scholarly journals High-throughput genotyping of a full voltage-gated sodium channel gene via genomic DNA using target capture sequencing and analytical pipeline MoNaS to discover novel insecticide resistance mutations

2019 ◽  
Author(s):  
Kentaro Itokawa ◽  
Koji Yatsu ◽  
Tsuyoshi Sekizuka ◽  
Yoshihide Maekawa ◽  
Osamu Komagata ◽  
...  
Keyword(s):  
Amino Acid ◽  
Insecticide Resistance ◽  
Sodium Channel ◽  
Genomic Dna ◽  
Genomic Structure ◽  
Voltage Gated Sodium Channel ◽  
Pyrethroid Insecticides ◽  
Voltage Gated ◽  
Probe Set ◽  
Capture Sequencing

AbstractInsects’ voltage-gated sodium channel (VGSC) is the primary target site of pyrethroid insecticides. Various amino acid substitutions in the VGSC protein are known to confer insecticide resistance and are selected under insecticide pressure. In the genome, the VGSC gene consists of more than 30 exons sparsely distributed across a large genomic region, which often exceeds 100 kbp. Due to this complex genomic structure of gene VGSC, it is usually challenging to genotype full coding nucleotide sequences (CDSs) of VGSC from individual genomic DNA (gDNA). In this study, we designed biotinylated oligonucleotide probes via annotated CDSs of VGSC of Asian tiger mosquito, Aedes albopictus. The probe set effectively concentrated (>80,000-fold) all targeted regions of gene VGSC from pooled barcoded Illumina libraries each constructed from individual A. albopictus gDNAs. The probe set also captured all homologous VGSC CDSs except tiny exons from the gDNA of other Culicinae mosquitos, A. aegypti and Culex pipiens complex, with comparable efficiency by virtue of the high conservation of VGSC at the nucleotide level. Furthermore, we developed an automated bioinformatic analysis pipeline to genotype VGSC after capture sequencing—MoNaS (Mosquito Na+ channel mutation Search)—which conducts mapping of reads, variant calling, and variant annotation for nonsynonymous mutations. The proposed method and our bioinformatic tool should facilitate the discovery of novel amino acid variants conferring insecticide resistance on VGSC and population genetics studies on resistance alleles with respect to the origin, selection, and migration of both clinically and agriculturally important insect pests.

scholarly journals Molecular Insight into the Knockdown Resistance (kdr) in the Voltage Gated Sodium Channel (vgsc) Gene of the Main Dengue Vector, Aedes aegypti (Diptera: Culicidae) and the Discovery of Novel Regional Specific Point Mutation A1007G in Malaysia.

2021 ◽  
Author(s):  
Mas Azlin M. Akhir ◽  
Mustafa F. F. Wajidi ◽  
Sébastien Lavoué ◽  
Ghows Azzam ◽  
Izhan Shahrin Jaafar ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Sodium Channel ◽  
Vector Control ◽  
Resistance Mechanism ◽  
Point Mutations ◽  
Voltage Gated Sodium Channel ◽  
Pyrethroid Insecticides ◽  
Voltage Gated ◽  
Control Programmes ◽  
Pirimiphos Methyl

Abstract Background: Characterization of the insecticide resistance mechanism imparts the society with the information on the evolutionary process involved in the adaptation of Aedes aegypti mosquito to environmental changes. Investigating the phenotypic status of the target mosquitoes, their resistance level as well as elucidating the genotypic profile provides information about the involvement of insecticide resistance mechanism, in terms of portraying the evolution of resistance in the field, to eventually managing vector control programmes. In this current study, we investigated the quantification responses for the phenotypic and genotypic resistance of Ae. aegypti population from different states in Malaysia. Methods: We tested insecticide susceptibility status of adult Ae. aegypti from populations of States of Penang, Selangor and Kelantan (Peninsular Malaysia) against 0.25% permethrin and 0.25% pirimiphos-methyl through WHO bioassay kit. Permethrin-resistant and permethrin susceptible samples were then genotyped for domains II and III in the voltage gated sodium channel (vgsc) gene using allele specific PCR (AS-PCR) for the presence of diagnostic single nucleotide mutations. AS-PCR results were then validated in sequencing these two domains to identify any possible additional point mutations. Results: Adult WHO bioassay revealed that populations of Ae. aegypti from these three states were highly resistant towards 0.25% permethrin and 0.25% pirimiphos-methyl. Genotyping results showed that three knockdown (kdr) mutations (i.e. S989P, V1016G and F1534C) were associated with pyrethroid resistance in these populations. We also report for the first time the presence of the A1007G mutation in Malaysian populations of Ae. aegypti.Conclusions: This study brings an insight on the occurrence and association of point mutations with insecticide resistance in Malaysian populations of Ae. aegypti. The results reveal the widespread of several kdr mutations in the field with the consequence to compromise the use of pyrethroid insecticides in vector control programmes. Knowledge on the distribution of target site resistance throughout Malaysia is vital to ensure the success of the insecticide-based vector control programme.

scholarly journals Molecular Insight into the Knockdown Resistance (kdr) in the Voltage Gated Sodium Channel (vgsc) Gene of the Main Dengue Vector, Aedes aegypti (Diptera: Culicidae) and the Discovery of Novel Regional Specific Point Mutation A1007G in Malaysia.

2020 ◽  
Author(s):  
Mas Azlin M. Akhir ◽  
Mustafa F. F. Wajidi ◽  
Sébastien Lavoué ◽  
Ghows Azzam ◽  
Izhan Shahrin Jaafar ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Sodium Channel ◽  
Vector Control ◽  
Environmental Changes ◽  
Resistance Mechanism ◽  
Point Mutations ◽  
Voltage Gated Sodium Channel ◽  
Pyrethroid Insecticides ◽  
Voltage Gated ◽  
Pirimiphos Methyl

Abstract Background: Characterization of the insecticide resistance mechanism imparts the society with the information on the evolutionary process involved in the adaptation of Aedes aegypti mosquito to environmental changes. Investigating the phenotypic status of the target mosquitoes, their resistance level as well as elucidating the genotypic profile provides information about the involvement of insecticide resistance mechanism, in terms of portraying the evolution of resistance in the field, to eventually managing vector control programs. In this current study, we investigated the quantification responses for the phenotypic and genotypic resistance of Ae. aegypti population from different states in Malaysia. Methods: We tested insecticide susceptibility status of adult Ae. aegypti from populations of States of Penang, Selangor and Kelantan (Peninsular Malaysia) against permethrin 0.25% and pirimiphos-methyl 0.25% through WHO bioassay kit. Permethrin-resistant and permethrin susceptible samples were then genotyped for domains II and III in the voltage gated sodium channel (vgsc) gene using allele specific PCR (AS-PCR) for the presence of diagnostic single nucleotide mutations. AS-PCR results were then validated in sequencing these two domains to identify any possible additional point mutations. Results: Adult WHO bioassay revealed that populations of Ae. aegypti from these three states were highly resistant towards Permethrin 0.25% and Pirimiphos-methyl 0.25%. Genotyping results showed that three knockdown (kdr) mutations (i.e. S989P, V1016G and F1534C) were associated with pyrethroid resistance in these populations. We also report for the first time the presence of the A1007G mutation in Malaysian populations of Ae. aegypti.Conclusions: This study brings an insight on the occurrence and association of point mutations with insecticide resistance in Malaysian populations of Ae. aegypti. The results reveal the widespread of several kdr mutations in the field with the consequence to compromise the use of pyrethroid insecticides in vector control programmes. Knowledge on the distribution of target site resistance throughout Malaysia is vital to ensure the success of the insecticide-based vector control program.

A mutation (L1014F) in the voltage-gated sodium channel of the grain aphid, Sitobion avenae , is associated with resistance to pyrethroid insecticides

2013 ◽  
Vol 70 (8) ◽  
pp. 1249-1253 ◽  
Author(s):  
Stephen P Foster ◽  
Verity L Paul ◽  
Russell Slater ◽  
Anne Warren ◽  
Ian Denholm ◽  
...  
Keyword(s):  
Sodium Channel ◽  
Sitobion Avenae ◽  
Voltage Gated Sodium Channel ◽  
Pyrethroid Insecticides ◽  
Voltage Gated ◽  
Grain Aphid

scholarly journals Modelling insecticide-binding sites in the voltage-gated sodium channel

2006 ◽  
Vol 396 (2) ◽  
pp. 255-263 ◽  
Author(s):  
Andrias O. O'Reilly ◽  
Bhupinder P. S. Khambay ◽  
Martin S. Williamson ◽  
Linda M. Field ◽  
B. A. WAllace ◽  
...  
Keyword(s):  
Sodium Channel ◽  
Binding Sites ◽  
Homology Model ◽  
Voltage Gated Sodium Channel ◽  
Pyrethroid Insecticides ◽  
Voltage Gated ◽  
Arthropod Species ◽  
State Dependent ◽  
Binding Cavity

A homology model of the housefly voltage-gated sodium channel was developed to predict the location of binding sites for the insecticides fenvalerate, a synthetic pyrethroid, and DDT an early generation organochlorine. The model successfully addresses the state-dependent affinity of pyrethroid insecticides, their mechanism of action and the role of mutations in the channel that are known to confer insecticide resistance. The sodium channel was modelled in an open conformation with the insecticide-binding site located in a hydrophobic cavity delimited by the domain II S4-S5 linker and the IIS5 and IIIS6 helices. The binding cavity is predicted to be accessible to the lipid bilayer and therefore to lipid-soluble insecticides. The binding of insecticides and the consequent formation of binding contacts across different channel elements could stabilize the channel when in an open state, which is consistent with the prolonged sodium tail currents induced by pyrethroids and DDT. In the closed state, the predicted alternative positioning of the domain II S4-S5 linker would result in disruption of pyrethroid-binding contacts, consistent with the observation that pyrethroids have their highest affinity for the open channel. The model also predicts a key role for the IIS5 and IIIS6 helices in insecticide binding. Some of the residues on the helices that form the putative binding contacts are not conserved between arthropod and non-arthropod species, which is consistent with their contribution to insecticide species selectivity. Additional binding contacts on the II S4-S5 linker can explain the higher potency of pyrethroid insecticides compared with DDT.

Sign in / Sign up

Export Citation Format

Share Document