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Transcriptomic analysis of s-methoprene resistance in the lesser grain borer, Rhyzopertha dominica, and evaluation of piperonyl butoxide as a resistance breaker
MARIA SAKKA
University of Thessaly
Corresponding Author
ORCiD: https://orcid.org/0000-0002-4624-0953
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: The lesser grain borer, Rhyzopertha dominica is a serious pest of stored grains. Fumigation and contact insecticides play a major role in managing this pest globally. While insects are developing genetic resistance to chemicals, hormonal analogues such as s-methoprene play a key role in reducing general pest pressure as well as managing pest populations that are resistant to fumigants and neurotoxic contact insecticides. However, resistance to s-methoprene has been reported in R. dominica with some reports showing a remarkable high resistance, questioning the use of this compound and other related analogues in grain protection. The current study attempts to identify possible molecular mechanisms that contribute in resistance to s-methoprene in R. dominica.
Results: Transcriptome analysis of resistant and susceptible strains of this pest species identified a set of differentially expressed genes related to cytochrome P450s, indicating their potential role in resistance to s-methoprene. Laboratory bioassays were performed with s-methoprene treated wheat grains in presence and absence of piperonyl butoxide (PBO), a cytochrome P450 inhibitor. The results indicate that PBO, when applied alone, at least at the concentration tested here, had no effect on R. dominica adult emergence, but has a clear synergistic effect to s-methoprene. The number of produced progeny decreased in presence of the inhibitor, especially in the resistant strain. In addition, we also identified CYP complement (CYPome) of R. dominica, annotated and analysed phylogenetically, to understand the evolutionary relationships with other species.
Conclusions: The information generated in current study suggest that PBO can effectively be used to break resistance to s-methoprene in R. dominica.
Keywords
piperonyl butoxide, s-methoprene, resistance, transcriptome analysis, Rhyzopertha dominica
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This is a list of supplementary files associated with this preprint. Click to download.
- Additionalfile1figureS1.jpg
Additional file 1, Figure S1: Mean mortality (±SEM) of Rhyzopertha dominica after 7, 14 and 21 days of exposure for the susceptible (Lab-S) and resistant (Met-R) strain for all the combinations tested (control, 0.01 mg/kg, 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, PBO, 0.01 mg/kg + PBO, 0.03 mg/kg + PBO, 0.1 mg/kg + PBO, 0.3 mg/kg + PBO for susceptible and control, 1 mg/kg, 3 mg/kg, 10 mg/kg, 30 mg/kg, PBO, 1 mg/kg + PBO, 3 mg/kg + PBO, 10 mg/kg + PBO, 30 mg/kg + PBO for resistant).
- Additionalfile2figureS2.pdf
Additional file 2, Figure S2: Principal components analysis of the transcript expression levels between the resistant (Met-R - blue) and susceptible (Lab-S - red) strains. The two strains are clearly different from each other, a prerequisite for downstream analyses.
- Additionalfile3TableS1.xlsx
Additional file 3, Table S1: Corresponding information regarding sequence identity of CYPs.
- Additionalfile4TableS2.xlsx
Additional file 4, Table S2: Details of all up- and down-regulated genes between the resistant Met-R and the susceptible Lab-S strains (padj <0.001, log2|FC| >2).
- Additionalfile5TableS3.xlsx
Additional file 5, Table S3, Amino acid sequences used in the phylogenetic study of the R. dominica P450s, presented in Fig. 4.
- Additionalfile6TableS4.xlsx
Additional file 6, Table S4: Primers used for the validation qPCRs.
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This preprint is under consideration at BMC Genomics. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Transcriptomic analysis of s-methoprene resistance in the lesser grain borer, Rhyzopertha dominica, and evaluation of piperonyl butoxide as a resistance breaker
MARIA SAKKA
University of Thessaly
Corresponding Author
ORCiD: https://orcid.org/0000-0002-4624-0953
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 4
Posted 30 Dec, 2020
No community comments so far
Editor assigned
On 13 Dec, 2020
Submission checks complete
On 13 Dec, 2020
Editor invited
On 13 Dec, 2020
No comments provided
Editorial decision: Minor revision
On 07 Dec, 2020
Editor assigned
On 28 Nov, 2020
Submission checks complete
On 28 Nov, 2020
Editor invited
On 28 Nov, 2020
No comments provided
Reviewer #1 agreed
On 21 Sep, 2020
Reviewer #2 agreed
On 21 Sep, 2020
Review #1 received
Received 21 Sep, 2020
Editor assigned
On 16 Sep, 2020
Reviewers invited
Invitations sent on 16 Sep, 2020
Submission checks complete
On 15 Sep, 2020
Editor invited
On 15 Sep, 2020
No comments provided
Editorial decision: Major revision
On 18 Jun, 2020
Review #2 received
Received 30 May, 2020
Reviewer #3 agreed
On 14 May, 2020
Reviewer #2 agreed
On 12 May, 2020
Review #1 received
Received 26 Apr, 2020
Reviewer #1 agreed
On 06 Apr, 2020
Reviewers invited
Invitations sent on 01 Apr, 2020
Editor assigned
On 31 Mar, 2020
Submission checks complete
On 19 Mar, 2020
Editor invited
On 19 Mar, 2020
First submitted
On 11 Mar, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Epigenetics & Genomics
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: The lesser grain borer, Rhyzopertha dominica is a serious pest of stored grains. Fumigation and contact insecticides play a major role in managing this pest globally. While insects are developing genetic resistance to chemicals, hormonal analogues such as s-methoprene play a key role in reducing general pest pressure as well as managing pest populations that are resistant to fumigants and neurotoxic contact insecticides. However, resistance to s-methoprene has been reported in R. dominica with some reports showing a remarkable high resistance, questioning the use of this compound and other related analogues in grain protection. The current study attempts to identify possible molecular mechanisms that contribute in resistance to s-methoprene in R. dominica.
Results: Transcriptome analysis of resistant and susceptible strains of this pest species identified a set of differentially expressed genes related to cytochrome P450s, indicating their potential role in resistance to s-methoprene. Laboratory bioassays were performed with s-methoprene treated wheat grains in presence and absence of piperonyl butoxide (PBO), a cytochrome P450 inhibitor. The results indicate that PBO, when applied alone, at least at the concentration tested here, had no effect on R. dominica adult emergence, but has a clear synergistic effect to s-methoprene. The number of produced progeny decreased in presence of the inhibitor, especially in the resistant strain. In addition, we also identified CYP complement (CYPome) of R. dominica, annotated and analysed phylogenetically, to understand the evolutionary relationships with other species.
Conclusions: The information generated in current study suggest that PBO can effectively be used to break resistance to s-methoprene in R. dominica.
Figure 1
Figure 2
Figure 3
Figure 4