The European grapevine moth Lobesia botrana (Denis & Shiffermüller 1776) is an economically important pest of the vine-growing areas worldwide. Chemical insecticides have been used for its control; however, its resistance status is largely unknown in many regions. We monitored the susceptibility of several L. botrana populations from Greece and Turkey. In addition, based on RNAseq transcriptome analysis, we identified and phylogenetically classify the cytochrome P450 genes of L. botrana, as well as analysed target site sequences and looked for the presence of known resistance mutations. Resistance against chlorantraniliprole, alpha-cypermethrin, spinetoram, etofenprox, and acetamiprid was very low (below 2.5-fold in all cases, compared to a reference strain from Greece) in all populations from Greece that were included in the study. However, resistance against indoxacarb (4–30-fold), spinosad (5–59-fold), and deltamethrin (18–30 fold) was detected in the L. botrana populations from Turkey, compared to a reference population from Turkey. De novo transcriptome assembly and manual annotation, and subsequent PCR-based analysis of insecticide target sequences (i.e. voltage-gated sodium channel – VGSC: target of pyrethroids and oxadiazines; nicotinic acetylcholine receptor subunit a6 – nAChR_α6: target of spinosad; ryanodine receptor – RyR: target of diamides; glutamate-gated chloride channel – GluCl: target of avermectins and; acetylcholinesterase – AChE: target of organophosphates) showed the absence of known resistance mutations in all specimens from both countries. Finally, the L. botrana CYPome (116 genes) was manually analysed and phylogenetically characterised, to provide resources for future studies that will aim the analysis of metabolic resistance.

A Palmer amaranth (Amaranthus palmeri S. Watson) population (KCTR: KS Conservation Tillage Resistant) collected from a conservation tillage field was confirmed with resistance to herbicides targeting at least six sites of action, including 2,4-D. The objectives of this research were using KCTR A. palmeri to investigate (1) the level of 2,4-D resistance, (2) 2,4-D absorption and translocation profiles, (3) the rate of 2,4-D metabolism compared with 2,4-D–tolerant wheat (Triticum aestivum L.), and (4) the possible role of cytochrome P450s (P450s) in mediating resistance. Dose–response experiments were conducted to assess the level of 2,4-D resistance in KCTR compared with susceptible plants, KSS (KS 2,4-D susceptible) and MSS (MS 2,4-D susceptible). KSS, MSS, and KCTR plants were treated with [14C]2,4-D to determine absorption, translocation, and metabolic patterns. Additionally, whole-plant dose–response assays were conducted by treating KCTR and KSS plants with P450 inhibitors (malathion, piperonyl butoxide [PBO]) before 2,4-D application. Dose–response experiments indicated a 6- to 11-fold 2,4-D resistance in KCTR compared with susceptible plants. No difference was found in percent [14C]2,4-D absorption among the populations. However, 10% less and 3 times slower translocation of [14C]2,4-D was found in KCTR compared with susceptible plants. Importantly, [14C]2,4-D was metabolized faster in KCTR than susceptible plants. At 24, 48, and 72 h after treatment (HAT), KCTR metabolized ∼20% to 30% more [14C]2,4-D than susceptible plants. KCTR plants and wheat generated metabolites with similar polarity. Nonetheless, at 24 HAT, ∼70% of [14C]2,4-D was metabolized in wheat, compared with only 30% in KCTR A. palmeri. Application of malathion before 2,4-D increased the sensitivity to 2,4-D in KCTR, suggesting involvement of P450s in mediating 2,4-D metabolism. However, no such impact of PBO was documented. Overall, this study confirms that enhanced metabolism is the primary mechanism of 2,4-D resistance in KCTR.

Quercetin is ubiquitous in terrestrial plants. The cotton bollworm Helicoverpa armigera as a highly polyphagous insect has caused severe crop losses. Until now, interactions between this pest and quercetin are poorly understood at the biochemical and molecular levels. In this study, we investigated the in vivo effects of quercetin on performance of cotton bollworm and on cytochrome P450 (P450) expression. Deleterious effects of quercetin on the performance of the cotton bollworm, including growth, survival, pupation and adult emergence were observed after oral administration of 3 and 10 mg g−1 quercetin to larvae since the third instar, whereas no significant toxic effect was found at 0.1 mg g−1 quercetin treatment. Piperonyl butoxide treatment enhanced the toxicity of quercetin. In vitro metabolism studies showed that quercetin was rapidly transformed by gut enzymes of fifth instar larvae of the cotton bollworm. qRT–PCR results revealed that the effect of quercetin on P450 expression was tissue- and dose-specific. Quercetin regulated P450 expression in a mild manner, and it could serve as P450 inducer (CYP337B1, CYP6B6) or repressor (CYP337B1, CYP6B7, CYP6B27, CYP9A14, CYP6AE11, and CYP4M7). These findings are important for advancing our understanding of the biochemical and molecular response of insects to plant toxins and have implications for a smart pest control.

Despite the continuous use of synthetic insecticides during the last two decades, the tea mosquito bug Helopeltis theivora Waterhouse still exists as the most destructive pest of tea in North East India. The susceptibility levels of the female sucking bug collected from conventional (synthetic insecticide treated: Terai and Dooars plain regions) and organic (synthetic insecticide untreated: low-altitude Darjeeling region) tea plantations of the northern part of West Bengal to two synthetic insecticides, quinalphos and cypermethrin, and the activity of three principal detoxifying enzymes were assayed. Compared with the susceptible Darjeeling population, the Terai and Dooars populations showed a resistance factor at the lethal concentrations for 50% level ranging from 547- to 2680.87-fold and from 3810- to 7480-fold for quinalphos and cypermethrin, respectively. General esterases (GEs), glutathione S-transferases (GSTs) and cytochrome P450-mediated mono-oxygenases (CYPs) also showed an increased activity in the Terai and Dooars populations compared with those from Darjeeling. Defence enzyme activity was enhanced by 15.4- and 17.6-fold for GEs, 1.8- and 1.9-fold for GSTs and 2.1- and 2.4-fold for CYPs in the synthetic insecticide-treated H. theivora populations when compared with the untreated Darjeeling populations. Electrophoretic analysis for GEs showed a higher level of expression for esterase I–VI isozymes in the Terai and Dooars populations when compared with that in the Darjeeling populations. This study reveals a reduced efficacy of quinalphos and cypermethrin against field populations of H. theivora, possibly due to enhanced activities of GEs, GSTs and CYPs. The findings may be used in developing integrated resistance management strategies that can help in the effective control of this major tea pest.