Effects of UVA on Flavonol Accumulation in Ginkgo biloba

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

 

Title: Effects of UVA on flavonoid accumulation in Ginkgo biloba

 

Comments to the authors.

I found that this manuscript does not follow the Author Guidelines (see below). I recommend the authors to read the Author Guidelines and check the entire manuscript before submission.

 

L51 Some flavonoids act as defense chemicals. Is this also the case for Gingko? If yes, give a brief explanation (with proper citation(s).

L34 & L78 Give precise scientific name for that particular cultivar “golden-leaved Gingko” (as in the tea cultivar in L363). Most readers are not familiar with Gingko. Explain the basic traits of that cultivar (e.g., why the color of their leaves differs from ordinary green Gingko leaves? Except for the color, are they similar to ordinary green Gingko?)

L90 What do they mean by “same white intensity”?

L61 Give scientific name (together with their common names) for any plant materials (e.g., “Chinse Kale baby leaves”). See Author’s Guideline of the journal.

L350 Why the family name “Brassicaceae” appears in italics? Only genus and species (subspecies) names are in italics.

L351 This scientific name should be in italics.

Comments on the Quality of English Language

N.A.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The reviewed manuscript, forests-2983591, is a good experimental study dedicated to the study of the influence of part of the UV spectrum on the production of flavonoids. The authors have used a modern experimental setup. Therefore, I believe that the paper will be interesting to readers and can be published in Forests after corrections according to the comments below.

Comments:

 

1. Please check the manuscript for typos (for example, missing spaces in lines 14, 53, 56, 58, etc.) and typing mistakes (line 64, 151, etc.).
2. Please check that all used abbreviations have full explanations at first mention (for example, UVA in line 12 and many others).
3. Use the equation editor for equation 1 (lines 124–125).
4. The structure of subchapter 2.3.4 is a little confusing.
5. Please check that the text size in the figures corresponds to the Instructions for authors.
6. The caption of Fig. 3 must be after the figure.
7. Add the reference to the website (line 202).

Comments on the Quality of English Language

Minor editing of the English language is required.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The study presents results on the effect of UVA treatment on flavonoid biosynthesis in leaves of Gingko biloba. Authors compared two varieties, the first with gold leaves and the second with green leaves. To explain UVA effect the content of three selected flavonols was determined, the expression of genes related to flavonoid biosynthesis and the transcriptomic sequencing was performed.

This is very huge and interesting study broadening the knowledge on subject investigated, however there are some aspects that should be improved or explain.

Specific comments:

1)      Abbreviations when used for the first time should be explained;

2)      Typing errors should be removed;

3)      Latin names of plants has to be written using Italic font;

4)      English language style should be improved;

5)      Too long sentences are used;

6)      Abstract:  at first it is mentioned that gingko flavonoids content was determined, but next only flavonols are mentioned – why?

Line 8: “….reaching the maximum in UVA-4..” – which is?

Line 12: “…The PAL, C4H,4CL...” what these abbreviations stand for?

Line 22: “.. TL, CKY, and CKL..” – what these abbreviations stand for?

Line 24: “…CHS, F3H, and FLS..” - what these abbreviations stand for?

Generally abstract is written for person familiar with the investigation describe in the study, whereas it should be a short and informative presentation of assumptions for this study and the most prominent results;

7)      Flavonol is not a synonymic name of flavonoids, this is cardinal error; if it is used in meaning of three investigated flavonols it should be corrected

8)      It is not proper to express total flavonoid content as a sum of three compounds multiplied by a 2.51, and why 2.51? LC-MS/MS analysis could reveal presence of other flavonoids;

9)      To express total flavonoid content spectrometric methods were developed, methods based on complexation of aluminium ions;

10)  Introduction:

Lines 51-53: “ Some… [10].” – by who?

Line 67: “effect of UVA supplementation…” – supplementation?

Line 69: “ added” - ?

11)  Materials and Methods:

Line 80: “..peeling the seeds in a 4°C..” – incomprehensive;

Lines 90-91: “2,4, and 6 UVA tubes (395-405 nm) were added..” - unclear - "were added"? further too long sentence;

Line 113: “buffer solution” – what kind of?

Line 122: “liquid chromatograms” - what is this?

Line 124: “mg/g” – g of what?

Section 2.3.3 and 2.3.4 are written as manuals not scientific text;

Line 132: “RIN” – what is this?

Line 143: “..USA). concerning its method.” – what does it mean?

Nothing is said about statistical evaluation of data;

12)  Results analysis – Results in enough

Lines 175-176: “…enzyme activities showed a tendency to increase and then decrease,..” – it means nothing, so what was the response? It must be better written;

Figure 2; Figure 3: “Golden-leaved” etc. – this is not proper English name;

Further what denotes the similar/different letters on the charts?

Lines 186-187: “..The flavonol and total flavonoid contents of green-leaved ginkgo showed a trend of increasing and then decreasing with the increase of UVA intensity, and the total flavonoid content of UVA-4 treatment was the largest, which was 20.69 mg/g.” - it means nothing, so what was the response? It must be better written;

Lines 191-192: “..and the total flavonoid content of green-leaved ginkgo increased and then decreased with the increase of UVA intensity, while the total flavonoid content in UVA-4 treatment was the largest, which was 26.3 mg/g.” - it means nothing, so what was the response? It must be better written;

Figure 9; Figure 12: what is the Y axis legend?

13)  Discussion:

Lines 338-339: “showed  a trend of rising followed by a decline, and the enzyme activities were greatest in the UVA-4 treatment.” – it means nothing, so what was the response? It must be better written; the same remark for text in lines 352-353;

Lines 357-358: “Flavonoids are biosynthesized by the phenylpropane metabolic pathway, and PAL, C4H, and 4CL are the key enzymes of the flavonoid synthesis pathway, which regulate and control the biosynthesis of secondary metabolites[36].” – similar statement was done in lines 331-332 and reference [26] was used, why?

Lines 395-399: “In this experiment, we found that the transcription factors MYB (Gb_02997), bHLH (Gb_05320), bZIP (Gb_00122), and NAC (Gb_13200, Gb_37720) showed the same trend of total flavonoids of ginkgo, which might pos-itively regulate the synthesis of ginkgo flavonols, while MYB (Gb_09986), bHLH (Gb_ 32351), ERF (Gb_10817, Gb_34849) and other transcription factors were in the opposite trend of total ginkgo flavonoids and may negatively regulate the synthesis of ginkgo flavonols.” – the same trend? What does it mean?

14)  Conclusions:

Lines 402-408: “The enzyme activities of the phenylpropane pathway and the flavonoid contents of the whole sibling hybrid offspring of G. biloba showed significant differences under different intensities of UVA treatment. The enzyme activities of PAL, C4H, and 4CL exhibited a pattern of initially rising and then declining with increasing UVA intensities, and the enzyme activities were the largest in the UVA-4 treatment. The contents of quercetin, kaempferol, and isorhamnetin along with total flavonoids tended to increase and then decrease with the increase of UVA intensity, and the flavonols and total flavonoids were the largest in the UVA-4 treatment.” - so what was the response? It must be better written;

 

Comments on the Quality of English Language

English language style should be improved.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

See the attachment

Comments for author File: Comments.pdf

Comments on the Quality of English Language

English language is pretty poor. Manuscript needs thorough revision.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

The following aspects of the texts still requires improvements:

1.     Latin names of plants has to be written using Italic font;

2.     It has not been explain why the sum of flavonoid glycosides had been multiplied by 2.51;

3.     The usage of “total” in context of concentration of three investigated compounds is unacceptable, it need to be corrected;

4.     Section 2.3.1 is still written as an manual which is unacceptable in scientific texts; further “certain mass” – it cannot be certain mass but it should be given e.g. 1 g, 100 mg etc;

5.     “As shown in Figure 3, with the increase in UVA intensity, the enzyme activities of PAL, C4H, and 4CL of yellow-leaf ginkgo mutant showed a tendency to increase and then decrease….” – this manner of results description is unacceptable, it need to be improved, also in other parts of the text;

6.     Figure 9, Figure 12: “ordinate”? the axis legends need to be added;

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

Title of the manuscript: Effects of UVA on flavonol accumulation in Ginkgo biloba

Over all, the manuscript has been revised keeping in view of the Referee’s comments.

However, there are still Latin names for example Brassica napus, which have not been put in Italics.

 

For enzyme units, it is written every where umols (It should be µmols, not umols). Insert the symbol micro.

Comments on the Quality of English Language

English language has been improved. However, subtle changes are necessary.

Author Response

Thank you very much for taking the time to review this manuscript. I have made changes to the Latin name format and enzyme activity units. [These changes can be found - at lines 211, 220, and 427.]