Transcriptomic and Metabolomic Analyses Provide New Insights into the Response of Strawberry (Fragaria × ananassa Duch.) to Drought Stress

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

I checked your manuscript and described comments below.

Strawberry (Fragaria × ananassa Duch.) is sensitive to drought, so it is important to analyze the expression of genes related to this. This paper provides a very good analysis.

I think it would be better to consider the following points.

1.       I think it would be easier to understand what type of strawberry it is if I included "strawberry (Fragaria × ananassa Duch.)" in the title.

2.       I think there are many people in the world who don't know where "Zhangji" is. I think it would be better to include a map.

3.       If there is anything special about "Zhangji" Strawberry (Fragaria × ananassa Duch.), I think it would be a good idea to describe it.

4.       MetaboAnalysis 4.0 is an old software. We recommend you use the latest version, MetaboAnalysis 6.0.

5.       I don't know what makes the Fragaria × ananassa Duch. produced in "Zhangji" different from others. If there is something different about this, I think it would be better to cooperate a little more.

I don't think this paper has new various major mistakes or grammatical problems.

Author Response

Thank you for your valuable suggestions, we have made the following revisions:

Question 1-3: "Zhangji" (Fragaria × ananassa Duch.) strawberry is an early-maturing variety that is hybridized with the Nyome variety and originated in Shizuoka, Japan. It was introduced to Donggang City, Liaoning Province in 1996. "Zhangji" strawberry is one of the main varieties in the greenhouse. It is large in size and small in deformation, which is suitable for pot experiments. And it is less studied. We added information about "Zhangji" strawberry in the introduction section and the materials and methods section;

Question 4: We have made revisions based on your comments, re-examined the results shown, and made corrections and revisions;

Thank you again for your valuable comments, please check our revised manuscript;

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Comments and remarks:

 The manuscript presented for review studied the drought resistance of cultivated strawberry (Fragaria × ananassa Duch.) to clarify physiological changes in strawberry during the drought response. Knowing the mechanisms of this response is of high importance in atempt for the breeding of drought-resistant strawberry varieties. Up to now, the molecular regulation of drought stress was revealed in wild strawberry (Fragaria nilgerrensis), but for the cul tivated strawberry varieties, little information is about the physiological and molecular mechanism of its response to different degrees of drought treatment. To clarify cited mechanisms, in this study transcriptomic sequencing and mass spectrometry methods were used.

The manuscript is constructed according to requirements of “Horticulturae”.The applied research methods are suitable for achieving the research objective. The results are clearly presented and illustrated in good quality figures and are statistically supported. The statements made are based on the results obtained.

 Although some remarks and recommendations should be made:

Introduction

-          Line 34: “Strawberry ….”  to put on a new line, and add the information given in “Discussion” /line 268/:”.. is one of the most well-received fruits, because of its sweet smell and rich nutrients”. Thus, this passage would sound like this: "Strawberry, a member of the Rosaceae family, is one of the most well-received fruit plants for its sweet aroma and rich nutrients. It is a shallow-rooted plant that requires loose, permeable and airy soils.”

-          The sentence in lines 49-52: “Bio-stress and …. adaptation [8].” To be moved at line 34, avter : “ ….of plants[2].”

-          Line 45-46: to give the meaning of ABA and ICE 

Materials and Methods

-          In the part describing the “Daily transpiration:” /lines 89-93/, the citation of the reference – “[11]” have to be puted in line 90: “…water 89 balance method [11]: …”

-          The same applies to the paragraphs of lines 128-133, 134-137 and 167-176, as follows:

-lines 128-133: the citation of the reference – [13] have to be puted in line 128: “..DEseq software [13]:..”, and the sentence “The overall distribution of DEGs in sam-128 ples was visualized using a volcano map.” Have to be moved at the end of paragraph, in line 133]

-lines 134-137: the citation of the reference – [14] have to be puted in line 134: “… using Blast2GO software [14],…”

-lines 167-176: the citation of the reference – [15] have to be puted in line 167: “Metabolic extracts [15]…”

Results

-          3.1. Effects of Drought on the Physiology of Strawberry- twe two firs sentences of this part have to be removed because repeat information given in “Introduction” and “Discussion”.

Discussion

-          The first paragraph of this section /lines 268-274/ have to be omitted because contains more general information, and discussion element in it.

-          Since in “Introduction” is noted that were carried out investigation on the molecular regulation of drought stress in wild strawberry, it is appropriate to draw here a parallel with the result received in present study on the tivated strawberry.

 In conclusion the manuscript is recommended to be published in “Horticulturae”.

 

Author Response

First of all, thank you very much for your valuable suggestions. We have made the following modifications:

We affirm that the paragraph statement you understand in line 34 is correct. We have revised and corrected the literature problems you mentioned, including the introduction and materials and methods.Duplicate information in the results and discussion section, and deletions and changes that have been made in the results section.

Thank you again for your valuable comments, please check our revised manuscript;

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Article horticulturae-3062427-peer-review-v1: Transcriptomic and Metabolomic Analyses Provide New Insights into the Response of Strawberry to Drought Stress.

 

1-      The introduction could be improved, and incorporate additional references:

 

When referring to main cultivated varieties; You could mention the varieties, and specify whether you are referring to the world or the authors' country. Include references and updated production statistics from relevant Agriculture agencies.

Also in the aim of the research that is being reported, specify whether the study is in the field, in a nursery, and other specific characteristics of the crop studied.

2-      Section 2.1. Experimental Design

 

Potted "Zhangji" strawberry (Fragaria × ananassa Duch.) was taken as the test material.

 

Please, indicate the origin of the plant material, and include, if possible, some certification that it is the species you mention.

 

3-      Section 2-       Section 2.1. Experimental Design

Improved paragraph between lines 66-81, page 2.

 

Mention if the developed protocol is yours or if it has already been developed or suggested by other authors, in which case include a bibliographic reference.

 

Briefly explain what it means:

"conventional water supply treatment were set".

 

4-      2.4.1. Preparation of the Metabolomic Standards

 

Include data referring to the company and country of origin from which the  24 hormone standards used were purchased.

 

5-      Section 2.4.2. Metabolites Extraction

 

Include data referring to the analytical quality, as well as company and country of origin from which the acetonitrile used were purchased.

 

6-      Include as supplementary material, a table with HPLC MS quantification of the 24 hormones.

7-      Paragraph 311-317, page 10

Mention what type of fruits the studies were carried out on.

 

8-      Include in the discussion a more specific paragraph on the contribution of

Cao et al.(2022) had revealed the molecular regulation of drought stress in wild strawberry (Fragaria nilgerrensis) with integrated transcriptome and methylome analysis[9]

 

Considering the suggestions, the paper should be considered for acceptance.

Comments on the Quality of English Language

Moderate editing of English language required

Author Response

Thanks for your valuable suggestions, we have made the following modifications:

Question 1-2: We have added the information about Zhangji strawberries in the introduction and the materials and methods section, and introduced the advantages and disadvantages of the selected varieties of Zhangji strawberries.

Questions 4-5, 7-8: We have supplemented the source information of the material in question.

Question 3: The relevant content has been added to the article.

Question 6: The table with HPLC MS quantification of the 24 hormones have been supplemented material in the form of attachments.

Thank you again for your valuable comments, please check our revised manuscript;

Author Response File: Author Response.pdf