CRISPR‐mediated mutagenesis of the odorant receptor co‐receptor (<i>Orco</i>) gene disrupts olfaction‐mediated behaviors in <i>Bactrocera dorsalis</i>

Xu, Li; Jiang, Hong‐Bo; Tang, Kai‐Yue; Yan, Ying; Schetelig, Marc F.; Wang, Jinjun

doi:10.1111/1744-7917.12997

Cited by 18 publications

(39 citation statements)

References 43 publications

(62 reference statements)

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“…BdorOrco −/− mutants do not respond to 1-octen-3-ol in EAG experiments and show a significant change in oviposition behavior in our previous study 20 . Given that Orco is strictly required for olfactory cue perception in insects, oviposition behavior guided by 1-octen-3-ol must be mediated by ORs.…”

Section: Discussionmentioning

confidence: 53%

“…Each gRNA was synthesized using the GeneArt Precision gRNA Synthesis Kit (Invitrogen) and purified using the GeneArt gRNA Clean-up Kit (Invitrogen). Embryos were microinjected as previously described 20 . Purified gRNA and Cas9 protein from the GeneArt Platinum Cas9 Nuclease Kit (Invitrogen) were mixed and diluted to final concentrations of 600 and 500 ng/µL, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Similarly, HassOR31 was found to guide egg-laying behavior of Helicoverpa assulta in response to Z-3-hexenyl butyrate released by host plants 13 . Furthermore, we found that deleting the olfactory receptor co-expressed receptor ( BdorOrco ) gene using the CRISPR/Cas9 system abolished 1-octen-3-ol induced oviposition preference in female B. dorsalis 20 . The previous study has also shown directly that BdorOR13a binds to 1-octen-3-ol in vitro 16 , but no in vivo behavioral data has been presented thus far.…”

Section: Introductionmentioning

confidence: 99%

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Two odorant receptors regulate 1-octen-3-ol induced oviposition behavior in the oriental fruit fly

Jiang

et al. 2023

Commun Biol

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The oriental fruit fly Bactrocera dorsalis (Hendel) is a notorious pest of fruit crops. Gravid females locate suitable oviposition sites by detecting host plant volatiles. Here, we demonstrate that 1-octen-3-ol, a volatile from mango, guides the oviposition behavior of female flies. Two odorant receptors (BdorOR7a-6 and BdorOR13a) are identified as key receptors for 1-octen-3-ol perception by qPCR analysis, heterologous expression in Xenopus laevis oocytes and HEK 293 cells followed by in vitro binding assays, as well as CRISPR/Cas9 genome editing in B. dorsalis. Molecular docking and site-directed mutagenesis are used to determine major binding sites for 1-octen-3-ol. Our results demonstrate the potential of 1-octen-3-ol to attract gravid females and molecular mechanism of its perception in B. dorsalis. BdorOR7a-6 and BdorOR13a can therefore be used as molecular targets for the development of female attractants. Furthermore, our site-directed mutagenesis data will facilitate the chemical engineering of 1-octen-3-ol to generate more efficient attractants.

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Section: Discussionmentioning

confidence: 53%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Two odorant receptors regulate 1-octen-3-ol induced oviposition behavior in the oriental fruit fly

Jiang

et al. 2023

Commun Biol

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“…The four choice olfactometer assay was carried out to detect the olfactory preference of treated flies ( β -cypermethrin, 10 ng/fly) to 1-octen-3-ol (Sigma-Aldrich, St Louis, MO, USA), methyl eugenol (Acros, Morris Plains, NJ, USA) and ethyl acetate (Sigma, St Louis, MO, USA) followed the published method [ 27 ] with slight modification. The apparatus consisted of a central chamber connected to four 50 mL cylindrical glass bottles.…”

Section: Methodsmentioning

confidence: 99%

Sublethal Dose of β-Cypermethrin Impairs the Olfaction of Bactrocera dorsalis by Suppressing the Expression of Chemosensory Genes

Chen

Lei

et al. 2022

Insects

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The oriental fruit fly, Bactrocera dorsalis, is one of the most destructive fruit insect pests. β-cypermethrin has been widely used in the orchard to control this major insect. Based on the resistance monitoring in 2011, B. dorsalis developed significant resistance against β-cypermethrin in fields. This indicated that the B. dorsalis has been exposed to sublethal concentrations of β-cypermethrin in the field for a long time. Thus, it is urgent to understand the sublethal effects of β-cypermethrin on this fly to guide the rational use of an insecticide. According to the olfactory preference assays and electroantennogram (EAG) recording, the B. dorsalis after β-cypermethrin exposure (LD30 = 10 ng/fly) severely decreased the ability to perceive the tested odorants. Moreover, we then performed quantitative real-time PCR and found the chemosensory genes including odorant receptor co-receptor (BdorORco) and ionotropic receptor co-receptors (BdorIRcos) were obviously suppressed. Our results demonstrated that the sublethal dose of β-cypermethrin impairs the olfaction of the pest insects by suppressing the expression of chemosensory genes (BdorORco and BdorIRcos), which expanded our knowledge of the sublethal effects of the pesticide on insects.

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“…We expect this approach to become the gold standard for demonstrating the functional relevance of specific gene products in chemical sensation. Indeed, CRISPR-mediated mutants in Orco have already been reported for several insect pests [ 6 , 114 , 115 , 116 , 117 ].…”

Section: Crispr (Clustered Regularly Interspaced Short Palindromic Re...mentioning

confidence: 99%

Recent Insights into Insect Olfactory Receptors and Odorant-Binding Proteins

Smith

2022

Insects

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Human and insect olfaction share many general features, but insects differ from mammalian systems in important ways. Mammalian olfactory neurons share the same overlying fluid layer in the nose, and neuronal tuning entirely depends upon receptor specificity. In insects, the olfactory neurons are anatomically segregated into sensilla, and small clusters of olfactory neurons dendrites share extracellular fluid that can be independently regulated in different sensilla. Small extracellular proteins called odorant-binding proteins are differentially secreted into this sensillum lymph fluid where they have been shown to confer sensitivity to specific odorants, and they can also affect the kinetics of the olfactory neuron responses. Insect olfactory receptors are not G-protein-coupled receptors, such as vertebrate olfactory receptors, but are ligand-gated ion channels opened by direct interactions with odorant molecules. Recently, several examples of insect olfactory neurons expressing multiple receptors have been identified, indicating that the mechanisms for neuronal tuning may be broader in insects than mammals. Finally, recent advances in genome editing are finding applications in many species, including agricultural pests and human disease vectors.

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CRISPR‐mediated mutagenesis of the odorant receptor co‐receptor (Orco) gene disrupts olfaction‐mediated behaviors in Bactrocera dorsalis

Cited by 18 publications

References 43 publications

Two odorant receptors regulate 1-octen-3-ol induced oviposition behavior in the oriental fruit fly

Two odorant receptors regulate 1-octen-3-ol induced oviposition behavior in the oriental fruit fly

Sublethal Dose of β-Cypermethrin Impairs the Olfaction of Bactrocera dorsalis by Suppressing the Expression of Chemosensory Genes

Recent Insights into Insect Olfactory Receptors and Odorant-Binding Proteins

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