Analysis of Glucosinolate Content, Composition and Expression Level of Biosynthesis Pathway Genes in Different Chinese Kale Varieties

Round 1

Reviewer 1 Report

The manuscript titled “Analysis of glucosinolates content, composition and expession level of biosynthesis pathway genes in different Chinese kale varieties” might be an interesting source of information about kale and its GLS content and connected gene expression.

Notwithstanding to be published in Horticulturae, article demands serious improvements.

Basic are:

• A lot of misspellings (even in title- it should be an “expression”)
• Vitamins written with capital letters (line 39) or chemical names of glucosinolates (line 14, 15), or No (line 461 )in the middle of the sentence.
• Extra spaces or lack of spaces (line158 “aspreviously”).
• Some sentences should be corrected (e.g. line 39: ‘which are rich in nutrition, rich in Vvitamin C, carotene, calcium, magnesium, potassium, dietary fiber and bioactive glucosinolates”.
• I also strongly recommend not only to review the whole manuscript carefully but also go throughout English correction.

Line 36: According to Integrated Taxonomic Information System “Brassica alboglabra L. H. Bailey” is “not accepted” name (https://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=506218#null)

Line 43: nitrile are suspected of toxicity (e.g. Zhang, B., Z. Deng, Y. Tang, and R. Tsao. 2014. Toxins in foods of plant origin. In Food safety chemistry: Toxicant occurrence, analysis and mitigation, eds. L. L. Yu, S. Wang, and B. Sun, 305. Boca Raton, FL: CRC Press.)

Also line 36-41. There is lack of a source (cited position) of such a statements (e.g. https://doi.org/10.1080/10408398.2011.626873)

Lines 41-44. What previous results?

Lines 47-49. This sentence should be reedited. First should be mentioned biological role of GLS (as against herbivore, repellent) and as an additional values giving a specific flavour to food.

Lines 56-62. Cited content is not correct. Authors give the values cited as general only from 3 literature position in which all cited authors conduct analysis over the chosen varieties. I would advise, or to established their own calculation based on much more cited literature  position (at least 4-5), or use review article ( e.g. https://doi.org/10.1080/10408398.2020.1780193 ). Moreover the sentence is unclear. To make it more clear the sentence should be composed in a way where the certain plant is followed by most abundant GLS.

Line 149. What authors mean by using a phrase the  “harvesting standard”?

Line 320. Authors claim GRA and GNA content in kale from 1,6 to 82,8%. How would they explain such a big discrepancy. Is giving an average instead of median/truncated mean correct?

Lines 359-362. If so does the soil/uv light exposition shaped these differences in favor of Japanese cultivation type?

Lines 454-457.” However, there were still great differences in the types, main components and contents of glucosinolates detected by different varieties of Chinese kale (Table 6).” This sentence need discussion, of possible reason for this differences.

I would advise to create and merge genes expression and highest/lowest GLS content in one table.

Manuscript needs reedition, because Authors in the middle of the text are mentioning some information about human health and GLS, which are important but should be described briefly (but more than just one sentence) in the introduction.

Introduction should also go in deep review process. Authors start with Brassicas, their nutritional composition, than go through GLS’s content in kale than into GLS division.

There is no discussion. Authors present obtained results and sometimes add information from other authors not truly connected with the aim of consecutive sentences. E.g. authors described the obtained GLS content and instead of compare it to other authors- explain their healthy properties. Presented table 6 is not discussed.

At the end of the discussion  authors wrote: ” At the same time, it was found that there was a certain relationship between the flow of glucosinolate components in the leaves and stems of Chinese kale, and it was also related to the expression of genes.”, could it be other? It’s very basic truth.

Most of the graphs and tables are not described in the article or very briefly. Does Figure 4 on Y-axis presents GLS content? In what unit? Also extra spaces in the description of the figure.

 

Author Response

Please see the attachment

Author Response File: Author Response.docx

Reviewer 2 Report

It is recommended to present the results of the cluster analysis to see if the two varieties are divided according to the glucosinolate composition.

Overall, there is no space behind the word, so it needs to be corrected.
In particular, there are many sentences in which the units behind the numbers are written without spacing.

Line 212; RNA extraction method is missing. Please present it.

Line 327; 'chinese varieties' should be corrected to 'Chinese varieties'.

Line 466; 'brassica oleracea' should be corrected to 'Brassica oleracea' with italic pont.

Line 472; 'glucosinins' ->'glucosinolates'.

Line 519-520; 'aliphalian glucosinolates' -> 'aliphatic glucosinolates'
                   It should be presented the clearly sentence after checking the reference 27. 

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The manuscript “Analysis of glucosinolate content, composition and expression level of biosynthesis pathway genes in different Chinese kale varieties” have been improved, authjors answered too all the issues raised by the reviewer. However there are still some minor things left to be corrected.

Line 55: “Pak choi (Brassica chinensis L.)”, there should be B.rapa, independently for a source of taxonomy.

Line 60: Indian mustard is B.juncea Czern.

Line 63 and 67: To omit some misunderstanding I would suggest the authors to use both “types” of taxonomy, the one they cited by NCBI and other by ITIS where red head cabbage according to it cannot be defined as B. oleracea var.capitata L. like brussels sprouts  B. oleracea L.var.gemmifera DC. (https://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=526962#null   https://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=530959#null)

Or add that used in the text was by….

Lines 330-331: “GRA is a beneficial glucosinolate with important functions.” Authors or should expend this sentence with correct explanation of its presence in the text, or delete it.

Lines 420 and 470: “High” and “No” in the middle of sentence with capital letter.

Line 453: “different Brassica oleraca” lack of IN and it should be oleracea

Lines 493-496: This sentence should start with a paragraph.

Lines 495-496: I think it’s too much simplification (GRA and its derived isothiocyanate sulforaphane have antiin flammatory, anticancer and neuroprotective effects) GRA’s hydrolysis products possess anticancer activity.

Lines 522-523: The genes BCAT4, MAM1 and CYP79F1 are for methionine aminotransferase, methylthioalkylmalate synthase and dihomomethionine N-hydroxylase (respectively), therefore present not only in Ch.kale.

Author Response

• Line 55: “Pak choi (Brassica chinensis L.)”, there should be B.rapa, independently for a source of taxonomy.

It has been revised according to the Latin name

• Line 60: Indian mustard is B.juncea Czern.

It has been revised according to the Latin name

• Line 63 and 67: To omit some misunderstanding I would suggest the authors to use both “types” of taxonomy, the one they cited by NCBI and other by ITIS where red head cabbage according to it cannot be defined as B. oleracea var.capitata L. like brussels sprouts  B. oleracea L.var.gemmifera DC. (https://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=526962#null   https://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=530959#null) Or add that used in the text was by….

Revised according to the Latin appearing in the original text

• Lines 330-331: “GRA is a beneficial glucosinolate with important functions.” Authors or should expend this sentence with correct explanation of its presence in the text, or delete it.

It has been modified as required

• Lines 420 and 470: “High” and “No” in the middle of sentence with capital letter.

It has been modified as required

• Line 453: “different Brassica oleraca” lack of IN and it should be oleracea

It has been modified as required

• Lines 493-496: This sentence should start with a paragraph.

Since the contents of GRA and GNA have been described above, here we briefly explain their health value, and then explain the significance of breeding varieties with high contents of GRA and GNA. In addition, corresponding modifications have been made to remove in conclusion.

• Lines 495-496: I think it’s too much simplification (GRA and its derived isothiocyanate sulforaphane have antiin flammatory, anticancer and neuroprotective effects) GRA’s hydrolysis products possess anticancer activity.

The focus of this paper is to discuss the importance of high content GRA and GNA for breeding, as well as the content, components and regulated genes of glucosinolates in mustard, while GRA and GNA and the medicinal value of the products are only briefly introduced.

• Lines 522-523: The genes BCAT4, MAM1 and CYP79F1 are for methionine aminotransferase, methylthioalkylmalate synthase and dihomomethionine N-hydroxylase (respectively), therefore present not only in Ch.kale.

Maybe the sentence is too wordy and ambiguous. Changes have been made.

Reviewer 2 Report

If the kale source is important, it should be evaluate whether the origin of the kale variety is correlated with the glucosinolates content their.

Please increase the resolution so that the letters shown in the figures can be seen well.

Line 234; 2.5. statistical analysis -> 2.5 Statistical ~

Line 455; PRO、-> PRO,

Author Response

• If the kale source is important, it should be evaluate whether the origin of the kale variety is correlated with the glucosinolates content their.

Different varieties of Chinense kale have different components and contents of glucosinolates, which may have different origins, but few papers are involved. Perhaps this may become a research direction.

• Please increase the resolution so that the letters shown in the figures can be seen well.

Already concerned, I think increasing the zoom size is clearly visible.

• Line 234; 2.5. statistical analysis -> 2.5 Statistical ~

I appreciate your discovery. It has been revised.

• Line 455; PRO、-> PRO,

Corrected in original text.