Aspartic Acid-Based Nano-Copper Induces Resilience in Zea mays to Applied Lead Stress Via Conserving Photosynthetic Pigments and Triggering the Antioxidant Biosystem
Neda Dalir
Round 1
Reviewer 1 Report
The study focuses on evaluating the enhancing lead stress resilience in Zea mays by the application of different concentrations of aspartic acid-based nano-copper. There are too much data that authors tried to express the increasing and decreasing trend and also the amount of these changes, but the discussion was not enough for answering the research question. It contains some basic analyses and I have not found any new scientific advancement in this study. Besides, the manuscript is written in poor English and should be revised by an English-speaking person before submitting it for a detailed revision. I do not recommend the publication of the current manuscript.
General comments:
Abstract:
The abstract provides a clear and concise overview of the study. The information is presented in a logical order, starting with the background and objectives, followed by the methods and results, and concluding with the findings. I would recommend revising the abstract to include more specific details about the methodology, including experimental design and statistical analysis.
Line 24: " Plant growth ", P should be written in lower case.
Introduction:
The introduction provides a solid overview of the topic, presenting the potential benefits of nanotechnology in agriculture and the challenges posed by heavy metal pollutants. However, there is a need for a more comprehensive literature review, including specific references, to support the statements made. Clarifying the terminology and improving the coherence of the sentences would enhance the clarity of the introduction and provide a solid foundation for the study.
Material and Methods:
Please provide more information on the rationale and methodology behind the selection of the concentrations used for the treatments. As the authors mentioned in the manuscript, Asp-CuNPs act both as plant growth regulators and plant growth inhibitors, depending upon their concentration. The results showed that Asp-CuNPs act as PGR at concentration 1 μg/mL ≤ X ≤10. How about lower and upper concentrations? It is the main question that authors should answer.
Results:
The resulting description is not concise enough, some unnecessary information should not be included in the result section. Remove what is already obvious in the tables and graphs. Focus on trends and patterns. The results could be much shorter. Besides the quality of graphs, figures, and tables are not good enough and should be improved.
Discussion:
The discussion section was loose and should be carried out based on the hypotheses and findings of the paper.
Conclusion:
The conclusion did not highlight the study's main finding. The chosen concentration treatments were in the range in which Asp-CuNPs would act as PGR, but the authors didn’t provide any evidence of how and up to which level this impact would be continued and when they act as PGI.
The manuscript is written in poor English and should be revised by an English-speaking person before submitting it for a detailed revision.
Author Response
Dear editor and reviewer,
Thank you so much for reviewing our manuscript by providing your quality time, valuable comments/ recommendations. Your comments has really improved the quality and structure of our manuscript. Our team members have extensively/ carefully reviewed the manuscript and properly addressed all suggestions/recommendations point by point as suggested by worthy reviewer. Further, all authors have extensively reviewed the English language of the manuscript and made extensive changes/ improvements wherever needed to further improve the structure, quality and grammar of the manuscript. Now, we hope that the revised manuscript is highly improved. We have again critically reviewed the manuscript for typos errors and other mistakes. If you recommend further suggestions, let us know, we will be very happy to address.
Comment: The study focuses on evaluating the enhancing lead stress resilience in Zea mays by the application of different concentrations of aspartic acid-based nano-copper. There are too much data that authors tried to express the increasing and decreasing trend and also the amount of these changes, but the discussion was not enough for answering the research question. It contains some basic analyses and I have not found any new scientific advancement in this study. Besides, the manuscript is written in poor English and should be revised by an English-speaking person before submitting it for a detailed revision. I do not recommend the publication of the current manuscript.
General comments:
Reply: The discussion has been revised with the addition of content related to the mechanism of action of amino acid in altering plant physiology. Further the detrimental mode of action of Pb on biochemistry and physiology of plant has also been, which will help in answering the research question. Further, the manuscript has been extensively reviewed by all authors to further improve the language, typo/grammatical mistakes that were important to be rectified. We hope that now the manuscript has been significantly improved in its final version.
Abstract:
The abstract provides a clear and concise overview of the study. The information is presented in a logical order, starting with the background and objectives, followed by the methods and results, and concluding with the findings. I would recommend revising the abstract to include more specific details about the methodology, including experimental design and statistical analysis.
Line 24: "Plant growth", P should be written in lower case.
Reply: The Abstract once again reviewed/ revised several times with details about the methodology, including experimental design and statistical analysis were added. Further in Line 24: "Plant growth ", P is changed to lower case.
Introduction:
The introduction provides a solid overview of the topic, presenting the potential benefits of nanotechnology in agriculture and the challenges posed by heavy metal pollutants. However, there is a need for a more comprehensive literature review, including specific references, to support the statements made. Clarifying the terminology and improving the coherence of the sentences would enhance the clarity of the introduction and provide a solid foundation for the study.
Reply: The introduction has been carefully reviewed and revised as per suggestion. Specific references, to support the statements provided in the introduction. Further, cohesion/ connections have made among different paragraph to further improve the structure and quality of manuscript.
Material and Methods:
Please provide more information on the rationale and methodology behind the selection of the concentrations used for the treatments. As the authors mentioned in the manuscript, Asp-CuNPs act both as plant growth regulators and plant growth inhibitors, depending upon their concentration. The results showed that Asp-CuNPs act as PGR at concentration 1 μg/mL ≤ X ≤10. How about lower and upper concentrations? It is the main question that authors should answer.
Reply: For the optimization of dose of Asp-CuNPs, experiment was conducted using a wide variety of doses (ranging from 0.1 to 100 µg/mL). The experimental doses were decided based on the results of dose optimization. It was observed the doses below 0.5 µg/mL has no effect on germination while doses higher to 20 µg/mL had inhibitory effect. Based on dose optimization experiment, the experimental doses of 1.0, 5.0 and 10 µg/mL were selected for further experimentation. As dose optimization wasn’t the part of the project so the data hasn’t been included but summarizing in the statement “Asp-CuNPs act both as plant growth regulators and plant growth inhibitors, depending upon their concentration”
Results:
The resulting description is not concise enough, some unnecessary information should not be included in the result section. Remove what is already obvious in the tables and graphs. Focus on trends and patterns. The results could be much shorter. Besides the quality of graphs, figures, and tables are not good enough and should be improved.
Reply: Thanks for your deep review and interest in our manuscript. The result section has also been revised/ improved as suggested, extra/ unnecessary data has been removed from the revised manuscript.
Discussion:
The discussion section was loose and should be carried out based on the hypotheses and findings of the paper.
Reply: The discussion section has also been strengthened by addition of mode of action of Pb and amino acids on plant physiology from the recent available literature. Thanks for your deep review and interest in our manuscript. Your comment has really improved our manuscript. Highly appreciated.
Conclusion:
The conclusion did not highlight the study's main finding. The chosen concentration treatments were in the range in which Asp-CuNPs would act as PGR, but the authors didn’t provide any evidence of how and up to which level this impact would be continued and when they act as PGI.
Reply: For the optimization of dose of Asp-CuNPs, experiment was conducted using a wide variety of doses (ranging from 0.1 to 100 µg/mL). The experimental doses were decided based on the results of dose optimization. It was observed the doses below 0.5 µg/mL has no effect on germination while doses higher to 20 µg/mL had inhibitory effect. Based on dose optimization experiment, the experimental doses of 1.0, 5.0 and 10 µg/mL were selected for further experimentation. As dose optimization wasn’t the part of the project so the data hasn’t been included but summarizing in the statement “Asp-CuNPs act both as plant growth regulators and plant growth inhibitors, depending upon their concentration”
Comments on the Quality of English Language
The manuscript is written in poor English and should be revised by an English-speaking person before submitting it for a detailed revision.
Reply: The manuscript has been carefully reviewed several times by all authors for English proficiency, grammar and sentence structure Language has been improved. Now English is highly improved.
Reviewer 2 Report
Respected authors,
Greetings!
The issues of this paper is worth studing, and the design is reasonable too. However this manuscript needs modifing because of some questions. My suggestion is the following.
1. In the section of 2.2.5, the enzymes are introduced in the sequence of SOD, POD and APX.
2.In the section of materials and methods, add the table of the combinations of the factorial treatments.
3. The tables in the section of results and analysis need modifing according to the sample of table which I give you, and add some contents of results which are displayed at the bottom of the sample table.
4. In figures 6-10, change the bars into the curves.
5. Add the indicators of Chla/b and Chl a+b.
6. In the section of results and analysis ,add the analysis of the results, and not only the elaboration of the results.
7.In the section of discussion, focus on the reason of the avoidance of the injures of lead stress by applying Asp-CuNPs.
8. In the section of conclusion, the last sentence will be preserved, and the other sentences will be deleted, then add the contents of the signifcance of the study in this paper.
Comments for author File: 
Comments.pdf
Author Response
Dear editor and reviewer,
Thank you so much for reviewing our manuscript by providing your quality time, valuable comments/ recommendations. Your comments has really improved the quality and structure of our manuscript. Our team members have extensively/ carefully reviewed the manuscript and properly addressed all suggestions/recommendations point by point as suggested by worthy reviewer. Further, all authors have extensively reviewed the English language of the manuscript and made extensive changes/ improvements wherever needed to further improve the structure, quality and grammar of the manuscript. Now, we hope that the revised manuscript is highly improved. We have again critically reviewed the manuscript for typos errors and other mistakes. If you recommend further suggestions, let us know, we will be very happy to address.
- In the section of 2.2.5, the enzymes are introduced in the sequence of SOD, POD and APX.
Reply: The sequence is made inline to the suggestion. Thanks for correction.
- In the section of materials and methods, add the table of the combinations of the factorial treatments.
Reply: Table is added as suggested. We highly appreciate your time and interest in review process.
- The tables in the section of results and analysis need modifying according to the sample of table which I give you, and add some contents of results which are displayed at the bottom of the sample table.
Reply: tables are redesigned as per suggestions. Thanks for your time. Highly appreciated.
- In figures 6-10, change the bars into the curves.
Reply: the mentioned format is not supported by the program used.
- In the section of results and analysis, add the analysis of the results, and not only the elaboration of the results.
Reply: Results section has been are revised as per suggestion.
- In the section of discussion, focus on the reason of the avoidance of the injures of lead stress by applying Asp-CuNPs.
Reply: discussion points are added for how amino acids including aspartic acid modulating physiology of plants under stress condition.
- In the section of conclusion, the last sentence will be preserved, and the other sentences will be deleted, then add the contents of the significance of the study in this paper.
Reply: Conclusion has been carefully and comprehensively revised/ reviewed/ improved as per suggestions by worthy reviewers.
Reviewer 3 Report
Here below is my revision for the paper entitled 'Aspartic acid-based nano-copper induces resilience in Zea mays to applied lead stress via conserving photosynthetic pigments and triggering antioxidant Biosystem'
1/The abstract is fine
2/The introduction is poor since the lacks in previous studies has not been highlighted efficiently
3/Mterials and methods section is not well documented regarding the used materials and equipements
Regarding the infra red there is no range or measurements and resolution as well
No references for many sections and analysis such as leaf protein content and extraction of antioxidant enzymes among others
No references for phenol content determination and Pb concentrations
4/Results
the section is fine but table 2 is missing statistics and there is many figures I suggest to authors to use the most important
5/Discussion section should be improved
6/conclusion: this section lacks where future studies should focus based on the actual findings.
English language is clear
Author Response
Dear editor and reviewer,
Thank you so much for reviewing our manuscript by providing your quality time, valuable comments/ recommendations. Your comments has really improved the quality and structure of our manuscript. Our team members have extensively/ carefully reviewed the manuscript and properly addressed all suggestions/recommendations point by point as suggested by worthy reviewer. Further, all authors have extensively reviewed the English language of the manuscript and made extensive changes/ improvements wherever needed to further improve the structure, quality and grammar of the manuscript. Now, we hope that the revised manuscript is highly improved. We have again critically reviewed the manuscript for typos errors and other mistakes. If you recommend further suggestions, let us know, we will be very happy to address.
1/The abstract is fine
Reply: Thanks a lot for your kind words and interest in reviewing our manuscript. Highly appreciated.
2/The introduction is poor since the lacks in previous studies has not been highlighted efficiently
Reply: The introduction is revised by adding the available literature on the subject matter to further improve the quality of the manuscript.
3/Materials and methods section is not well documented regarding the used materials and equipments
Reply: this section has also been revised as per suggestion.
Regarding the infra red there is no range or measurements and resolution as well
Reply: The range and equipment model has been added to the revised manuscript. Your feedback is of high value to our manuscript.
No references for many sections and analysis such as leaf protein content and extraction of antioxidant enzymes among others
Reply: Appropriate/ most relavent references have been added to the methodologies followed.
No references for phenol content determination and Pb concentrations
Reply: Relavent references have been added at their appropriate positions.
4/Results
The section is fine but table 2 is missing statistics and there is many figures I suggest to authors to use the most important.
Reply: in table 2, now table 3, the interval mean analysis (means with 95% confidence intervals) was used instead of ANOVA. The number of figures are already reduced to possible level by merging multiple parameters into a single figure.
5/Discussion section should be improved
Reply: Discussion section has been critically reviewed and improved and supported with relavent references.
6/conclusion: this section lacks where future studies should focus based on the actual findings.
Reply: Conclusion section has been carefully revised/ reviewed/ improved in the revised manuscript.
Comments on the Quality of English Language
English language is clear
Reply: Thanks for your kind words and point by point review. Highly appreciated.
Reviewer 4 Report
The work is interesting but needs minor improvements, such as:
1. the formulas in the paper are incorrectly written, e.g. line 181 is Na2SO4, whereas it should be Na2SO4, because the number of atoms of a given element in the formula should be written as a subscript. Please correct the formula notations throughout the paper.
2. which spectrophotometer was used for the determination phenol content and according to which method (specific name(s) ) the determination was carried out.
3.what, please specify the particular model of instrument specific atomic absorption spectroscopy [AAS].
4. according to which method (name(s)) the sugars were analysed, and which spectrophotometer was used please specify the particular model.
5. line 135 should be Chl
6. kindly ask for a more detailed description of the determination of chlorophyll a and b and of carotenoids, so that another scientist reading the paper can reproduce the methodology described.
Once these errors have been taken into account, the paper can be published in the journal Sustainability
Author Response
Dear editor and reviewer,
Thank you so much for reviewing our manuscript by providing your quality time, valuable comments/ recommendations. Your comments has really improved the quality and structure of our manuscript. Our team members have extensively/ carefully reviewed the manuscript and properly addressed all suggestions/recommendations point by point as suggested by worthy reviewer. Further, all authors have extensively reviewed the English language of the manuscript and made extensive changes/ improvements wherever needed to further improve the structure, quality and grammar of the manuscript. Now, we hope that the revised manuscript is highly improved. We have again critically reviewed the manuscript for typos errors and other mistakes. If you recommend further suggestions, let us know, we will be very happy to address.
Comments and Suggestions for Authors
The work is interesting but needs minor improvements, such as:
- the formulas in the paper are incorrectly written, e.g. line 181 is Na2SO4, whereas it should be Na2SO4, because the number of atoms of a given element in the formula should be written as a subscript. Please correct the formula notations throughout the paper.
Reply. The formula notations are corrected throughout the paper wherever needed.
- which spectrophotometer was used for the determination phenol content and according to which method (specific name(s) ) the determination was carried out.
Reply. Proper citation has been added to the methodology to properly support it.
3.what, please specify the particular model of instrument specific atomic absorption spectroscopy [AAS].
Reply: Model names and maker name has been added to the revised manuscript as suggested by worthy reviewers.
- According to which method (name(s)) the sugars were analysed, and which spectrophotometer was used please specify the particular model.
Reply: Appropriate citation has been added.
- line 135 should be Chl
Reply: The word “Chl” has been corrected in the revised manuscript.
- kindly ask for a more detailed description of the determination of chlorophyll a and b and of carotenoids, so that another scientist reading the paper can reproduce the methodology described.
Reply: More detail has been added to the methodology section including the photosynthetic pigments.
Round 2
Reviewer 2 Report
Respected authors,
Greetings!
The manuscript had been modified almost according to the comments last time. Good job!
Author Response
Comment: The manuscript has been modified according to the comments last time. Good job.
Response: Dear reviewer and editorial board,
Thanks for your kind words and appreciation.
Your comments have really improved the structure and quality of our manuscript.
