Pesticide Resistance and Related Mutation Frequencies of Tetranychus urticae in Hainan, China

Round 1

Reviewer 1 Report

Comments to the Author

 

This paper reports a study on pesticide resistance and related mutation frequencies of Tetranychus urticae in Hainan, China. The originality of the research is good and results are useful. However, some revisions as listed below are needed before it can be published.

 

1) 3.2. Resistance of ~

What is the level (criterion or ratio) for low, medium, and high resistance?

 

2) 3.3. Genotype and ~

It is not enough to express as a frequency, because you did a general PCR. Please do a real-time PCR experiment or change the title.

 

3) Please check the references again. The form is not correct.

 

 

Other comments are noted on the manuscript.

Comments for author File: Comments.pdf

Author Response

This paper reports a study on pesticide resistance and related mutation frequencies of Tetranychus urticae in Hainan, China. The originality of the research is good and results are useful. However, some revisions as listed below are needed before it can be published.

1. 2. Resistance of ~
   What is the level (criterion or ratio) for low, medium, and high resistance?
   Response: The criterion for resistance level was presented in original manuscript, please see lines 136-139 in revision.
2. 3. Genotype and ~
   It is not enough to express as a frequency, because you did a general PCR. Please do a real-time PCR experiment or change the title.
   Response: After careful consideration, we changed the subheading to “mutation frequencies of target sites”.
3. Please check the references again. The form is not correct.
   Response: Done.

Other comments are noted on the manuscript.
Response: Other comments on the manuscript have been confirmed or supplemented.

Reviewer 2 Report

The manuscript in reference describes the determination of the resistance to seven pesticides in cucurbit crop-derived field populations of T. urticae (n = 5), collected in Hainan in 2021. Although the study is very local, the manuscript has relevant information and results that will be interesting for readers. However, some issues should be addressed prior to further consideration.

1.       Detailed scrutiny should be performed throughout the manuscript to revise certain grammar and stylistic issues.

2.       Line 68: Phaseolus vulgaris in italics

3.       Line 70: was each subsample collected on the same day? How long is the difference? collecting dates? Was each subsample collected in the same plant group or crop? This information should be clarified in the manuscript.

4.       Line 71. More details about host plants should be provided. For instance, cultivation practice, crop size, pesticide application, etc., since these factors are relevant to the conditions of the collected mite population.

5.       Line 103: 30 l? what does mean “l”? Revise.

6.       Revise in detail the M&M section. Some experimental details are missing to ensure outcome reproducibility. For instance, brand, model, and grade of reagents, solvents, materials, and instruments must be provided. In addition, details about sequencing should also be clarified.

7.       Line 108: More details on determining mutation frequencies should also be expanded for readers, specifically to obtain Figure 1.

8.       Lines 120 and 122: LC₅₀ instead of LC50_.

9.       Lines 167-169: Revise this text since it seems to be a typo.

10.   The discussion section can be improved since it is highly descriptive and even speculative. In addition, it can be even reorganized, possibly by subheadings.

11.   A conclusion section should be generated, I mean, the conclusion section should be outside of the discussion section.

12.   Lines 263-266: These conclusions are not entirely supported by results, since the number of populations (vide infra) is very low. I consider important to give an adequate scope of these results.

 

13.   Finally, although the study seems to be adequately developed from the experimental point of view, my main criticism of this manuscript is related to the number of populations of T. urticae evaluated since it is very low and not enough to give the final authors’ conclusions. Therefore, the manuscript has an approach drawback that must be clarified for readers. From this fact, the scope of the manuscript should be re-oriented to afford better scientific quality and merit. For instance, it should be determined or attempted to establish a plausible relationship between the populations and the details of the crops where mite populations were obtained (which, in fact, those details are missing), whose host conditioning is the main factor for this mite to win resistance or not. Accordingly, the discussion, apart from expanding the descriptions of obtained results, should be oriented towards discussing the results from a mechanistic point of view and the relationship between the evaluated populations and their detailed origin since, I repeat, the number of mite populations is very low to draw better, more far-reaching conclusions.

Author Response

1. The manuscript in reference describes the determination of the resistance to seven pesticides in cucurbit crop-derived field populations of T. urticae (n = 5), collected in Hainan in 2021. Although the study is very local, the manuscript has relevant information and results that will be interesting for readers. However, some issues should be addressed prior to further consideration.

   1. Detailed scrutiny should be performed throughout the manuscript to revise certain grammar and stylistic issues.
      Response: We read the manuscript carefully and made revisions on the certain grammar and stylistic issues.
   2. Line 68: Phaseolus vulgaris in italics.
      Response: Done.
   3. Line 70: was each subsample collected on the same day? How long is the difference? collecting dates? Was each subsample collected in the same plant group or crop? This information should be clarified in the manuscript.
      Response: The collection dates are not exactly the same, which was added into the Table 1. Each subsample was collected from the different plant of the same species. This information was added into the 2.1 section in the revision.
   4. Line 71. More details about host plants should be provided. For instance, cultivation practice, crop size, pesticide application, etc., since these factors are relevant to the conditions of the collected mite population.
      Response: More details about host plants were provided. See lines 74-78.
   5. Line 103: 30 l? what does mean “l”? Revise.
      Response: Sorry for the stylistic error and we have made revision.
   6. Revise in detail the M&M section. Some experimental details are missing to ensure outcome reproducibility. For instance, brand, model, and grade of reagents, solvents, materials, and instruments must be provided. In addition, details about sequencing should also be clarified.
      Response: Details about brand, model of instruments and sequencing were added.
   7. Line 108: More details on determining mutation frequencies should also be expanded for readers, specifically to obtain Figure 1.
      Response: The mutation frequencies was calculated by the formula. Detailed information on the formula for calculating mutation frequency and the software used for Figure 1 were added. See lines 139-141 and 144-145 in revision. 
   8. Lines 120 and 122: LC₅₀ instead of LC50.
      Response: Done.
   9. Lines 167-169: Revise this text since it seems to be a typo.
      Response: We deleted this typo error.
   10. The discussion section can be improved since it is highly descriptive and even speculative. In addition, it can be even reorganized, possibly by subheadings.
       Response: Thanks for the comments and we made revisions on the discussion section. See lines 206-214. We didn’t added subheadings after careful consideration and referring to the published papers in Horticulturae.
   11. A conclusion section should be generated, I mean, the conclusion section should be outside of the discussion section.
       Response: We've separated the conclusion from the discussion.
   12. Lines 263-266: These conclusions are not entirely supported by results, since the number of populations (vide infra) is very low. I consider important to give an adequate scope of these results.
       Response: We’ve modified the conclusion statement to specify the scope of the results and make it more scientific and rigorous.
   Finally, although the study seems to be adequately developed from the experimental point of view, my main criticism of this manuscript is related to the number of populations of T. urticae evaluated since it is very low and not enough to give the final authors’ conclusions. Therefore, the manuscript has an approach drawback that must be clarified for readers. From this fact, the scope of the manuscript should be re-oriented to afford better scientific quality and merit. For instance, it should be determined or attempted to establish a plausible relationship between the populations and the details of the crops where mite populations were obtained (which, in fact, those details are missing), whose host conditioning is the main factor for this mite to win resistance or not. Accordingly, the discussion, apart from expanding the descriptions of obtained results, should be oriented towards discussing the results from a mechanistic point of view and the relationship between the evaluated populations and their detailed origin since, I repeat, the number of mite populations is very low to draw better, more far-reaching conclusions.
   Response: Thanks for the comments. We agree with the reviewer’s concerning on the “the low number of populations of T. urticae evaluated”. According to the suggestions, we raised this point in the discussion and specified the scope for the results, in order to make the conclusion more scientific and rigorous. Please see lines 288-295.

Reviewer 3 Report

Comments to the editor and authors 

Horticulturae #1778823

 

Editor and author(s),

 

The manuscript exhibits promising results, which can attract an audience to the journal. The research involves a toxicity assessment of seven pesticides against the target species (Tetranychus urticae), as well as resistance levels in contrast to the laboratory susceptive population. The authors undertake laboratory testing on five two-spotted spider mite field populations, including mutation frequency in the GluCl1, GluCl3, VGSC, Ace, Cytb, and PSST genes.

 

The paper is well-written and simple to understand. Five T. urticae populations from Hainan, China, were tested and shown to have high to extremely high resistance to abamectin and bifenthrin, as well as medium to high resistance to pyridaben, profenofos, and cyflumetofen. On the other hand, four of the five populations were either vulnerable to or had only limited resistance to bifenazate and B-azolemiteacrylic. These findings are important for pest management programs because they emphasize the importance of pesticide rotation and monitoring of control failure and resistance.

 

I have identified some points that require minor reviews by the authors, which are below.

 

Specific comments

 

I recommend that the authors use keywords that aren't in the title. Perhaps pest management, mite control, and chemical control.

 

The introduction emphasizes the protagonists in the work and demonstrates the possible significance of the research. Only the remarks on each pesticide's action mode were not included. I think that may be of interest to readers.

 

Abamectin-Glutamate-gated chloride channel (GluCl) activators

Profenofos-Acetylcholinesterase (AChE) inhibitors

Bifenthrin-Sodium channel modulators

Pyridaben-Mitochondrial complex I electron transport inhibitors

Bifenazate-Mitochondrial complex III electron transport inhibitors

Cyflumetofen-Mitochondrial complex II electron transport inhibitors

B-azolemiteacrylic-inhibition effects on mitochondrial respiratory chain complexes II

 

Lines 59: Remove "We also determine"

 

The materials and methods are detailed and easy to understand. I suggest including the pesticide concentrations utilized.

 

Line 68: Italic "Phaseolus vulgaris"

 

The results and discussion are remarkably interesting and easy to understand. In general, the authors support their results with other references. Furthermore, some pest management advice is presented, which is very interesting.

 

Lines 167-168. Remove "Proof of Theorem 1. Text of the proof." Note that the phrase "of Theorem 1" is optional if it is clear which theorem is being referred to. Always finish a proof with the following symbol. "

 

References should be double-checked. There are journals' names without abbreviations (see numbers 1, 13, 17, 27, 31, 32) and without italics (see number 35).

 

I hope that the comments improve your manuscript,

All the best.

reviewer

 

Author Response

Editor and author(s),

The manuscript exhibits promising results, which can attract an audience to the journal.The research involves a toxicity assessment of seven pesticides against the target species (Tetranychus urticae), as well as resistance levels in contrast to the laboratory susceptive population. The authors undertake laboratory testing on five two-spotted spider mite field populations, including mutation frequency in the GluCl1, GluCl3, VGSC, Ace, Cytb, and PSST genes.

The paper is well-written and simple to understand. Five T. urticae populations from Hainan, China, were tested and shown to have high to extremely high resistance to abamectin and bifenthrin, as well as medium to high resistance to pyridaben, profenofos, and cyflumetofen. On the other hand, four of the five populations were either vulnerable to or had only limited resistance to bifenazate and B-azolemiteacrylic. These findings are important for pest management programs because they emphasize the importance of pesticide rotation and monitoring of control failure and resistance.
Response: Thanks for the positive comments.

I have identified some points that require minor reviews by the authors, which are below.

 

Specific comments

1. I recommend that the authors use keywordsthat aren't in the title. Perhaps pest management, mite control, and chemical control.
   Response: We change the keywords to Tetranychus urticae; resistance monitoring; chemical control; Hainan
2. The introduction emphasizes the protagonists in the work and demonstrates the possible significance of the research. Only the remarks on each pesticide's action mode were not included. I think that may be of interest to readers.

Abamectin-Glutamate-gated chloride channel (GluCl) activators

Profenofos-Acetylcholinesterase (AChE) inhibitors

Bifenthrin-Sodium channel modulators

Pyridaben-Mitochondrial complex I electron transport inhibitors

Bifenazate-Mitochondrial complex III electron transport inhibitors

Cyflumetofen-Mitochondrial complex II electron transport inhibitors

B-azolemiteacrylic-inhibition effects on mitochondrial respiratory chain complexes II
Response: We added the action mode for some pesticides according to the reviewer’s advice, please see lines 50-55.

3. Lines 59: Remove "We also determine"
   Response: Done.
4. The materials and methods are detailed and easy to understand. I suggest including the pesticide concentrations utilized.
   Response: Thanks for the comments. In the bioassay, the resistance of five T. urticae field populations were determined against 7 pesticides, individually. The concentrations utilized in this section are original data, so we think it’s unnecessary to display the pesticide concentrations. Of course the LC₅₀ values are exhibited in the manuscript.
5. Line 68: Italic "Phaseolus vulgaris".
   Response: Done.
6. The results and discussion are remarkably interesting and easy to understand. In general, the authors support their results with other references. Furthermore, some pest management advice is presented, which is very interesting.
   Response: Thanks for the positive comment.
7. Lines 167-168. Remove "Proof of Theorem 1. Text of the proof." Note that the phrase "of Theorem 1" is optional if it is clear which theorem is being referred to. Always finish a proof with the following symbol. "
   Response: Done.
8. References should be double-checked. There are journals' names without abbreviations (see numbers 1, 13, 17, 27, 31, 32) and without italics (see number 35).
   Response: Done.

I hope that the comments improve your manuscript,

All the best.