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PONE-D-23-28830Induction of agarwood formation by artificial inoculation with fungi: Isolation and screening of fungi promoting agarwood formation in Aquilaria sinensis treesPLOS ONE

Dear Dr. chuang,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Dec 04 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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We look forward to receiving your revised manuscript.

Kind regards,

Niraj Agarwala, Ph.D.

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Partly

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

Reviewer #1: No

Reviewer #2: N/A

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Overall Comment

The publication doesn’t meet the standards of the PLOSone journal. The manuscript needs major refinement at usage of English, Data and its appropriate presentation and scientific elaboration and novelty of the study needs to be highlighted upon in the manuscript.

1. Very poorly written manuscript, major revision at writing level needed. Statements made are not complete, very undecipherable, data is basic going by standards of PLOSone, presentation needs major changes to make it suitable for publication. Methodology part is only lucid and elaborately written, all other parts need major revision.

2. There have been many publications on artificial inoculation in A sinensis recently (Ma et al., 2021, Jalil et al 2021 etc.), what is the innovative method used in the study needs to be highlighted.

3. The comparative analysis of the data in form of a discussion on the new knowledge gained in background all the existing knowledge is totally lacking. That makes the manuscript sound vague though the data has the potential to make a good read. No scientific reasoning for the experimentation could be found in the text. The conclusion is confusing, what is being concluded is not clear at all.

4. Background and sound scientific basis for carrying out the experimentation is not reflected by current version of the manuscript which needs to be majorly addressed upon.

Specific comments

For Abstract

1. Overall issues with English hugely noted, like the statement in abstract “ In this study, different aroma-making treatments were applied to Aquilaria sinensis plantation pure forests to isolate and purify the dominant fungi on the tree surface and discolored wood, and the strains were identified by molecular biology.

2. Why were stains screened for resistance using benzylacetone? Explanation needed.

3. “The Trichoderma sp. and Neurospora sp. samples yielded ethyl ether extracts of 3.62% and 4.04%, respectively” what significance does this statement has? Statement that make characteristic contribution to the results should to be highlighted in abstract.

4. The sum of the relative contents of the aromatic components was 30.1% and 32.86%, respectively. What significance does this statement has? Statement that make characteristic contribution to the results need to be highlighted in abstract.

5.The relative contents of sesquiterpenes were 10.21% and 11.19%. Trichoderma sp., Neurospora sp. were able to effectively induce the production of agarwood within 6 months, and their good effect on agarwood formation and shortening the time of agarwood formation. Not clear what new information/ knowledge or method has been created by this study.

6. What does the statement “It provides effective support for the expansion of the fungal library that promotes agarwood formation” substantialize or conclude? Choice of word usages have diluted the real meaning of the statement.

From main text

7.What is Introduce? Should be Introduction.

8.Its trunk does not produce agarwood if there is no external damage, only after natural or man-made injuries such as slashes, injuries and diseases, insects and ants, lightning strikes, etc….is a incomplete sentence

9.Agarwood is mildly warm in nature. How??

10.Since when Agarwood has been used as Precious spice? Please quote references.

11.Historically, China's agarwood resources are very rich, with the discovery of the medicinal value of agarwood ……………………….. field of Aquilaria sinensis has been very little, was sporadic distribution. Is a very complex statement and doesn’t make a good read. This needs simplification and usage of proper English to convey the core thought.

12. There have been many publication on artificial inoculation in A sinensis recently, a comparative account of the same would have been more interesting read which could have highlighted upon the advantages of the method devised in the study.

13.What is stained wood?

14.Why Benzylacetone was selected as a resistance indicator needs to be elaborated? What added advantage it confers to the study is not apparent.

15.Justification for carrying out the study in not adequate in view of the available already.

16.The method used is amalgamation of too many method Physical, heat, electric current creating a state of over stress to the plant , where within under humid condition any fungal pathogen attack on the bark is possible. The fungus growing may not necessarily be endofungal community specific to the plant. A statement or clarification in this regard would be welcome by readers.

17.The prepared sample was weighed 3 g wet weight into a 50 mL centrifuge tube and 15 mL of ether was added to extract the sample ………………………………….to obtain the ether extract, the extraction was repeated three times, then the extracts were combined and weighed in a fume hood after drying and the volatile oil content was calculated. How was it calculated? Statement also needs restructuring to make it easy to understand and read.

18.Table 1 showing that benzylacetone produced varying degrees of inhibition on all 26 isolated fungal strains can be a shown as supplementary data.

19.What is LV, H and CK need to be introduced to readers more properly in a one statement.

20.The discoloured wood was subjected to chemical compositional identification and the yield of ethyl ether extract was 3.62%, 4.04% and 1.94% for LV, H and CK respectively. Its GC-MS total ion flow (Figure 5). What does this statement mean?

21.“signifying the relative stability of the sesquiterpenes” How? Elaborate.

22.Which lacked any of the 3 major active chemical components of Agarwood. What are these major active chemicals name them here.

23.The comparative analysis of the data in form of a discussion on the new knowledge gained in background all the existing knowledge is totally lacking.

24.Discussion of results is missing. That makes the manuscript sound vague. There is no reference to previous work in same field. There is basic data but very poorly presented. No scientific reasoning for the experimentation could be found in the text.

25.Table 1 and 2 should be made supplementary data.

26.Figure 3 should be made supplementary data.

27.Figure legends for Figure 4 and 5 poorly presented. Figure legend for 4 is missing.

28.The conclusion is confusing. What is the conclusion of the study?

29.“The fungi selected in this paper have good effects in promoting Agarwood production and shortening Agarwood formation time in Aquilaria sinensis trees” How? Prove the statement by comparing with previous available data needed in text.

30. Fusarium and Trichoderma are popular choices for inoculation, the study should list out the novelty of the present work more clearly.

31. Referencing for study not adequate.

Reviewer #2: Title: Induction of agarwood formation by artificial inoculation with fungi: Isolation and screening of fungi promoting agarwood formation in Aquilaria sinensis trees

Summary: In order to efficiently and accurately screen Aquilaria sinensis aroma-promoting fungi, expand the library of Aquilaria sinensis aroma-promoting fungi, and shorten the aroma-forming cycle of Aquilaria sinensis. Present results provide effective support for the expansion of the fungal library that promotes agarwood formation. The authors attempted a good approach to perform the following study. Following are my suggestions for further improvements.

Review comments:

1. Abstract need to be more summarized with the actual study results.

2. The introduction section needs to be adhered to with the proposed title. Also needs to be referred to relevant published literature. The following published articles might be helpful in the enhancements of the introduction section.

- Production of volatile compounds by a variety of fungi in artificially inoculated and naturally infected Aquilaria malaccensis. Current Microbiology, 79(5), 151.

- An is gene-mediated molecular detection of fungi associated with natural and artificial agarwood from Aquilaria malaccensis. Journal of microbiology, biotechnology, and food sciences, e9465-e9465.

- Fungi mediated agarwood (A. malaccensis) production and their pharmaceutical applications: A systematic review. International Journal of Plant-Based Pharmaceuticals, 2(2), 261-270.

3. The novelty of the work needs to be highlighted. The objectives of the work can be elaborated for reproduction of the work.

4. Methods need to be reproducible. The authors provided too descriptive methods. I suggest shortening them and extra parts can be moved to the results section.

5. Also, the methods can be separated into sub-topics for ease of access. Like GCMS analysis and the different shock treatments.

6. For the GCMS analysis following published literature might be useful such as Gas chromatography analysis of the microwave-aided extracted agarwood oil from physically induced Aquilaria malaccensis trees in Northern Thailand. Maejo International Journal of Energy and Environmental Communication, 4(3), 52-55.

7. In the methods, the authors mentioned the PCR amplification, but in the results, I am unable to find the results related to it. No gel images or DNA analysis results were presented.

8. If the authors plan to include Table 3 can be moved to supplementary information. AS Figure 5 shows the chromatographs.

9. The results need to be enhanced with the help of literature. The discussion section is not comprehensive enough to refer to published articles for improvements.

10. The grammatical errors and the English language need to be refined.

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Reviewer #1: Yes: Sofia Banu

Reviewer #2: No

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PONE-D-23-28830R1真菌的分离和筛选以增强沉香树的形成PLOS ONE

Dear Dr. chuang,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. 

==============================

Please submit your revised manuscript by Feb 01 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Niraj Agarwala, Ph.D.

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

**********

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

Reviewer #1: No

Reviewer #2: N/A

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: No

Reviewer #2: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: No

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Overall Comment:

Though the Authors took the effort to modify the manuscript. Still there is a huge scope to modify it further to make it a much appreciable piece of work. A major revision is still needed to make it a scientifically presented manuscript.

From the previous comments 3 comments where not answered adequately

1. A thorough English check of the manuscript

2. Comments 15 Not adequately answered. A more insightful explanation needed for the amalgamation of too many methods (Physical, heat, electric current creating a state of over stress to the plant). Justification on the role of the pretreatment needed to be elaborated more scientifically in discussion

3. Comment 19: Not adequately answered.

Abstract:

1. Agarwood is formed when Aquilaria sinensis trees are harmed naturally or by humans: Modify the statement.

2. In this study, dominant fungal species were isolated and purified from the surfaces and electroshock holes of Aquilaria sinensis trees by fire and electroshock treatments: Modify the statement

3. In agarwood products formed by using Trichoderma sp. And Neurospora sp., the proportions of ether extracts were 50.22% and 48.71%, respectively, exceeding the 10% threshold specified by the Chinese Pharmacopoeia: Modify to make it applicable to International Readers too. What is agarwood products? Its just raesin deposition.

4. Fungal application shortened the time of natural agarwood formation: If you have applied fungal inoculum how is its natural infection, its induced formation. Modify the statement.

Introduction:

5. Fungi promoting the formation of agarwood are firstly tolerant to some extent to the active ingredients of agarwood: Quote the reference for the statement.

6. In this study, pretreatment was carried out before exposing the Aquilaria sinensis trees to fungi. Moreover, isolated fungi were screened for resistance…………………… The study would expand the library of fungi known for promoting agarwood formation: These are concluding lines used by author in conclusion also. These cannot be part of Introduction segment.

7. The method presented here is able to narrow down the range of fungi that promote agarwood formation, and reduce the time and monetary cost of screening these fungi. The study would expand the library of fungi known for promoting agarwood formation: Two contradictory statements made in same para, Modify.

Materials and methods

8. Aquilaria sinensis needs to be italic

9. The discolored wood of Aquilaria sinensis was collected after treatment by the electric shock pretreatment for 3 months

10. 1.5ml should be 1.5 ml. Follow international norms Value space SI Units.

11. -20℃ should be -20 ℃.

12. 5cm should be 5 cm. Follow this throughout the manuscript.

13. Fig.2 Map of the dominant colony on the surface of the tree and the dripping and sampling position of the bacterial liquid: Should be Fig.2. Picture of the of the dominant colony on the surface of the tree, the dripping and sampling position of the fungal liquid.

14. Evidences in form of Photographs are not adequate, some more detailing needed, e.g. pictures of inoculation with GI tag pictures etc.

Results

1. Using molecular identification, the dominant fungal strains on the surface of Aquilaria sinensis were initially identified as Trichoderma sp. and Neurospora sp., which were labelled as LV and H, respectively: Please provide the evidences of identification eg. Amplification of the PCR products (gel images), Phylogenetic analysis etc as supplementary data

2. The red and green fungal colonies that covered the burnt parts of the trees after the fire treatment (Fig 2) were attributed to the decay and removal of other microorganisms from the trees due to fire, resulting in the vigorous growth of Trichoderma sp. and Neurospora sp.: Is a very very vague statement. Either remove it or provide evidences for stating the same.

3. The sentences “The red and green fungal colonies that covered the burnt parts of the trees after the fire treatment………………………………. intrinsic factors (such as chemical compositions) and extrinsic factors (such as the climate, light, rainfall and soils) may affect the parasitism and diversity of endophytic fungi in agarwood” are not results need to be moved to discussion section.

4. In the sentence “Promotion of agarwood formation in Aquilaria sinensis by artificial inoculation of benzyl acetone-tolerant fungi” Aquilaria sinensis needs to be in italics.

5. The sentences “The low sesquiterpene content in the agarwood……………………………………absence of chromone components in agarwood samples may be due to insufficient induction time” are explanations to the results obtained and would be appropriate if are moved to discussion section.

6. Figure legends are poorly presented

Discussion:

1. After injection, the fungal solution is mixed into the trunk sap, and then diffused into other parts of tree: Is the statement correct?

Conclusion:

1. Needs further modification to highlight upon the novelty of the work. The continuity between the statements is missing, which the authors need to improve upon majorly. The conclusion needs to be rewritten.

Reviewer #2: The authors successfully responded to the comments raised. The revised version is acceptable for publication.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachments

Attachment

Submitted filename: Comments for Manuscript_PLOS 15_12_2023.docx

Attachment

Submitted filename: Comments for Manuscript_PLOS 15_12_2023 (2).docx

PONE-D-23-28830R2Isolation and screening of fungi for enhanced agarwood formation in Aquilaria sinensis treesPLOS ONE

Dear Dr. chuang,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Sorry for the delay in getting the reviewers report regarding the manuscript. The reviewers recommend some minor revisions to the manuscript for acceptance. Please address the reviewers comment.

Please submit your revised manuscript by May 10 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Niraj Agarwala, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

**********

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

Reviewer #1: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Abstract:

1. Agarwood is not produced naturally; it only forms after natural or man-made damage to Aquilaria sinensis. Replace man-made damage with suitable wording

Introduction:

1. The naturally occurring agarwood has long outstripped the market demand, and at the end of the..Replace outstripped with outrun

2. Benzyl acetone is a representative compound of the aromatic content in agarwood……….the statement needs to be in context. Here it is just a hanging statement.

Results

1. Screening of benzyl acetone-tolerant fungi : a certain extent, the isolated and purified fungal strains were screened for tolerance to benzyl acetone.

2. GC-MS detection of agarwood samples ……where is th evidence for the same provided.

3. The agarwood products…..use statement “The agarwood”

4. Fig.3 Agarwood samples…………Reasin deposition seen in figure is very very less than claimed in the text. Please explain.

5. For the query Using molecular identification, the dominant fungal strains on the surface of Aquilaria sinensis were initially identified as Trichoderma sp. and Neurospora sp., which were labelled as LV and H, respectively: Please provide the evidences of identification eg. Amplification of the PCR products (gel images), Phylogenetic analysis etc as supplementary data

6. The alignment information of the sequencing results has been provided which I think is not adequate, please provide the gel images etc.

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Reviewer #1: Yes: Sofia Banu

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PONE-D-23-28830R3Isolation and screening of fungi for enhanced agarwood formation in Aquilaria sinensis treesPLOS ONE

Dear Dr. chuang,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Additional Editor Comments:

Dear authors,

I can see that most of the queries raised by the reviewers are now resolved. In my view the article is almost ready for publication, some minor modifications will enhance the readability and scientific merit of the article.

1. Figure 2 and Figure 3 descriptions can be more specific. For figure 2, specifically mention which inoculation method results in agarwood formation shown in the photograph. For figure.3 specifically mention the type of sample used for GC MS analysis for each. Not in the form of Note. If abbreviations are used then also provide full form in figure captions.

2. Regarding methodology used for GC-MS analysis reference 20 is cited, for which it is mentioned that full text is only available in Chinese. Give complete details of GC-MS methods used in the study in the manuscript

I hope my inputs will be useful to improve your manuscript. Best wishes

Regards

Niraj

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Isolation and screening of fungi for enhanced agarwood formation in Aquilaria sinensis trees

PONE-D-23-28830R4

Dear Dr. Liu,

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Academic Editor

PLOS ONE

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