RNA interference of odorant receptor CquiOR114/117 affects blood-feeding behavior in Culex quinquefasciatus

Wu, Qun; Li, Chunxiao; Liu, Qin-mei; Guo, Xiaoxia; Shi, Qingming; Zhang, Hengduan; Dong, Yan-De; Xing, Dan; Zhang, Yingmei; Liao, Chenghong; Han, Qian; Zhao, Tongyan

doi:10.1016/j.actatropica.2020.105343

Cited by 15 publications

(15 citation statements)

References 15 publications

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“…One microgram of messenger RNA in a 20-µl reaction volume was reverse transcribed into complementary DNA (cDNA) in accordance with the Prime Script RT Reagent Kit with genomic DNA Eraser protocol under the following conditions: 37 °C for 15 min, 85 °C for 5 s, followed by a 4 °C hold. Ribosomal protein L8 gene [ 33 ] and 18S ribosomal RNA gene [ 26 ] were used as the internal control genes for calibration. The primers and probes used for the target and internal control genes were synthesized based on the sequences given in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

Deltamethrin interacts with Culex quinquefasciatus odorant-binding protein: a novel potential resistance mechanism

Shen

Wang

et al. 2022

Parasites Vectors

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Background Odorant-binding proteins (OBPs) play important roles in many physiological processes of mosquitoes. Previous high-throughput sequencing studies have revealed that some OBPs of Culex quinquefasciatus might be involved in the development of resistance to insecticides. Methods Based on the results of sequencing analyses, the OBP28 gene was selected for evaluation in this study. Three laboratory strains of Cx. quinquefasciatus [susceptible strain (SS), deltamethrin-resistant strain 1 (HN) and deltamethrin-resistant strain 2 (RR)] were first examined by using the Centers for Disease Control and Prevention bottle bioassay, after which the expression level of the OBP28 gene in the susceptible and deltamethrin-resistant strains was determined by real-time quantitative polymerase chain reaction. The OBP28 gene in deltamethrin-resistant strain RR was silenced using RNA interference technology. The expression level of OBP28 and the resistance level were tested in the silenced strain and control strain after microinjection of double-stranded RNA for a 48-h interference period. Four field-collected strains (henceforth ‘field strains’) of Cx. quinquefasciatus were also examined for their resistance to deltamethrin and levels of OBP28 expression. Finally, a correlation analysis between deltamethrin resistance and gene expression was carried out for all seven strains, i.e. the four field strains and the three laboratory strains. Results In the bioassay, the mortality of SS, HN and RR was 100%, 21.33% and 1.67%, respectively. The relative expression levels of OBP28 in strains HN and RR were 6.30- and 6.86-fold higher, respectively, than that of strain SS. After silencing of the OBP28 gene, the mortality of strain RR was 72.20% and that of the control strain 26.32%. The mortality of strain RR increased significantly after interference compared to that of the control strain. There was a negative correlation between OBP28 gene expression and mortality in adult mosquitoes after exposure to deltamethrin. Conclusions To our knowledge, this study shows for the first time a correlation between the expression of a gene coding for OBP and insecticide resistance in mosquitoes. The potential resistance mechanism that was elucidated provides a new target gene for the surveillance of resistance in mosquitoes. Graphical abstract

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Section: Methodsmentioning

confidence: 99%

Deltamethrin interacts with Culex quinquefasciatus odorant-binding protein: a novel potential resistance mechanism

Shen

Wang

et al. 2022

Parasites Vectors

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“…CmPBF, CqPBF v.s.CqNBF and CmPBF v.s.CmNBF. Some genes that were eligible but had been validated in previous studies were excluded [29,30]. Finally we selected 14 olfactory genes and 2 housekeeping genes (RPL8 and 18S) to verify the accuracy of the results obtained herein.…”

Section: Qpcr Verification Of Identified Olfactory Genesmentioning

confidence: 99%

Screening of olfactory genes related to blood-feeding behaviors in Culex pipiens quinquefasciatus and Culex pipiens molestus by transcriptome analysis

Gao

Yang

et al. 2022

PLoS Negl Trop Dis

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Background Culex pipiens quinquefasciatus Say (Cx. quinquefasciatus) and Culex pipiens form molestus Forskal (Cx. molestus) in the Culex pipiens complex group show considerable differences in host seeking, blood feeding, mating behavior and in vector competence. Blood-feeding mosquito behaviors are closely related to their olfactory gene expression and olfactory gene repertoire composition. Comparing olfactory genes between these two subspecies with significantly different blood-feeding behaviors can support further research on the molecular mechanism of the Culex pipiens complex olfactory sensory system, providing a new approach for determining candidate attractant or repellent compounds. Methods Non-blood-feeding (NBF) and post-blood-feeding (PBF) olfactory system transcriptomes of the two subspecies were sequenced, and the biological functions of their differentially expressed genes were described by bioinformatics analysis. A quantitative polymerase chain reaction (qPCR) was applied to validate the RNA-seq data. The roles of particular olfactory receptors in Cx. quinquefasciatus blood-feeding behaviors were evaluated by RNAi. Results Five, 7, 24, and 3 Cx. quinquefasciatus-specific OBPs, Cx. molestus-specific OBPs, Cx. quinquefasciatus-specific ORs and Cx. molestus-specific ORs were identified, respectively. The majority of selected ORs were consistent with the predicted transcriptome sequencing results after qRT-PCR validation. OR5 was expressed only in Cx. quinquefasciatus, and OR65 was the only gene upregulated after blood feeding in Cx. molestus. The blood-feeding rates of the OR5 and OR78 dsRNA groups were significantly lower (4.3%±3.1% and 13.3%±11.5%) than those of the enhanced green fluorescence protein (EGFP) group (64.5%±8.7%). Conclusion Most OBPs and ORs were expressed in both subspecies but showed divergence in expression level. OR5 and OR65 might be species-specific expressed genes that regulate the olfactory behaviors of Cx. quinquefasciatus and Cx. molestus, respectively. The RNA interference of OR5 and OR78 could inhibit the blood-feeding behavior of Cx. quinquefasciatus, providing new targets for screening effective repellent compounds to control mosquito-borne diseases effectively and efficiently.

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“…The qRT-PCR results showed that OR27, OR30, OR62, OR86, OR91 and OR117 were signi cantly upregulated after blood feeding, so these genes were presumed to be related to the search for suitable oviposition sites based on oviposition attractants by mosquitoes to breed their offspring. Wu [29] found that CquiOR114/117 was upregulated after blood feeding in Cx. quinquefasciatus.…”

Section: Discussionmentioning

confidence: 99%

Screening for Odorant Receptor Genes Expressed in Aedes Aegypti Involved in Blood-Feeding and Oviposition Behaviors

Zhao

et al. 2021

Preprint

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Background: Aedes aegypti is one of the most important vector worldwide, and its survival and reproductive processes depend heavily on the olfactory system. In this study, the expression levels of all odorant receptor (OR) genes of Ae. aegypti were explored in different physiological periods to identify olfactory genes that may be associated with mosquito blood sucking and searching for oviposition sites.Methods: Four groups, consisting of Ae. aegypti males (M), pre-blood-feeding females (F), post-blood-feeding females (B) and post-oviposition females (O), were established. A total of 114 pairs of primer targeting all OR genes were designed based on the whole genome of Ae. aegypti. The expression of OR genes was evaluated by real-time fluorescence quantitative PCR for relative quantification and the comparison of differences between groups.Results: A total of 53 differentially expressed OR genes were identified between males and females in Ae. aegypti antennae. And 8, 5 and 13 differentially expressed OR genes were identified before versus after blood feeding, before versus after oviposition and post-blood-feeding versus post-oviposition, respectively. Meanwhile, 16 OR genes were significantly differentially expressed in multiple physiological periods of mosquitoes.Conclusions: A large number of ORs with significant intergroup differences and high expression levels were screened in this study, including OR75, OR88, OR110 and OR115 and so on. Some of these genes are reported for the first time, providing possible targets for the development of mosquito control pathways based on the olfactory system.

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RNA interference of odorant receptor CquiOR114/117 affects blood-feeding behavior in Culex quinquefasciatus

Cited by 15 publications

References 15 publications

Deltamethrin interacts with Culex quinquefasciatus odorant-binding protein: a novel potential resistance mechanism

Deltamethrin interacts with Culex quinquefasciatus odorant-binding protein: a novel potential resistance mechanism

Screening of olfactory genes related to blood-feeding behaviors in Culex pipiens quinquefasciatus and Culex pipiens molestus by transcriptome analysis

Screening for Odorant Receptor Genes Expressed in Aedes Aegypti Involved in Blood-Feeding and Oviposition Behaviors

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