[EXSCINDED]

PONE-D-19-19801

Regulation of nitrogen availability results in changes in grain protein content and grain storage subproteomes in barley (Hordeum vulgare L.)

PLOS ONE

Dear Dr Xu,

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.

Although the research provided valuable information to understand the mechanisms for N regulation, the results were more descriptive and lack conclusions. It would be useful to have a detail comparison with the previous results. For this kind of research, detail information of soil nutrients is a must in the paper. The English  also need be thoroughly  edited  

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We look forward to receiving your revised manuscript.

Kind regards,

Chengdao Li, PhD

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. We noticed you have some minor occurrence of overlapping text with the following previous publication(s), which needs to be addressed:

https://onlinelibrary.wiley.com/doi/full/10.1111/tpj.12881

In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed.

Additional Editor Comments (if provided):

The MS reports different protein expression under different nitrogen supply. The research provided valuable information to understand the mechanisms for N regulation. The results were more descriptive and lack conclusions. It would be useful to have a detail comparison with the previous results. For this kind of research, detail information of soil nutrients is a must in the paper. The English need be thoroughly edited

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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: Yes

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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: The manuscript “Regulation of nitrogen availability results in changes in grain protein content and grain storage subproteomes in barley (Hordeum vulgare L.) ” by Xu et al. describes comparison proteomics analysis of barley grain grown in both low and high nitrogen level conditions. The differential expression proteins were identified by MS. This detailed analysis of grain subproteomes provides information on how barley GPC may be controlled by nitrogen supply. Thus, the manuscript need minor revised.

1. The English style should be revised by native English speaker.

2. P20 Line 382, the authors referred “A total of eight protein spots representing five hordein proteins were differentially expressed in Yangsimai 3 and Naso Nijo, which could contribute to the difference in GPC of mature barley seeds. Why? The authors should be further discuss.

3. The introduction should explain the aim of the study.

4. References should be in uniform format as required by Plos one publication.

Reviewer #2: The authors presented grain protein content and grain storage subproteomes in barley under different N supply. Data were well presented. Following are my comments to the MS:

1. The MS is a bit too descriptive and lacks major conclusion. The mechanisms underline the changes need to be discussed, i.e. many differentially expressed proteins were detected (the same as previously reported) but their roles in causing the difference between two varieties were not discussed.

2. Not sure if it's appropriate to compare the results with previously reported QTL.

3. can the authors provide soil test results (how much N in the soil before N application)?

4. Don't think they can use “biological" before replicates if their replicates are from a single experiment.

5. A conclusion paragraph is needed at the end of Discussion.

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Reviewer #1: No

Reviewer #2: No

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Regulation of nitrogen availability results in changes in grain protein content and grain storage subproteomes in barley (Hordeum vulgare L.)

PONE-D-19-19801R1

Dear Dr. Xu,

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

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

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With kind regards,

Chengdao Li, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The paper can be accepted for publication. However, there are still a few spelling and grammar errors, e.g. Hordeum not Hordeums. Please have a final read and correct all the errors along with any remaining formatting changes. 

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

**********

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

**********

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: I have no further comments apart from a few spelling errors, i.e. Hordeums vulgare L.should be Hordeum vulgare L.

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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 #2: No