Impact of Pesticide Resistance on Toxicity and Tolerance of Hostplant Phytochemicals in Amyelois Transitella (Lepidoptera: Pyralidae).

For some polyphagous insects, adaptation to phytochemically novel plants can enhance resistance to certain pesticides, but whether pesticide resistance expands tolerance to phytochemicals has not been examined. Amyelois transitella Walker (navel orangeworm) is an important polyphagous pest of nut and fruit tree crops in California. Bifenthrin resistance, partially attributable to enhanced cytochrome P450 (P450)-mediated detoxification, has been reported in an almond-infesting population exposed to intense pesticide selection. We compared the toxicity of bifenthrin and three phytochemicals-chlorogenic acid, and the furanocoumarins xanthotoxin and bergapten-to three strains of A. transitella: pyrethroid-resistant R347 (maintained in the laboratory for ∼10 generations), fig-derived FIG (in the laboratory for ∼25 generations), and CPQ-a laboratory strain derived from almonds ∼40 years ago). Whereas both Ficus carica (fig) and Prunus dulcis (almond) contain chlorogenic acid, furanocoumarins occur only in figs. Both R347 and FIG exhibited 2-fold greater resistance to the three phytochemicals compared with CPQ; surprisingly, bifenthrin resistance was highest in FIG. Piperonyl butoxide, a P450 synergist, increased toxicity of all three phytochemicals only in CPQ, implicating alternate tolerance mechanisms in R347 and FIG. To test the ability of the strains to utilize novel hostplants directly, we compared survival on diets containing seeds of Wisteria sinensis and Prosopis pallida, two non-host Fabaceae species; survival of FIG was highest and survival of R347 was lowest. Our results suggest that, while P450-mediated pesticide resistance enhances tolerance of certain phytochemicals in this species, it is only one of multiple biochemical adaptations associated with acquiring novel hostplants.

Characterization of Field-Derived Drosophila suzukii (Diptera: Drosophilidae) Resistance to Pyrethroids in California Berry Production.

The spotted-wing drosophila, Drosophila suzukii (Matsumura), is a global economic pest of berry crops and stone fruit. Since management of this pest primarily relies on calendar insecticide applications, and field-derived resistance to spinosad has already been documented in California caneberry production, there is significant concern for development of resistance to other insecticides. In this study, susceptibility of D. suzukii populations collected from caneberry and strawberry fields to two pyrethroids, zeta-cypermethrin, and bifenthrin, was assessed in 2019 and 2020. Resistance to both pyrethroids were observed in flies from all sampling sites. For flies collected from caneberries in 2019, the LC50 values ranged from 4.5 to 5.2 mg liter-1 with RR50s ranging from 7.5- to 8.7-fold. Our 2020 assays showed that susceptibility of flies to the discriminating dose of zeta-cypermethrin decreased significantly as the season progressed. For flies collected from strawberries in 2020, the LC50s ranged from 19.0 to 36.1 mg liter-1 and from 30.3 to 90.7 mg liter-1 for zeta-cypermethrin and bifenthrin, respectively. The RR50 values varied from 19.0- to 36.1-fold for zeta-cypermethrin and from 15.9- to 47.7-fold for bifenthrin. This study is the first report of field-derived pyrethroid resistance in D. suzukii from two major California berry production areas. Adoption of informed insecticide resistance management practices would be essential to prolong the efficacy of products available to control D. suzukii. Future molecular work is needed to unravel the underlying genetic mechanisms conferring the observed pyrethroid resistance and to develop robust diagnostics that can inform integrated pest management of this pest.

Spatio-temporal Variation of Spinosad Susceptibility in Drosophila suzukii (Diptera: Drosophilidae), a Three-year Study in California's Monterey Bay Region.

Spinosyn insecticides are widely used in conventional berry production, and spinosad is regarded as the most effective insecticide for managing Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), spotted-wing drosophila, in organic berry crops. Following the 2017 identification of spinosad resistance in caneberry fields in the Watsonville area, Santa Cruz Co., California, we conducted a study to examine the seasonal and annual susceptibility of D. suzukii over a three-year period. Adult flies were collected from two conventional and two organic caneberry fields in the Monterey Bay region, California, at 'early', 'middle', and 'late' time points during the 2018-2020 growing seasons, and their susceptibility to spinosad was assessed. Results demonstrated that spinosad susceptibility in the D. suzukii field populations generally decreased during the fruit production season (from June through November), and over consecutive seasons. LC50 values of adults from the conventional sites were determined to be as high as 228.7 mg l-1 in 2018, 665.6 mg l-1 in 2019, and 2700.8 mg l-1 in 2020. For the organically managed fields, LC50s of adults were as great as 300.0 mg l-1 in 2018, 1291.5 mg l-1 in 2019, and 2547.1 mg l-1 in 2020. Resistance ratios based on the LC50 values were as high as 10.7-, 13.2-, and 16.9-fold in 2018, 2019, and 2020, respectively. These results should serve as a caution for growers in other production areas, facilitate informed choice of insecticides used in D. suzukii management, and emphasize the need to develop effective insecticide resistance management strategies for this insect.

Differential regulation of cytochrome P450 genes associated with biosynthesis and detoxification in bifenthrin-resistant populations of navel orangewom (Amyelois transitella).

Pyrethroid resistance was first reported in 2013 for the navel orangeworm, Amyelois transitella, but the genetic underpinnings of pyrethroid resistance are unknown. We investigated the role of cytochrome P450 monooxygenases (P450s) belonging to the CYP3 and CYP4 clans using colonies derived from individuals collected in 2016 from almond orchards in two counties. One colony (ALM) originated from an almond orchard in Madera County with no reported pyrethroid resistance and the second colony (R347) originated from the same Kern County orchard where pyrethroid resistance was first reported. We used high-throughput quantitative real-time PCR (qRT-PCR) analyses of 65 P450s in the CYP3 and CYP4 clans of A. transitella to identify P450s induced by bifenthrin and associated with pyrethroid resistance. Nine P450s were constitutively overexpressed in R347 compared to ALM, including CYP6AE54 (11.7-fold), belonging to a subfamily associated with metabolic pesticide detoxification in Lepidoptera and CYP4G89 (33-fold) belonging to a subfamily associated with cuticular hydrocarbon (CHC) synthesis and resistance via reduced pesticide penetrance. Cuticular hydrocarbons analysis revealed that R347 produced twice as many total CHCs in the egg and adult stages as ALM. Topical toxicity bioassays for R347 determined that egg mortality was reduced at low bifenthrin concentrations and larval mortality was reduced at high concentrations of bifenthrin compared to ALM. Our discovery of both changes in metabolism and production of CHCs for R347 have implications for the possible decreased efficacy of other classes of insecticide used to control this insect. The threat of widespread pyrethroid resistance combined with the potential for cross-resistance to develop through the mechanism of reduced penetrance warrants developing management strategies that facilitate insecticide passage across the cuticle.