PONE-D-21-30327Distribution and pathogenicity of  Beauveria bassiana  in soil with earthworm action and feedingPLOS ONE

Dear Dr. Xie,

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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Yulin Gao

Academic Editor

PLOS ONE

****************************************Dr. Yulin GaoProfessorDepartment of EntomologyInstitute of Plant Protection(IPP)Chinese Academy of Agricultural Sciences(CAAS)2# West Yuan Ming Yuan RoadHaidian District, Beijing, 100193, P.R, ChinaOffice: 01062815930Mobile: 13552643313E-mail: gaoyulin@caas.cn****************************************

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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: This study assessed the impact of the earthworm Eisenia fetida affecting on Beauveria bassiana, including its distribution in soil and pathogenicity to Atrijuglans hetaohei Yang. The result showed that action and feeding of earthworm may promote distribution of B. bassiana TST05 in soil, but decreased the viable spores of B. bassiana. The viable spores of B. bassiana TST05 passed through earthworm gut still had certain germination ability and higher pathogenicity to the insects. This study is important for guiding the scientific application of B. bassiana in biological control of pests. However, several questions should be addressed.

1.Beauveria bassiana TST05 strain that have been identified in the previous studies in line 67-79 do not need to be described in detail in Result section.

2.The statistical difference of fungal colony numbers between layers should been marked in Fig. 1 C and the statistical analyses between the CK group and the treatment group should been labelled in Fig. 5.

3. The discussion section need to be streamlined and logic, e.g. the content same as the result section be deleted and reasonable discussion.

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PONE-D-21-30327R1Distribution and pathogenicity of  Beauveria bassiana  in soil with earthworm action and feedingPLOS ONE

Dear Dr. Yingping Xie,

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 31 July. 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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• 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'.
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• 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,

Mei Li

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: (No Response)

Reviewer #2: All comments have been addressed

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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: Yes

Reviewer #2: Partly

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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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

Reviewer #2: Yes

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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

Reviewer #2: No

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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: (No Response)

Reviewer #2: Results in this study may be of concern in soil biological interaction because authors provide data on the effects of earthworm action on the distribution of B. bassiana and its pathogenicity to target insects. However, I have got many concerns and there are so many basic errors that I cannot recommend this MS for publication.

CONCERNS:

1. Why the earthworms were maintained in sterilized soil with 40 cm soil thickness? Eisenia fetida is epigeic (0-20cm) and rarely you will find it burrowing in deep soil.

2. The introduction section is poor and do not justify your aims. Line 40-42, why emphasize entomopathogenic nematodes? Line 50, how could the author draw a conclusion that earthworm is a nonhost organism of B. bassiana, is there any evidence? Line 55-57, the sentence is confused. Line 57-62, the life history of insect A. hetaohei should be deleted. Line 64-68, more like within the scope of method. Line 69-77, more like the results of previous study. In this section, please clarify: what gap are you trying to tackle? And why it is important to study the effect of earthworm on distribution and pathogenicity of Beauveria bassianain?

3. The Methods section are not well justified. Line 125, 131, etc., what is the status of sampled soil (dry or wet)? Line 119-120, how about the light condition during culture period? The effective counting concentration is the dilution concentration corresponding to the plate colony count of 20 to 200. Line 126, all samples use the same dilution ratio of 10,000, the CFU of B. bassiana in CK is scientifically inaccurate (Fig.1C). In addition, what is the diameter of the Petri dish? The addition of one milliliter mixture for each plate is excessive.

4. The soil column used in Fig.1a is not described in Method.

5. Line 219-220, is it due to the dilution ratio of 10,000 is too large?

6. Line 155-157, does the midgut material contain intestinal tissue? What components contain in the supernatant? Results in 3.2 showed that great damage occurred in the midgut of the earthworm, and the colonies was conducted based on the midgut contents of the earthworms (without excreting casting). Why collect midgut material after excreted casting? Line 159, Why incubated temperature set as 4°C, far below the ambient temperature for earthworm growth?

7. I found quite difficult to discuss inhibitory effect of the midgut fluid on the spore germination in Fig.3 since the plate is seriously contaminated.

8. The MS requires language editing (really !). Some sentences are awkward. (Line 92; Line 103-104, and many more…)

BASIC ERRORS:

1. Corresponding Author name (see Manuscript Draft in submitted PDF)

2. Latin name (A. hetaohei Yang and A. hetaohei; E. fetida and E. foetida; full name and abbreviation of the Latin name)

3. The use of Arabic numerals. They should never be used at the beginning of sentences. (Line 167)

4. Keywords should not be words found in the title. More, exceed the required number of keywords.

5. Segmentation is confusing (Line 179-191)

6. And many more ...

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Reviewer #1: Yes: Yin Jiao

Reviewer #2: No

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Distribution and pathogenicity of  Beauveria bassiana  in soil with earthworm action and feeding

PONE-D-21-30327R2

Dear Dr. Xie,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Mei Li

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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

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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: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: 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

Reviewer #2: Yes

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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

Reviewer #2: 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: The manuscript has been significantly improved throughout the review process, so i think the revised manuscript is suitable for publication in Plos One.

Reviewer #2: (No Response)

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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: Yes: Jiao Yin

Reviewer #2: No

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