Potential Biopesticides from Seed Extracts: A Sustainable Way to Protect Cotton Crops from Bollworm Damage

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The article, "Potential Biopesticides from Seed Extracts: A Sustainable Way to Protect Cotton Crops from Bollworm Damage," explores the inhibitory effects of seed extracts from various plant species on digestive alpha-amylase activity in the cotton bollworm, Helicoverpa armigera. Here is a critical review of the article:

- Strengths:

1. The article is well-structured, with a clear introduction, detailed methods, and a thorough discussion of the results.

2. The study explores the potential of biopesticides, a relevant and vital topic in the context of sustainable agriculture.

3. The use of statistical analysis adds credibility to the results.

4. The abstract provides a clear and concise overview of the study, its objectives, methods, and results. It highlights the potential of seed extracts as biopesticides, crucial for capturing the attention of readers interested in crop protection.

-Weaknesses:

1. The article could benefit from more specific details regarding the extraction and purification processes for the proteinaceous inhibitors. Providing information on equipment, solvents, and specific procedures would enable better replication of the study.

2. The connection between the experimental findings and the practical implications for cotton crop protection could be further emphasized in the discussion.

3. A more detailed discussion of the mode of action of the inhibitors present in the seed extracts is needed. Exploring the biochemical processes involved would enhance the scientific rigor of the study.

4. The article should suggest specific areas for future research or experiments that can build upon the current findings. For example, investigating the stability of these inhibitors under field conditions or conducting experiments to assess their safety to non-target organisms.

5. If genetic engineering for crop protection is suggested, the article should provide more details about the feasibility, potential challenges, and safety considerations. Explaining how this can be practically achieved and addressing the potential implications for the environment and human health would be beneficial.

6. The article should offer a more comprehensive discussion of the limitations and challenges of the study. Acknowledging any potential biases, constraints, or external factors that might have affected the results is essential.

7. In the context of developing biopesticides, the article should elaborate on the potential environmental impact and safety of using these seed extracts. Discussing the ecological consequences and addressing any concerns regarding non-target organisms would add depth to the research.

Author Response

- Strengths:

1. The article is well-structured, with a clear introduction, detailed methods, and a thorough discussion of the results.
2. The study explores the potential of biopesticides, a relevant and vital topic in the context of sustainable agriculture.
3. The use of statistical analysis adds credibility to the results.
4. The abstract provides a clear and concise overview of the study, its objectives, methods, and results. It highlights the potential of seed extracts as biopesticides, crucial for capturing the attention of readers interested in crop protection.

 

I am deeply grateful for your kindness and the positive remarks you shared about the article's strengths. Your words have provided us with much-needed encouragement.

-Weaknesses:

The yellow-highlighted portions in the article's text pertain to the comments that you graciously provided.

1. The article could benefit from more specific details regarding the extraction and purification processes for the proteinaceous inhibitors. Providing information on equipment, solvents, and specific procedures would enable better replication of the study.

Line 133-138

Thanks for your highly valuable feedback, which greatly enhances the quality of the manuscript. In the materials and methods section, we have included additional information on the extraction process of plant extracts and the utilization of ammonium sulfate salt in various fractions. We intended to simplify these aspects, making them more accessible and less complex for future use by other researchers.

2. The connection between the experimental findings and the practical implications for cotton crop protection could be further emphasized in the discussion.

Line 333

Thank you for your precise and comprehensive understanding of the utilization of plant extracts, in the section on the "application of plant extracts, benefits, and limitations", we aim to highlight various studies that have explored experimental findings and practical implications of using plant extracts specifically in cotton plants.

3. A more detailed discussion of the mode of action of the inhibitors present in the seed extracts is needed. Exploring the biochemical processes involved would enhance the scientific rigor of the study.

The first and second paragraph of discussion: line 229-241

Plant extracts are used as natural alternatives to synthetic pesticides in pest management. They can exert their influence in a variety of ways, including repellency, toxicity, and growth control. Plant extracts that repel pests by emitting strong scents or tastes discourage bugs from feeding or laying eggs. Toxic plant extracts contain bioactive substances that affect the nervous system, enzyme action, or the cuticle of the insect, causing death or decreased function. Plant extracts that regulate growth disrupt hormonal balance or molting processes, leading to aberrant growth, sterility, or developmental stoppage.

Based on various studies, alpha-amylase inhibitory proteins can be divided into 7 distinct groups based on their protein structure. However, the majority of these groups exhibit minimal or no inhibitory activity. Interestingly, two of these groups have been found to possess a dual function, effectively inhibiting both alpha-amylase and protease enzymes.

4. The article should suggest specific areas for future research or experiments that can build upon the current findings. For example, investigating the stability of these inhibitors under field conditions or conducting experiments to assess their safety to non-target organisms.

At the end of the section "application of plant extracts, benefits, and limitations" we added this part. Line 402-418

Future research directions in the field of plant extract-based pesticides should focus on investigating the stability of plant extract inhibitors under field conditions and assessing their safety to non-target organisms. Specifically, studies should evaluate the stability of bioactive compounds in plant extracts under varying environmental conditions and determine the degradation rates of plant extract inhibitors in different formulations. Additionally, toxicity studies should be conducted to assess the impact of plant extract inhibitors on beneficial insects and other non-target organisms, while investigating the sub-lethal effects and potential interactions with beneficial organisms. Furthermore, the application of nanotechnology in this field should be explored to enhance the efficacy and delivery of plant extract inhibitors. Investigations into the development of nanoformulations, such as nanoemulsions or nanocapsules, can improve the stability and controlled release of plant extracts. Additionally, the potential of nanomaterials as carriers for plant extract compounds should be explored to enhance their efficacy and targeted delivery to pests. Furthermore, it is crucial to assess the safety and environmental impact of nanomaterial-based formulations on non-target organisms for sustainable pest management strategies. By integrating nanotechnology into plant extract-based pesticides, we can unlock new possibilities for effective and environmentally friendly crop protection solutions.

5. If genetic engineering for crop protection is suggested, the article should provide more details about the feasibility, potential challenges, and safety considerations. Explaining how this can be practically achieved and addressing the potential implications for the environment and human health would be beneficial.

Line 334-350

Thank you for your valuable contribution, which has greatly enhanced both the manuscript and the ensuing discussion. As highlighted in the instances you have cited, the aforementioned contents were incorporated within the initial section of the “application of plant extracts”. Resistance to transgenic plants has been reported in several research; plant extracts are one potential remedy; they are included in the article's content. With the exception of transferring bacterial crystal proteins and boosting plants' and their secondary metabolites' and antioxidative resistance against pests, our research has not found any studies on genetically modifying plants to obtain their extracts for insect control.

 

6. The article should offer a more comprehensive discussion of the limitations and challenges of the study. Acknowledging any potential biases, constraints, or external factors that might have affected the results is essential.

Line 383-391

All of your valuable suggestion added in the section of "application of plant extracts, benefits, and limitations"

7. In the context of developing biopesticides, the article should elaborate on the potential environmental impact and safety of using these seed extracts. Discussing the ecological consequences and addressing any concerns regarding non-target organisms would add depth to the research.

At the line 391-401

Biopesticides, which encompass bacteria, fungi, viruses, plant extracts, nematodes, and other biological agents, are not exclusively derived from natural sources. However, their effectiveness is enhanced through the incorporation of preservatives, carriers, and companions, which prevent their degradation and depletion in the environment. These formulations must possess characteristics such as affordability, environmental friendliness, minimal risk to natural enemies, and efficacy at the intended site of action. By combining biopesticides with other management strategies, integrated pest management can be effectively employed to suppress pest populations below economically damaging levels while minimizing adverse effects on non-target organisms and preserving ecosystem integrity.

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The paper entitled “Potential Biopesticides from Seed Extracts: A Sustainable Way to Protect Cotton Crops from Bollworm Damage”, authored by Masoud Chamani, Narjes Askari, Reza Farshbaf Pourabad, Ali Chenari Bouket, Tomasz Oszako and Lassaad Belbahri, presents the results that can be considered a contribution to formulation of new biopesticides. The manuscript is not acceptable for publication in its current form. The main comments regarding this manuscript are listed below.

·        Abstract: “Graminae” and “Leguminosae” should be written in italic (Latin names should be in italic);

·        I guess the R.H. is relative humidity, however it is not necessary to use this abbreviation in abstract. The same comment stays for L:D (please define L:D).

·        The last sentence in the Abstract should be re-written. It is not quite understandable at first glance.

·        The authors listed so many results in the abstract, however one key sentence is missing: further recommendations and investigations that can arise from this study.

·        Introduction: line 44, “Studies revealed that plants or other organism…” Which studies are the authors referring to? The references are missing.

·        The citations must be uniform. The authors mostly used the numerical citations in the text, however in line 85 there are the citations in the form of surnames. I guess that should be the references 13, 14, 15 and 16).

·        The last paragraph in Introduction section must be extended. The authors should list the main aims of the study, as well as explicitly mention the studies on which their investigation relies on.

·        Line 101: Please do not start the sentence with the number.

·        Line 116: It is not clear what are “four concentrations” since the authors provided the ranges of concentrations, such as 0-30%, 30-50%, etc.

·        Line 130: Bradford methods should be cited (the number of the reference is missing).

·        Line 136: Why did you choose those pH values? What was the reference?

·        Figure 3: the graph should be changed in the form of Scatter plot.

·        The results are nicely and thoroughly discussed and described. I suggest the addition of separate paragraph or subsection regarding limitations of the present results and further recommendations.

Comments on the Quality of English Language

There are some minor corrections that should be done.

Author Response

The paper entitled “Potential Biopesticides from Seed Extracts: A Sustainable Way to Protect Cotton Crops from Bollworm Damage”, authored by Masoud Chamani, Narjes Askari, Reza Farshbaf Pourabad, Ali Chenari Bouket, Tomasz Oszako and Lassaad Belbahri, presents the results that can be considered a contribution to formulation of new biopesticides. The manuscript is not acceptable for publication in its current form. The main comments regarding this manuscript are listed below.

The contents of the manuscript that are highlighted in green correspond to the valuable comments you provided.

• Abstract: “Graminae” and “Leguminosae” should be written in italic (Latin names should be in italic);

Thank you for your care in writing scientific names. In scientific writing, it is customary to write the genus and species names in italic format, while the family names are written in non-italic format.

• I guess the R.H. is relative humidity, however it is not necessary to use this abbreviation in abstract. The same comment stays for L:D (please define L:D).

Line 22-23

Thank you very much for your careful reading of this manuscript, the points you mentioned have been applied

• The last sentence in the Abstract should be re-written. It is not quite understandable at first glance.

Thank you for your attention to clearing up this ambiguity, the sentence you mentioned has been rewritten:

Nanotechnology offers several ways to enhance biopesticides, which are a solution for making plant extract usage more efficient.

• The authors listed so many results in the abstract, however one key sentence is missing: further recommendations and investigations that can arise from this study.

This important point that you mentioned was added in another form in the abstract text:

The exploration of plant-based pesticides, in conjunction with the incorporation of nanotechnology and other scientific fields, offers a wide range of prospects for further investigation.

• Introduction: line 44, “Studies revealed that plants or other organism…” Which studies are the authors referring to? The references are missing.

It was added

• The citations must be uniform. The authors mostly used the numerical citations in the text, however in line 85 there are the citations in the form of surnames. I guess that should be the references 13, 14, 15 and 16).

Line 87

Thank you for your attention and thank you for pointing this out. This item has been corrected Referencing was done with Mendeley and the name was not converted into a number in this way.

• The last paragraph in Introduction section must be extended. The authors should list the main aims of the study, as well as explicitly mention the studies on which their investigation relies on.

Line 93-101

It was added before the main aims paragraph, but it was discussed in discussion part in detailed.

• Line 101: Please do not start the sentence with the number.

Line 113

It was corrected

• Line 116: It is not clear what are “four concentrations” since the authors provided the ranges of concentrations, such as 0-30%, 30-50%, etc.

The concentrations of ammonium sulfate salts are expressed as a percentage spectrum in enzyme inhibition because this inhibition is a gradual process. At low concentrations of ammonium sulfate salts, the enzyme inhibition is partial and the enzyme activity does not decrease significantly. With increasing concentrations of ammonium sulfate salts, the enzyme inhibition becomes stronger and the enzyme activity gradually decreases.

In enzyme inhibition, ammonium sulfate salts prevent the enzyme from binding to the substrate by binding to the enzyme itself. At low concentrations of ammonium sulfate salts, only a small number of enzyme molecules bind to the salts, and the enzyme activity does not decrease significantly. With increasing concentrations of ammonium sulfate salts, the number of enzyme molecules binding to the salts increases, and the enzyme activity gradually decreases.

Therefore, expressing the concentrations of ammonium sulfate salts as a percentage spectrum in enzyme inhibition reflects the gradual nature of this process. Expressing the concentrations of ammonium sulfate salts as a percentage spectrum in enzyme inhibition is a standard method. This method provides accurate information about the extent of enzyme inhibition at different concentrations of ammonium sulfate salts.

One of the examples is this paper:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7695795/

• Line 130: Bradford methods should be cited (the number of the reference is missing).

Line 158

It was corrected

• Line 136: Why did you choose those pH values? What was the reference?

Numerous studies have confirmed that the ideal pH for butterfly digestive enzymes is alkaline, ranging from 7 to 10. As a result, most enzyme activities and inhibitions are expected to occur within this pH range. However, acidic pHs are also considered because they can provide insights into how enzyme activity changes when the pH is lowered. Therefore, this study included two acidic pHs and two alkaline pHs.

Line 322 and another study in below:

https://onlinelibrary.wiley.com/doi/abs/10.1002/ps.3911

• Figure 3: the graph should be changed in the form of Scatter plot.

Line 225

The suggested improvement you made for enhancing the quality of the article has been implemented, resulting in the desired change. We appreciate your contribution in helping us make the necessary adjustments.

• The results are nicely and thoroughly discussed and described. I suggest the addition of separate paragraph or subsection regarding limitations of the present results and further recommendations.

Based on your input, it has been noted that this particular case shares similarities with another reviewer's perspective. As a result, we have incorporated a dedicated section titled " Application of plant extract, benefits, and limitations " at the end of the discussion section. This section encompasses all the cases that you and the previous reviewer had in mind, ensuring comprehensive coverage of the topic.

 

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript has been revised by the authors in accordance with the provided suggestions.

Reviewer 2 Report

Comments and Suggestions for Authors

The authors corrected the manuscript according to the suggestions. Therefore, I recommend its publication.