PONE-D-23-27285Genetic differentiation at extreme latitudes in the socially plastic sweat bee Halictus rubicundusPLOS ONE

Dear Dr. Boulton,

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"Many thanks to Stephen Quinn (community park manager, Belfast city council), Ross Watson (Woodland trust, Migdale and Ledmore), Jeff Cherrington (lead ranger, Boscastle to Morwenstow, National Trust), John Keast (Bodmin commons council), Robert Clark (landowner at Bodmin), Martin Wright (common land and village greens registration officer, natural environment and open spaces service, Cornwall council), Lynne Jones (St. Neot Parish council) and South West Water for facilitating access to field sites. This work is part of a project that received funding from the European Research Council (ERC) under the European Horizon's 2020 research and innovation programme (grantagreement no. 695744). RB was funded by a Wageningen Graduate School Postdoctoral Talent fellowship and a BBSRC discovery fellowship."

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Additional Editor Comments: 

The manuscript requires comprehensive revision for the improvement of the draft. 

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: Yes

Reviewer #2: Partly

Reviewer #3: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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: No

Reviewer #3: No

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: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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: The paper presents the results of a study aiming to understand whether differences in phenology due to

environmental conditions could limit gene flow in Halictus rubicundus bees. My impression is that the

data are interesting and, with a few adjustments, would make a nice paper in PLOS ONE. Follow my

comments:

1) In general, in the introduction, I think it misses a description of the types of barriers, in addition to

the distance between populations, that could influence bee gene flow. The authors cite the Irish Sea and

temperature differences, but between the south and north of the UK, there are possibly other barriers

that could be better described. My knowledge about UK geography is poor, just like many readers

abroad. So, the detail of this point is important to strengthen the work.

2) Lines 43–44: “Halictus rubicundus (Hymenoptera: Halictidae) is a locally abundant species with a

broad geographic range”. Where are the bees locally abundant? In the north of the globe? In UK? I

understand your point, but starting the text with this information without giving a basis to understand it

doesn’t seem right.

3) Lines 205-207: “Migdale was the most genetically different from all other populations (Bodmin:

FST = 0.061; Nei’s = 0.618. Boscastle: FST = 0.059; Nei’s = 0.625. Belfast: FST = 0.054; Nei’s =

0.491)”. I suggest adding the values of Migdale at this point in the text to facilitate the comparison.

4) Lines 207-210: “Bees collected in the south of mainland Britain (Bodmin and Boscastle) were more

genetically similar to bees across the Irish Sea (Belfast) than they were to bees collected in the North of

mainland Britain in Migdale (Bodmin: FST = 0.046; Nei’s = 0.504. Boscastle: F ST = 0.044; Nei’s =

0.503).” Again, please add the values of all points cited in the text.

Reviewer #2: Article number PONE-D-23-27285 “Genetic differentiation at extreme latitudes in the socially plastic sweat bee Halictus rubicundus” carried results that need revision for the improvement of the draft. Moreover, the conclusion portion should be further developed based on the findings and future implications of the study. Some suggested changes are in the comments portion to revise and improve the manuscript. Please find suggested corrections, Reference writing, journal-style format, author’s instructions, use of abbreviations and missing information for revision.

Abstract: Abstract is lengthy, please be precise in the abstract writings.

Line 45: Please recheck the abbreviations H. rubicundus” and write complete scientific name at the start of the sentence.

Line 45-48: Please rephrase the sentence

Line 48: (the number of generations per year)” remove the sentence

Line 53-54: (a single generation per 54 year)” please remove the sentence.

Line 54: see also Hogendoorn and Leys 1997” please follow the author’s instructions and journal style format while writing the references” this may be recheck throughout the manuscript

Figure 1 may be placed as supplementary figure 1 (figure taken from Schürch et al. 2016) while mentioning the Figure S1 in the text.

Material and methods

The authors collected samples in 2019-2020 and conducted analysis? There is no data found regarding the background of the samples and even the generational data?

Line 160: HWE?

Line 249: to the” remove from the sentence

Please add a precise conclusion

Please write the references in the text and in the reference portion according to the author's instructions and Journal style/formatting. Please double-check for typos and inconsistencies in Journal style/formatting/ authors instructions, double spaces, spellings of the words, English vocabulary, missing italics, scientific names, missing information

Reviewer #3: This article presents a study on the genetic differentiation seen in different populations of Halictus rubicundus in the UK. The paper addresses an interesting topic and is well written; I don’t have comments about the analyses or the interpretation of the results. I feel the discussion could be fleshed out a bit, however, and so I recommend it for publication in PLOS One pending the minor revisions to the discussion outlined below.

The results of the study uncover genetic differentiation between populations in the north and the south of the UK and the authors state that their results “improve understanding of barriers to gene flow”. While the authors suggest that this differentiation may be caused by both phenological and /or physical barriers to gene flow, the study does not appear to have made any attempt to separate these factors. To truly understand the barriers to gene flow, and thus the potential consequences of climate change or habitat degradation on these populations, separating phenological barriers from physical barriers seems essential. Perhaps it was not possible in the present study…but how could this be done in a future study? To highlight geographical barriers - would it be useful to increase the sampled nests to include populations from along the north-south gradient of the UK? If the Scottish Highlands truly represent a barrier to gene flow, then populations in the Scottish lowlands should also be quite different from the Migdale populations. The Irish Sea was not found to have been a total barrier to gene flow (Belfast populations overlapped quite a bit with the Cornish and, to a lesser degree, with the Migdale). Maybe this should be discussed in more detail, given what we know about dispersal capacities of species of bees across bodies of water? Or maybe there’s another explanation for this overlap, i.e. similar alleles that have been retained in different populations despite reproductive isolation? And to look at phenological barriers to gene flow - could one take into consideration voltinism when comparing genetic differentiation (i.e. is there more gene flow between populations that are either univoltine or bivoltine, and less among populations with different social structure?)?

Finally, to the question "Have the authors made all data underlying the findings in their manuscript fully available?", I answered no because I couldn't find any mention of where the microsat markers generated in the study were stored...

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: Yes: Jesse Litman

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Attachments

Attachment

Submitted filename: PONE-D-23-27285_review.pdf

PONE-D-23-27285R1Genetic differentiation at extreme latitudes in the socially plastic sweat bee Halictus rubicundusPLOS ONE

Dear Dr. Boulton,

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.

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

ACADEMIC EDITOR: Please revise the draft according to comments of the reviewers  

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

Please submit your revised manuscript by Apr 25 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.

We look forward to receiving your revised manuscript.

Kind regards,

Bilal Rasool, PhD

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.

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: (No Response)

Reviewer #3: 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: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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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

Reviewer #3: 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

Reviewer #3: Yes

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6. Review Comments to the Author

Reviewer #1: I found the revised manuscript much easier to read than the previous version. Particularly, the results seems clear to me now. I think this is an interesting study worth publishing.

Reviewer #2: The revised article, PONE-D-23-27285R1, "Genetic differentiation at extreme latitudes in the socially plastic sweat bee Halictus rubicundus" has improved after revision; however, some issues still need to be verified before the draft is officially considered.

The authors feel that “figure 1 is essential to the introduction, as a big part of this paper is about the life cycle of H. rubicundus and how differences could influence phenology and gene flow. Therefore, we believe that figure 1 is an important visual aid to set up the study and so we have left it as a main figure.......” Kindly recheck and consult for the journal's policies with the managing editor. "It is okay with me if the journal policies permit the reproduction of figure 1 from the earlier literature.

The authors should also recheck the formatting guidelines for tables. Journal-style formatting and adjustments should be made as needed.

Line 380: Reference [2] please recheck if this reference is relevant to the text (if necessary)

Could you please confirm that the references in the discussion section follow the numerical sequence, as they appear to be out of order? (Make changes if necessary.)

Please write the references in the text and in the reference portion according to the author's instructions and Journal's style and formatting. Please double-check for typos and inconsistencies in Journal style formatting, author's instructions, double spaces, spellings of the words, English vocabulary, missing italics, scientific names, and missing information etc.

Reviewer #3: The authors have addressed the comments from three different reviewers and I feel the manuscript is now ready for publication in PLOS One.

**********

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

Reviewer #3: No

**********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.

Genetic differentiation at extreme latitudes in the socially plastic sweat bee Halictus rubicundus

PONE-D-23-27285R2

Dear Dr. Rebecca,

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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If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Bilal Rasool, PhD

Academic Editor

PLOS ONE

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 #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 #2: Partly

**********

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

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 #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 #2: Yes

**********

6. Review Comments to the Author

Reviewer #2: The article is much improved after revisions, however, formatting of figures and tables according to journal style may be considered in proofreading.

**********

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 #2: No

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