Co-Expression of a Homologous Cytochrome P450 Reductase Is Required for In Vivo Validation of the Tetranychus urticae CYP392A16-Based Abamectin Resistance in Drosophila

Abstract

Overexpression of the cytochrome P450 monooxygenase CYP392A16 has been previously associated with abamectin resistance using transcriptional analysis in the two-spotted spider mite Tetranychus urticae, an important pest species worldwide; however, this association has not been functionally validated in vivo despite the demonstrated ability of CYP392A16 to metabolize abamectin in vitro. We expressed CYP392A16 in vivo via a Gal4 transcription activator protein/Upstream Activating Sequence (GAL4/UAS) system in Drosophila melanogaster flies, driving expression with detoxification tissue-specific drivers. We demonstrated that CYP392A16 expression confers statistically significant abamectin resistance in toxicity bioassays in Drosophila only when its homologous redox partner, cytochrome P450 reductase (TuCPR), is co-expressed in transgenic flies. Our study shows that the Drosophila model can be further improved, to facilitate the functional analysis of insecticide resistance mechanisms acting alone or in combination.

Frequencies and mechanisms of pesticide resistance in Tetranychus urticae field populations in China

“…Consistent with that possibility, the high abamectin resistance of the XY‐SX population in this study was predicted to be associated with increased activities of detoxification enzymes. In addition, overexpression of P450, GST, CarEs and UGTs have been suggested to contribute to abamectin resistance in T. urticae (Ahn et al ., 2014; Choi et al ., 2020; Dermauw et al ., 2020; Riga et al ., 2020; Wang et al ., 2020). The P450 genes CYP389 and CYP392 were previously associated with the development of abamectin resistance in spider mites (Riga et al ., 2014; Xu et al ., 2021; Xu et al ., 2021), besides the GST ( TuGSTd10 , TuGSTd14 , and TuGSTm09) and UGT201D3 (Dermauw et al ., 2013; Riga et al ., 2015; Wang et al ., 2020).…”

Functional characterization of cytochrome P450s associated with pyrethroid resistance in the olive fruit flyBactrocera oleae

“…At the same time the in vitro assays suggest that these enzymes are not involved in the detoxification of other classes of insecticides, such as the neonicotinoids thiacloprid and acetamiprid, also used for B. oleae control, as all of the enzymes were found inactive against these molecules. This indicates that these enzymes may have a specialized role in pyrethroid resistance, similar to CYP6A51 from Ceratitis capitata , which was found to metabolize only deltamethrin and lambda-cyhalothrin, but not spinosad and malathion (Tsakireli et al, 2019) (Tsakireli et al, 2019), as well as the CYP392A11 and CYP392E10 from Tetranychus urticae which metabolize only METIs and spirodiclofen, respectively (Demaeght et al, 2013; Riga et al, 2020, 2015, 2014) (Demaeght et al, 2013; Riga et al, 2020, 2015, 2014). This is in contrast to other P450s, which have a broader catalytic role, such as the CYP6G1 from Drosophila melanogaster , which is capable of metabolizing both the organochlorine DDT and the neonicotinoid imidacloprid (Joussen et al, 2008) (Joussen et al, 2008), or the CYP6CM1 from Bemisia tabaci , which is capable of metabolizing neonicotinoids, pyridine azomethin and pyriproxyfen (Karunker et al, 2009; Nauen et al, 2013; Roditakis et al, 2011) (Karunker et al, 2009; Nauen et al, 2013; Roditakis et al, 2011).…”

Functionally characterized arthropod pest and pollinator cytochrome P450s associated with xenobiotic metabolism

“…T. urticae CYP392E10 has been validated for xenobiotic detoxification using in vitro systems (Demaeght et al, 2013). Additionally, T. urticae CYP392A11 is involved in insecticide metabolism, demonstrated by the use of in vitro systems, D. melanogaster heterologous expression (genome modification), and also RNAi (Riga et al, 2015;Xu et al, 2021), while CYP392A16 was expressed in vitro, to prove its catalytic activity against insecticides (Riga et al, 2014) and genome modification techniques were also explored (Riga et al, 2020);…”

“…In vivo functional validation of the role of P450s to xenobiotic tolerance and resistance has been facilitated by the RNA interference (RNAi) approach (Bai-Zhong et al, 2020;Ding et al, 2013;Gao et al, 2016;Kalsi and Palli, 2017;Mao et al, 2007;Pang et al, 2016; as well as Drosophila transformation tools (Daborn et al, 2012;Manjon et al, 2018;Riga et al, 2020;Samantsidis et al, 2020;Troczka et al, 2019;Tsakireli et al, 2019;Zhu et al, 2010;Zimmer et al, 2018)and the recent advances in genome editing technology in non-model organisms Zuo et al, 2021).…”

“…(M). aeneus CYP6BQ23 (Samantsidis et al, 2020), CYP321A16 and CYP332A1 from S. exigua (Bo et al, 2020), M. persicae CYP6CY3 (Bass et al, 2013), N. lugens CYP6ER1 (Pang et al, 2016;Zimmer et al, 2018), T. urticae CYP392A11 and CYP392A16 (Riga et al, 2020(Riga et al, , 2015, A. mellifera CYP9Q2 and CYP9Q3 (Manjon et al, 2018), B. terrestris CYP9Q4 and CYP9Q6 (Manjon et al, 2018;Troczka et al, 2019).…”