Effect of Heat Stress on Root Architecture, Photosynthesis, and Antioxidant Profile of Water Spinach (Ipomoea aquatica Forsk) Seedlings

Round 1

Reviewer 1 Report

Article 1

Effect of heat stress on root architecture, photosynthesis, and antioxidant profile of water spinach (Ipomoea aquatica Forsk) seedlings

 

 

INTRODUCTION

L:40. 10ºC. ¿? Or 1ºC ¿?

 

MATERIALS AND METHODS

L: 103-105 Are the units correct?

L:107 Supescripts in units

L:119 Spectrophotometer specifications

L:150 indicate the specifications of the kit

L:155 specifications of the equipment used, spectrophotometer?

L:170, 174, 177 units, check and correct throughout the document

 

RESULTS

I suggest reordering the topics and subtopics so that they are more congruent with the order in which the materials and methods are presented.

 

Figure 2. Improve the name of the figure so that the components of the figure, mainly section G, are understood.

 

Figure 6. Improve the name of the figure, indicate what the images per row refer to.

 

Figure 7. Improve the name of the figure, indicate what the images per column refer to.

 

L:288 sub indixes

 

DISCUSSION

 

I suggest that topics and subtopics be included so that there is congruence with the order in which the results are presented.

 

L:317 improve the wording of this line, since it seems that in the present research we worked with tomato.

 

In general, they suggested deepening the discussion by explaining at the level of metabolic pathways of the plant.

 

CONCLUSIONS

Missing

Author Response

Effect of heat stress on root architecture, photosynthesis, and antioxidant profile of water spinach (Ipomoea aquatica Forsk) seedlings

INTRODUCTION

L:40. 10ºC. ¿? Or 1ºC ¿?

Response back: Its 1 ºC

MATERIALS AND METHODS

L: 103-105 Are the units correct?

Response back: We have corrected the Units.

L:107 Supescripts in units

Response back: We have removed typos and corrected subscripts.

L:119 Spectrophotometer specifications

Response back: We have added spectrophotometer specifications,

L:150 indicate the specifications of the kit

Response back: We have mentioned kit procedure. Kit word was used, its phosphate buffer solution.

L:155 specifications of the equipment used, spectrophotometer?

Response back: We have added spectrophotometer specifications.

L:170, 174, 177 units, check and correct throughout the document

Response back: We have corrected all units and also corrected throughout the draft.

RESULTS

I suggest reordering the topics and subtopics so that they are more congruent with the order in which the materials and methods are presented.

Response back: We have reordered the result section as previous mentioned material method section.

Figure 2. Improve the name of the figure so that the components of the figure, mainly section G, are understood.

Response back: We have explained the figure legend from A to G.

Figure 6. Improve the name of the figure, indicate what the images per row refer to.

Response back: Per row refer to various zoom views of leaf cell structure, importantly, various temperature indicated vertically.

Figure 7. Improve the name of the figure, indicate what the images per column refer to.

Response back: Per Colum refer to various zoom views of leaf cell paraffin image, importantly, various temperature indicated in a row.

L:288 sub indixes

Response back: We have corrected.

DISCUSSION

I suggest that topics and subtopics be included so that there is congruence with the order in which the results are presented.

Response back: We have reordered the discussion section as previous presented result and material method section.

L:317 improve the wording of this line, since it seems that in the present research we worked with tomato.

Response back: We have revised the sentence, thank you for your attention.

In general, they suggested deepening the discussion by explaining at the level of metabolic pathways of the plant.

Response back: Thank you for your suggestion. In this experiment we are focusing on heat stress work, our team also doing deep study on this plant, we are analyzing the data, specially metabolomic analysis, we will consider your suggestion in our next study. Thank you for your kind remarks,

CONCLUSIONS

Missing

Response back: We have added the conclusion section.

Reviewer 2 Report

Dear authors

The following modifications are required:

Abstract

ü  This section is generally written poorly. It is written in an uncomplicated manner. This section should include the most important findings of the study. As a result, this section should be improved. This manuscript contains no novelties.

ü  For this section, the authors should stick to the title. The authors should adhere to the sequence and order of writing in this section as specified in the title.

ü  Before describing the goal, the authors must define the issue in a single line and explain why they chose this approach to study this research.

ü  No information about the type of experimental design and its component is available in this manuscript.

ü  L19-20 (The results showed that no significant difference was seen between the control (25 0C) and 30 0C for some traits): The name of traits should be should be mentioned

ü  The authors should be provided some measured or quantitative data

ü  The authors should provide the decreasing percentage in all physiological traits under severe heat stress conditions

ü  The authors should provide percentage of increasing or decreasing in biochemical traits under heat stress conditions

ü  The authors should present a decisive conclusion derived from the research in the final line of the abstract and provide a single line of future prospects.

Keywords

ü  The keywords did not accurately reflect the content of the manuscript, and the words in the title should not be used as keywords. As a result, the keyword structure must be modified.

Introduction

ü  In the introduction section, the authors should be listed after the title. This section's content should adhere to the order of writing in the materials, method, and results sections.

ü  No information about the impact of heat stress on the expression of genes is available

ü  No information about the impact of heat stress on the root structure is available

ü  Along with the hypothesis statement, the authors should include some lines about the knowledge gap addressed by their research.

ü  At the end, the authors should include a novelty statement. What new things have the authors done or correlated in this study in comparison to previous ones?

ü  The general and specific aim should be inserted

Materials and methods

ü  The units should be corrected in all parts of this manuscript

ü  The procedure of determination of chlorophyll should be detailed

ü  The procedures of leaf gas exchange and chlorophyll fluorescence should be written in detail

ü  All abbreviation should be written in full name

ü  The chemical structure of some chemical compounds should be checked and corrected

ü  The detail of statistical data analysis should be inserted

Results and discussion

ü  In general, this section is not clearly written and should be modified. It is better to authors that they show their results as the increasing or decreasing percentage of traits under heat condition, compared to control plants

ü  The authors claimed in the statistical data analysis that they used LSD, but in the caption of the table and figure, they mentioned DMR, so the authors should double-check it.

ü  The authors should be provided some quantitative data

ü  The captions of all tables and figures should be modified and improved

ü  The data of Figures 9 and 10 should be subjected to statistical analysis

ü  The discussion is weak and should be improved because most of the sentences are written as the literature review rather than interpretation of results

Conclusion

ü  The authors should summarize the most important findings from this study. Furthermore, additional works pertaining to this research should be included as future works.

 

 

 

Extensive correction is needed

 

Author Response

Dear authors

The following modifications are required:

Abstract

ü  This section is generally written poorly. It is written in an uncomplicated manner. This section should include the most important findings of the study. As a result, this section should be improved. This manuscript contains no novelties.

Response back: We have revised the abstract and try to improve the abstract by adding key findings of the study.

ü  For this section, the authors should stick to the title. The authors should adhere to the sequence and order of writing in this section as specified in the title.

Response back: We have revised the abstract.

ü  Before describing the goal, the authors must define the issue in a single line and explain why they chose this approach to study this research.

Response back: We have explained in a single line why we choose the study. Thank you for your kind remarks. Line 16-17.

ü  No information about the type of experimental design and its component is available in this manuscript.

Response back: We have explained in the material method section, heading statistical analysis, Its abstract, in this short paragraph we explain background and key findings of the study.

ü  L19-20 (The results showed that no significant difference was seen between the control (25 0C) and 30 0C for some traits): The name of traits should be should be mentioned

Response back: We have mentioned the traits name,

ü  The authors should be provided some measured or quantitative data

Response back: We have provided the quantitative data in the result section; it is hard to explain all quantitative in abstract section. Because of multiple number of treatments with multiple number of parameters. Abstract word limit exceeds, so that’s why we are agree with your remarks, we have explained in the result section. Thank you for your remarks.

ü  The authors should provide the decreasing percentage in all physiological traits under severe heat stress conditions

Response back: Thank you for your question, as I mentioned in the previous questions, its is hard to explain all decreasing percentage in an abstract section, we have explained in the result section. Thank you for understanding.

ü  The authors should provide percentage of increasing or decreasing in biochemical traits under heat stress conditions

Response back: We have explain increasing and decreasing values of biochemical traits in a result section, We are really sorry, because of words limit, its hard to explain all percentage value explain in a abstract section.

ü  The authors should present a decisive conclusion derived from the research in the final line of the abstract and provide a single line of future prospects.

Response back: We have provided the final conclusion line and also added single line of future prospects.

Keynote: Dear reviewer, thank you for your time and thank you for reviewing our manuscript. We are understanding you raise a good and valid question for the abstract section. From our end we are try to revise the abstract section, but its very hard and difficult to add quantitative data and also added increasing and decreasing values of enzymes data, already abstract word around 261 words, its too long abstract, so we have added all quantitative data in the result section. Thank you for understanding. Thank you for your remarks.

Keywords

ü  The keywords did not accurately reflect the content of the manuscript, and the words in the title should not be used as keywords. As a result, the keyword structure must be modified.

Response back: We have modified the structure of the keywords.

Introduction

ü  In the introduction section, the authors should be listed after the title. This section's content should adhere to the order of writing in the materials, method, and results sections.

Response back: We have reordered the section content, as other reviewer also raised same questions, we have reordered the whole chapter sub section, same explain in material method, results and as well as discussion.

ü  No information about the impact of heat stress on the expression of genes is available

Response back: In this study, we only studied physiological, biochemical analysis. In the future study, we are already conducted transcriptomic analysis, and gene expression analysis. We will consider your suggestion for our future study. In this study, only focus on physiological analysis, that’s why not added information related gene expression,

ü  No information about the impact of heat stress on the root structure is available

Response back: We have added the data, effect of heat stress on root architecture. We have also present the information in the discussion section, we have presented our results and match with previous studies.

ü  Along with the hypothesis statement, the authors should include some lines about the knowledge gap addressed by their research.

Response back: We have added the lines at the end of introduction.

ü  At the end, the authors should include a novelty statement. What new things have the authors done or correlated in this study in comparison to previous ones?

Response back: We have added the novelty lined at the end of draft in conclusion lines,

ü  The general and specific aim should be inserted

Response back: We have revised the aims of the study, at the end of introduction.

Materials and methods

ü  The units should be corrected in all parts of this manuscript

Response back: We have corrected all units in all parts of this manuscript.

ü  The procedure of determination of chlorophyll should be detailed

Response back: We have explained the procedure in details in relevant section.

ü  The procedures of leaf gas exchange and chlorophyll fluorescence should be written in detail

Response back: We have explained the procedure in details in relevant section.

ü  All abbreviation should be written in full name

Response back: We have explained all abbreviation at first use.

ü  The chemical structure of some chemical compounds should be checked and corrected

Response back: We have checked and removed all typos, thank you for your attention.

ü  The detail of statistical data analysis should be inserted

Response back: We have revised detail data analysis.

Results and discussion

ü  In general, this section is not clearly written and should be modified. It is better to authors that they show their results as the increasing or decreasing percentage of traits under heat condition, compared to control plants

Response back: We have presented all results in a percentage value, expect fresh weights, oxidative damage and antioxidant enzymes analysis, now I have also revised the remaining results and explained by increasing and decreasing value. Further, oxidative stress biomarkers and enzymes analysis was done by time and each time 5 time, its too long results, that’s why we present the general results in a result section.

ü  The authors claimed in the statistical data analysis that they used LSD, but in the caption of the table and figure, they mentioned DMR, so the authors should double-check it.

Response back: It was typos, we have removed all typos, thank you for correction.

ü  The authors should be provided some quantitative data

Response back: We have provided quantitative data in each result section.

ü  The captions of all tables and figures should be modified and improved

Response back: We have modified the figure and table legends.

ü  The data of Figures 9 and 10 should be subjected to statistical analysis

Response back: Thank you for your question, actually number treatment and number of time data was too much that’s why we did not added statistical analysis data, that shows the significant difference among treatment on different time.

ü  The discussion is weak and should be improved because most of the sentences are written as the literature review rather than interpretation of results

Response back: We have improved the discussion section.

Conclusion

ü  The authors should summarize the most important findings from this study. Furthermore, additional works pertaining to this research should be included as future works.

Response back: We have added the conclusion section at the end of manuscript.

Reviewer 3 Report

Dear authors:

First of all I apologize for my comments, I am not specialist on this field research. Although when I read the abstract I thought that the paper will be very interesting, I changed my mind reading the other sections.

In my opinion the research is not well design. Of course you can talk about stress but I think that you cannot try to correlate to climate change... Of course the temperature of the earth is increasing year by year, but your experiment is not real… I think you should considered the night and day. The plants will not be at the same temperature 96 hours. In fact I think that your experiment it should be very interesting including a break time, maybe 10 or 12 hours with 19 or 20ºC. That could be considered as a night and you can check if they could recover the values or if the stress effects are permanent after beginning.

However, if you rewrite the paper focus it only in the stress effects, it could be interested although unreal because the plants will be ever under that conditions…

In the “Experimental setup” section you mentioned that “There were four replications of each treatment” but later in the results you say that the samples were analyzed for triplicate. I suppose that it is correct but why you have not use the four samples that you could get?

The section “Secondary metabolites” I think that you should not make a section that is bibliographic. Besides, I think that an experiment of a paper should be replicated without checking other bibliography….

In the section “Oxidative stress biomarkers and antioxidant enzymes analysis” why you do not order the absorbance values measures according to alphabetic order of the activities used?

With respect to the bibliography I could not check all of them but in some cases, the title of the paper does not correspond with the topic you are explaining in the text.

 

Comments for author File: Comments.pdf

Author Response

Dear authors:

First of all I apologize for my comments, I am not specialist on this field research. Although when I read the abstract, I thought that the paper will be very interesting, I changed my mind reading the other sections.

Response back: Thank you for your time and thank you reviewing our manuscript.

In my opinion the research is not well design. Of course you can talk about stress but I think that you cannot try to correlate to climate change... Of course the temperature of the earth is increasing year by year, but your experiment is not real… I think you should considered the night and day. The plants will not be at the same temperature 96 hours. In fact I think that your experiment it should be very interesting including a break time, maybe 10 or 12 hours with 19 or 20ºC. That could be considered as a night and you can check if they could recover the values or if the stress effects are permanent after beginning.

However, if you rewrite the paper focus it only in the stress effects, it could be interested although unreal because the plants will be ever under those conditions…

Response back: Thank you for your comment. We are agreeing with your question and we will consider your suggestion, we will apply in our upcoming articles, by adjust day and night temperature. In the experiment we focused effect of continuous temperature stress on water spinach plant.  Second, before designing of this study, we study previous literature and then designed our study, mostly researcher studies prolonged heat stress on water spinach I am sharing Doi of article (https://doi.org/10.1186/s12864-020-06953-9)

In the “Experimental setup” section you mentioned that “There were four replications of each treatment” but later in the results you say that the samples were analyzed for triplicate. I suppose that it is correct but why you have not use the four samples that you could get?

Response back: Thank you for your correction, it was typo, four replication was used in whole experiment.

The section “Secondary metabolites” I think that you should not make a section that is bibliographic. Besides, I think that an experiment of a paper should be replicated without checking other bibliography….

Response back: We have explained the procedure. Thank you for your suggestion.

In the section “Oxidative stress biomarkers and antioxidant enzymes analysis” why you do not order the absorbance values measures according to alphabetic order of the activities used?

Response back: We have reordered the absorbance value according to your suggestion.

With respect to the bibliography I could not check all of them but in some cases, the title of the paper does not correspond with the topic you are explaining in the text.

Response back: Thank you for your attention, we have cross check and revise it and remove all typos.

Thank you for your and thank you for reviewing the manuscript.

 

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors made the requested corrections

Reviewer 2 Report

The authors have been addressed all comments

moderate correction is needed