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Article
Peer-Review Record

Osmanthus fragrans Ethylene Response Factor OfERF1-3 Delays Petal Senescence and Is Involved in the Regulation of ABA Signaling

Forests 2024, 15(9), 1619; https://doi.org/10.3390/f15091619 (registering DOI)
by Gongwei Chen 1,2,†, Fengyuan Chen 1,†, Dandan Zhang 1, Yixiao Zhou 1, Heng Gu 1, Yuanzheng Yue 1, Lianggui Wang 1 and Xiulian Yang 1,*
Reviewer 1: Anonymous
Reviewer 2: Anonymous
Reviewer 3:
Forests 2024, 15(9), 1619; https://doi.org/10.3390/f15091619 (registering DOI)
Submission received: 9 August 2024 / Revised: 7 September 2024 / Accepted: 11 September 2024 / Published: 14 September 2024
(This article belongs to the Section Genetics and Molecular Biology)

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

There are some major suggestions aiming to further improve this paper in terms of its readability, ease of understanding, clarity, and presentation. Therefore, the paper needs major modifications. Below are the comments, which led to this decision.

 

Check and use SI units in the manuscript, such as min instead of minutes, d instead of days, etc.

Line 168:  change MgCl2 to MgCl2

Authors should add "Experimental design and data analysis" in the "Material and Method" section.

Figure 2: Authors should add images of five flower development stages along with ERF1-3 expression data.

Figure 6: Add scale bar to Figure 6A

Line 357-358: Not relevant to the content of the experiment.

Line 391: Italicize "A. thaliana"

Line 422: Add related references for this sentence.

The discussion part is basic and poor. It is expected that the discussion section of the article is written strongly.

The conclusion part needs to be rewritten more precisely with significant future research direction.

Generally, overexpression of ethylene response factor (ERF) genes or high expression levels of ERFs genes in plant tissues can cause early flower senescence. ERFs are key components in the ethylene signaling pathway, which regulates various aspects of plant growth and development, including flower senescence. When ERF genes are overexpressed or up-regulated, they can enhance the plant's sensitivity to ethylene or mimic ethylene signaling, leading to accelerated tissue senescence. This means that flowers may be senescent sooner than they would under normal conditions. The overexpression of specific ERF genes has induced early senescence in several flower species, demonstrating their role in this process. What do you think about this? And compare with your results.

Comments on the Quality of English Language

Minor editing of English language required.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Major:

1) The most important point about this manuscript (Ms) is that it is difficult to understand the results as presented. So far, I cannot say any significant comments on the essence of the paper until the authors correct the following:

a)     Carefully reread the entire text of the Ms and pay attention to quoting figures and tables - for example, there is no citation of Fig. 1, also Fig. 7 and 8 are not quoted according to the sequence indicated in the Ms text.

b)    Increase the quality of all figures.

c)     Increase the font size in Fig. 1, 4, 5, 6, 7, and 8.

d)    Improve all legends for figures, e.g. explain all used abbreviations, plant material, etc.

e)     This Ms contains misprints, mistakes in English grammar and in the writing style. I recommend that the authors should use some help of a native English speaker or send the Ms to an English Editing Service that proofreads scientific writing.

2) How many OfERF genes does a sweet osmanthus Osmanthus fragrans have? Can the authors add them to the Fig. 1a?

3) Authors should include the GeneBank accession number of the used OfERF1-3 gene.

4) Where did the transcript data in Fig. 2 come from?

 

Minor:

5) Line 142: correct “E. coli” to “Escherichia coli” and in italic.

6) Line 156: correct “Nicotiana tabacum” to “N. tabacum”.

7) Line 172: correct “minutes” to “min”. The same remark for all other “minutes” in Ms text.

8) Line 199: correct “E. coli” in italic.

9) Include scale bar in Fig. 4, 6,

Comments on the Quality of English Language

This Ms contains misprints, mistakes in English grammar and in the writing style. I recommend that the authors should use some help of a native English speaker or send the Ms to an English Editing Service that proofreads scientific writing.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

This manuscript is dedicated to investigating the role of ethylene response factor OFERF1-3 in the petal wilting process of the Osmanthus fragrans plant. The document is well-written and relevant to the field of research, and is presented in a logical and organized manner. The cited references are current and mostly recent publications. The manuscript is based on solid scientific principles, and the methods employed are diverse and appropriate for testing the hypothesis. The authors have thoroughly explored the function of the gene using state-of-the-art techniques in molecular biology. Results from the study are supported by the details provided in the methods section. Figures, tables, and diagrams are consistent with the data presented, and are easy to understand. However, some of the plant photos could be improved in terms of quality.The data presented throughout the manuscript is interpreted consistently, and the results are presented in a manner that makes them interesting to read. The conclusions drawn from the evidence and arguments presented are consistent with each other.

 

How long can tissues be stored at -80 degrees Celsius without compromising RNA quality? 

Why was the S4 time point chosen for DNA extraction in Chapter 2.3? 

How can we explain the fact that OfERF1-3, which is normally associated with petal senescence, has significantly higher expression at the S5 time point, but its overexpression also extends the flowering period? 

The materials and methods section doesn't specify which generation of transgenic plants was used. Was there a homozygous stage? If not, it's not clear whether the transformation is complete and the overexpression stable. 

In line 157, please provide a link to the relevant method. 

Figure 4a: can you please clarify what the signatures under the plants represent and whether they are duplicated in the caption? 

Figure 6: what does this image show? Does it show that the cut parts of the plant survive longer? How is aging related to this? What does 1300 refer to?

I would like to see plans for future work. Will the findings from this study be translated into practical applications, and if so, in which area(s)? Alternatively, will we see a continuation of this comprehensive research in the next publication?

You transformed plants using an empty vector. In which specific section of the manuscript did you compare the transgenes to this vector?

Figure 6 indicates that flowering was delayed, rather than the flowering period being extended. Was this the outcome you were aiming for?

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript looks better than the previous version. Accept in present form.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Major:

1) As in the first review – the most important point about this manuscript (Ms) is that it is difficult to understand the results as presented. So far, I cannot say any significant comments on the essence of the paper until the authors correct the following:

a)     Carefully reread the entire text of the Ms and pay attention to quoting figures and tables - for example, there is no citation of Table S3. Also, Table. S4 and 2 are not quoted according to the sequence indicated in the Ms text.

b)    I did not find Table. S4 and S2 in the supplementary material.

c)     Increase the quality of Fig. 1, and 4.

d)    In Fig. 4a, can authors submit photos not every 3 days, but every 6? To make the photos bigger.

e)     Increase the font size in Fig. 1.

f)      Improve all legends for figures, e.g. explain all used abbreviations, plant material, etc., e.g. in Fig. 2, explain FKPM and origin of the transcriptome data.

g)    This Ms contains misprints, mistakes in English grammar and in the writing style. I recommend that the authors should use some help of a native English speaker or send the Ms to an English Editing Service that proofreads scientific writing.

 

2) Authors should include the GeneBank accession number of the used OfERF1-3 gene and transcriptome from Fig. 2. Include the Ms line number in the reply.

3) Why do the expression data in transgenic tobacco plants (Fig. 3) repeat the expression of OfERF1-3 gene in petals of Osmanthus fragrans? Which promoter was used?

4) Improve explanation of the obtaining of the transgene tobacco plants (Line 155-163).

Comments on the Quality of English Language

This Ms contains misprints, mistakes in English grammar and in the writing style. I recommend that the authors should use some help of a native English speaker or send the Ms to an English Editing Service that proofreads scientific writing.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

I would like to extend my sincere thanks for the responses provided. I have no additional questions at this time, and I wish to express my appreciation for the excellent work done, particularly the challenging aspect involving the two-hybrid yeast library.

Following the improvement of image quality, editing of text, provision of comprehensive responses to questions, and supplementary information, the manuscript has acquired special significance for the scientific community and has become more appealing to readers. Moreover, the presentation of material has become more coherent and logical.

However, I would still recommend that the authors use the T3 generation of transgenic homozygous plants for future studies on gene function, as this is essential to achieving the desired outcome. The use of T1 plants can also be employed, but if the purpose is to produce a plant with desired properties, it is advisable to use the T3 generation.

We look forward to continuing our research in a future article!

Best regards,

Dr. Ogneva

 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

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