Integrated Volatile Metabolome and Transcriptome Analyses Provide Insights into the Formation of Benzenoid–Phenylpropanoid Aroma Substance Eugenol in the Rosa hybrida ‘Lanxing’ Flowering
Round 1
Reviewer 1 Report
The study of the genes responsible for the biosynthesis of secondary metabolites, especially volatile compounds, is a very difficult task. Dozens of genes and transcription factors can be involved in the process of biosynthesis. The difficulty may also lie in the fact that some of the genes may be specific to only one plant species. The authors conducted a large and varied study. Unfortunately, there are a number of comments.
All tables are missing in the Supplementary, except for S2.
121 - indicate the amount of the sample in digitals
128 - indicate which solvent was used
130 - specify "scan mode:san"?
211 - more accurately detected, not identified. You have not established the structure of all substances
What parameters were included in the K-mean method? Not listed under Materials and Methods
In Figure 3, indicate on the "y" axis the content units
313- what does "All these pathways were related to energy and matter, suggesting the variety ‘Lanxing’ flowering process was also an energy and matter metabolism process" mean. All metabolic processes are related to matter and energy, not only these.
346 - where is the information about the HMM domain?
It follows from Figure 7b that, in terms of expression level, the RcEGS28 gene is more suitable than the RcEGS26 gene. Perhaps that is why good results were obtained only with the RcEGS32 gene.
426 - on line -211 - it is indicated that 328 volatile substances were identified (detected) by HS-SPME GS-MS / MS methods, and 1046 were already identified (detected) in the Discussion section. Where is the real data?
444, 446 - references, according to the rules
470, 476 - in vivo, in vitro italic
Author Response
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Author Response.docx
Reviewer 2 Report
Optional: in title if you add the name rose, hybrid rose or R. chinensis?? etc. the add specie name
Abstract
ok
Introduction
Write about ‘Lanxing’. variety/ specie/land race etc. its history, breeding or in other sense why you choose this variety.
You discuss R. hybrid in introduction, were is R. chinensis.
Materials and Methods
Start from plant material, origin, history, breeding how it come to your station
what was the criteria/morphology/differences with others you selected this variety on the basis of scent?
Sample preparation
The variety ‘Lanxing’ possess typical benzenoid-phenylpropanoid volatiles scent characteristic.????? what is concerned here??? still not detected in your research.
For stage 3, the flower tissue of sepal, leaf, style, anther, and petal were sampled and placed in liquid nitrogen, immediately. did not discuss any where in manuscript.
RNA extraction
You started with the samples? what material you use sapeles, petals etc.????
3.2. Transcriptome alterations of the variety ‘Lanxing’ during flowering
Is this differential gene expression??? you compared with other roses. is this sample differences. or stander.
Figure 7.
variety ‘Lanxing’ flowering. is this for flowering or scent.
3.5. Identification of key eugenol synthetic enzyme gene
petal developmental process was in agreement with the transcriptome data (Figure 8a, b)???
3.6. Identification of transcription factor participated in eugenol synthesis process
In order to identify the specific transcription factor about structural gene????
Figure 9.
during the variety ‘Lanxing’ flowering.???
Discussion
purple blue variety ‘Lanxing’ as material?? did not mention anywhere.
Last line Rosa hybrid.??? R. chinensis?? which one or what is the difference.
Improve english making proper sentences: is, are, was and were mistakes.
Author Response
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Author Response File: Author Response.docx
Reviewer 3 Report
Reviewer comments
Journal: Forests (ISSN 1999-4907)
Manuscript ID: forests-2602195
Type: Article
Manuscript Title: "Integrated volatile metabolome and transcriptome analyses provide insights into the formation of benzenoid-phenylpropanoid aroma substance eugenol in the variety 'Lanxing' flowering"
In the current work, the authors wrote an article about “Integrated volatile metabolome and transcriptome analyses provide insights into the formation of benzenoid-phenylpropanoid aroma substance eugenol in the variety 'Lanxing' flowering”
Although this study is very good and the manuscript is clear and well written, it is more suitable for other MDPI journals such as Horticulture, Agriculture, Molecules (ISSN 1420-3049), [Foods (ISSN 2304-8158), Special Issue "Essential Oils: Chemistry and Food Applications"], Biomolecules. However, the topic is nice and the results are of interest for the scientific community. The study is well-designed and the methods are clear. The figures are prepared in a good quality and the results and discussions are well-structured. However, the text needs a minor revision before publication.
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The comments:
1- L 16, L 22, L 25, L 92, L99, L 440……….: “we and Our” Do not write the personal pronouns in the entire manuscript
2- The results need further clarification in the abstract section
3- Keywords: please capitalize the first letter.
4- L 37: “rosa” correct to “rose”.
5- From line 42 to line 53: The biosynthesis of Phenylpropanoids part is more suitable in the discussion section, so, pls remove it from the introduction section and rewrite it in the discussion.
6- In the introduction: the paragraph that started with Eugenol (L 55), the authors have to jointed or explain the relationship between Phenylpropanoids and Eugenol.
7- In L 59: “EGS” and L 66 “MYB” The usage of abbreviation should be used after the full term. Please be consistent with usage of all abbreviations. Pls revise the abbreviations in the whole parts of MS.
8- L 60: “Ocimum basilicum and Clarkia breweri” italic.
9- L 79-81: please add relevant reference.
10- L 89: “Torreya grandis” Italic.
11- L 89: “Sorghum, and Potato” Write the scientific names.
12- Please briefly describe the results in the last paragraph of introduction.
13- L 104: “The variety” the variety of What??.
14- All the results were separated without a storyline or connection between each other, please revise and improve.
15- L 141: “A total of 33 eligible RNA samples were obtained for transcriptome analysis and qRT-PCR, BUT, in line 145, A total of 18 transcriptome profiles (6 stages × 3 replicates) have been deposited in ….. .”Are all 33 samples used OR only 18 samples?. Thus, where other 15 samples were used?
16- L 194-196: “Three biological replicates were performed for each sample, and three technique replicates was conducted for each biological replicate. What is the mean of technique replicates?” This means that 9 replicates were used for each sample?. Please be clear with this.
17- Figure 3: the (X) axis, add (1, 2, 3 ……. 25). In (b), (Y) axis, the first number (5.0e+7) is correct?. I think it is (0.5e+7)
18- Fig. 3 and 8: Authors should do statistical analysis to indicate the significance amount the treatment, then, add letters of significance above columns and indicate these differences in the results with df, F, P values.
19- L 198: “the variety ‘Lanxing’” delete
20- L 256: “of the variety ‘Lanxing’” delete.
21- L 288: “the variety ‘Lanxing’” delete.
22- L 321: “in the variety ‘Lanxing’” delete
23- Figure (5 a). unclear
24- L 507: “Rosa hybrida” italic.
25- L 524:”Clarkia breweri and C. concinna” italic.
26- L 529: Clarkia breweri and Petunia hybrida. Italic.
27- L 532: “llicium anisatum” Italic.
28- L 569: “Rosa chinensis” Italic.
29- L 580: “Rosa rugosa” Italic.
30- L 582-583: “Rosa 582 odorata and R. chinensis” Italic.
31- L 588: “Xanthoceras sorbifolium” Italic.
32- Please, make an effort to synthesize the text avoiding redundancies and repetitions in the discussion section.
33- The manuscript contains some typo errors; please revise it very carefully. A carful revision for the English Grammar is required.
34- Summarizing, this is a good study, which certainly merits a publication after a minor revision.
Comments for author File: Comments.docx
Minor editing of English language required
Author Response
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Author Response File: Author Response.docx
