Tomato Chlorosis Virus (ToCV) Infection Induced the Resistance of Bemisia tabaci to Two Insecticides: Pyrethroids and Flupyradifurone

Round 1

Reviewer 1 Report

Dear Dr. Zhang,

your manuscript Zhang et al., titled "Tomato chlorosis virus (ToCV) infection induced the resistance of Bemisia tabaci to two insecticides pyrethroids and flypyradifurone" presents very interesting data. Such a multitrophic study on  interactions between plants, plant pathogens, and pest insects, and the consequences for crop production are much needed.

Your manuscript certainly merits publication after some revision. In the following I will detail my comments and suggestions:

 

Introduction

very good and in my opinion sufficiently exhaustive text on B. tabaci, some more information on the problem of insecticide resistance in pest insects in general and in "in-door-pests" especially would improve the introdution

 

Material and methods

line 55: state exact location where the study was conducted 

line 59: state origin of the clones used

line 95: state all concentrations for both insecticides tested and the relevance of those for practical application purposes

line 97: should say 25 insects of mixed sex (?) instead of "heads"

line 96 and 98: what is a "finger tube"?

line 103-104: unclear, please rewrite, English is confusing

line 105, 109, 114: give detailed methodology, just citing the instruction of each kit is not sufficient here for the reader, especially since the kits are maybe not availabe outside of China?

line 120: how many insects did you collect from cotton?

line 129: do you consider three replication are sufficient to for such a study?

I would suggest at least one more for statistical reasons.

 

Results

Table 2: 1 or 2 digits after point are sufficient, most have three, one has 2-please adjust to journal format

give insecticide concentrations tested as well in this table

 

Figure 2 and Fig. 3: text on both axis is very small and hard to read, also axis are of different range - please change into uniform axes for reason of comparison

 

Discussion

line 209: pest resistance ... "is"... instead of was

line 215-216: delete

line 217-260: re-write this complete section, entire phrases are difficult or not at all understandable; the discussion lacks comparison of your findings on B. tabaci virus-induced resistance to similar interactions concerning other pest insects- are there any? if so, please detail and discuss in relation with your results

line 248-250: very interesting result with regard to ecological and epidemic consequences, please discuss in detail, this discussion is currently completely missing; compare to similar studies for example: phytoplasmas and plant hoppers on apples (Meyer and Gross)

line 260: what do you mean by "insects injected arboviruses"? re-write!

 

Conclusion

Re-write entire section, not possible to fully understand what you wanted to tell the readers

 

References

all references deal with B. tabaci - are there no other pest insects of interest for your study? other cases of plant pathogen-pest interactions that influence resistance? 

also newer literature should be included if available

 

 

 

 

 

 

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

Review of the manuscript entitled “Tomato chlorosis virus (ToCV) infection induced the resistance of Bemisia tabaci to two insecticides pyrethroids and flupyradifurone”

 

The paper addresses the important problem of B. tabaci resistance to insecticides and makes a contribution on the relationship between genes and resistance. It is a valuable attempt to explain the reasons behind resistance to bifenthrin and flupyradifurone.

Below are comments and questions to particular sections of the paper

Throughout the paper I propose the notation B. tabaci MED. instead of MED B. tabaci.

2. Materials and Methods

2.1. Tomato, cotton and B. tabaci

Analysing “Tomato plants at the three-true-leaf stage were injected with an infections ToCV 58 cDNA clone, and 0.5 ml of the infectious cDNA clone was injected into each plant to 59 achieve systemic infection [17].”

Is the correct procedure not better presented in this publication: Orílio, A. F., Fortes, I. M., & Navas-Castillo, J. (2014). Infectious cDNA clones of the crinivirus Tomato chlorosis virus are competent for systemic plant infection and whitefly-transmission. Virology, 464, 365-374.

 

2.2. Pyrethroids and flupyradifurone bioassay of viruliferous and viruliferous MED B. tabaci

In this sentence it should be: “non-viruliferous B. tabaci MED”

 

Sentences: “The leaf dish was embarrassed 96 with ager, fits, stuffed with cotton plugs, marked them, and about 25 heads of B. tabaci eaten on infected tomatoes for 48 h were been blow into the finger tube. The finger tube containing B. tabaci were been put in the light incubator. The temperature was set to 26 ± 99 2 °C, the humidity was set to 70 % ± 10 %, and the light cycle was 16 h : 8 h（L:D）. After 100 48 h, the survival of B. tabaci was checked.” This passage is difficult for me to understand. It seems to me that two issues have been mixed up. Please pay attention to it and correct it.

 

2.3. Metabolic enzyme assays

In sentence “by comparing the O.D. value of the 107 samples to the standard curve.”

Instead of O.D. - write in full words

 

Figure 1. - there is no reference in the text of the paper to Figure 1.

 

2.5. Statistical analyses

There is no mention of the Post-hoc test in the methodology. As I see it it is Tukey's HSD. In my opinion it should say Tukey's HSD with α = 0.05. This should be corrected in figure 2 and 3.

 

3. Results

3.1. Bioassays

Table 2. Susceptibility of MED B. tabaci adults to pyrethroids and flupyradifurone

Stock abbreviations are given incorrectly in the table. The legend below the table is incomplete.

 

3.2. Activities of metabolic enzymes

The statistical description seems partly inadequate. As I understand it, different letters above the columns indicate statistical differences. Therefore, the notation seems somewhat troublesome: GST: There was no particularly significant difference. However, this is not an error.

 

3.3. Expression patterns of detoxification-related P450, GST and CarE

When the authors write about "gene LOC109038667 expression" there is the sentence: "Quantitative analysis showed that:" I have a question, was this analysis (Quantitative analysis) described in material I of the method?

 

4. Discussion

“Among them, the gene CYP6CX4 has been significantly increased, and the expression of LOC109038667 after indoor sensitive populations obtains ToCV 48 h was even close to the healthy field population.” It would be of value to explain why GSTs2 is not included in this compilation? The amount of GSTs2 did indeed increase after 48h exposure to ToCV. Please note that earlier the authors wrote: „Studies had found that the enhancement of metabolic enzyme activity was considered an important mechanism for B. tabaci resistance, the overexpression of cytochrome P450 (P450) related gene CYP6CX4 and glutathione S-transferase (GST) related gene CSTs2 also played crucial roles in the flupyradifurone resistance of B. tabaci; carboxylesterase (CarE) played an important role in the resistance of B. tabaci to pyrethroids [14-16].”

 

5. Conclusions

To sum up, the research showed that there was a mutually beneficial and mutually beneficial relationship

 

References

Reference to items 26 and 27 is missing from the text of the manuscript

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

there are only some very minor changes in the footnote of table 2 and on the table that are listed on the pdf  attached

Comments for author File: Comments.pdf

Author Response

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Reviewer 4 Report

Authors proved that insect-born plant virus ToCV  show the possibility of enhancing insect-born insecticide resistance.

Author Response

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Author Response File: Author Response.docx