Invertase Plays a Vital Role in Regulating Wound-Induced Accumulation of Phenolic Compounds in Carrot Tissue
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript is interesting, useful and relevant for the field. I have made comments in the attached pdf.
The Materials and Results sections do not mention whether whole carrots or CK carrots are the control in the second experiment. This makes the description and interpretation of results confusing and difficult to follow. Please correct this.
Comments for author File: 
Comments.pdf
The Results and Conclusion sections need revising and rephrasing to correct language mistakes.
Author Response
In this revised version, we have accepted most of the reviewers’ suggestions and explained reviewers’ comments. The major revisions of the manuscript have been marked in red color. We appreciate the reviewers' constructive comments, the quality of our manuscript has been improved significantly after the revision.
Specific changes addressing Reviewer 1’ Point-by-Point comments are as follows:
Reviewer #1: General concepts comments:
The manuscript is interesting, useful and relevant for the field. I have made comments in the attached pdf.
The Materials and Results sections do not mention whether whole carrots or CK carrots are the control in the second experiment. This makes the description and interpretation of results confusing and difficult to follow. Please correct this.
The Results and Conclusion sections need revising and rephrasing to correct language mistakes.
Response: As the reviewer mentioned, the missing information in the Materials and Results easily makes the relevant part of results or discussion confusing and difficult to follow. We have considered this suggestion carefully and add the descriptions in lines 110-112.
Other comments:
- Line 105: What do you mean by individuals: carrot pieces, polypropylene containers? Please be specific.
Response: Follow the reviewer’s suggestion, to make it clear, we have changed “individuals” to “whole carrots” in this sentence (line 105).
- Line 106: Please mention which type of carrot is considered the control for this experiment.
Response: The cubed carrots immersed in distilled water were used as control. We have added the descriptions in lines 110-112.
- Line 108: Please correct this.
Response: Thanks for this kind suggestion, it was a mistake here. We have changed ‘regents’ to ‘reagents’ in this sentence (line 108).
- Lines 184-185: Do you mean "completely randomized design"? Please correct the phrase.
Response: Follow the reviewer’s suggestion, we have changed ‘absolutely haphazard design’ to ‘completely randomized design’ in this sentence (lines 186-187).
- Line 201: Please correct this.
Response: Thanks for this kind suggestion, it was a mistake here. We have changed ‘was peaked’ to ‘peaked’ in this sentence (line 203).
- Lines 223-224: Please rephrase.
Response: Follow the reviewer’s suggestion, we have rewritten this sentence (lines 225-226).
- Line 257: The writing is too small and difficult to read. Please make these graphs larger and easier to understand.
Response: To make the Figure 4 more clear, we have replaced it in line 258.
- Line 302: Figure 7b shows glucose level to be higher in whole carrots than in any of the treated samples.
Response: Fructose, glucose, and sucrose were all water-soluble sugars. In this study, immersing the cubed carrots in water induced the initial loss of these sugars and the lower levels of sugars were observed in cubed carrots, as compared with whole carrots. A similar phenomenon was also observed in total soluble phenolics. Unlike with the sharp increase of phenolics content, the increase of glucose derived mainly from the degradation of sucrose was relatively limited. Besides, the glucose was utilized in many metabolic processes like respiration. The production rate, the utilization rate and the soaking treatment combined contributed to making glucose level in cubed carrots was lower than that in whole carrots.
- Line 313: Please correct this to figure 7.
Response: Thanks for this kind suggestion, it was a mistake here. We have changed ‘Figure 6’ to ‘Figure 7’ (line 315).
- Line 371: Please explain this acronym.
Response: Follow the reviewer’s suggestion, we added the description behind the acronym of CsINV (lines 375-376).
- Lines 414-415: Please rephrase.
Response: Follow the reviewer’s suggestion, we have rewritten this sentence (lines 419-420).
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe manuscript describes a simple study on the role of wounding in the accumulaton of phenolics in carrot. The issue of increase in phenolics in result of wounding is not new and it has been described in many species including carrot (https://www.sciencedirect.com/science/article/abs/pii/S0308814612001495). This study enriches existing knowledge on the relation between wounding and sucrose degradation.
In general paper is well written. I am curious Authors response to a question weather the sucrose degradation after wounding to favor the phenolics to a certain level will affect the sensory properties important to customers.
Comments for author File: Comments.pdf
small adjustments needed
Please change the phrase 'consumption of sucrose' to ' degradation of sucrose' in whole manuscript.
Author Response
In this revised version, we have accepted most of the reviewers’ suggestions and explained reviewers’ comments. The major revisions of the manuscript have been marked in red color. We appreciate the reviewers' constructive comments, the quality of our manuscript has been improved significantly after the revision.
Specific changes addressing Reviewer 2’ Point-by-Point comments are as follows:
Reviewer #2: General concepts comments:
The manuscript describes a simple study on the role of wounding in the accumulaton of phenolics in carrot. The issue of increase in phenolics in result of wounding is not new and it has been described in many species including carrot (https://www.sciencedirect.com/science/article/abs/pii/S0308814612001495). This study enriches existing knowledge on the relation between wounding and sucrose degradation.
In general paper is well written. I am curious Authors response to a question weather the sucrose degradation after wounding to favor the phenolics to a certain level will affect the sensory properties important to customers.
Please change the phrase 'consumption of sucrose' to ' degradation of sucrose' in whole manuscript.
Response:
As a physiological response to wounding, the synthesized phenolics in wounded carrots were ultimately utilized for the production of lignin or suberin. Accompanied by lignification, the white blush occurred in the surface of wounded carrots. Based on this, the biosynthesis of phenolics in wounded carrots adversely affected the sensory properties of carrots. In this study, due to the relatively high humidity, the sensory properties of wounded carrots were acceptable for customers. Actually, the threshold of sensory properties should be well considered. The correlation between phenolics level and sensory properties should be established. That’s interesting and meaningful. Thanks for this kind suggestion. We will do this experiment in the future.
In addition, follow the reviewer’s suggestion, “consumption of sucrose” was changed to “degradation of sucrose” in whole manuscript.
Other Comments:
- Line 85: How do you think-the length of storage of roots would affect the results?
Response: As illustrated in the figure below, the concentration of phenolic compounds in carrots differed with the position of tissue. In my opinion, keeping the uniform size of carrot was beneficial to eliminate the impact of individual difference.
(Title: Peel removal improves quality without antioxidant loss, through wound-induced phenolic biosynthesis in shredded carrot. http://dx.doi.org/10.1016/j.postharvbio.2016.07.004)
- Line 111: cite which authors.
Response: Follow the reviewer’s suggestion, the information of referenced research was added in this sentence (line 113).
- Line 153: List which sugars you evaluated.
Response: Follow the reviewer’s suggestion, the sugars evaluated in the present study were listed in line 154.
- Line 184: Remove and replace with 'completly randomized.
Response: Follow the reviewer’s suggestion, we have changed ‘absolutely haphazard’ to ‘completely randomized’ in this sentence (line 186).
- Line 193: rephrase as it is repetition.
Response: Follow the reviewer’s suggestion, we have rewritten this sentence (line 195).
- Line 341: Replace by 'fruit subjected to wounding.
Response: Follow the reviewer’s suggestion, we have changed ‘pitaya fruit suffered from wounding’ to ‘pitaya fruit subjected to wounding’ in this sentence (line 345).
- Line 343: Please change the wording consumption to more proper one.
Response: Follow the reviewer’s suggestion, we have changed ‘consumption’ to ‘loss’ in this sentence (line 347).
- Line 368: please change the wording to more proper one.
Response: Follow the reviewer’s suggestion, we have changed ‘suffered from’ to ‘with’ in this sentence (line 372).
- Lines 384-388: Different font size?
Response: Follow the reviewer’s suggestion, we used the same font size (lines 389-393).
- Line 404: Simple sugars? total? please insert.
Response: Follow the reviewer’s suggestion, we have changed ‘sugars’ to ‘soluble sugars’ in this sentence (lines 409-410).
- Line 414: Strongly.
Response: It was a mistake. Follow the reviewer’s suggestion, we have changed ‘strong’ to ‘strongly’ in this sentence (line 419).
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsQuestions:
1. why the sugar content did not decrease over time? if they are the carbon sources for phenolic biosynthesis Figures 2A and 2B.
2. why the control dipped in water showed lower accumulation of phenolics?
This publication should be considered in the introduction/discussion:
https://doi.org/10.1016/j.afres.2023.100303
Author Response
In this revised version, we have accepted most of the reviewers’ suggestions and explained reviewers’ comments. The major revisions of the manuscript have been marked in red color. We appreciate the reviewers' constructive comments, the quality of our manuscript has been improved significantly after the revision.
Specific changes addressing Reviewer 3’ Point-by-Point comments are as follows:
Reviewer #3: General concepts comments:
- Why the sugar content did not decrease over time? if they are the carbon sources for phenolic biosynthesis Figures 2A and 2B.
Response: We have considered this question carefully. After wounding, the changes of fructose and glucose were determined by two factors. On one hand, new fructose and glucose were generated as a result of the degradation of sucrose; On the other hand, the fructose and glucose could be utilized in many metabolic processes like respiration. When the decomposition rate of sucrose faster than the utilization rate of reducing sugar, the increase of reducing sugar content was observed.
- Why the control dipped in water showed lower accumulation of phenolics?
Response: A similar result was observed in previous study that washing the shredded-carrots on water induced lower production of total phenolic compounds (Cross-talk between signaling pathways: The link between plant secondary metabolite production and wounding stress response. https://doi.org/10.1038/srep08608). As interpreted by the author, the partial removal of a primary signal that triggers the production of total phenolic compounds in wounded plants, such as extracellular ATP which has been proposed as the primary signal produced at the site of injury that trigger NADPH oxidase activity and thus ROS production. The control dipped in water resulted in the partial removal of primary signals like ATP, thereby reducing the accumulation of phenolics.
- This publication should be considered in the introduction/discussion:
https://doi.org/10.1016/j.afres.2023.100303
Response: The helpful information from this publication was adopted in the section of Discussion (lines 325-327). And this publication was added in the section of References.
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for AuthorsThe authors presented the article describing the correlation between wounding and activation of sucrose-cleaving enzymes, invertases. The authors clearly showed that wounding of carrot tissue activated invertases and, at the same time, upregulated the expression of most analyzed invertase encoding genes. The findings presented in this well organized and nicely written work are interesting and also provide some new insights into the role of invertase regulation during wounding and accumulation of phenolic compounds in carrot.
Introduction part is correctly written with sufficient literature data. The whole experimental methods used in this work are nicely described with all sufficient details. The obtained results, discussed sufficient and in details, are followed by summary conclusion.
I have only one remark considering results section. In this part of manuscript during the explanation of obtained results, and during the comparison of results determined in some treatment(s) and control, the authors in some part of the text used “times higher” as well as “% higher”. I stated these parts of manuscript directly in the original submitted version. This should be uniform through the whole text manuscript.
Comments for author File: Comments.pdf
Author Response
In this revised version, we have accepted most of the reviewers’ suggestions and explained reviewers’ comments. The major revisions of the manuscript have been marked in red color. We appreciate the reviewers' constructive comments, the quality of our manuscript has been improved significantly after the revision.
Specific changes addressing Reviewer 4’ Point-by-Point comments are as follows:
Reviewer #4: General concepts comments:The authors presented the article describing the correlation between wounding and activation of sucrose-cleaving enzymes, invertases. The authors clearly showed that wounding of carrot tissue activated invertases and, at the same time, upregulated the expression of most analyzed invertase encoding genes. The findings presented in this well organized and nicely written work are interesting and also provide some new insights into the role of invertase regulation during wounding and accumulation of phenolic compounds in carrot. Introduction part is correctly written with sufficient literature data. The whole experimental methods used in this work are nicely described with all sufficient details. The obtained results, discussed sufficient and in details, are followed by summary conclusion. I have only one remark considering results section. In this part of manuscript during the explanation of obtained results, and during the comparison of results determined in some treatment(s) and control, the authors in some part of the text used “times higher” as well as “% higher”. I stated these parts of manuscript directly in the original submitted version. This should be uniform through the whole text manuscript.
Response: Thanks for this kind suggestion. As the reviewer mentioned, the uniform of description was really important. We have used "times higher" uniformly in the section of Results.
Other Comments:
- Line 198: The authors should use the same comparation system in comparison to control. In this sentence the authors used "times" to explain comparation to control values but further the authors used the percent... please uniform om one or another way.
Response: Follow the reviewer’s kind suggestion, we have used "times higher" uniformly in the section of Results (line 200).
- Line 201: Here the authors used % in order to compare the results with the control... previously the authors used "times". Please uniform...
Response: Follow the reviewer’s kind suggestion, we have used "times higher" uniformly in the section of Results (line 203).
- Line 233: Here also... times or percent... please unifrom...
Response: Follow the reviewer’s kind suggestion, we have used "times higher" uniformly in the section of Results (line 235).
- Line 253: times or percent... please uniform...
Response: Follow the reviewer’s kind suggestion, we have used "times higher" uniformly in the section of Results (line 255).
- Line 309: percent ot times... please uniform...
Response: Follow the reviewer’s kind suggestion, we have used "times" uniformly in the section of Results (line 311).
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe paper can be published in current form.
