtaining live measurements due to the current conditions of the experimental materials' growing environment. Specifically, the conditions necessary for accurate physiological assessments were not fully met, rendering live measurements unfeasible at this time. We emphasize that while we recognize the significance of physiological data in elucidating the mechanisms underlying plant responses to environmental stimuli, the constraints imposed by the experimental conditions prevented us from including these measures in our analysis.
We hope that these explanations adequately address your concerns. Thank you once again for your valuable feedback, and we look forward to your continued guidance throughout the revision process.
Q7：
The discussion is too long, it is scattered. It is necessary to reorganize it trying to make more punctual references to the hydrangea
R7：
In response to your suggestion, we have made significant revisions to the discussion section. While we haven't listed all the modifications here, they are clearly marked in red in the revised document.
Specifically addressing your concern about the length and organization of the discussion, we have restructured it to include punctual references to hydrangea throughout. This reorganization involves the introduction of subheadings, which serve to delineate different aspects of the discussion and enhance its coherence.
We believe that these enhancements have significantly improved the readability and comprehension of the manuscript. We hope that you find the revised version satisfactory and suitable for publication.
Thank you once again for your valuable feedback and consideration. Best regards.
Sincerely yours,
Yingmin Lyu
Reviewer 2 Report
Comments and Suggestions for AuthorsLi et al has reported the manuscript “The Molecular Biology Analysis for the Growing and Development of Hydrangea macrophylla 'Endless Summer' under Different Light and Temperature Conditions”. The topic is generally of interest to the scientific community and industry. The manuscript describes bioinformatics investigations and qPCR in different conditions.
I see some aspects which should be improved before it is accepted for publication.
Comments:
1) line 29, what is meaning of the phrase “additional energy substances”?
2) Line 438, what is “I” ?
3) Submit transcriptome data in NCBI and mention its accession number in manuscript.
4) Please provide RNAseq validation data by qPCR.
4) Fig 6. should be related with KEGG, not GO. Please describe Fig 6 in its legend (Qvalue, [0,1,3), [1,3,2] codes …etc.
5) Figure A1, Figure A2 , Table A2, is missing.
Please write the manuscript carefully.
Comments on the Quality of English LanguageNeed to be improved.
Author Response
Dear Reviewer:
Thank you very much for your hard work to review my manuscript.We have carefully revised our manuscript in response to your feedback and provided detailed responses to each comment. The following are our responses to the reviewer's comments.
Q1：
line 29, what is meaning of the phrase “additional energy substances”?
R1：
Regarding your inquiry about the phrase "additional energy substances" in line 29, we appreciate the opportunity to clarify and elaborate on this concept.
In our study, we investigated the genetic mechanisms underlying flowering in plants, particularly focusing on the role of various genes involved in hormone signal transduction and energy metabolism. As you rightly pointed out, the expression of genes such as BGLUs (Beta-glucosidases) and TPSs (Trehalose-6-phosphate synthases) provides additional energy substances to support flowering.
To elaborate, Beta-glucosidases (BGLUs) are enzymes involved in the breakdown of complex carbohydrates, such as cellulose, into simpler sugars like glucose, which serve as essential energy sources for plant growth and development. Trehalose-6-phosphate synthases (TPSs), on the other hand, are enzymes responsible for the synthesis of trehalose-6-phosphate, a critical metabolite involved in energy metabolism and stress response in plants.
During the transition to flowering, plants undergo significant metabolic changes to allocate resources towards reproductive processes. The upregulation of BGLUs and TPSs genes during this stage facilitates the mobilization and utilization of stored carbohydrates, thereby providing the necessary energy substrates to support floral initiation and development.
In summary, the expression of BGLUs and TPSs genes contributes to the provision of energy substrates essential for flowering, highlighting the intricate interplay between hormonal regulation and metabolic processes in plant development.
We hope this clarification adequately addresses your query and enhances the clarity and understanding of our manuscript.
Q2：
Line 438, what is “I” ?
R2：
Thank you for bringing the typo at line 438 to our attention. Upon review, we found that the letter “I” was inadvertently included due to an error. We have thoroughly checked the manuscript to correct this and similar issues to ensure the clarity and professionalism of our text. We apologize for any confusion this may have caused.
Q3：
Submit transcriptome data in NCBI and mention its accession number in manuscript.
R3：
Thank you for your valuable feedback. We have now submitted the transcriptome data to the National Center for Biotechnology Information (NCBI), and the corresponding accession number is PRJNA1109561.
Q4：
Please provide RNAseq validation data by qPCR.
R4：
In response to your request for RNAseq validation data by qPCR, in the supporting information (Figure S1), we have provided fluorescence quantitative results as obtained through qPCR analysis. These data serve as a validation of the RNAseq findings presented in the main manuscript.
Q5：
Fig 6. should be related with KEGG, not GO. Please describe Fig 6 in its legend (Qvalue, [0,1,3), [1,3,2] codes …etc.
R5：
We sincerely apologize for the oversight regarding Fig 6 and the inappropriate utilization of KEGG instead of GO. We have duly rectified these issues in the revised manuscript.
Regarding Fig 6, it now incorporates KEGG-related data as suggested. In the legend of Fig 6, Qvalue represents the adjusted p-value, indicating the significance of the findings. The values within the brackets [0,1,3) denote the range of one parameter, while [1,3,2] signifies another set of parameters, each crucial for understanding the depicted data.Furthermore, to ensure better understanding, we have included an explanatory note in manuscript.
Q6：
Figure A1, Figure A2 , Table A2, is missing.
R6：
We have carefully considered your comments regarding the missing figures and table and have made the necessary revisions.
In response to your concern, we have included the missing figures and table in the supporting information section of the manuscript. Specifically, we have added Figure S1, Figure S2, and Table S2 to provide the complete set of data and visual representations necessary for a comprehensive understanding of our study.
Thank you once again for your constructive feedback, which has undoubtedly strengthened the quality of our manuscript. We are confident that the inclusion of Figure S1, Figure S2, and Table S2 will enhance the comprehensiveness and clarity of our study for the benefit of the readership.
Q7：
Please write the manuscript carefully.
R7：
In response to your feedback regarding the need to "write the manuscript carefully," we assure you that we are committed to upholding the highest standards of academic writing in our work. To ensure clarity, coherence, and precision, we have taken several measures during the manuscript preparation process:
Thorough Literature Review: We have conducted an extensive review of the existing literature relevant to our study, ensuring that our research is situated within the broader context. By synthesizing previous findings and identifying gaps in knowledge, we have positioned our study to make a meaningful contribution to the field.
Clear Objectives and Methodology: Our manuscript clearly outlines the objectives of our research and the methodology employed to achieve these objectives. We have provided detailed descriptions of our research design, data collection methods, and analytical techniques, allowing readers to understand the rationale behind our approach and the validity of our findings.
Precise and Concise Writing: We have paid meticulous attention to the clarity and conciseness of our writing, ensuring that each sentence contributes directly to the overall coherence of the manuscript. Through careful editing and revision, we have eliminated unnecessary jargon and verbosity, making the text more accessible to a diverse audience of readers.
Thorough Proofreading: Prior to submission, our manuscript underwent multiple rounds of proofreading to identify and correct any grammatical errors, typos, or inconsistencies in formatting. We have strived to present our research findings in a polished and professional manner, reflecting our commitment to excellence in scholarship.
Q8：
Comments on the Quality of English Language
Need to be improved.
R8：
Thank you for taking the time to review our manuscript. We appreciate your feedback regarding the quality of the English language, and we have made the necessary improvements as suggested to ensure clarity and coherence in language usage.
We believe that these enhancements have significantly improved the readability and comprehension of the manuscript. We hope that you find the revised version satisfactory and suitable for publication.
Thank you once again for your valuable feedback and consideration. Best regards.
Sincerely yours,
Yingmin Lyu
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsRows 40-43: At present, horticulturists have cultivated (five) four categories of hydrangea, including Hydrangea. macrophylla, Hydrangea arborescens, Hydrangea paniculata, Hydrangea quercifolia and Hydrangea serrata.
Author Response
Dear Reviewer:
Thank you very much for your hard work to review my manuscript.We have carefully revised our manuscript in response to your feedback and provided detailed responses to each comment. Below are our responses to your comments.
Q1：
Rows 40-43: At present, horticulturists have cultivated (five) four categories of hydrangea, including Hydrangea. macrophylla, Hydrangea arborescens, Hydrangea paniculata, Hydrangea quercifolia and Hydrangea serrata.
R1:
We apologize for the spelling errors, which now have been corrected as per your advice. Thanks for your correction.
Best regards.
Sincerely yours,
Yingmin Lyu
E-mail: [email protected]
Reviewer 2 Report
Comments and Suggestions for Authors1] Please mention accession number- PRJNA1109561 in appropriate place in the manuscript.
2] In Figure S1, Please provide information regarding house keeping gene and show differential gene expression in terms of Log2FC, but not in terms of FPKM or RPKM value.
3] Please include explanation of Fig 6 in its legend in concise way.
Comments on the Quality of English Languageneed to be improved.
Author Response
Dear Reviewer:
Thank you very much for your hard work to review my manuscript. We have carefully revised our manuscript in response to your feedback and provided detailed responses to each comment. Below are our responses to your comments.
Q1:
Please mention accession number-PRJNA1109561 in appropriate place in the manuscript.
R1:
We sincerely appreciate your thorough review and constructive feedback. The accession number is now included in Rows 215-216 of the manuscript.
Q2：
In Figure S1, Please provide information regarding house keeping gene and show differential gene expression in terms of Log2FC, but not in terms of FPKM or RPKM value.
R2：
Thank you for your thoughtful review of our manuscript. We sincerely apologize for any lack of clarity in our description of regarding house keeping gene.
We have mentioned in Row 191 of the manuscript that the house keeping gene is RPL34 and the RT-qPCR primer sequence for RPL34 is included in Table S1. We hope this explanation clarifies the information regarding house keeping gene.
For another revision, Figure S1 now illustrates differential gene expression using Log2 FC.
Q3:
Please include explanation of Fig 6 in its legend in concise way.
R3：
We appreciate your valuable feedback regarding enhance the legend of Figure 6. We have accordingly added a simple and clear explanation to the legend in Figure 6.
Q4:
Comments on the Quality of English Language need to be improved.
R4:
Thank you for your careful consideration of our manuscript.
We apologize that our previous revisions fell short of your expectations. This time, we have made more extensive modifications to the manuscript to ensure linguistic accuracy.
Thank you for your attention to these points, and we remain open to any further feedback or suggestions you may have.
Best regards.
Sincerely yours,
Yingmin Lyu
E-mail: [email protected]
