Differential Metabolite Analysis of Anthocyanins in Variously Colored Petal Types During Different Developmental Stages of Sophora japonica L.
, Tao Sun 1,†, Yanjuan Mu 1, Yan Wang 1, Yi Han 1, Yanguo Sun 1, Xinhui Li 1,2, Kongan Xie 3, Shuxin Zhang 1,2, Chenxia Song 1,4 and Yizeng Lu 1,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe topic of this research is highly interesting, reporting on the differential metabolites analysis of anthocyanins in variously colored petal types during different developmental stages of Sophora japonica L. The manuscript presents an insightful and valuable analysis; however, there are several areas that require attention before it can be considered for publication in the journal Horticulturae. The detailed corrections are as follows:
1. It seems there are some errors in the species name formatting in the title. The species name should be italicized as Sophora japonica L.
2. There appear to be additional errors in species name formatting in lines 81, 84, 90, and others. Please carefully review and correct these inconsistencies.
3. It would be helpful to explain the rationale for using Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) to analyze anthocyanin-related metabolites. Highlighting the advantages of this method, especially in comparison to other metabolomic techniques mentioned earlier, would provide greater clarity to readers.
4. The title for Figure S2 is missing. Including this title will improve the completeness and readability of the supplementary materials.
5. It would be beneficial to thoroughly proofread the manuscript to address any typographical or formatting errors that may have been overlooked.
Author Response
Introduction need to provide sufficient background and include all relevant references, it must be improved.
Response:We have carefully revised the introduction and added the relevant background information.
- It seems there are some errors in the species name formatting in the title. The species name should be italicized as Sophora japonica L.
Response: Original manuscript has been modified.
- There appear to be additional errors in species name formatting in lines 81, 84, 90, and others. Please carefully review and correct these inconsistencies.
Response: We have thoroughly reviewed and made revisions throughout the manuscript.
- It would be helpful to explain the rationale for using Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) to analyze anthocyanin-related metabolites. Highlighting the advantages of this method, especially in comparison to other metabolomic techniques mentioned earlier, would provide greater clarity to readers.
Response: We have added the missing relevant section in the introduction.
- The title for Figure S2 is missing. Including this title will improve the completeness and readability of the supplementary materials.
Response: The title has been added to the manuscript.
- It would be beneficial to thoroughly proofread the manuscript to address any typographical or formatting errors that may have been overlooked.
Response: We have carefully proofread the entire manuscript.
Reviewer 2 Report
Comments and Suggestions for AuthorsDear Authors.
I have studied the presented article with great interest; this article contains a large volume of labor-intensive mass-spectrometric studies, which in itself is of great value. Without a doubt, this article is quite worthy of publication in such a respectable Journal.
Using UPLC-MS/MS, 36 anthocyanins and 9 flavonoids were identified in the QB, YB, and LGB petal types at the flower bud phase, initial flowering phase, full bloom phase, and final flowering phase in "AM". Based on petal color phenotypic analysis, OPLS-DA, and different metabolite enrichment, ten metabolites, including peonidin-3-O-rutinoside, etc., were identified as key substances distinguishing the differences of the color between YB and LGB petals compared to QB. These findings could enhance the understanding of the biometrical mechanisms underlying petal coloration in S. japonica and clarify the anthocyanin biosynthetic pathway.
However, some maxims and conclusions made by the authors require additional clarification, possibly the use of additional literature interpreting the results obtained.
1. Authors write: In S. japonica "AM", the coloration of QB differed from that of YB and LGB, with QB being yellowish-white, and YB and LGB exhibiting light purplish-red hues. However, the mechanisms underlying these color differences remain unclear.
A very controversial statement. Additional clarifications are required, possibly using additional literature interpreting the results obtained.
2. Authors write: Although the delphinidin-like anthocyanins were more prevalent in QB than in YB and LGB, delphinidin-3-O-arabinoside and dephinidin-3-O-sophoroside were undetectable at the full bloom and final f flowering phases, indicating that their synthesis was completed in the earlier phases and their accumulation thereafter was limited.
Additional clarifications are required, possibly using additional literature interpreting the results obtained.
Author Response
- Authors write: In S. japonica "AM", the coloration of QB differed from that of YB and LGB, with QB being yellowish-white, and YB and LGB exhibiting light purplish-red hues. However, the mechanisms underlying these color differences remain unclear.
A very controversial statement. Additional clarifications are required, possibly using additional literature interpreting the results obtained.
Response: The relevant section has been further explained and revised in the manuscript.
- Authors write: Although the delphinidin-like anthocyanins were more prevalent in QB than in YB and LGB, delphinidin-3-O-arabinoside and dephinidin-3-O-sophoroside were undetectable at the full bloom and final f flowering phases, indicating that their synthesis was completed in the earlier phases and their accumulation thereafter was limited.
Additional clarifications are required, possibly using additional literature interpreting the results obtained.
Response: We have provided further explanation and made revisions in the manuscript.
Reviewer 3 Report
Comments and Suggestions for AuthorsDear authors, I have some comments about the manuscript:
line 4: Sophora japonica should be italic
lines18-22: too long sentence. It is unclear whether they studied Sophora japonica or the AM variety . The meaning of AM is not clarified .
line 84 Helianthus annuus L. should be italic.
line 90 S. japonica should be italic. Please describe the variant AM.
Figure 1: The picture has low resolution.
lines 118-122 The colorimetric assay should be better described.
line 156. Section 2.5: Metware Database (MWDB) needs a reference.
line160 Multiple Reaction Monitoring (MRM) please describe and add relevant references.
line 162 Analyst 1.6.3 and MultiQuant 162 3.0.3 software lease describe and add relevant references.
line 176-178 Orthogonal Partial Least Squares 176 Discriminant Analysis (OPLS-DA), Hierarchical Cluster Analysis 177 (HCA), and Principal Components Analysis (PCA), please describe these methods and add relevant lireferences.
line 180 S. japonica should be italic.
line 182 Variable Importance in Projection (VIP) , please add references.
line 187-190: These are sentences from the journal template.
lines 224-225 The correlation analysis is not mentioned in the experimental section.
lines 243-244 The caption of Figure 2 completely lacks information about the content; it should be rewritten.
line 248 Description of the model coefficients should be described on the experimental section.
line 255 The score plot (figure S2) of OPLS-DA model should be reported in the main text.
line 268 The VIP values should be reported in a Table.
line 270 Cluster analysis is not mentioned in the experimental section.
lines 310-311 The symbols and characters in Figure 3 are too small ,making it very difficult to read and observe the content of the figure .
lines 379-390 The authors should discuss their findings, these sentences should be in the introduction section.
lines 399-424 The authors should discuss their findings, these sentences should be in the introduction section.
lines 474-47 5 These are sentences from the journal template.
lines 495-502 These are sentences from the journal template.
lines 516 Figure S2 lacks of a caption.
The experimental section lacks a comprehensive description of the methods and appropriate bibliographical references. The figures are not adequately described, and the captions do not provide the necessary information to explain the content. The font styles and line spacing used in certain sections of the manuscript are inconsistent. There are sentences from the journal's template.
My suggestion is to pay more attention to detail and thoroughly revise the presentation of the results.
Author Response
The methods need to be adequately described, it must be improved.
Response: We have supplemented the methods section and made revisions to the original manuscript.
The results need to be clearly presented, it must be improved.
Response: We have revised the relevant sections in the original manuscript.
line 4: Sophora japonica should be italic
Response: Original manuscript has been modified.
lines18-22: too long sentence. It is unclear whether they studied Sophora japonica or the AM variety . The meaning of AM is not clarified .
Response: The manuscript has been revised to “Flower color serves as a vital ornamental feauture of landscape plants, Sophora japonica L. mutant ‘AM’ exhibits the different colors from the common S. japonica., ‘AM’ presenting light purple-red wing and keel and yellowish-white flag petals, while common S. japonica is yellow and white”.
line 84 Helianthus annuus L. should be italic.
Response: Original manuscript has been modified.
line 90 S. japonica should be italic. Please describe the variant AM.
Response: The manuscript has been revised,.
Figure 1: The picture has low resolution.
Response:The original image has been modified and re-uploaded.
lines 118-122 The colorimetric assay should be better described.
Response: Original manuscript has been modified.
line 156. Section 2.5: Metware Database (MWDB) needs a reference.
Response: The relevant reference has been added to the manuscript.
line160 Multiple Reaction Monitoring (MRM) please describe and add relevant references.
Response: The relevant reference has been added to the manuscript.
line 162 Analyst 1.6.3 and MultiQuant 162 3.0.3 software lease describe and add relevant references.
Response: The relevant reference has been added to the manuscript.
line 176-178 Orthogonal Partial Least Squares 176 Discriminant Analysis (OPLS-DA), Hierarchical Cluster Analysis 177 (HCA), and Principal Components Analysis (PCA), please describe these methods and add relevant lireferences.
Response: Original manuscript has been modifiedand the relevant reference has been added to the manuscript.
line 180 S. japonica should be italic.
Response: Original manuscript has been modified.
line 182 Variable Importance in Projection (VIP) , please add references.
Response: The relevant reference has been added to the manuscript.
line 187-190: These are sentences from the journal template.
Response: Original manuscript has been modified.
lines 224-225 The correlation analysis is not mentioned in the experimental section.
Response:A description of the correlation analysis has been added to the manuscript.
lines 243-244 The caption of Figure 2 completely lacks information about the content; it should be rewritten.
Response: Original manuscript has been modified.
line 248 Description of the model coefficients should be described on the experimental section.
Response: The relevant experimental section has been revised.
line 255 The score plot (figure S2) of OPLS-DA model should be reported in the main text.
Response: Figure S2 has been corrected to Figure 3.
line 268 The VIP values should be reported in a Table.
Response: Original manuscript has been modified.
line 270 Cluster analysis is not mentioned in the experimental section.
Response: The relevant description of cluster analysis has been added to the experimental section.
lines 310-311 The symbols and characters in Figure 3 are too small ,making it very difficult to read and observe the content of the figure .
Response: The original image has been revised.
lines 379-390 The authors should discuss their findings, these sentences should be in the introduction section.
Response: Original manuscript has been modified.
lines 399-424 The authors should discuss their findings, these sentences should be in the introduction section.
Response: Original manuscript has been modified.
lines 474-47 5 These are sentences from the journal template.
Response: Original manuscript has been modified.
lines 495-502 These are sentences from the journal template.
Response: Original manuscript has been modified.
lines 516 Figure S2 lacks of a caption.
Response: A title has been added to the manuscript.
The experimental section lacks a comprehensive description of the methods and appropriate bibliographical references. The figures are not adequately described, and the captions do not provide the necessary information to explain the content. The font styles and line spacing used in certain sections of the manuscript are inconsistent. There are sentences from the journal's template.
Response: We have carefully organized the experimental section and supplemented the relevant references. We have corrected problematic images and re-uploaded them. We have also revised the details of the manuscript.
Round 2
Reviewer 3 Report
Comments and Suggestions for AuthorsThe referees have addressed all my issues. I recommend publication
