The resistance status of Tetranychus urticae Koch and Lygus hesperus Knight sourced from California strawberry fields was evaluated in laboratory bioassay experiments. Adult female T. urticae sourced from a miticide naïve population were exposed to a range of miticide active ingredient concentrations mixed with acetone in contact bioassay experiments. LC50 concentrations for the active ingredients fenpyroximate, abamectin, bifenazate, and spiromesifen as well as LC90 concentrations for the active ingredients fenpyroximate, abamectin, and bifenazate were derived from this susceptible population. A total of ten field populations of T. urticae, five from the Santa Maria growing region and five from the Ventura/Oxnard growing region, were exposed to these concentrations and mortality response was compared with the susceptible population to determine the resistance status of T. urticae on a per-field basis. Resistance detection occurred less often in the LC50 concentration groups compared to their respective LC90 concentrations. Resistance to the LC50 concentration of spiromesifen could not be detected in any field population. Resistance to the LC90 concentrations of fenpyroximate and abamectin was detected in three field populations from Santa Maria and four populations from Ventura/Oxnard. Resistance to the LC90 concentration of bifenazate was detected in three populations from Ventura/Oxnard and in no populations from Santa Maria. Lygus hesperus were exposed to formulated products registered in California strawberries after being separated into three groups based on life stage. Instars one through three were considered small, instars four and five were considered large, and winged individuals were considered adults. The small and large groups were exposed to the maximum field labeled rate of Rimon® (a.i. = novaluron). Adults were exposed to the maximum field labeled rate of Actara®, Brigade®, and Sivanto®, the active ingredients of which are thiamethoxam, bifenthrin, and flupyradifurone respectively. A total of 14 field populations were screened for resistance, six collected from the Santa Maria growing region in 2021, and eight collected in 2022, five of which came from the Santa Maria region, and three from the Salinas/Watsonville region. Resistance was detected by comparing the mortality response of an insecticide naïve population of L. hesperus and field collected populations in residual laboratory bioassay experiments. Resistance to Rimon® in the small group was effectively detected in all populations in 2021 and 2022. Resistance to Rimon® in the large group was detected in four populations in 2021 and five populations in 2022. Resistance to Actara® and Brigade® was effectively detected in all populations in 2021 and 2022. Resistance to Sivanto® was effectively detected in four populations in 2021 and two of three populations in 2022. A full submersion assay was utilized for the second half of Sivanto® trials in 2022 since mortality response of the susceptible population was low in the residual group. Five of five populations tested with the modified assay were considered effectively resistant.

Cultivated tomato plants are extremely susceptible to the two-spotted spider mite Tetranychus urticae Koch. Selection for pest resistance is usually a crucial step required to achieve successful genetic resistance transfer from wild into cultivated tomato genotypes. S. habrochaites LA2329, a wild relative of tomato, is highly resistant to arthropods. Its resistance has been attributed to the presence of a high density of type IV and type VI trichomes and abundant production of 7-epi-zingiberene, a sesquiterpene hydrocarbon. The interspecific backcross hybrids used in this research were derived from the cross between the wild relative tomato, S. habrochaites LA2329, and the cultivated tomato, S. lycopersicum ‘Zaofen 2’ (ZH2). This population has been directly selected for type IV trichome density and zingiberene. The arthropod resistance status of the backcross hybrids was unknown when this research was initiated. Thus, the main objective of the research was to verify the transfer of arthropod resistance from S. habrochaites to cultivated tomato. The effects of glandular trichome densities and leaf zingiberene contents on spider mite behavior and biology were also explored. Also, the chemical composition of the trichome secretions in the wild tomato donor is segregating for presence and abundance of sesquiterpenoids related to zingiberene. The bioactivity of these sesquiterpenoids was explored in this research. To evaluate the relative bioactivities of zingiberene alcohol and 7-epizingiberene, extracted from glandular trichomes of Solanum habrochaites accession LA2329, as well as alpha-zingiberene obtained from ginger oil, these were purified by silica gel chromatography and bioassayed with two-spotted spider mites, Tetranychus urticae Koch (Acari: Tetranychidae) using a bean leaf disc bioassay. Zingiberene alcohol was most efficacious and alpha-zingiberene, was least efficacious, while the efficacy of 7-epizingiberene was intermediate. Thus, tomato breeders should consider introgression of the genes responsible for the oxidation of 7-epizingiberene into zingiberene alcohol to potentially improve the spider mite resistance of cultivated tomato. Also, it is possible that this compound may be exploited as eco-biopesticide approach for integrated pest management against a broad spectrum of herbivorous pests. To verify transfer of arthropod resistance, a bioassay utilizing whole leaves was employed. Nine hybrids (BC3F3 and BC3F4) were chosen for this bioassay, based on variation of type IV trichome density and zingiberene concentration among the hybrids. The experiment also included three susceptible and three resistant control plants. Mite responses on some of the hybrids were similar to those on the resistant wild donor parent, S. habrochaites, as indicated by number of leaflet surfaces infested by mites, degree of mite webbing and feeding damage. Egg density on four backcross hybrids was similar to that on the S. habrochaites resistant controls. Based these results, we concluded that resistance had been successfully transferred from the wild accessions to the hybrids by deployment of backcrossing and indirect selection. There was a significant negative correlation of almost all mite behavioral and biological responses with Type IV trichome density and zingiberene content. This bioassay illuminated behavioral variations of mites associated with presence or absence of leaf compounds and glandular trichome densities. Also, the results support the idea that introgression of type IV trichomes and zingiberene has led to effective spider mite resistance. In another bioassay-based experiment to verify transfer of resistance, seven interspecific backcross hybrids (BC3F2), the resistant parent LA2329, and two susceptible cultivated tomato lines, the recurrent parent ZH2 and ‘Small Roma’, were used in thumbtack bioassays. Mite movement was measured by imaging bioassayed leaves at 15, 20, 30, 45, and 60 min intervals. In addition to confirming transfer of spider mite resistance, other objectives included determination of the relative contributions of type IV and VI trichome densities and leaf compounds to mite behavior over time intervals. Our findings confirmed the transfer of mite repellency from the wild resistant parent to advanced backcross hybrids. Several backcross hybrids performed similarly to the wild donor parent, displaying shorter distances traveled on the leaves after 15 and 30 min. The type IV and type VI trichome densities as well as zingiberene contents had a significant positive correlation with the number of spider mites remaining on tack. There was a significant negative correlation of type IV density and zingiberene concentration with the total distance travelled by mites for both the abaxial and adaxial surfaces across most time intervals. Stepwise multiple regression analysis showed that the type IV trichome density was the most critical factor, and zingiberene content was a secondary factor across over most time intervals. T. urticae remained longer on the thumbtack heads and traveled shorter distances on the leaf surface of the wild donor parent LA2329 and the interspecific hybrids compared to S. lycopersicum leaves. These results indicated that introgression of genetic resistance, especially repellence, against spider mite from the wild relative into cultivated tomato varieties has been successfully achieved. In conclusion, trichome type IV and/or zingiberene content has been successfully transferred from the wild relative into interspecific tomato hybrids, and the hybrids show significant adverse impact on spider mite behavior and/or biology in whole leaf and thumbtack bioassays. Type IV trichome density is the most crucial factor in mite deterrence while zingiberene seemed to be a second key factor across most of time durations for both surfaces. Collectively, several backcross hybrids had similar leaf characteristics to the S. habrochaites LA2329, also may be a potential source of resistance to other insect pests.