Sublethal concentrations of clothianidin affect fecundity and key demographic parameters of the chive maggot, Bradysia odoriphaga.

Bradysia odoriphaga is a major insect pest that infests Chinese chive in northern China. Clothianidin is a second-generation neonicotinoid insecticide that is commonly used against B. odoriphaga. In this study, the effect of sublethal clothianidin concentrations (LC5 and LC10) on key biological characteristics of B. odoriphaga was investigated using an age-stage, two-sex life table method. Bioassays results showed that clothianidin exhibited high toxicity against B. odoriphaga with LC50 of 1.898 mg L-1 following 24 h exposure. The developmental duration of larvae was significantly increased when exposed to the LC5 (0.209 mg L-1) and LC10 (0.340 mg L-1) of clothianidin. No significant effects were observed on the pupal stage, adult pre-oviposition period (APOP), total pre-oviposition period (TPOP), and mean longevities of male and female. The oviposition period and fecundity of B. odoriphaga were reduced in clothianidin-treated groups. Moreover, key demographic parameters, including the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0), were significantly decreased by the LC5 and LC10 of clothianidin, while no effects were noted on mean generation time (T). Overall, this study showed that sublethal concentrations of clothianidin have a detrimental effect on B. odoriphaga developmental period, fecundity, and life table parameters. Therefore, clothianidin has the potential to suppress the population of B. odoriphaga even at sublethal concentrations.

Acetamiprid resistance and fitness costs of melon aphid, Aphis gossypii: An age-stage, two-sex life table study.

The melon aphid, Aphis gossypii is a globally distributed crop pest with a wide host range. The intensive use of insecticides against this insect over several years has led to develop resistance against many insecticides including acetamiprid. Understanding the relationship between acetamiprid resistance and fitness of A. gossypii is essential to limit the spread of the resistant population in the field. In this study, age-stage, two-sex life table approach was used to investigate these relationships in the lab. Results showed that resistant strain (Ace-R) had a reduced fitness (relative fitness = 0.909) along with significantly decreased adult longevity, fecundity, net reproductive (R0), mean generation time (T) and gross reproductive rate (GRR). Compared to the susceptible strain (Ace-S), the pre-adult period and total pre-oviposition period (TPOP) were also significantly shorter in Ace-R strain. Moreover, the expression profiles of EcR, JHBP, JHAMT, JHEH, USP and Vg genes supposed to be involved in insect reproduction and development were analyzed using Quantitative Real Time PCR. The EcR, JHBP, JHAMT and USP genes were up-regulated, Vg gene was down-regulated while the mRNA level of JHEH gene was statistically same in the Ace-R strain compared to the Ace-S strain. Collectively, this study provides the occurrence and magnitude of fitness costs of A. gossypii against acetamiprid resistance and could be helpful to manage the resistance evolution in field populations.

Characterization and functional analysis of two acetylcholinesterase genes in Bradysia odoriphaga Yang et Zhang (Diptera: Sciaridae).

Two acetylcholinesterase genes (Boace1 and Boace2) were cloned from Bradysia odoriphaga, a devastating soil pest that mainly damages Chinese chives. The Boace1 encodes BoAChE1 protein consisting of 696 amino acid residues, while Boace2 encodes BoAChE2 containing 638 amino acids. Phylogenetic analysis showed that Boace1 and Boace2 are appeared to be distinct clusters. The gene expression patterns at different development stages and various body parts tissues were examined, and their biological functions were characterized by RNA interference and analog docking prediction. The results showed that both Boace genes were expressed in all developmental stages and examined tissues. The transcript level of Boace2 was significantly higher than Boace1 in all tested samples, and Boace1 was found most abundant in the head while Boace2 was highly expressed in the fat body of B. odoriphaga. The silencing of Boace1 and Boace2 significantly decreased the AChE activity of 36.6% and 14.8% separately, and increased the susceptibility of B. odoriphaga to phoxim, with 60.8% and 44.7% mortality. Besides, overexpression and gene duplication of Boace1 were found in two field resistant populations, and two major mutations, A319S and G400V, were detected in Boace1. Moreover, the docking results revealed that BoAChE1 had a higher affinity towards organophosphorus than BoAChE2. It is concluded that Boace2 is the most abundant ace type in B. odoriphaga, while both Boace play vital roles. Boace1 might play a major neurological function and more likely be the prime target for insecticides, while Boace2 might play some important unidentified roles.

Sublethal effects of beta-cypermethrin modulate interspecific interactions between specialist and generalist aphid species on soybean.

In agroecosystems, plant-pest interactions are at the basis of complex food webs, which can be affected by both biotic and abiotic factors. In the present study, we evaluated the impact of the insecticide beta-cypermethrin on interspecific interactions between the specialist aphid Aphis glycines and the generalist aphid Aulacorthum solani on soybean. Aphis glycines showed higher fecundity than A. solani on soybean and the aphids caused unbalanced reduction in population growth on each other. A sublethal concentration of beta-cypermethrin (LC5 for A. glycines) stimulated the reproduction of A. glycines but it did not impact the fecundity of A. solani. However, the LC5 of beta-cypermethrin enhanced the interspecific inhibition of fecundity between the two aphid species. Moreover, the two species showed different spatial distribution on soybean seedlings. Aphis glycines mainly aggregated on the stem of soybean plant while A. solani colonized soybean leaves. The LC5 of beta-cypermethrin drove A. solani migrating from soybean leaves to stems independently of interspecific competition. Aphis glycines facilitated A. solani colonization on soybean plant through impacting host susceptibility, and vice versa. Nevertheless, such facilitated colonization-induced susceptibility could be modulated through exposure to the LC5 of beta-cypermethrin. These findings hinted that the pyrethroid insecticide beta-cypermethrin has the potential to mediate the interspecific competition between specialist and generalist aphids (at the sublethal concentration of LC5), and that it could influence aphid population growth and community structure in soybean crops. This knowledge could contribute to rationalize application of insecticides and to optimize Integrated Pest Management in soybean.

Fitness costs in chlorfenapyr-resistant populations of the chive maggot, Bradysia odoriphaga.

The chive maggot, Bradysia odoriphaga (Yang and Zhang) is an economically important insect pest, affecting many key vegetables, including Chinese chive, especially in northern China. Chlorfenapyr, a halogenated pyrrole insecticide that interferes with mitochondrial oxidative phosphorylation is widely used against B. odoriphaga. In this study, we evaluated selection-induced resistance to chlorfenapyr and fitness costs in B. odoriphaga. The results showed that B. odoriphaga developed 43.32-fold resistance after continuous exposure to chlorfenapyr for over 10 consecutive generations. The life-history traits of chlorfenapyr-resistant and susceptible strains were compared using age-stage, two-sex life table approach. No significant effects were observed on the longevity and pre-adult period. However, reduction in the total pre-oviposition period (TPOP) and fecundity (eggs/female) were observed in the resistant strain. Moreover, the demographic parameters such as intrinsic rate of increase (r), net reproductive rate (R0) and finite rate of increase (λ) were also decreased significantly in the resistant strain compared to the susceptible strain. These results showed the potential of B. odoriphaga to develop resistance against chlorfenapyr under continuous selection pressure. Furthermore, there was a fitness cost linked with chlorfenapyr resistance in B. odoriphaga. We conclude that a better knowlegde on the trade-off at play between resistance degree and fitness cost could be crucial for developing further management of B. odoriphaga in China.

Functional analysis of cytochrome P450 genes linked with acetamiprid resistance in melon aphid, Aphis gossypii.

Cytochrome P450 monooxygenases (P450s) are highly conserved multifunctional enzymes that play crucial roles in insecticide resistance development. In this study, the molecular mechanisms of P450s in acetamiprid resistance development to melon aphid, Aphis gossypii was investigated. Acetamiprid resistant (32.64-fold resistance) population (Ace-R) of A. gossypii was established by continuous selection with acetamiprid for 24 generations. Quantitative Real Time PCR was carried out to analyze the expression of P450 genes in both acetamiprid resistant (Ace-R) and susceptible (Ace-S) strains. Result showed that nine genes (CYP6CY14, CYP6DC1, CYP6CZ1, CYP6DD1, CYP6CY5, CYP6CY9, CYP6DA1, CYP6CY18, and CYP6CY16) of CYP3 clade, four genes (CYP302A1, CYP315A1, CYP301A1, and CYP314A1) of CYP2 clade, two genes (CYP4CK1, CYP4G51) of CYP4 clade and three genes (CYP306A1, CYP305E1, CYP307A1) of mitochondrial clade (Mito clad) were significantly up-regulated, in Ace-R compared to Ace-S strain. Whilst CYP4CJ2 gene from (CYP4 clade) was significantly down-regulated in Ace-R strain. Furthermore, RNA interference-mediated knockdown of CYP6CY14, CYP6DC1, and CYP6CZ1 genes significantly increased the sensitivity of Ace-R strain to acetamiprid. Taken together, this study showed that P450 genes especially CYP6CY14, CYP6DC1 and CYP6CZ1 are potentially involved in acetamiprid resistance development in A. gossypii. This study could be useful to understand the molecular basis of acetamiprid resistance mechanism in A. gossypii.

Thiamethoxam induces transgenerational hormesis effects and alteration of genes expression in Aphis gossypii.

Insecticide induced-hormesis, a bi-phasic phenomenon characterized by low dose stimulation and high dose inhibition following exposure to insecticide, is crucial to insect pest resurgence. In this study, the effects of low or sublethal concentrations of thiamethoxam on biological traits and genes expression were investigated for Aphis gossypii Glover following 72 h exposures. Leaf-Dip bioassay results showed that thiamethoxam was very toxic against adult A. gossypii with an LC50 of 1.175 mg L-1. The low lethal (LC15) and sublethal (LC5) concentrations of thiamethoxam significantly reduced longevity and fecundity of the directly exposed aphids. However, stimulatory effects on pre-adult stage, longevity, and fertility were observed in the progeny generation (F1) of A. gossypii, when parental aphids (F0) were exposed to LC15 of thiamethoxam. Subsequently, biological traits such as intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) increased significantly to F1 individuals due to LC15 treatment. No significant responses were observed for LC5 of thiamethoxam. The LC15 of thiamethoxam significantly increased the expression level of vitellogenin and ecdysone receptors genes in progeny generation, while no effects were observed for treatment with LC5. Additionally, the expression levels of P450 genes including CYP6CY14, CYP6CZ1, CYP6DC1, CYP6CY9, and CYP6DD1 were up-regulated in the exposed aphids. Taken together, our results show the hormetic effects of thiamethoxam on F1 individuals, which might be due to the intermittent changes in expression of genes involved in fertility, growth and insecticide detoxification in A. gossypii.

UDP-glucosyltransferases potentially contribute to imidacloprid resistance in Aphis gossypii glover based on transcriptomic and proteomic analyses.

The cotton aphid, Aphis gossypii Glover, is a destructive global crop pest. Control of A. gossypii has relied heavily on the application of chemical insecticides. The cotton aphid has developed resistance to numerous insecticides, including imidacloprid, which has been widely used to control cotton pests in China since the 1990s. Our objective was to investigate the potential role of UDP-glycosyltransferases (UGTs) in imidacloprid resistance based on transcriptomic and proteomic analyses of field-originated imidacloprid-resistant (IMI_R) and -susceptible (IMI_S) A. gossypii clones. The transcriptomic and proteomic analyses revealed that 12 out of 512 differentially expressed genes and three out of 510 differentially expressed proteins were predicted as UDP-glycosyltransferase (UGT). Based on quantitative real-time PCR analysis, nine UGT genes, UGT343A4, UGT344A15, UGT344A16, UGT344B4, UGT344C7, UGT344C9, UGT344N4, UGT 24541, and UGT7630, were up-regulated in the IMI_R clone compared to the IMI_S clone. Meanwhile, UGT344A16, UGT344B4, UGT344C7, and UGT344N4 were overexpressed at the protein level based on western blot analysis. Furthermore, knockdown of UGT344B4 or UGT344C7 using RNA interference (RNAi) significantly increased sensitivity to imidacloprid in the IMI_R clone. In conclusion, UGTs potentially contributed to imidacloprid resistance in A. gossypii originating from cotton-growing regions of China. These results provide insights into the way we study insecticide resistance in cotton aphids.

Aphid (Myzus persicae) feeding on the parasitic plant dodder (Cuscuta australis) activates defense responses in both the parasite and soybean host.

Dodders (Cuscuta spp.) are shoot holoparasites, whose haustoria penetrate host tissues to enable fusion between the parasite and host vascular systems, allowing Cuscuta to extract water, nutrients and other molecules from hosts. Aphids are piercing-sucking herbivores that use specialized stylets to feed on phloem sap. Aphids are known to feed on Cuscuta, but how Cuscuta and its host plant respond to aphids attacking the parasite was unknown. Phytohormone quantification, transcriptomic analysis and bioassays were performed to determine the responses of Cuscuta australis and its soybean (Glycine max) hosts to the feeding of green peach aphid (GPA; Myzus persicae) on C. australis. Decreased salicylic acid levels and 172 differentially expressed genes (DEGs) were found in GPA-attacked C. australis, and the soybean hosts exhibited increased jasmonic acid contents and 1015 DEGs, including > 100 transcription factor genes. Importantly, GPA feeding on C. australis increased the resistance of the soybean host to subsequent feeding by the leafworm Spodoptera litura and soybean aphid Aphis glycines, resulting in 21% decreased leafworm mass and 41% reduced aphid survival rate. These data strongly suggest that GPA feeding on Cuscuta induces a systemic signal, which is translocated to hosts and activates defense against herbivores.

Sublethal and hormesis effects of beta-cypermethrin on the biology, life table parameters and reproductive potential of soybean aphid Aphis glycines.

Beta-cypermethrin has long been recommended as an effective pesticide to control the soybean aphid, Aphis glycines Matsumura, a serious pest in soybean crops. Besides acute toxicity, it leads to changes in life history traits of A. glycines, notably its reproductive potential. This study has assessed the effects of five sublethal concentrations (0.625, 1.25, 2.5, 5 and 10 µg/L) of beta-cypermethrin on different life history traits of A. glycines. Exposure to these concentrations caused shorter oviposition period and reduced adult longevity. The strongest stimulatory effect on aphid reproduction was achieved when exposed to a higher sublethal beta-cypermethrin concentration (5 µg/L). Net reproduction rate (R 0 ), intrinsic rate of increase (r m ) and finite rate of increase (λ) were significantly higher than that of the control, increasing by 20.58, 4.89 and 2.06%, respectively. We found no significant difference in mean generation time (T) between the treatment of 5 µg/L beta-cypermethrin and the control. However, when the concentration increased to 10 µg/L, the reproduction behavior was restrained and the mean generation time (T) was shortened, resulting in significant decrease in R 0 and T by 16.58 and 3.83%, respectively. In conclusion, a sublethal concentration (5 µg/L) of beta-cypermethrin triggered the strongest hormesis on A.glycines, thus providing valuable knowledge on the sublethal effects of this insecticide on soybean aphids. Hormesis may be one of the mechanisms underlying pest resurgences, and better knowledge would enable a more effective use of insecticides in Integrated Pest Management programs.

Both point mutations and low expression levels of the nicotinic acetylcholine receptor ß1 subunit are associated with imidacloprid resistance in an Aphis gossypii (Glover) population from a Bt cotton field in China.

Aphis gossypii Glover is a destructive pest of numerous crops throughout the world. Although the expansion of Bt cotton cultivation has helped to control some insect pests, the damage from cotton aphids has not been mitigated. The evolution of aphid resistance to imidacloprid has made its chemical control more difficult since its introduction in 1991. Field populations of A. gossypii that were collected from different transgenic (Bt) cotton planting areas of China in 2014 developed different levels of resistance to imidacloprid. The IMI_R strain has developed high resistance to imidacloprid with the resistance ratio >1200-fold. Compared with the susceptible IMI_S strain, the IMI_R strain also developed a high level cross resistance to sulfoxaflor and acetamiprid. The limited synergism with either PBO or DEF suggests that resistance may be due to the site mutation of molecular target rather than to enhanced detoxification. Three target-site mutations within the nicotinic acetylcholine receptor (nAChR) ß1 subunit were detected in the IMI_R strain. The R81T mutation has been reported to be responsible for imidacloprid resistance in A. gossypii and M. persicae. Both V62I and K264E were first detected in A. gossypii. These point mutations are also present in field populations, suggesting that they play a role in the resistance to imidacloprid. Furthermore, the expression level of transcripts encoding ß1 subunit was decreased significantly in the IMI_R strain compared with the IMI_S strain, suggesting that both point mutations and the down-regulation of nAChR ß1 subunit expression may be involved in the resistance mechanism for imidacloprid in A. gossypii. These results should be useful for the management of imidacloprid-resistant cotton aphids in Bt cotton fields in China.

Sublethal and transgenerational effects of sulfoxaflor on the biological traits of the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae).

Sulfoxaflor is a novel insecticide belonging to sulfoximine chemical class that can be used to control sap-feeding insects, notably Aphis gossypii Glover. In addition to its acute toxicity, it is also important to consider the possible sublethal effects when establishing a comprehensive understanding of the toxicity of a new insecticide. We assessed the effects of a low lethal concentration (LC25) of sulfoxaflor on biological parameters of A. gossypii adults (F0) and subsequent transgenerational effects, i.e., on the progeny (F1 generation). The data were analyzed using an age-stage life table procedure. The results showed that the longevity and fecundity were not significantly affected by the LC25 of sulfoxaflor in the F0 or F1 generations. In addition, no significant differences were observed on the developmental time of each instar, the adult pre-oviposition period, and on the longevity of F1 individuals. However, the duration of their pre-adult stage and total pre-oviposition period, as well as their mean generation time were significantly increased. These observed effects affected aphid demographic traits; the survival rate, the intrinsic rate of increase (r i ), the finite rate of increase (λ), the net reproductive rate (R0), and the gross reproduction rate (GRR) of the F1 individuals (i.e., from F0 mothers) were significantly lower compared to the control. Our results showed that sublethal effects of sulfoxaflor significantly slowed down A. gossypii population growth; they indicated that effects of sulfoxaflor might be increased (beyond lethal effect) through sublethal effects when concentrations decreased in sulfoxaflor-treated areas after initial application in field.

Novel (E)-ß-Farnesene Analogues Containing 2-Nitroiminohexahydro-1,3,5-triazine: Synthesis and Biological Activity Evaluation.

In order to discover novel eco-friendly compounds with good activity for aphid control, (E)-ß-farnesene (EßF), the main component of the aphid alarm pheromone, was chosen as the lead compound. By introducing a 2-nitroimino-hexahydro-1,3,5-triazine moiety (abbreviated NHT) to replace the unstable conjugated double bond system of EßF, a series of novel EßF analogues containing the NHT moiety were synthesized via the reaction of substituted NHT rings with (E)-1-chloro-3,7-dimethylocta-2,6-diene. All the compounds were characterized by ¹H-NMR, (13)C-NMR, IR, and high resolution mass spectroscopy (HRMS). The bioassay results showed that all the analogues displayed different repellent and aphicidal activities against green peach aphid (Myzus persicae). Particularly, the analogue 4r exhibited obvious repellent activity (repellent proportion: 78.43%) and similar aphicidal activity against M. persicae (mortality: 82.05%) as the commercial compound pymetrozine (80.07%). A preliminary structure-activity relationship (SAR) study was also performed, which offered valuable clues for the design of further new EßF analogues.

Sublethal and hormesis effects of imidacloprid on the soybean aphid Aphis glycines.

The soybean aphid, Aphis glycines Matsumura, is a major pest in soybean crop. Current management of this pest relies mainly on insecticides applications, and the neonicotinoid imidacloprid has been proposed as an effective insecticide to control A. glycines in soybean field. Imidacloprid at lethal concentrations not only exerts acute toxicity to A. glycines, but also cause various biological changes when aphids are chronically exposed to lower concentrations. In this study, we assessed the effects of a low-lethal (0.20 mg L(-1)) and two sublethal (0.05 and 0.10 mg L(-1)) imidacloprid concentrations on various A. glycines life history traits. Aphid exposure to 0.20 mg L(-1) imidacloprid caused slower juvenile development, shorter reproductive period, and reduced adult longevity, fecundity and total lifespan. Stimulatory effects, i.e. hormesis, on reproduction and immature development duration were observed in aphids exposed to the lower sublethal imidacloprid concentrations. Consequently, the net reproduction rate (R 0) was significantly higher than in the control aphids. These findings stress the importance of the actual imidacloprid concentration in its toxicological properties on A. glycines. Therefore, our results would be useful for assessing the overall effects of imidacloprid on A. glycines and for optimizing integrated pest management programs targeting this pest.

Quantification of γ-aminobutyric acid in the heads of houseflies (Musca domestica) and diamondback moths (Plutella xylostella (L.)), using capillary electrophoresis with laser-induced fluorescence detection.

A novel method was developed for quantifying the levels of γ-aminobutyric acid (GABA) in the heads of houseflies (Musca domestica) and diamondback moths (Plutella xylostella (L.)), using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). The GABA in sample was derivatized with 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) prior to CE-LIF analysis. In total, 32 mmol/L borate buffer, at pH 9.2 and containing 5.3 mmol/L ß-cyclodextrin (ß-CD) and 10.4 mmol/L sodium dodecyl sulfate (SDS), was determined to be the optimum CE background electrolyte (BGE) for GABA analysis. The detection limit of GABA was 0.016 µmol/L. The relative standard deviations (RSDs) of the migration time and peak area of GABA were 1.78 and 4.93%, respectively. The average recoveries of 0.97, 3.88, and 5.83 µmol/L of GABA, each added to the head sample of housefly, ranged from 88.9 to 110.5%. This method is simple and applicable to GABA assays of the heads of insects. With this newly developed CE-LIF method, the amounts of GABA in the heads of houseflies (M. domestica) and diamondback moths (P. xylostella (L.)) were measured. The results are relevant to the understandings of some insecticides and insecticide-resistance mechanisms in pests.

V101I and R81T mutations in the nicotinic acetylcholine receptor ß1 subunit are associated with neonicotinoid resistance in Myzus persicae.

BACKGROUND: The peach-potato aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), is a major pest worldwide. The intensive use of insecticides has led to the development of resistance against neonicotinoid insecticides. The R81T mutation in the nicotinic acetylcholine receptor (nAChR) beta1 subunit is considered a crucial mechanism adaptation to neonicotinoid resistance in M. persicae and Aphis gossypii. RESULTS: Resistance-related mutations (R81T and V101I) were detected in the imidacloprid-resistant M. persicae AH19 population. The V101I mutation is reported for the first time. The V101I and R81T mutations existed separately, indicating that the two mutations evolved independently. Imidacloprid resistance in the AH19 population was stable without insecticide exposure. Four mutant strains were selected from the population with stable resistance. The resistance of the AH19-T, AH19-I, and AH19-TI strains to imidacloprid, thiamethoxam, and dinotefuran was significantly increased compared with the AH19-W strain. Synergism bioassays showed that the inhibition of three detoxification enzymes did not affect imidacloprid resistance in the AH19-T and AH19-I strains. Expression of nAChR ß1 subunits in the AH19-W, AH19-T, and AH19-I strains remained unchanged. CONCLUSION: The V101I mutation is associated with neonicotinoid resistance in M. persicae. The resistance of the AH19-T and AH19-I strains to neonicotinoids appears to be mainly due to the R81T and V101I mutations, whereas these mutations, together with changes in the cytochrome P450 monooxygenases and nAChR expression may be responsible for the development of neonicotinoid resistance in the AH19-TI strain. © 2021 Society of Chemical Industry.

Impact of Imidacloprid Resistance on the Demographic Traits and Expressions of Associated Genes in Aphis gossypii Glover.

Imidacloprid is one of the most widely used neonicotinoid insecticides to control sap-sucking insect pests, including Aphis gossypii. The intensive application of chemical insecticides to A. gossypii led to the development of resistance against several insecticides, including imidacloprid. Therefore, it is crucial to understand the association between imidacloprid resistance and the fitness of A. gossypii to limit the spread of the resistant population under field contexts. In this study, we used the age-stage, two-sex life table method to comprehensively investigate the fitness of imidacloprid resistant (ImR) and susceptible strains (SS) of melon aphids. Results showed that ImR aphids have prolonged developmental stages and decreased longevity, fecundity, and reproductive days. The key demographic parameters (r, λ, and R0) were significantly reduced in ImR strain compared to SS aphids. Additionally, the molecular mechanism for fitness costs was investigated by comparing the expression profile of juvenile hormone-binding protein (JHBP), juvenile hormone epoxide hydrolase (JHEH), juvenile hormone acid O-methyltransferase (JHAMT), Vitellogenin (Vg), ecdysone receptor (EcR), and ultraspiracle protein (USP) supposed to be associated with development and reproduction in insects. The results of RT-qPCR showed that EcR, JHBP, JHAMT, JHEH, and Vg genes were downregulated, while USP was statistically the same in ImR A. gossypii compared to the SS strain. Together, these results provide in-depth information about the occurrence and magnitude of fitness costs against imidacloprid resistance that could help manage the evolution and spread of A. gossypii resistance in field populations.

Characterization of the insecticide detoxification carboxylesterase Boest1 from Bradysia odoriphaga Yang et Zhang (Diptera: Sciaridae).

BACKGROUND: In insects, carboxylesterases (CarEs) are enzymes involved in the detoxification of insecticides. However, the molecular mechanism of CarE-mediated insecticide metabolism in Bradysia odoriphaga, a serious agricultural pest, remains unclear. The aim of this study is to investigate the detoxification process of malathion, bifenthrin, and imidacloprid by B. odoriphaga carboxylesterase (Boest1). RESULTS: An alpha class CarE gene Boest1 was cloned from B. odoriphaga. The results of real-time quantitative polymerase chain reaction showed that Boest1 is up-regulated with age during the larval stage, and the level of transcription of Boest1 is higher in the midgut and Malpighian tubule than in other tissues. The expression level of Boest1 was significantly increased after exposure to malathion and bifenthrin. Recombinant BoEST1 expressed in vitro showed high catalytic activity toward α-naphthyl acetate, which was substantially inhibited by malathion and triphenyl phosphate. The in vitro metabolism assays showed that BoEST1 demonstrates hydrolytic capacity toward malathion and bifenthrin but not imidacloprid. The binding free energy analysis indicates that BoEST1 has a higher affinity for malathion and bifenthrin than imidacloprid. CONCLUSION: These results suggest that BoEST1 plays a role in the breakdown of insecticides and may be involved in the development of resistance in the Chinese chive pest B. odoriphaga; our findings also provide data for better pest management and perspectives for new pesticides development. © 2021 Society of Chemical Industry.

Resistance Mechanisms of Sitobion miscanthi (Hemiptera: Aphididae) to Malathion Revealed by Synergist Assay.

A resistant strain (MRS) of Sitobion miscanthi was cultured by continuous selection with malathion for over 40 generations. The MRS exhibited 32.7-fold resistance to malathion compared to the susceptible strain (MSS) and 13.5-fold, 2.9-fold and 4.8-fold cross-resistance for omethoate, methomyl and beta-cypermethrin, respectively. However, no cross-resistance was found to imidacloprid in this resistant strain. The realized heritability for malathion resistance was 0.02. Inhibitors of esterase activity, both triphenyl phosphate (TPP) and S,S,S,-tributyl phosphorotrithioate (DEF) as synergists, exhibited significant synergism to malathion in the MRS strain, with 11.77-fold and 5.12-fold synergistic ratios, respectively, while piperonyl butoxide (PBO) and diethyl maleate (DEM) showed no significant synergism in the MRS strain. The biochemical assay indicated that carboxylesterase activity was higher in MRS than in MSS. These results suggest that the increase in esterase activity might play an important role in S. miscanthi resistance to malathion. Imidacloprid could be used as an alternative for malathion in the management of wheat aphid resistance.

Slow resistance evolution to neonicotinoids in field populations of wheat aphids revealed by insecticide resistance monitoring in China.

BACKGROUND: The wheat aphids, Rhopalosiphum padi and Sitobion miscanthi, are serious agricultural insect pests of many crops. Neonicotinoid insecticides are commonly used as alternatives to organophosphate and pyrethroid insecticides for controlling wheat aphids. RESULTS: Long-term monitoring of resistance to imidacloprid and acetamiprid in R. padi and S. miscanthi was carried out between 2007 and 2019. For this study, 344 specimens of the two wheat aphids were collected from field populations found in the main wheat production areas in China, from 2007 to 2019. In R. padi, the fluctuation in resistance was 14.7 times for imidacloprid and 1.4 times for acetamiprid; in S. miscanthi, it was 9.7 times for imidacloprid and 6.5 times for acetamiprid. CONCLUSION: Our results demonstrated that both R. padi and S. miscanthi tended to have higher resistance to imidacloprid compared with acetamiprid. However, it is difficult for wheat aphids to develop a high level of neonicotinoid resistance given the pest control practices used in China. These results should be useful for the biorational application and resistance management of neonicotinoid insecticides. © 2021 Society of Chemical Industry.