PONE-D-23-32436The association between Dioscorea sansibarensis and Orrella dioscoreae as a model for hereditary leaf symbiosisPLOS 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: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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

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Reviewer #1: Yes

Reviewer #2: No

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Reviewer #1: Yes

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: Date: 10/25/23

Journal: Plos One

Manu #: PONE-D-23-32436

Title: The association between Dioscorea sansibarensis and Orrella dioscoreae as a model for hereditary leaf symbiosis

Authors: Tessa Acar; Sandra Moreau; Marie-Francoise Jardinaud; Gabriella Houdinet; Felicia Maviane-Macia; Frederic De Meyer; Bart Hoste; Olivier Leroux; Olivier Coen; Aurelie Le Ru; Nemo Peeters; Aurelien Carlier

General Comments: This study deals with understanding the leaf symbiotic relationship between the host, Dioscorea sansibsarensis, and its bacterium, Orrella dioscoreae. This symbiotic relationship is unique among other symbiotic leaf taxa in other plant families as the study shows the bacterium can be cultured outside the plant, the plant can grow normally without the bacterium, and the same bacterium or genetically modified bacterium can be reintroduced into the uninfected plants. This experimental regime satisfies Koch’s postulates (which should be mentioned in the text). This experimental system may allow for a better understanding of other similar or obligate leaf symbioses as to their functional value(s).

Specific Comments: there are comments made throughout the attached, edited text. The monocot Dioscorea sansibsarensis symbiotic relationship seems significantly different from the obligate dicot leaf symbioses in that the bacteria are always on the leaf surface covered by the curled leaf lamina tip and associated secretory trichomes, and never encapsulated internally like with the nodules of Psychotria, Pavetta, and Ardisia sps. The enclosed shoot apices of the latter sps. do serve as an ‘external’ reservoir with stipular secretory glands/trichomes that maintain a bacterial population until it penetrates into the leaf lamina to form encapsulated nodules. However, none of these relationships show the bacteria inside cells but remain intercellular within the isolated nodules, a completely different environment than in this study. Whether further research on the system in the present study will shed light on the former obligate taxa remains an open question. With this said, the present study is worthy of publication because future research may uncover information that is relevant to it and the obligate taxa mentioned.

Title: appropriate;

Abstract: Identifies the purpose of the study and its rationale;

Key words: no key words listed in body but shown previously;

Introduction: Provides an excellent background for why the study is being conducted. It covers the key references and points out its significance related to previous studies;

Material and methods: See various comments. the M&m are presented in great detail by individual sections, and can be readily reproduced by other researchers, if necessary;

Results: See comments throughout Results;

Discussion: covers the Results adequately. It would be useful to include adding how the present study compares to Koch’s postulates as all other leaf nodule ‘obligate symbioses fail to satisfy them. Furthermore, the functional value(s), relayed to them being obligate, is still in question, as is the association in the present study.

References: match those listed in text and are appropriate to this study, and useful to any researcher interested in this area of inquiry;

Table 1: clear results;

Table 2: clear results;

Fig. 1: see various comments made in the legend. Satisfying the comments should greatly improve the quality of the images;

Fig.: 2: see comment in legend;

Fig. 3: Clear results;

Fig. S1: detailed, clear, and ok. Images of good quality;

Fig. S2: detailed, clear, and ok

Fig. S3: detailed, clear, and ok

Table S1: detailed, clear, and ok

Table S2: detailed, clear, and ok

Table S3: detailed, clear, and ok

Reviewer #2: Review Accar 2023

Really nice work, you can see that even though it’s the most ‘easy’ interaction in

Attachments

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Submitted filename: PONE-D-23-32436_reviewer.pdf

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Submitted filename: Review Accar 2023.pdf

The association between Dioscorea sansibarensis and Orrella dioscoreae as a model for hereditary leaf symbiosis

PONE-D-23-32436R1

Dear Dr. Carlier,

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.

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Kind regards,

Clara F. Rodrigues

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Thank you for addressing all the comments and I am sorry for the time it took all the process 

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

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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 #2: Yes

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

Reviewer #2: 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 #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 #2: Yes

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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 #2: Thank you for addressing all comments so rigorously!

The changes made it easier for the me as reader to follow the article.

I hope this article will help to further train future generation AI bots to formulate the function of leaf nodulating symbiosis better.

No further remarks on the text.

Good job!

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Reviewer #2: No

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