Bioactivities of scent gland chemicals from Mictis fuscipes Hsiao (Hemiptera: Coreidae) on Solenopsis invicta Buren (Hymenoptera: Formicidae).

BACKGROUND: Given the chemical diversity within stink bugs scent glands, they can be convenient models for bioprospecting novel pest control products. Preliminary behaviour observations indicated that adult Mictis fuscipes stink bugs secrete liquid droplets when defending against Solenopsis invicta fire ants, killing them within minutes. Hence, this study aimed to analyse the chemical composition of the metathoracic scent gland secretions of M. fuscipes adults, as well as assess their biological activities against fire ants. RESULTS: Bioassaying fire ants against secretions of several local stink bugs confirmed that the defensive secretions of two Mictis species are significantly more lethal, where M. fuscipes was the most lethal. Volatiles chromatography analysis indicated the secretions of female and male M. fuscipes stink bugs contains 20 and 26 components, respectively, chiefly hexanoic acid and hexyl hexanoate. Five compounds were consistently present in the secretion of female adults: hexyl hexanoate, hexanoic acid, hexyl acetate, hexyl butyrate, and eugenol. These yielded a strong electrophysiological antennal (EAD) response from S. invicta workers, female alates and males, where hexyl acetate showed the strongest response. The combination of these five compounds proved strongly repellent to S. invicta. When tested singly, hexanoic acid, hexyl butyrate, hexyl hexanoate, and eugenol were repellent to S. invicta, but hexyl acetate seemed slightly attractive. Additionally, the same mixture of five components exhibited strong contact and fumigant toxicity towards S. invicta workers, eugenol being the strongest. CONCLUSION: Defensive chemicals of M. fuscipes exhibit robust biological activity against S. invicta and could inspire the development of biopesticides. © 2024 Society of Chemical Industry.

Cry51Aa Proteins Are Active against Apolygus lucorum and Show a Mechanism Similar to Pore Formation Model.

Reduced insecticide spray in crop fields due to the widespread adoption of Bacillus thuringiensis (Bt) crops has favored the population increases of mirid bugs. Cry51Aa proteins are new types of Bt proteins that belong to aerolysin-like ß pore-forming proteins with insecticidal activity against hemipteran and coleopteran pests. Here, we studied the activity of Bt Cry51Aa1 and Cry51Aa2 against Apolygus lucorum, an emerging pest in cotton, and their mechanism of action. Cry51Aa1 exhibited almost 5-fold higher toxicity than Cry51Aa2 with LC50 of 11.87 and 61.34 µg/mL, respectively. Protoxins could be activated both in vitro, by trypsin and midgut contents, and in vivo, by A. lucorum midgut. Both Cry51Aa protoxins were processed in two steps, producing pre-activated (∼30 kDa) and final activated (∼25-28 kDa) proteins. Cry51Aa proteins bound to a 25 kDa midgut protein, and Cry51Aa2 showed 2 times higher binding affinity than Cry51Aa1. Incubating Cry51Aa proteins with midgut homogenate resulted in toxin oligomers of 150-200 kDa. Our findings provide a theoretical basis for using Cry51Aa proteins to control A. lucorum and a better understanding of their mode of action.

Insecticide resistance by a host-symbiont reciprocal detoxification.

Insecticide resistance is one of the most serious problems in contemporary agriculture and public health. Although recent studies revealed that insect gut symbionts contribute to resistance, the symbiont-mediated detoxification process remains unclear. Here we report the in vivo detoxification process of an organophosphorus insecticide, fenitrothion, in the bean bug Riptortus pedestris. Using transcriptomics and reverse genetics, we reveal that gut symbiotic bacteria degrade this insecticide through a horizontally acquired insecticide-degrading enzyme into the non-insecticidal but bactericidal compound 3-methyl-4-nitrophenol, which is subsequently excreted by the host insect. This integrated "host-symbiont reciprocal detoxification relay" enables the simultaneous maintenance of symbiosis and efficient insecticide degradation. We also find that the symbiont-mediated detoxification process is analogous to the insect genome-encoded fenitrothion detoxification system present in other insects. Our findings highlight the capacity of symbiosis, combined with horizontal gene transfer in the environment, as a powerful strategy for an insect to instantly eliminate a toxic chemical compound, which could play a critical role in the human-pest arms race.

Extracts of Euphorbia nivulia Buch.-Ham. showed both phytotoxic and insecticidal capacities against Lemna minor L. and Oxycarenus hyalinipennis Costa.

Many phytochemicals can affect the growth and development of plants and insects which can be used as biological control agents. In this study, different concentrations of crude, hexane, chloroform, butanol, and aqueous extracts of Euphorbia nivulia Buch.-Ham., an endemic plant of the Cholistan desert in South Punjab of Pakistan, were analysed for their chemical constituents. Their various concentrations were also tested for their phytotoxic and insecticidal potential against duckweed, Lemna minor L., and the dusky cotton bug, Oxycarenus hyalinipennis Costa. various polyphenols, i.e., quercetin, gallic acid, caffeic acid, syringic acid, coumaric acid, ferulic acid, and cinnamic acid were detected in different concentrations with different solvents during the phytochemical screening of E. nivulia. In the phytotoxicity test, except for 100 µg/mL of the butanol extract gave 4.5% growth regulation, no phytotoxic lethality could be found at 10 and 100 µg/mL of all the extracts. The highest concentration, 1000 µg/mL, of the chloroform, crude, and butanol extracts showed 100, 63.1, and 27.1% of growth inhibition in duckweed, respectively. In the insecticidal bioassay, the highest O. hyalinipennis mortalities (87 and 75%) were recorded at 15% concentration of the chloroform and butanol extracts of E. nivulia. In contrast, the lower concentrations of the E. nivulia extracts caused the lower mortalities. Altogether, these findings revealed that E. nivulia chloroform extracts showed significant phytotoxicity while all the extracts showed insecticidal potential. This potential can be, further, refined to be developed for bio-control agents.

Sex Pheromone of the Alfalfa Plant Bug, Adelphocoris lineolatus: Pheromone Composition and Antagonistic Effect of 1-Hexanol (Hemiptera: Miridae).

The sex pheromone composition of alfalfa plant bugs, Adelphocoris lineolatus (Goeze), from Central Europe was investigated to test the hypothesis that insect species across a wide geographical area can vary in pheromone composition. Potential interactions between the pheromone and a known attractant, (E)-cinnamaldehyde, were also assessed. Coupled gas chromatography-electroantennography (GC-EAG) using male antennae and volatile extracts collected from females, previously shown to attract males in field experiments, revealed the presence of three physiologically active compounds. These were identified by coupled GC/mass spectrometry (GC/MS) and peak enhancement as hexyl butyrate, (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal. A ternary blend of these compounds in a 5.4:9.0:1.0 ratio attracted male A. lineolatus in field trials in Hungary. Omission of either (E)-2-hexenyl-butyrate or (E)-4-oxo-2-hexenal from the ternary blend or substitution of (E)-4-oxo-2-hexenal by (E)-2-hexenal resulted in loss of activity. These results indicate that this Central European population is similar in pheromone composition to that previously reported for an East Asian population. Interestingly, another EAG-active compound, 1-hexanol, was also present in female extract. When 1-hexanol was tested in combination with the ternary pheromone blend, male catches were reduced. This compound showed a dose-response effect with small doses showing a strong behavioral effect, suggesting that 1-hexanol may act as a sex pheromone antagonist in A. lineolatus. Furthermore, when (E)-cinnamaldehyde was field tested in combination with the sex pheromone, there was no increase in male catch, but the combination attracted both males and females. Prospects for practical application are discussed.

Learning can be detrimental for a parasitic wasp.

Animals have evolved the capacity to learn, and the conventional view is that learning allows individuals to improve foraging decisions. The parasitoid Telenomus podisi has been shown to parasitize eggs of the exotic stink bug Halyomorpha halys at the same rate as eggs of its coevolved host, Podisus maculiventris, but the parasitoid cannot complete its development in the exotic species. We hypothesized that T. podisi learns to exploit cues from this non-coevolved species, thereby increasing unsuccessful parasitism rates. We conducted bioassays to compare the responses of naïve vs. experienced parasitoids on chemical footprints left by one of the two host species. Both naïve and experienced females showed a higher response to footprints of P. maculiventris than of H. halys. Furthermore, parasitoids that gained an experience on H. halys significantly increased their residence time within the arena and the frequency of re-encounter with the area contaminated by chemical cues. Hence, our study describes detrimental learning where a parasitoid learns to associate chemical cues from an unsuitable host, potentially re-enforcing a reproductive cul-de-sac (evolutionary trap). Maladaptive learning in the T. podisi-H. halys association could have consequences for population dynamics of sympatric native and exotic host species.

Selectivity of mycoinsecticides and a pyrethroid to the egg parasitoid Cleruchoides noackae (Hymenoptera: Mymaridae).

Plants of the genus Eucalyptus, cultivated in many countries, have great importance for the world economy. In Brazil, this culture occupies a total of 5.7 million hectares, but native and exotic insect pests can reduce its productivity. Thaumastocoris peregrinus Carpintero & Dellapé (Hemiptera: Thaumastocoridae), an exotic Australian pest, damages Eucalyptus plants. Biological control using the egg parasitoid Cleruchoides noackae Lin & Huber (Hymenoptera: Mymaridae), Heteroptera predators and entomopathogenic fungi, such as Beauveria bassiana and Metarhizium anisopliae, have potential for managing T. peregrinus. Chemical insecticides, including bifenthrin and acetamiprid + bifenthrin, also control this insect. The compatibility of chemical and biological control methods favors integrated pest management. The objective of this study was to evaluate the selectivity of commercial products based on B. bassiana, M. anisopliae and the chemical bifenthrin on the parasitoid C. noackae and its parasitism on T. peregrinus eggs. The selectivity test followed the standards recommended by the International Organization for Biological Control (IOBC). Beauveria bassiana has selectivity to parasitism as well as viability, but was slightly harmful to C. noackae adults; M. anisopliae was innocuous to adults and to the viability of the offspring of this parasitoid, but it reduced the parasitism rate; and bifenthrin did not show selectivity to this parasitoid.

Thiamethoxam Differentially Impacts the Survival of the Generalist Predators, Orius insidiosus (Hemiptera: Anthocoridae) and Hippodamia convergens (Coleoptera: Coccinellidae), When Exposed via the Food Chain.

Insect predators are seldom considered during toxicological trophic assessments for insecticide product development. As a result, the ecological impact of novel insecticides on predators is not well understood, especially via the food chain, i.e., when their prey is exposed to insecticides. Neonicotinoids are systemic insecticides widely used in agriculture to control herbivorous insects, but their effects on predatory insects via the food chain have not been well characterized. In this study, we documented the time-course effects of the neonicotinoid thiamethoxam on the survival of two predators, the insidiosus flower bug Orius insidiosus (Say) and the convergent lady beetle Hippodamia convergens Guérin-Méneville, when preying upon the aphids Aphis glycines Matsumura (Hemiptera: Aphididae), Aphis gossypii Glover (Hemiptera: Aphididae), and Myzus persicae (Sulzer) (Hemiptera: Aphididae). Aphids were exposed to thiamethoxam-treated or untreated plants every week over the course of 5 wk. After transferring aphids to Petri dishes, predators were allowed to feed on aphids. We found that the survival of the insidiosus flower bug, but not the convergent lady beetle, was reduced after consuming aphids reared on thiamethoxam-treated plants compared to untreated plants. Survival reduction of the insidiosus flower bug was observed only during the first weeks after thiamethoxam application; no reduction occurred 28 d after treatment or beyond. These results demonstrate that a systemic application of thiamethoxam could be compatible with convergent lady beetles and insidiosus flower bugs, if the time of predator release does not coincide with thiamethoxam activity. These findings are critical for the development of future pest control programs that integrate biological and chemical control.

Respiration, predatory behavior and prey consumption by Podisus nigrispinus (Heteroptera: Pentatomidae) nymphs exposed to some insecticides.

Podisus nigrispinus Dallas (Heteroptera: Pentatomidae) preys on insect pests in eucalyptus plantations where it can be exposed to insecticides used in pest control. The effect of insecticides on non-target natural enemies requires further study. The objective of the present study was to evaluate the side-effects of Bacillus thuringiensis (Bt), permethrin, tebufenozide and thiamethoxam on third instar nymphs of the predator P. nigrispinus in the laboratory. The toxicity of insecticides for this insect was determined by estimating their lethal concentrations. Podisus nigrispinus behavior after exposure to insecticides was analyzed using a video tracking system and the respiratory rate with a respirometer. Prey/nymph consumption was assessed after 24 h of starvation. The preference of P. nigrispinus nymphs, for prey treated or not with the insecticides, was evaluated in free choice tests. The insecticides Bt [LC50 = 1.10(0.83-1.46) mg mL-1], permethrin [LC50 = 0.25(0.17-0.34) mg mL-1], tebufenozide [LC50 = 5.71(4.17-7.57) mg mL-1] and thiamethoxam [LC50 = 0.04(0.02-0.06) mg mL-1] are toxic to P. nigrispinus nymphs. Bt and the insecticides tebufenozide, permethrin and thiamethoxam reduced the respiratory rate of P. nigrispinus. The insecticides permethrin, tebufenozide and thiamethoxam affect the locomotion of this insect's nymphs. Prey treated with Bt, permethrin and thiamethoxam are less preferred by P. nigrispinus. The survival of the nymphs of this predator was 93.3%, 66.7%, 56.6%, 0% and 0% in the control, tebufenozide, Bt, permethrin and thiamethoxam treatments, respectively. In addition, the reduction of prey consumption, treated with neurotoxic insecticides, reduces the predatory potential of this natural enemy. Bt and tefubenozide present low toxicity for P. nigrispinus, but the neurotoxic products have low compatibility with this natural enemy and, therefore, are not recommended, with this predator in the management of forest insect pests.

A Laboratory Diet-Overlay Bioassay to Monitor Resistance in Lygus lineolaris (Hemiptera: Miridae) to Insecticides Commonly Used in the Mississippi Delta.

A laboratory, diet-overlay pesticide bioassay was developed using a susceptible population of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), to study its susceptibility to neonicotinoid, sulfoxamine, organophosphate, and pyrethroid insecticides (thiamethoxam, sulfoxaflor, acephate, and permethrin, respectively). The diet-overlay bioassay was compared to the traditional glass-vial surface residue bioassay. We measured LC50 values by feeding tarnished plant bug adults known doses of insecticides dispensed on top of diet in a 10% solution of honey water for thiamethoxam and 10% acetone in water solutions for permethrin, acephate, and sulfoxaflor. Both the diet-overlay and glass-vial bioassays used dose-response (mortality) regression lines to calculate LC50 values for each insecticide at 6-, 24-, 48-, and 72-h post-exposure. Data variability from the glass-vial bioassay was higher for permethrin, sulfoxaflor, and thiamethoxam than the diet-overlay bioassay, for all evaluation times. In contrast, there was lower variability among replicates to acephate in the glass-vial assay compared to the diet-overlay assay. Control mortalities observed on diet-overlay bioassay were lower (0-5%) than those observed on the glass-vial bioassay (4-27%). The use of green beans, floral-foam, rolling glass vials, and insect handling made the existing standard method tedious to manipulate and difficult to handle large numbers of individuals. The nonautoclaved solid diet provides an opportunity to significantly reduce cost and variability associated with procedures of other bioassay methods. In general, the baseline data provide a basis for future comparison to determine changes in resistance over time.

Sublethal and transgenerational effects of sulfoxaflor on the demography and feeding behaviour of the mirid bug Apolygus lucorum.

Sulfoxaflor, the first commercially available sulfoximine insecticide, has been used for the control of sap-feeding insect pests such as plant bugs and aphids on a variety of crops. However, its sublethal effects on the mirid bug Apolygus lucorum, one of the key insect pests of Bt cotton and fruit trees in China, have not been fully examined. Here, we evaluated the demography and feeding behaviour of A. lucorum exposed to sulfoxaflor. The leaf-dipping bioassay showed that the LC10 and LC30 of sulfoxaflor against 3rd-instar nymphs of this insect were 1.23 and 8.37 mg L-1, respectively. The LC10 significantly extended the nymphal duration and decreased the oviposition period by 5.29 days and female fecundity by 56.99% in the parent generation (F0). The longer duration of egg, 5th-instar nymphs, preadult, and male adult longevity were observed in the F1 generation (F1) at LC10. At the LC30, the duration of egg and 1st-instar nymph, female adult longevity, and oviposition period of the F1 were significantly shorter, while the nymphal duration in the F0 and duration of 5th-instar nymphs, preadult survival rate, and male adult longevity in the F1 significantly increased. The net reproductive rate (R0), intrinsic rate of increase (r), and finite rate of increase (λ) in the F1 were not significantly affected by these two concentrations, whereas the mean generation time (T) was lower at the LC30. Additionally, the probe counts and cells mixture feeding time were markedly lengthened by the LC10 and LC30, respectively, when A. lucorum nymphs exposed to sulfoxaflor fed on Bt cotton plants without insecticides. These results clearly indicate that sulfoxaflor causes sublethal effects on A. lucorum and the transgenerational effects depend on the tested concentrations.

Sublethal and transgenerational effects of acetamiprid and imidacloprid on the predatory bug Orius sauteri (Poppius) (Hemiptera: Anthocoridae).

The predatory bug Orius sauteri (Poppius) is currently one of the most important beneficial arthropods in Northeast Asia and used as a biological control agent of several small pest arthropods including Frankliniella occidentalis (Pergande). Two neonicotinoid chemical insecticides, acetamiprid and imidacloprid, mainly used in China as chemical control on F. occidentalis, although applied at sublethal concentrations in the field or greenhouse to protect beneficial arthropods, still may affect the predator O. sauteri. The objective of present work is to assess the long-term effects of 24-h exposure time to these two insecticides on the life-cycle of O. sauteri at application rates similar to the laboratory 24-h LC10, LC20 and LC30 of O. sauteri. Results showed that acetamiprid and imidacloprid at all tested concentrations significantly decreased the fecundity of O. sauteri females, while the effect of acetamiprid was higher than that of imidacloprid. Moreover, the oviposition period and longevity of O. sauteri to both insecticides shortened. The sublethal effects on the first progeny (F1 generation) were also found to increase nymphal mortality, shorten adult longevity and reduce fecundity. However, all treatments of acetamiprid and imidacloprid at the concentration of LC10, LC20 and LC30 caused no significant effect on the developmental time of different nymphal stages and sex ratio of the F1 generation. This paper is the first one that assesses the compatibility between neonicotinoid insecticides and O. sauteri, and shows that the application of acetamiprid and imidacloprid likely interferes with the population dynamic of O. sauteri.

Identification of Brassicadiene, a Diterpene Hydrocarbon Attractive to the Invasive Stink Bug Bagrada hilaris, from Volatiles of Cauliflower Seedlings, Brassica oleracea var. botrytis.

Brassicadiene, a novel tricyclic diterpene hydrocarbon, was identified by a combination of mass spectrometry, microchemical tests, and analysis of NMR spectra. The compound constitutes >90% of the volatile organic compounds produced by cauliflower seedlings, Brassica oleracea var. botrytis. The invasive stink bug Bagrada hilaris is strongly attracted to brassicadiene, providing a mechanism for this herbivore, which specializes on cruciferous plants, to locate its hosts in a nutrient-rich and vulnerable stage.

TRPA1 modulates noxious odor responses in Lygus hesperus.

Lygus hesperus isa key pest of many economically important crops across western North America. Central to many aspects of the lives of these insects is chemical signalling, with identified roles in host plant selection, aggregation and passive mate guarding. The development of novel monitoring and control approaches for this insect will rely on a sound understanding of how these cues are perceived and processed, and their impact on behavior. Towards this end, we investigated allyl isothiocyanate, cinnamaldehyde and citronellal, compounds that are noxious repellents to other insects. We found that L. hesperus avoided areas containing the three compounds and that exposure induced increases in movement velocity and duration in both nymphs and adults. This suggests these compounds may work as repellents. To better understand the underlying physiology of this response, RNA interference by dsRNA injection was used to inhibit the expression of two chemosensory-associated proteins, the odorant receptor co-receptor (Orco) and the transient receptor potential A (TRPA1) channel. While knockdown of Orco did not change the reaction of adult females to citronellal, TRPA1 silencing effectively eliminated the induced increase to movement, suggesting a chemoperceptory role in citronellal detection.

Analysis of Differentially Expressed Transcripts in Apolygus lucorum (Meyer-Dür) Exposed to Different Temperature Coefficient Insecticides.

The existence of a temperature effect of insecticides frustrated the control of the green plant bug Apolygus lucorum (Meyer-Dür). Previous studies mostly focused on the application of insecticides, but the underlying mechanism remains incompletely understood. Here, we report a transcriptome profiling of A. lucorum treated by three kinds of temperature coefficient insecticides (TCIs) (positive TCI: imidacloprid, negative TCI: b-cypermethrin and non-effect TCI: phoxim) at 15 °C, 25 °C and 35 °C by using next- and third-generation RNA-Seq methods. A total of 34,739 transcripts were annotated from 277.74 Gb of clean data. There were more up-regulated transcripts than down-regulated transcripts in all three kinds of TCI treatments. Further Venn diagrams indicate the regulatory transcripts and regulatory modes were different at the three temperatures. The responses to imidacloprid involved more detox and stress response transcripts such as cytochrome P450 (CYP450), carboxylesterase (CarE) and catalase (CAT) at 35 °C, which was the case for beta-cypermethrin at 15 °C. UDP-glucuronyltransferase (UGT) and heat shock protein (HSP) transcripts were heavily involved, and thus deserve particular note in the temperature effect of insecticides. This high-confidence transcriptome atlas provides improved gene information for further study on the insecticide temperature effect related physiological and biochemical processes of A. lucorum.

Sublethal and transgenerational effects of dinotefuran on biological parameters and behavioural traits of the mirid bug Apolygus lucorum.

The mirid bug, Apolygus lucorum, has become a major pest of many crops and fruit trees since the widespread adoption of Bt cotton in northern China. Neonicotinoid insecticides, such as dinotefuran, applied to control this pest may show sublethal effects, but evidence for such effects is lacking. Here, we investigated the sublethal and transgenerational effects of dinotefuran on biological parameters and feeding behavioural traits of A. lucorum using the age-stage, two-sex life table and electrical penetration graphs (EPGs), respectively. The LC10 and LC30 of dinotefuran against 3rd-instar nymphs of A. lucorum were 14.72 and 62.95 mg L-1, respectively. These two concentrations significantly extended the development duration from 3rd-instar nymph to adult in parent generation (F0). LC30 also increased the oviposition period and male adult longevity and reduced nymphal survival rate in the F0. For offspring generation (F1), the egg duration, preadult duration, and total preoviposition period were significantly lower at LC10 than in the control, and the egg duration, duration of 4th-instar nymphs, preadult duration, oviposition period, and fecundity were also decreased at LC30. However, the four demographic parameters of F1 generation, namely, net reproductive rate (R0), intrinsic rate of increase (r), finite rate of increase (λ), and mean generation time (T), were not affected by dinotefuran. The significant differences in the number of probes and duration of each of four feeding waveforms failed to be detected when A. lucorum nymphs treated by dinotefuran feed on Bt cotton plants without insecticide exposure. Overall, the dinotefuran concentrations tested here have sublethal, but no transgenerational impacts on A. lucorum.

Histopathological and cytotoxic changes induced by spinosad on midgut cells of the non-target predator Podisus nigrispinus Dallas (Heteroptera: Pentatomidae).

Broad-spectrum insecticides used in pest control are a risk for non-target insects. Their compatibility to the insecticide spinosad, used in agriculture and forestry as a biological control tool, needs to be evaluated. Podisus nigrispinus Dallas (Heteroptera: Pentatomidae) is a predatory bug used in the pest management of agricultural and forest systems where spinosad is also frequently applied. The aim of this study was to evaluate the toxicity, histopathology and cytotoxicity in midgut cells of P. nigrispinus exposed to spinosad. The toxicity test was performed to determine the lethal concentrations of spinosad after exposure by ingestion. The histopathology and cytotoxicity caused by spinosad were analyzed in the three midgut regions (anterior, middle and posterior) of P. nigrispinus during different exposure periods. Spinosad, at low concentrations, was toxic to P. nigrispinus [LC50 = 3.15 (3.02-3.26) µg.L-1]. Cell degeneration features such as cytoplasm vacuolization, chromatin condensation and release of cell fragments to the midgut lumen were observed in this organ. Cell death via apoptosis was found in the three midgut regions of this predator after exposure to the insecticide. Spinosad is toxic to P. nigrispinus, and causes histological and cytological damage followed by cell death in the midgut, suggesting a dangerous effect on a beneficial non-target insect.

Genetic analysis and molecular detection of resistance to chlorpyrifos mediated by the A216S substitution in acetylcholinesterase-1 in the plant bug Apolygus lucorum.

The green plant bug Apolygus lucorum is a major pest of Bacillus thuringiensis cotton in China. Previously, we reported that chlorpyrifos resistance in a laboratory-selected strain of A. lucorum (BZ-R) is associated with the homozygosis of an allele in the ace-1 gene encoding an alanine to serine substitution at position 216 of acetylcholinesterase-1. Here we describe the results of crosses between the resistant BZ-R strain (41-fold to chlorpyrifos) and the unselected susceptible BZ-S strain homozygous for the wild type alanine allele at position 216. Resistance to chlorpyrifos was inherited as a semi-dominant trait mainly controlled by a single autosomal gene and co-segregates strongly but not completely with the serine substitution in ace-1. Synergism bioassays and enzyme assays showed that minor contributions to resistance are also made by enhanced cytochrome P450 and carboxylesterase activities. A survey of 25 field populations from five Chinese provinces showed strong positive correlations between 50% lethal concentration against chlorpyrifos and S216 allele and genotype frequencies, although the most tolerant populations still only show 40%-50% S216 allele frequencies. The results above provide important information for designing effective resistance monitoring and management strategies for A. lucorum in China.

Development of efficient RNAi in Nezara viridula for use in insecticide target discovery.

Stink bugs are an emerging pest in many regions of the world but their molecular biology is still poorly understood. While several transcriptomes are available, the lack of validated gene manipulation tools like RNA interference (RNAi) in species such as the southern green stinkbug Nezara viridula precludes the characterization of individual genes in vivo. Such tools are particularly useful in performing high-throughput screens to search for essential genes that can be prioritized as potential insecticide targets. Here, we developed and optimized an efficient RNAi in N. viridula for use in insecticide target discovery and beyond. The visible marker Sex combs reduced and the essential gene Actin were used to verify the usability and efficiency of RNAi by microinjection at both the adult and nymphal stages, respectively, with nymphal approach presenting significant advantages. Following validation, RNAi was then used to measure lethality following the knockdown (KD) of two genes that are known insecticide targets, Chitin synthase, and Acetyl-CoA carboxylase. The KD of each gene resulted in >75% corrected mortality. These results indicate that RNAi is an effective tool in N. viridula and set a benchmark to evaluate potential targets in future RNAi screens aimed at insecticide target discovery.

Resistance to a chemical pesticide increases vulnerability to a biopesticide: Effects on direct mortality and mortality by predation.

Pesticide mixtures are increasingly used to fight pest species that developed resistance to pesticides. To assess the pesticide control efficiency and to reduce ecological damage to non-target species, it is important to quantify the effect of these mixtures and compare them with the effect of their single pesticides on pest species, non-target species and their predator-prey interactions. We studied the effects of the chemical pesticide chlorpyrifos (CPF), the biopesticide Bacillus thuringiensis israelensis (Bti) and their mixture both on the direct mortality and on the mortality by predation. We focused on larvae of a CPF-resistant and a non-resistant strain of the vector mosquito Culex quinquefasciatus and its predator, the pygmy backswimmer Plea minutissima. In the CPF-Bti mixture, both pesticides interacted antagonistically for direct mortality. Exposure to the mixture caused equal direct mortality and equal mortality by predation in both strains. As expected, exposure to CPF resulted in less direct mortality and less mortality by predation in the CPF-resistant mosquito strain compared to the non-resistant strain. Notably, Bti caused a higher mortality in the mosquito larvae of the CPF-resistant strain compared to the non-resistant strain. Furthermore, the predator killed more mosquito larvae of the resistant strain compared to the non-resistant strain when exposed before to Bti alone. These observations identify a novel cost of resistance to a chemical pesticide in terms of increased vulnerability to a biopesticide.