Effects of Different Levels of Physical Damage Combined with Fungal Induction on Agarwood Formation

Round 1

Reviewer 1 Report

The research article presented by Weiwei Zhao and co-authors titled “Effects of different levels of physical damage combined with fungal induction on agarwood formation” is relevant to Forests Journal. The study is well-founded and well-organized; the experiments were made correctly by using adequate methods, and the obtained results were well presented and discussed. The paper has both fundamental and practical value and will attract the interest of readers. Application of fungi isolated from trees is of special interest and has a huge potential as an eco-friendly approach for good-quality agarwood formation.

However, there are specific comments to the authors are outlined below that are necessary to improve the manuscript:

Material and Methods. The detailed information should be provided. How did you isolate fungus? Is it endophytic fungus? From which material, trees? Did you use healthy trees or infected? And which fungus exactly was selected for further experiments, Latin names?  (i.e., in Table 1). If even it is a technical secret the name of fungus should be given.

Line 13. ‘sinesis’ must be italicized.

Line 16. Use ‘microbial strains’ instead of ‘strains”.

Lines 18-19. ‘A. sinensis’ must be italicized.

Line 33. ‘therefore’ must begin with a capital letter.

Line 35. ‘(IUCN)’ give the full name.

Line 36. ‘CITES’ give the full name.

Line 64. Must be ‘endophytic’ not ‘endo-phytic’.

Line 66. ‘which’ must begin with a capital letter. Probably, it will be better to rephrase the whole sentence (lines 66-68).

Line 70. ‘sinesis’ must be italicized. Please, check everywhere in the text.

Line 72. ‘Fusarium solani’ must be italicized.

Line 73. ‘Aspergillus penicillioides’ must be italicized.

Line 82. Put a dot instead of a comma before ‘However’.

Lines 84. “baimuxinal and agarotetro” do not italicize the name of these compounds.

There are many typographical and stylistic errors throughout the text (including missing spaces). Check please.

Best wishes

Author Response

Dear Editor and Reviewers:

We sincerely appreciate reviewer for your valuable comments and suggestions. The review comments are laid out below in bold and specific concerns have been numbered. The point-to-point responses to the reviewer's comments are marked in red and presented following.

1. Material and Methods. The detailed information should be provided. How did you isolate fungus? Is it endophytic fungus? From which material, trees? Did you use healthy trees or infected? And which fungus exactly was selected for further experiments, Latin names?  (i.e., in Table 1). If even it is a technical secret the name of fungus should be given.                                                                                         Response 1: Thank you very much for pointing out this important issue. We agree with your opinion. The original text has been revised and improved to include a detailed description of the selected materials, the method of isolation and identification, and the steps of induction operation, which are elaborated in (line 153 to line 228, page 3 to page 4). The material utilized is agarwood produced under natural conditions, without the use of artificial induction technology, as indicated in (line 127 to line 129, page 2). Three types of fungi exhibiting high biological activity were chosen, and their specific names have been included in the methods for identification and isolation, as outlined in Table 1. Please consult (line 202 to line 204, page 3 and line 211 to line 217, page 4) for further details.
2. Line 13. ‘sinesis’ must be italicized.                                                       Response 2: We were really sorry for our careless mistakes. Thank you for your reminder. We have thoroughly proofread the entire text and made the necessary corrections. Please refer to the details provided at (line 22, page 1; line 108, page 3; line 339; line 371, page 9; line 403, page 11; line 408, page 11; line 437, page 12; line 467, page 13; line 519, page 14).
3.  Line 16. Use ‘microbial strains’ instead of ‘strains”.                      Response 3: Thank you for bringing this to our attention. The    correction has been made and can be found on (line 18，page1).
4. Lines 18-19. ‘A. sinensis’ must be italicized.                                     Response 4: Thank you for bringing this to our attention. We have thoroughly proofread the entire text and made the necessary corrections. Please refer to the details provided at (line 16, page 1; line 22, page 1; line 108, page 3; line 408, page 11).
5. Line 33. ‘therefore’ must begin with a capital letter.                        Response 5: We feel sorry for our carelessness. In our resubmitted manuscript, the typo is revised. Thanks for your correction. as outlined in the (line 57, page 2).
6. Line 35. ‘(IUCN)’ give the full name.                                                Response 6: We appreciate your attention to this matter, and we have provided the complete name of the (IUCN) as the International Union for the Conservation of Nature and Natural Resources. See (line 60, page 2).
7. Line 36. ‘CITES’ give the full name.                                                  Response 7: We appreciate your attention to this matter, and we have provided the complete name of the CITES as the Convention on International Trade in Endangered Species of Wild Fauna and Flora. See (line 62, page 2).
8. Line 64. Must be ‘endophytic’ not ‘endo-phytic’                                  Response 8: Thank you for bringing attention to the issue in the article. We feel sorry for our carelessness. We have thoroughly reviewed the entire text for similar problems and have made comprehensive revisions to the phrases or sentences with similar issues throughout the text, as outlined in (line 114, page 3; line 525, page 14).
9. Line 66. ‘which’ must begin with a capital letter. Probably, it will be better to rephrase the whole sentence (lines 66-68).                         Response 9: We sincerely appreciate the valuable comments. We have re-written this part according to the Reviewer’s suggestion: The researchers introduced endophytic fungal mycelia or endophytic fungal culture supernatants, which were isolated and purified from agarwood, into the trunks of healthy Aquilaria trees in order to simulate the pathological conditions observed in wild Aquilaria trees through biological stress. see (line 95 to line 99, page 2 to page 3).
10. Line 70. ‘sinesis’ must be italicized. Please, check everywhere in the text.                                                                                                         Response 10: Thank you for bringing this to our attention. We have thoroughly proofread the entire text and made the necessary corrections. Please refer to the details provided at (line 22, page 1; line 108, page 3; line 339; line 371, page 9; line 403, page 11; line 408, page 11; line 437, page 12; line 467, page 13; line 519, page 14).
11. Line 72. ‘Fusarium solani’ must be italicized.                                        Response 11: Thank you very much for pointing out this formatting issue, we have proofread the full text and corrected it, see details at (line 106, page 3).
12. Line 73. ‘Aspergillus penicillioides’ must be italicized.                       Response 12: Thank you very much for pointing out this formatting issue, we have proofread the full text and corrected it, see details at (line 108, page 3).
13.  Line 82. Put a dot instead of a comma before ‘However’.               Response 13: We sincerely thank the reviewer for careful reading. As suggested by the reviewer, we have proofread the full text and corrected it.
14. “baimuxinal and agarotetro” do not italicize the name of these compounds.                                                                                         Response 14: Thank you very much for pointing out this formatting issue, we have proofread the full text and corrected it, see details at (line 121, page 3; line 699 to line 701, page 18).                                                               Response to Comments on the Quality of English Language                     Point 1: There are many typographical and stylistic errors throughout the text (including missing spaces). Check please.                              Response 1：Thank you for reviewing our manuscript and providing valuable comments. You have pointed out many typographical and stylistic errors in the manuscript, including missing spaces, and requested that this be checked. We appreciate your correction and apologize for the typographical and stylistic errors in the manuscript. We carefully checked the entire manuscript, including typographical and stylistic issues, and revised and corrected the errors to improve its quality and readability. In the revised manuscript, we check for stylistic issues and correct inappropriate expressions to ensure that the style and language of the manuscript conforms to the norms of academic writing, as well as that the manuscript is typographically correct, with appropriate use of spaces, and so on. We are very grateful for all the suggestions made by the reviewers and have revised and improved the manuscript. We deeply apologize for the inconvenience caused to the reviewers and the journal team. Please allow me to say sorry as well as to express our sincere thanks once again, we have followed the reviewers' suggestions with comments to ensure that the revised manuscript meets the journal's requirements in terms of typography and style. In the meantime, we look forward to receiving your approval again after the revision.                                                                                                                                                                                                                                                                                                               Additional detailed response information can be found in the attached word version.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript titled "Effects of different levels of physical damage combined with fungal induction on agarwood formation" is novel and informative and discusses some specific strategies for improving the quality of agarwood. However, the work needs to be rectified due to some of its shortcomings: 

1. In the abstract, there is no quantitative or numerical data. Please include significant results concisely.

2. Line 35: (IUCN) Red List: Remove brackets

3. Line 63-65: "In recent years, as the mechanism of agarwood production has been extensively studied, researchers have noted that endophytic fungal communities differ in different qualities of agarwood, biological induction methods have also been developed". Please rephrase the sentence.

4. Line 66-68: "which mycelium or fungal endophyte culture supernatants gathered from the agarwood contaning A. sinensis that produces agarwood is inoculated into a healthy A. sinensis trunk to mimic the pathological induction of wild agarwood to form agarwood." The sentence is incomplete, and the spelling of contaning is incorrect. Please rewrite and correct spelling.

5. Line 77: "Aspergillus penicillioides" should be Italic

6. Line 206: "software for independent samples t-test and one-way ANOVA one-way ANOVAs to de-": one-way ANOVA repeated Please delete repeated one.

7. Fungal Isolation and Identification methods are not in the methodology sections. Please mention it in detail.

8. No fungus name is provided in the text; only symbols are used.

9. Figure 7B: Quality is too low and invisible. Please improve its quality.

10. Possible reasons behind the secondary metabolite production are not properly discussed in the Discussion section. 

11. Please discuss the enzymatic activities to degrade the cellulose/lignin/hemicellulose activities by fungal studies.

12.  To improve the discussion section, please follow and cite the latest relevant research by Tripathi et al., 2023 to provide a comprehensive analysis. "Tripathi, S.N.; Sahney, M.; Tripathi, A.; Pandey, P.; Jatav, H.S.; Minkina, T.; Rajput, V.D. Elucidating the Anatomical Features, Adaptive and Ecological Significance of Kopsia fruticosa Roxb. (Apocynaceae). Horticulturae 2023, 9, 387) ."

13. The document contains several grammatical and punctuation errors that should be fixed. Before considering publication, the author must thoroughly edit and improve the text's coherence and clarity.

14. References must be formatted according to journal guidelines and consistent. Please carefully format references following journal guidelines.

The document contains several grammatical and punctuation errors that should be fixed. Before considering publication, the author must thoroughly edit and improve the text's coherence and clarity.

Author Response

Dear Reviewer:

We sincerely appreciate the editor and all reviewers for your valuable comments and suggestions. The review comments are laid out below in bold and specific concerns have been numbered. The point-to-point responses to the reviewer's comments are marked in red and presented following.

1. In the abstract, there is no quantitative or numerical data. Please include significant results concisely.                                 Response 1: Thank you for your review and feedback on our abstracts. Your point about the abstract needing to be in the summary, without quantitative or numerical data, has been brought to our attention. In our revised manuscript, we have made the needed changes to improve and present the manuscript in a concise and accurate manner with important results in order to make the abstract more informative and to better summarize the important findings of our study (line 11 to line 41, page 1). Meanwhile, we have revised and improved the abstract to give readers a comprehensive understanding of the background, purpose, methodology, and key findings of our study. Thank you again for your guidance on our abstract, and we look forward to your satisfaction in the revised manuscript. 
2. Line 35: (IUCN) Red List: Remove brackets                                       Response 2: We sincerely thank the reviewer for careful reading. As suggested by the reviewer, we have corrected the “(IUCN) Red List” into “the Red List of the International Union for Conservation of Nature and Natural Resources” (line 60, page 2).
3. Line 63-65: "In recent years, as the mechanism of agarwood production has been extensively studied, researchers have noted that endophytic fungal communities differ in different qualities of agarwood, biological induction methods have also been developed". Please rephrase the sentence.                             Response 3: Thank you very much for the reviewers' comments on the quality improvement of the paper, we have revised and rewritten the sentence as: In recent years, extensive research has been conducted on the production mechanism of the agarwood, leading to the gradual development of bio-induced methods (line 93, page 2). We hope that the revision will be recognized. Thank you again for your comments and suggestions.
4. Line 66-68: "which mycelium or fungal endophyte culture supernatants gathered from the agarwood contaning A. sinensis that produces agarwood is inoculated into a healthy A. sinensis trunk to mimic the pathological induction of wild agarwood to form agarwood." The sentence is incomplete, and the spelling of contaning is incorrect. Please rewrite and correct spelling.                                                                                               Response 4: Thank you very much for the reviewers' comments on the quality improvement of the paper, we have revised and rewritten the sentence as: The researchers introduced endophytic fungal mycelia or endophytic fungal culture supernatants, which were isolated and purified from agarwood, into the trunks of healthy Aquilaria trees in order to simulate the pathological conditions observed in wild Aquilaria trees through biological stress (line 95 to line 99, page 2 to page 3). we hope that the revision will be recognized. Thank you again for your comments and suggestions.
5. Line 77: "Aspergillus penicillioides" should be Italic                       Response 5: Thank you very much for pointing out this formatting issue, we have proofread the full text and corrected it, see details at (line 108, page 3).
6.  Line 206: "software for independent samples t-test and one-way ANOVA one-way ANOVAs to de-": one-way ANOVA repeated Please delete repeated one.                                               Response 6: We were really sorry for our careless mistakes. Thank you for your reminder. We have corrected it to read " All experimental data were preprocessed with Excel and analyzed using SPSS 22.0 software using a t-test and a one-way ANOVA to determine the significant differences between treatments. Statistical differences were considered significant at P < 0.05. "(line 303 to line 306, page 7).
7. Fungal Isolation and Identification methods are not in the methodology sections. Please mention it in detail.                         Response 7: We express our gratitude to the reviewers for their valuable feedback. we have elaborated on the methods of fungal isolation and identification in the manuscript, and here we did not list the changes but marked in red in the revised paper. See (line 169 to line 204, page 4 to page 5).
8. No fungus name is provided in the text; only symbols are used.  Response 8: We express our gratitude to the reviewers for their valuable feedback. In the manuscript we have proceeded to add the names of the three fungi, Trichoderma maatroviride, Fusarium solani, and Lasiodiplodia theobromae. and expressed them in tables and notes (line 203, page 5; line 235 to line 238, page 6). 
9. Figure 7B: Quality is too low and invisible. Please improve its quality.                                                                                                 Response 9: Thank you very much for this suggestion, we have improved the quality of the images and added them to the revised manuscript. If there are any other modifications we could make, we would like very much to modify them and we really appreciate your help (line 532, page 15).
10. Possible reasons behind the secondary metabolite production are not properly discussed in the Discussion section.               Response 10: Thank you very much for your review and valuable comments on our manuscript. According to your suggestions, we will add the discussion section of the paper, such as adding secondary metabolite production and categorization, and reviewing the relevant literature for discussing the effect on secondary metabolite production after fungal  to analyze our findings more comprehensively and to compare and discuss them fully with related literature to ensure the depth and comprehensiveness of the paper, and here we did not list the changes but marked in red in the revised paper. See (line 564 to line 601, page 16). We also look forward to your reviewing our manuscript again.
11.  Please discuss the enzymatic activities to degrade the cellulose/lignin/hemicellulose activities by fungal studies.  Response 11: Thank you very much for your review and valuable comments on our manuscript. According to your suggestions, we have added the information about the effect of fungi on the activities of degradative enzymes such as cellulose, lignin and hemicellulose, for example, the effect of fungal infiltration process on plant cell wall, etc. in our manuscript, and we have made full comparisons and discussions with the related literatures in order to analyze our findings more comprehensively and to make sure that the paper is in-depth and comprehensive. And here we did not list the changes but marked in red in the revised paper. See (line 584 to line 601, page 16). We also look forward to your reviewing our manuscript again.
12. To improve the discussion section, please follow and cite the latest relevant research by Tripathi et al., 2023 to provide a comprehensive analysis. "Tripathi, S.N.; Sahney, M.; Tripathi, A.; Pandey, P.; Jatav, H.S.; Minkina, T.; Rajput, V.D. Elucidating the Anatomical Features, Adaptive and Ecological Significance of Kopsia fruticosa Roxb. (Apocynaceae). Horticulturae 2023, 9, 387)."                                                                                          Response 12: Thank you very much for reviewing our paper and for your valuable suggestions. We have taken note of the latest relevant study by Tripathi et al. and will cite and discuss it accordingly in the revised manuscript. And here we did not list the changes but marked in red in the revised paper. See (line 602 to line 650, page 17). We also look forward to your reviewing our work again.
13. The document contains several grammatical and punctuation errors that should be fixed. Before considering publication, the author must thoroughly edit and improve the text's coherence and clarity.                                                                                 Response 13: Thank you for your careful review and valuable comments on our paper. We apologize for the incorrect grammar and punctuation, and we have sought out Dr. Xiaofei Li and Dr. Peng Zhang, both of whom have expertise in the field, for potential problems with the manuscript, to make corrections and to reflect edits and corrections in the revised manuscript in order to improve the coherence and clarity of the text. We sincerely appreciate your guidance and attention, and we will do our best to enhance the quality of the paper through revisions in anticipation of your further review of the improved version.
14. References must be formatted according to journal guidelines and consistent. Please carefully format references following journal guidelines.                                                                            Response 14: Thank you for your review and feedback on our manuscript. We take your guidance on reference formatting very seriously. In the revised version, we will carefully format the references in strict accordance with the journal guidelines to ensure that the references are formatted in accordance with the journal requirements and remain consistent. We take your suggestions seriously and strive to ensure the accuracy and standardization of references in the expectation that you will be satisfied with our revision (line 777 to line 947, page23 to page 27). We sincerely thank you for your guidance and attention, and we look forward to receiving more valuable suggestions for the improved version.

4. Response to Comments on the Quality of English Language

Point 1: The document contains several grammatical and punctuation errors that should be fixed. Before considering publication, the author must thoroughly edit and improve the text's coherence and clarity.

Response 1: Thank you for your review and valuable comments on our paper. You mentioned that the English style of the text needs to be improved and that you have already done the work of text editing on some of them, which we greatly appreciate. And, we have also sought out experts in the field to revise the manuscript to further refine and improve the linguistic presentation of the article to ensure that the language is more fluent, accurate, and in line with the norms of academic writing.

Author Response File: Author Response.pdf

Reviewer 3 Report

The Authors describe a procedure to induce artificially the production of agarwood in Aquilaria trees by a combination of wounding and inoculation with fungi recovered from trees producing agarwood naturally.

Before giving a detailed revision of the article I have to evidentiate a basic criticism. The Authors did not describe the formulation they used to induce the production of agarwood and did not indicate the name of fungi they isolated from naturally produced agarwood and used to inoculate the trees to induce artificially the production of agarwood. They say this is a secret. I suppose when they say it a secret they plan to patent the formulation or they already patented it. In the latter case, in my opinion  they have to indicate at least which is the patent num,ber or the registered trade name of the formulation. By contrast, if they are planning to submit the formulation for a patent registration I think it is too early to publish a paper in a scientific journal or at least the Authors have to indicate the species of fungi they identified. Otherwise the article can not be accepted as methods can not be reproduced or analyzed. I let the Editor to decide.

Overall the study is interesting. 

There is a criticism in the experimental design: a second control is missing (holes made with fire, without fungi). However even without this second control results can be correctly interpreted.

The Abstract has to reflect the content of the text and be self-explanatory (I added a brief description on the definition of agarwood)

Lane 376. A fungus C is cited. Perhaps it is a mistake as in the rest of the text only Fungi A and B are mentioned.

There is no need of subheadings in the section Discussion.

The style of References should conform with the Author's instructions.

The quality of images and graphs is good.

For more tedailed criticisms and suggestion see notes in the text (attached PDF file)

Comments for author File: Comments.pdf

The English style of the text has to be improved (i made some text editings)

Author Response

Dear Reviewer:

On behalf of all the contributing authors, I would like to express our sincere appreciations of your constructive comments concerning our article entitled “Effects of different levels of physical damage combined with fungal induction on agarwood formation” (Manuscript No:2792110). These comments are all valuable and helpful for improving our article. According to the associate comments, we revised and improved the manuscript and added detailed procedures for the methods used in the study design to make our manuscript replicable and analyzable. In this revised version, changes to our manuscript were all highlighted within the document by using red-colored text. Point-by-point responses to the nice associate reviewer is listed below this letter.

1. Before giving a detailed revision of the article I have to evident ate a basic criticism. The Authors did not describe the formulation they used to induce the production of agarwood and did not indicate the name of fungi they isolated from naturally produced agarwood and used to inoculate the trees to induce artificially the production of agarwood. They say this is a secret. I suppose when they say it a secret, they plan to patent the formulation or they already patented it. In the latter case, in my opinion they have to indicate at least which is the patent number or the registered trade name of the formulation. By contrast, if they are planning to submit the formulation for a patent registration, I think it is too early to publish a paper in a scientific journal or at least the Authors have to indicate the species of fungi they identified. Otherwise, the article cannot be accepted as methods cannot be reproduced or analyzed. I let the Editor to decide.  

   Response 1: Thank you for your fundamental criticisms and concerns about our paper. You mentioned that the formulation used to induce agarwood production was not described in the manuscript, as well as the names of the fungi isolated from agarwood for inoculation, information that you felt was critical to ensure the reproducibility and analytical validity of the method. We apologize for your concerns and understand the importance you place on this critical information. We would like to provide a detailed explanation and response to the issues you have raised: Firstly, with regard to the use of the word 'secret', we recognize that this may have created a misunderstanding amongst our readers. In fact, the formulation used in our trials has been patented and licensed, but we wanted to continue to extend the series of studies on this formulation, and we apologize for not making this clear in the manuscript.

   In view of your suggestion, we have provided sufficient additional information on this in the revised manuscript, including the patents for the formulations (fungal names and ratios) that we have used and the treatment steps of the induction operation that we have referenced (line 210 to line 238, page 5). We will ensure that this information adequately demonstrates our research methodology in order to allow other researchers to replicate and analyze our experimental results to better meet the requirements of the journal and the needs of our readers. to ensure that the final revised version meets the requirements of the journal and we sincerely look forward to receiving your approval.
2.  There is a criticism in the experimental design: a second control is missing (holes made with fire, without fungi). However even without this second control results can be correctly interpreted.                                                              Response 2: Thank you for reviewing and criticizing our experimental design. Your criticism about the lack of a second control has caused us to think deeply. We understand that a rigorous experimental design needs to include multiple controls to ensure the reliability and validity of the results. Thank you again for your suggestion, and we intend to continue to extend to the relevant research on fire holes as a control group in the subsequent experiments and continue to optimize and improve the methodology of the artificial induction technique, so as to provide a reliable reference data base for the development of the agarwood industry. Compared with the control group without (fire punched holes, no fungi) in this test，the reasons mainly include: 1. We choose to inoculate into the trunk, which will cause damage to the tree, we exclude these physical damage factors. 2. We want to exclude the impact of physical punching caused by the tree, and compare the advantageous fungi chosen by D1 and D2 to inoculate into the tree, in the process of agarwood formation, the material changes and composition of the agarwood produced by the differences. 2. We want to exclude the damage to the tree caused by physical perforation, and compare the differences between direct inoculation of fungi and inoculation of fungi into the tree after burning, in terms of material changes in the process of agarwood formation and the composition of the agarwood wood produced, and select a more suitable joint induction method. We also look forward to your reviewing our manuscript again.
3. The Abstract has to reflect the content of the text and be self-explanatory （I added a brief description on the definition of agarwood）                                                                            Response 3: Thank you for your review and feedback on our abstracts. Your request that the abstracts need to reflect the content of the text and be self-explanatory has been brought to our attention. In our revised version, we have paid particular attention to your mention of adding a brief description of the definition of sedum and ensuring that important quantitative or numerical data are included to more accurately present the key findings of our study. So that readers can quickly understand our research findings (line 11 to line 41, page 1). Meanwhile, we have revised and improved the abstract to give readers a comprehensive understanding of the background, purpose, methodology, and key findings of our study. Thank you again for your guidance on our abstract, and we look forward to your satisfaction in the revised manuscript. 
4. Lane 376. A fungus C is cited. Perhaps it is a mistake as in the rest of the text only Fungi A and B are mentioned.        Response 4: We feel sorry for our carelessness. The errors in our resubmitted manuscript have been rectified (line 480, page 13). Thanks for your correction.  
5. There is no need of subheadings in the section Discussion.    Response 5: Thank you for reviewing our manuscript and for your valuable suggestions. You pointed out that subheadings are not needed in the "Discussion" section. In the revised version, we will remove the subheadings in the "Discussion" section according to your suggestion (line 563, page 16; line 651, page 17), so as to maintain the unity of the article style and the overall reading fluency.
6. The style of References should conform with the Author's instructions.                                                                                Response 6: Thank you for your review and feedback on our manuscript. We take your guidance on reference formatting very seriously. In the revised version, we will carefully format the references in strict accordance with the journal guidelines to ensure that the references are formatted in accordance with the journal requirements and remain consistent. We take your suggestions seriously and strive to ensure the accuracy and standardization of references in the expectation that you will be satisfied with our revision (line 777 to line 947, page 23 to page 27). We sincerely thank you for your guidance and attention, and we look forward to receiving more valuable suggestions for the improved version.  
7. For more tedailed criticisms and suggestion see notes in the text （attached PDF file）                                                  Response 7: Thank you very much for reviewing our paper and providing criticism and suggestions. We have carefully read and taken seriously the additional guidance you have given in the in-text annotations. In order to better understand and fully respond to your suggestions, we have downloaded the attached PDF file and analyzed each of the notes therein in detail. We will make the necessary changes and revisions to the paper based on these criticisms and suggestions. We express our sincere gratitude for the professional guidance and detailed annotations, which hold significant value in enhancing the manuscript quality. We will carefully address the content of each annotation, make the necessary revisions, and ensure that your valuable comments are reflected in the final version in anticipation of your satisfaction in the revised manuscript.

4. Response to Comments on the Quality of English Language

Point 1: The English style of the text has to be improved (i made some text editings).
Response 1: Thank you for your review and valuable comments on our paper. You mentioned that the English style of the text needs to be improved and that you have already done the work of text editing on some of them, which we greatly appreciate. And, we have also sought out experts in the field to revise the manuscript to further refine and improve the linguistic presentation of the article to ensure that the language is more fluent, accurate, and in line with the norms of academic writing.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Manuscript improved after revision, however the the references are not written properly as per the journals guidelines. Please carefully correct all the references. In addition discussion part can be improved, consider interpreting significant results in more detail and justify each with supporting literature.

Author Response

Dear Reviewer:

Thank you once again for your review of our manuscript and for providing constructive feedback on its revisions and improvements. On behalf of all the authors, I express our heartfelt gratitude to you for carefully pointing out the issues in our manuscript. Based on your suggested modifications, we have further improved and enhanced the discussed sections of our manuscript. Additionally, we have followed the journal guidelines to correctly format the references correctly, ensuring that our manuscript is readable and complete. We look forward to your review of our manuscript once again. In this revised version, changes to our manuscript were all highlighted within the document using red-colored text. Point-by-point responses to the nice reviewers are listed below this letter.

1. Point-by-point response to Comments and Suggestions for Authors

Comments 1: Manuscript improved after revision, however the the references are not written properly as per the journals guidelines. Please carefully correct all the references. In addition discussion part can be improved, consider interpreting significant results in more detail and justify each with. 

Response 1: Thank you very much for the careful review of our manuscript and for pointing out the issues and providing constructive feedback. We sincerely apologize for not correctly formatting the reference citations according to the journal guidelines, as you pointed out. We appreciate your attention to detail, and we have made the necessary modifications to adhere to the journal's reference citation requirements. Please see the specific changes in the revised manuscript (lines 846 to 1016, pages 24 to 28).

Furthermore, we greatly appreciate your constructive comments on the quality and readability of our manuscript. Based on your suggestions to improve the discussion section by providing more detailed explanations of significant results and demonstrating the rationality of each result, we sincerely thank you for your efforts to enhance the quality and readability of our manuscript. We have made revisions and improvements to the discussion section accordingly. Please see the specific changes in the revised manuscript (lines 570 to 777, pages 16 to 20). We hope that these revisions meet the requirements of the journal and your expectations.

Thank you once again for your valuable input, and we look forward to your review of the revised manuscript.

Reviewer 3 Report

the Authors removed maio flaws. However, after the second revision round there are still some minor text editings and few sentences to be rephrased as they are not clear (see notes in the text, attached PDF file).

Comments for author File: Comments.pdf

Text editinfs still needed

Author Response

Dear Reviewer:

Thank you once again for your constructive feedback on our manuscript. On behalf of all the authors, I express our heartfelt gratitude to you. Based on your suggested modifications and remarks, we have revised and improved our manuscript, including text editing and rewriting of certain passages to improve its readability and coherence. We look forward to your review of our revised version. In this revised version, changes to our manuscript were all highlighted within the document using red-colored text. Point-by-point responses to the nice reviewers are listed below this letter.

1. Point-by-point response to Comments and Suggestions for Authors

Comments 1: the Authors removed maio flaws. However, after the second revision round there are still some minor text editings and few sentences to be rephrased as they are not clear (see notes in the text, attached PDF file).

Response 1: Thank you very much for carefully reviewing our manuscript and providing feedback on the issues and constructive suggestions for modifications. We greatly appreciate the efforts you have made to improve the quality and readability of our manuscript based on your comments in the PDF. We have strictly followed the comments you provided in the manuscript during the revision process, and we hope that our modifications meet the requirements of the journal and your expectations. The specific text edits can be found in the revised version, highlighted in red. Regarding the passages where there were disagreements in expression, we have made the following modifications:

1. Abstract: As wild Aquilaria sinensis resources are exhausted and protected, China has established a huge number of plantations of Aquilaria trees and developing artificial induction techniques. However, the current output and quality of artificial induction technology have not yet met the expected results. It has been found that high-oil-containing agarwood may contain particular fungal stains associated with agar-wood production. To enhance the quality of agarwood, we recovered and characterized three highly active fungi from high-oil-containing agarwood, and inoculated them onto A. sinensis trees using two traditional physical methods. The results show that fungi extracted from high-oil-containing agarwood can effectively increase the yield and quality of agarwood. During the agarwood formation process, parenchyma cells, xylem rays, and axial parenchyma cells in the xylem gradually undergo apoptosis, thereby promoting the expansion of the color range of agarwood. Nine months after the treatment, the alcohol-soluble extract content in agarwood reached the standard specified in the Chinese Pharmacopoeia (10%), and the proportions of sesquiterpenes and chromones in each treatment were 55.82%, 58.31%, 62.65%, 70.97%, and 13.71%, respectively. These results indicate that fungal induction has a positive impact on the quality of agarwood. In addition, compared to drilling and fungus combined induction "burning holes and fungi" combined induction demonstrates better results and can further improve the yield and quality of agarwood.
2. all treated samples showed a significant increase in the abundance of starch granules. Among them, the F1 (Figure 2i) treated samples only contained a small amount of starch substances in the interfascicular cambium, and the F2(Figure 2l) treated samples had the lowest abundance of starch granules. This indicates that the combination treatment of burning holes and fungi has a stronger metabolic capacity and consumes a large amount of starch. The paragraph was amended to read: although the abundance of starch granules in all samples increased to varying extents, the samples treated with "burning holes and fungi" combined treatment still showed lower starch abundance compared to those treated with drilling holes and fungi combined treatment. Among them, the F1 (Figure 2i) treated only contained a small amount of starch material in the interfascicular cambium, while the F2 (Figure 2l) treated showed almost no visible abundance of starch granules. This indicates that the combined treatment induced by burning holes and fungi has a stronger metabolic capability and can consume a large amount of starches. (Line 357 to line 364, page 8 to page 9.)
3. This indicates that different fungi may need to be co-induced with their adapted damage to effectively increase the yield and quality of agarwood, further promoting the development of the agarwood industry. The paragraph was amended to read: This indicates that the utilization of various fungal strains alongside appropriate treatment methods is essential for the significant enhancement of both agarwood yield and quality. (Line 765 to line 767, page 19).

Based on the comments in the PDF revised manuscript provided by you, please refer to the red highlighted annotations in the revised version for other modifications. Thank you once again for your valuable input, and we look forward to your review of the revised manuscript.