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The raw data for the manuscript of "Rapid Detection of Pyrethroid Resistance-Associated Mutation L1014F in the Field Based on Loop-Mediated Isothermal Amplification (LAMP) Technique". &nbsp

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.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 β1 subunits in the AH19-W, AH19-T, and AH19-I strains remained unchanged.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.

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.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.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.

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.

A resistant strain (MRS) of

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.

Stimulatory effects (hormesis) of acetamiprid on demographical traits and vitellogenin gene expression of melon aphid, Aphis gossypii Glover, were examined. The longevity and fecundity of the parental aphids (F-0) were both reduced by a low lethal concentration of acetamiprid (LC15), while no effect was observed when exposed to a sublethal concentrations (LC5). However, acetamiprid-induced hormetic effects were observed in the (F)1 generation. The pre-adult developmental period and fertility of progeny aphids were increased by both concentrations (LC5 and LC15). Subsequently, key demographic parameters such as intrinsic rate of increase (r) and finite rate of increase (lambda) were increased by the LC5, and the mean generation time (T) and net reproductive rate (R-0) were enhanced by both concentrations of acetamiprid. Furthermore, both acetamiprid concentrations induced the expression level of Vg gene in the progeny generation. The higher mRNA transcript level of Vg might be translated into an increased reproduction of F-1 generation in A. gossypii. Based on the results, acetamiprid at low or sublethal concentrations may induce stimulatory effects on the subsequent generations of A. gossypii along with the increased transcription of Vg gene. These findings may help understanding the hormetic effects of acetamiprid in insects and could help optimizing the use of this insecticide against A. gossypii.

The melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae) is a major pest affecting plants of Cucurbitaceae family worldwide. Clothianidin has been broadly used for the control of this insect pest. In this study, we investigated the impact of clothianidin on the demographical traits and expression changes of vitellogenin and ecdysone receptors genes in A. gossypii after exposure to the low or sublethal concentrations (LC15 and LC5, respectively). The two concentrations of clothianidin did not affect the adult longevity and fecundity of parental aphids (F-0). However, the developmental duration of 3rd instar nymph, pre-adult stage, and total pre-ovipositional period (TPOP) were increased significantly in F-1 individuals when F o was exposed to the LC15 of clothianidin. In contrast, the adult longevity and fecundity of progeny generation were decreased by the LC15. Similarly, the net reproductive rate (R-0) and gross reproduction rate (GRR) were also reduced significantly in the LC15 treated group. However, no significant effects were observed for the intrinsic rate of increase (r), finite rate of increase (lambda) and mean generation time (T). Furthermore, the expression level of vitellogenin receptor gene significantly decreased by 24.9 and 56.9% in the progeny generation when F-0 was exposed with LC5 and LC15 of clothianidin, respectively. Moreover, the ecdysone receptor mRNA expression was also reduced by 29.2 and 46.1% in the LC5 and LC15 treated groups, respectively. These results indicated that clothianidin at low or sublethal concentrations adversely affect the development and reproduction of progeny generations and effects may be related to the down-regulation of VgR and EcR genes in A. gossypii.

Background The cotton aphid Aphis gossypii Glover is one of the most destructive insect pests. It has evolved resistance to numerous insecticides around the world due to the application of insecticides. Uridine diphosphate (UDP)-glycosyltransferases (UGTs) have been reported to potentially facilitate the detoxification process of imidacloprid and thiamethoxam in A. gossypii. Results In this study, the field populations of A. gossypii developed different levels of resistance to multiple insecticides. A UGT inhibitor, 5-nitrouracil, dramatically increased the toxicity of acetamiprid in resistant populations, moderately increased the toxicity of sulfoxaflor in the imidacloprid susceptible (IMI_S) population, and populations from Yuncheng in Shanxi Province (SXYC) and Jingzhou in Hubei Province (HBJZ), and increased the toxicity of bifenthrin in the IMI_S and HBJZ populations, but there was no synergism on omethoate or carbosulfan. Quantitative real-time PCR analysis revealed that UGT344B4 and UGT344C7 were overexpressed in all field populations, and UGT344N4 was overexpressed in the SDBZ and HBZJ populations. Furthermore, the suppression of UGT344B4 or UGT344C7 by RNA interference significantly increased the susceptibility to bifenthrin in the IMI_S population and the susceptibility to sulfoxaflor in the SXYC population. Conclusion These results suggested that UGTs are potentially involved in the detoxification of neonicotinoid, sulfoximine, and pyrethroid insecticides in A. gossypii. Furthermore, the overexpression of UGTs could be associated with insecticide resistance in field populations of A. gossypii. The results might be helpful for the management of insecticide resistance in field populations of A. gossypii. © 2019 Society of Chemical Industry.

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).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.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.

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.

International audience; In agmecosystems, 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 betacypermethrin. 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.

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.

Wheat (Triticum aestivum L.) is one of the major source of food worldwide. However, it suffers from numerous constraints. The aphids, particularly Rhopalosiphum padi (Linnaeus) and Sitobion avenae (Fabricius) species are among the major causes that severly affect wheat production acrossthe globe. To determine the population dynamics of aphids associated with wheat crop in Khyber Pakhtunkhwa, we examined the mean population of both species in the selected areas (Maho Dheri, Sarband, Kandare, Rag Narai and Khazana Dheri) of district Mardan in 2015. The mean population of R. padi and S. avenae per plant were recorded every week by counting the number of aphids on 5 plants picked up randomly from each selected wheat field using the diagonal methods. The results on current study showed that the infestation of both aphid species was started in the 1st week of February (05 standard weeks). The densities of aphids per plant increased as the vegetative growth proceeded and was highest till the mid of March (10th and 11th standard week). The declined in aphid populations for both species were started after mid of March and ended in April. The mean population of both aphid species was found low in the 1st week of February (05th standard week) that is probably due to the low temperature. The population density of aphids reached to the its peak in the second week of March due to suitable temperature for aphid infestation. Keywords: District Mardan; Population dynamics; Rhopalosiphum padi; Sitobion avenae and Wheat http://dx.doi.org/10.19045/bspab.2020.90004

BACKGROUND: The aphid alarm pheromone, (E)‐β‐farnesene (EβF), is a natural product secreted from the aphid cornicle as a signal to warn companions of danger. Odorant binding proteins (OBPs) are the vital targets in insect signal transduction pathways. To improve bioactivity of EβF as more economic and stable aphid control agents, EβF derivatives containing an active substructure, salicylic acid moiety, were designed, synthesized, and evaluated for their bioactivities in a structure–function study under laboratory conditions. RESULTS: EβF derivatives, (E)‐3,7‐dimethylocta‐2,6‐dien‐1‐yl‐2‐hydroxy‐3‐methylbenzoate and (E)‐3,7‐dimethylocta‐2,6‐dien‐1‐yl‐2‐hydroxy‐3‐methoxybenzoate showed outstanding aphid‐repellent activity at a dose of 5 μg against Acyrthosiphon pisum (repellency proportions of 67.3% and 71.2%, respectively) and Myzus persicae (repellency proportions of 80.0% and 74.4%, respectively) in laboratory. EβF and most of its derivatives bound strongly to ApisOBP9 with a higher affinity than those of the reported potential targets AphisOBP3 and ApisOBP7. The binding affinities to these three ApisOBPs were generally consistent with the in vivo aphid‐repellent activity. A molecular docking study suggested that the hydrophobic effect was crucial for the interactions between the derivatives and the OBPs. CONCLUSION: New EβF derivatives containing salicylic acid moiety and their repellent activity, binding mechanism with three potential OBPs are presented. A new OBP, ApisOBP9, was characterized as a potential EβF and EβF derivatives binding protein for the first time. The hydrophobic nature of these analogues is responsible for their activity. Two analogues 3b and 3e with outstanding aphid‐repellent activity could be new leads for aphid control agents.

Imidacloprid is a largely commercialized neonicotinoid insecticide, and it is used effectively against a variety of sap-sucking insect pests including melon aphid, Aphis gossypii. In this study, the inhibitory and stimulatory effects of low and sublethal concentrations of imidacloprid were investigated on two successive generations of A. gossypii. The initial bioassay results showed that imidacloprid exhibited high toxicity against adult melon aphids with a LC50 of 0.42 mg/l after 72 h exposure. The low lethal (LC15) and sublethal (LC5) concentrations of imidacloprid reduced adult longevity and fecundity in exposed aphids (F-0). By contrast, hormesis effects were observed on the biological traits of progeny generation (F-1) exposed to both concentrations of imidacloprid. Substantial increases were noted for the adult longevity and fecundity of F-1 aphids. Consequently, the demographic indexes such as intrinsic rate of increase (r) and finite rate of increase (lambda) were enhanced by both concentrations whereas the net reproductive rate (R-0) and gross reproduction rate (GRR) were increased only by the LC15 of imidacloprid. Nonetheless, the mean generation time (T) of F-1 A. gossypii was extended by the LC5 and LC15 of imidacloprid. These results indicated that low or sublethal concentrations of imidacloprid, while negatively affecting biological traits of directly exposed aphids, could induce hormesis effects in the progeny of exposed individuals in the aphid A. gossypii. Therefore, the occurrence of potential hormesis should be taken into consideration when using imidacloprid against aphid pests.

Chitin is a vital part of the insect exoskeleton and peritrophic membrane, synthesized by chitin synthase (CHS) enzymes. Chitin synthase 1 (CHS1) is a crucial enzyme in the final step of chitin biosynthetic pathway and consequently plays essential role towards insect growth and molting. RNA interference (RNAi) is an agent that could be used as an extremely target-specific and ecologically innocuous tactic to control different insect pests associated with economically important crops. The sole purpose of the current study is to use CHS1 as the key target gene against the cotton-melon aphid, Aphis gossypii, via oral feeding on artificial diets mixed with dsRNA-CHS1. Results revealed that the expression level of CHS1 gene significantly decreased after the oral delivery of dsRNA-CHS1. The knockdown of CHS1 gene caused up to 43%, 47%, and 59% mortality in third-instar nymph after feeding of dsCHS1 for 24, 48, and 72 h, respectively, as compared to the control. Consistent with this, significantly lower longevity (approximately 38%) and fecundity (approximately 48%) were also found in adult stage of cotton-melon aphids that were fed with dsCHS1 for 72 h at nymphal stage. The qRT-PCR analysis of gene expression demonstrated that the increased mortality rates and lowered longevity and fecundity of A. gossypii were attributed to the downregulation of CHS1 gene via oral-delivery-mediated RNAi. The results of current study confirm that CHS1 could be an appropriate candidate target gene for the RNAi-based control of cotton-melon aphids.

The chive maggot, Bradysia odoriphaga Yang and Zhang (Diptera: Sciaridae) is an economically important insect pest, affecting many important vegetables, including Chinese chive, especially in northern China. Clothianidin is a second-generation of neonicotinoid insecticide that is currently used for the management of B. odoriphaga. In the present study, we investigated the development of resistance after continuous laboratory selection and associated fitness costs in B. odoriphaga against clothianidin insecticide. The results showed that B. odoriphaga population developed 76-fold resistance, following exposure with clothianidin insecticide for 10 generations. Similarly, life table analysis showed that the fecundity, along with male and female longevity in a resistant strain of B. odoriphaga was decreased significantly compared with the susceptible strain. The developmental duration of larvae and pupae were increased dramatically in the resistant strain, while no effects were observed on egg hatchability. Furthermore, the demographic parameters such as the intrinsic rate of increase (r) (d(-1)), net reproductive rate (R-0) (offspring/individual) and finite rate of increase (lambda) (d(-1)) were found to be significantly decreased in the laboratory selected resistant population when compared with the susceptible strain of B. odoriphaga, in addition, only mean generation time (T) (d) increased significantly. These results suggest that B. odoriphaga responded quickly and develop a high level of resistance against clothianidin insecticide under continuous selection pressure. Additionally, the associated fitness costs may have direct implication for the management of B. odoriphaga against clothianidin.

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) β1 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 β1 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 β1 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.

P-glycoprotein (P-gp) exists in animals, fungi and bacteria and likely evolved as a defense mechanism against harmful substances. Here a cDNA (4054bp) encoding a putative P-glycoprotein gene from Helicoverpa armigera was cloned and named HaPgp1. This putative HaPgp1 sequence encoded a protein of 1253 amino acids with a molecular mass of approximately 137kDa. qPCR analyses demonstrated that the expression of HaPgp1 was significantly higher in 4th instar larvae when compared to other developmental stages. HaPgp1 transcripts were more abundant in the head and fat bodies than in other tissues. Compared with the control, the expression of HaPgp1 reach a peak at 12h after the treatment by 2-tridecanone in all tissues. However, the expression of HaPgp1 increased from 12h to 48h after treatment with abamectin in all tissues. Immunohistochemistry analyses also verified that 2-tridecanone and abamectin can induce the increase of HaPgp1 expression. RNAi of HaPgp1 significantly raised the mortality rate of larvae treated by 2-tridecanone and abamectin, as compared to control larvae fed with GFP dsRNA. These results illustrate the possible involvement of HaPgp1 as a component of the protective mechanisms to plant secondary chemicals such as 2-tridecanone and to certain classes of insecticides, like abamectin.

BACKGROUND The cotton aphid, Aphis gossypii (Glover), is a destructive pest that affects numerous crops throughout the world. Pyrethroid resistance has become endemic in A. gossypii populations in the cotton growing areas of China due to wide- spread application of insecticides. To assess the extent of pyrethroid resistance, bioassays were conducted on field populations collected from several cotton cultivation areas from 2010–2015. The frequency of a known resistance-associated sodium channel mutation (M918L) in A. gossypii was evaluated and the bioassay of bifenthrin with or without the synergist was performed to illuminate the mechanisms underlying resistance to pyrethroids. RESULTS The field populations exhibited very high levels of resistance to both beta-cypermethrin and deltamethrin. Pretreatment with synergists, DEF and PBO, significantly increased the toxicity of bifenthrin to cotton aphid populations collected from Bt cotton fields in China. Further, 96.8%-100% of individuals with the M918L mutation (including both RR and RS individuals) were observed in various populations, and only 2.8%-3.2% of individuals with wild-type homozygotes (SS) were detected. CONCLUSION The mutation M918L in the voltage-gated sodium channel along with detoxifying metabolism was contributed to the pyrethroid resistance in the field populations of Aphis gossypii from cotton growing regions of China. And insecticides with different modes of action should be recommended for the control of A. gossypii in the future.

In order to discover novel compounds with high-activity to control aphid, a series of novel ( E )- β -farnesene analogues containing 1,2,3-thiadiazole were designed and synthesized, and their structures were confirmed by IR, 1 H NMR, 13 C NMR, and HRMS (ESI). The stability of representative compounds was studied by HPLC and 1 H NMR techniques. Repellent activity results indicated that compounds 8h and 8j displayed 60.3% and 62.0% repellent rates, respectively. The aphicidal bioassay results showed that most analogues exhibited considerable aphicidal activity against Myzus persicae . Especially, analogues 8l , 8s and 8t exhibited high activity with LC 50 values of 33.4 μg/mL, 50.2 μg/mL and 61.8 μg/mL, respectively, which were higher than the lead compound ( E) - β -farnesene, but lower than commercial insecticide pymetrozine with a LC 50 of 7.1 μg/mL.

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.

The joint toxicity of chlorantraniliprole, a novel insecticide that acts on ryanodine receptors, and sinigrin, a natural plant defense compound from brassicaceous vegetables, to the larvae of Spodoptera exigua (Hübner) was determined in this paper. Additionally, the joint effects of the two compounds on cytochrome P450 enzyme activity and on the expression levels of mRNA of three P450 genes (including CYP9A9, CYP6B, and CYP4G37) and an NADPH cytochrome P450 reductase gene (HQ852049) were investigated. The toxicity of the mixture of chlorantraniliprole and sinigrin to fourth-instar S. exigua larvae was 1.60-fold higher than the toxicity of the chlorantraniliprole-only treatment after 24 h. Induced by chlorantraniliprole and sinigrin, the specific activity of the P450 O-deethylase was affected in a time-, dose-, and organ-specific manner in fifth-instar S. exigua larvae. The effects were more pronounced in the midgut than in the fat body. The specific activity of the P450 O-deethylase in almost all treatments increased at 12, 24, and 36 h posttreatment compared with that in the control. Based on real-time PCR analyses, the expression levels of the P450 genes CYP9A9, CYP6B, and CYP4G37 and the NADPH cytochrome P450 reductase gene HQ852049 in fifth-instar S. exigua larvae were induced by chlorantraniliprole and sinigrin, and the trends were similar to the specific activity of the P450 O-deethylase. Therefore, the CYP9A9, CYP6B, and HQ852049 in the tested genes were the most inducible genes that were expressed when the S. exigua larvae were exposed to chlorantraniliprole and sinigrin.

International audience; Clothianidin is a second-generation neonicotinoid insecticide, widely used against sap-sucking insect pest including melon aphid,Aphis gossypiiGlover (Hemiptera: Aphididae). This pest causes severe economic damage to Cucurbitaceae plants worldwide. In this study, we investigated clothianidin resistance development under continuous selection pressure. Moreover, the age-stage, two-sex life table approach was used to evaluate the impact of clothianidin resistance on the fitness ofA.gossypii. A clothianidin resistant strain (CT-R) with a 23.17-fold resistance level was developed from a susceptible strain (CT-S) after continuous selection for 24 generations. Life table results showed a significant reduction in the relative fitness (0.847) of CT-R strain compared to the CT-S strain ofA.gossypii. The developmental duration, oviposition days, total pre-oviposition period (TPOP), longevity, and fecundity of CT-R strain were found to be significantly lower when compared to CT-S strain. The demographic parameters, including the intrinsic rate of increase (r), finite rate of increase (lambda), net reproductive rate (R-0), and mean generation time (T) were also significantly decreased in CT-R strain compared to the CT-S strain. Both the reproductive and survival rates were affected by clothianidin resistance in CT-R strain compared with the CT-S strain ofA.gossypii. Overall, our results demonstrate that in-depth knowledge about the trade-off at play between resistance degree and fitness cost might be useful to design resistance management strategies againstA.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.

Background It is difficult to efficiently silence gene expression in some insects, probably because of the degradation of dsRNA by enzymes present in the gut and hemolymph post-oral feeding or body injecting of dsRNA. We previously developed a nanocarrier delivery system that can systemically deliver dsRNA into chewing mouthpart insects by oral feeding and efficiently silence gene expression. For the purpose of pest control in the field, there is a great demand to develop a spray method to apply dsRNA formulation. Results A formulation of the nanocarrier/dsRNA/detergent was developed and could be easily applied just by dropping it on the notum of the aphid. The formulation penetrated the body wall into the hemocoel and then spread into various tissues within 1 h. The delivered dsRNA efficiently silenced the target gene expression at a high knockdown effect (95.4%) and the aphid population was largely suppressed (80.5%). Conclusion A novel dsRNA formulation was developed with the help of a nanocarrier and detergent that can quickly penetrate the insect body wall and efficiently silence gene expression. The formulation may provide a fast and easy tool for gene silence in some tough insects and for pest control in the field. © 2019 Society of Chemical Industry.

International audience; Wheat is a major crop and plays a significant role in food security in China. Cereal aphids Rhopalosiphum padi L. and Sitobion avenae Fab. are widespread worldwide and considered as serious pests in the winter wheat belt of North China. Natural enemies are well known for their ability to control aphid species in a wide range of cropping systems. Imidacloprid is one of the most widely used insecticides for the management of cereal aphids. Nevertheless, insecticides are harmful to the environment and disrupt the biological services. A full understanding on the effects of insecticides, natural enemies and their interactions on the cereal aphids in the complex agroecosystem is quite crucial. Field studies were conducted in the two consecutive seasons 2014/2015 and 2015/2016 to address this question using exclusion cage techniques. Our data demonstrated strong effects of natural enemies, application of pesticide as well as their interactions on the population densities and growth rates of cereal aphids (R. padi and S. avenae) in the wheat field. The instantaneous rate of population growth of R. padi and S. avenae strongly declined by the application of imidacloprid, nevertheless, the parasitism rate was also significantly affected. We conclude that imidacloprid insecticide is still effective in suppressing cereal aphid populations in winter wheat fields of North China throughout the growing season, however, the negative impact on the biological control services needs to be considered with caution.

The insect kinins are a class of multifunctional insect neuropeptides present in a diverse variety of insects. Insect kinin analogs showed multiple bioactivities, especially, the aphicidal activity. To find a biostable and bioactive insecticide candidate with simplified structure, a series of N-terminal modified insect kinin analogs was designed and synthesized based on the lead compound [Aib]-Phe-Phe-[Aib]-Trp-Gly-NH 2 . Their aphicidal activity against the soybean aphid Aphis glycines was evaluated. The results showed that all the analogs maintained the aphicidal activity. In particular, the aphicidal activity of the pentapeptide analog X Phe-Phe-[Aib]-Trp-Gly-NH 2 (LC 50 = 0.045 mmol/L) was similar to the lead compound (LC 50 = 0.048 mmol/L). This indicated that the N-terminal protective group may not play an important role in the activity and the analogs structure could be simplified to pentapeptide analogs while retaining good aphicidal activity. The core pentapeptide analog X can be used as the lead compound for further chemical modifications to discover potential insecticides.

Insect kinin neuropeptides are pleiotropic peptides that are involved in the regulation of hindgut contraction, diuresis, and digestive enzyme release. They share a common C-terminal pentapeptide sequence of Phe(1)-Xaa(2)-Yaa(3)-Trp(4)-Gly(5)-NH2 (where Xaa(2) = His, Asn, Phe, Ser, or Tyr; Yaa(3) = Pro, Ser, or Ala). Recently, the aphicidal activity of insect kinin analogues has attracted the attention of researchers. Our previous work demonstrated that the sequence-simplified insect kinin pentapeptide analogue Phe-Phe-[Aib]-Trp-Gly-NH2 could retain good aphicidal activity and be the lead compound for the further discovery of eco-friendly insecticides which encompassed a broad array of biochemicals derived from micro-organisms and other natural sources. Using the peptidomimetics strategy, we chose Phe-Phe-[Aib]-Trp-Gly-NH2 as the lead compound, and we designed and synthesized three series, including 31 novel insect kinin analogues. The aphicidal activity of the new analogues against soybean aphid was determined. The results showed that all of the analogues exhibited aphicidal activity. Of particular interest was the analogue II-1, which exhibited improved aphicidal activity with an LC50 of 0.019 mmol/L compared with the lead compound (LC50 = 0.045 mmol/L) or the commercial insecticide pymetrozine (LC50 = 0.034 mmol/L). This suggests that the analogue II-1 could be used as a new lead for the discovery of potential eco-friendly insecticides.

The cotton aphid, Aphis gossypii (Glover), is a destructive pest that affects numerous crops throughout the world. Pyrethroid resistance has become endemic in A. gossypii populations in the cotton growing areas of China due to wide- spread application of insecticides. To assess the extent of pyrethroid resistance, bioassays were conducted on field populations collected from several cotton cultivation areas from 2010 to 2015. The frequency of a known resistance-associated sodium channel mutation (M918 L) in A. gossypii was evaluated and the bioassay of bifenthrin with or without the synergist was performed to illuminate the mechanisms underlying resistance to pyrethroids.The field populations exhibited very high levels of resistance to both beta-cypermethrin and deltamethrin. Pretreatment with synergists, DEF and PBO, significantly increased the toxicity of bifenthrin to cotton aphid populations collected from Bt cotton fields in China. Further, 96.8-100% of individuals with the M918 L mutation (including both RR and RS individuals) were observed in various populations, and only 2.8-3.2% of individuals with wild-type homozygotes (SS) were detected.The mutation M918 L in the voltage-gated sodium channel along with detoxifying metabolism was contributed to the pyrethroid resistance in the field populations of Aphis gossypii from cotton growing regions of China. And insecticides with different modes of action should be recommended for the control of A. gossypii in the future. © 2017 Society of Chemical Industry.

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.

A series of novel neonicotinoid analogues were designed and synthesized by introducing a hydrazide group into clothianidin. Their structures were confirmed by IR, 1 H NMR, and HRMS (ESI). Preliminary bioassay showed that some compounds, 5b and 5g , exhibited good activity against soybean aphids ( Aphis glycines ) at 100 mg L −1 . In addition, molecular docking with receptor was carried out to explain their different activity from clothianidin.

BACKGROUD The function of acetylcholinesterase (AChE) is to terminate synaptic transmission by hydrolysing the neurotransmitter acetylcholine (ACh) in the synaptic cleft, and thus it is an effective target for organophosphate (OP) and carbamate (CB) insecticides. RESULTS The transcript levels of the four Ace genes were dramatically suppressed by injection of their respective dsRNA in Rhopalosiphum padi and Sitobion avenae. However, the AChE activity changes in the Ace1 knockdown aphids were consistent with the significant transcript level changes of Ace1 genes in these aphids, but not for Ace2. Bioassay results indicated that the suppression of RpAce1 increased its susceptibilities to pirimicarb and malathion, and SaAce1 silencing also increased susceptibility to pirimicarb in S. avenae, whereas the knockdowns of RpAce2 and SaAce2 had a slight effect on their susceptibilities. The knockdown of Ace1 genes also caused significant reductions in fecundity in the aphids of their parental generation. CONCLUSIONS These results suggest that AChE1 is a predominant cholinergic enzyme and is the target of anticholinesterase insecticides in both R. padi and S. avenae. It also plays a non-cholinergic role in fecundity of these aphids. AChE2 may also be important for the toxicological function, although its importance appeared to be lower than that of AChE1. © 2014 Society of Chemical Industry

This study examined the acetylcholinesterase 1 gene (AChE1) in Plutella xylostella strains with different sensitivities to acephate. Multiple haplotypes of the gene were found in the field-collected strains including distinct haplotypes carrying one or both previously reported mutations (A298S and G324A). Moreover, sequencing results indicated the presence of duplicated copies of the gene in the field-collected strains. No correlation was found between copy numbers of AChE1 and levels of resistance to acephate suggesting that extensive AChE1 duplication is not a major resistance factor at least in some P. xylostella strains. Proportions of the A298S and G324A mutations showed no correlation with levels of resistance to acephate. This suggests that acephate resistance of P. xylostella is complex and cannot be evaluated based on the AChE1 copy number or proportions of the resistance mutations alone.

Two peracylated β-cyclodextrin derivatives, 2,3,6-tri-O-valeryl-β-cyclodextrin and 2,3,6-tri-O-octanoyl-β-cyclodextrin, were synthesized and used as capillary gas chromatography (CGC) chiral stationary phases. The two peracylated β-cyclodextrin derivatives showed enantioseparation abilities to some chiral compounds tested, such as enantiomers of methyl 2-chloropropionate, 5-hydroxy-4, 4-dimethyl-dihydrofuran-2-one, and methyl 2-hydroxypropionate. As CGC chiral stationary phases, the two peracylated β-cyclodextrin derivatives showed excellent enantioseparation abilities to methyl 2-hydroxypropionate (i.e., methyl lactate). On 2,3,6-tri-O-valeryl-β-cyclodextrin, the enantiomer separation factor and resolution of methyl lactate were 1.21 and 6.08, respectively, and on 2,3,6-tri-O-octanoyl-β-cyclodextrin, 1.22 and 5.49, respectively. The enantioseparation results of methyl lactate on the two peracylated β-cyclodextrin derivatives were better than on most reported β-cyclodextrin derivatives, including a commercial chiral column of permethylated β-cyclodextrin tested in this study. Results suggest that CGC methods using 2,3,6-tri-O-valeryl-β-cyclodextrin or 2,3,6-tri-O-octanoyl-β-cyclodextrin as chiral stationary phase could be used for enantioseparation and evaluating the enantiomeric optical purity of methyl lactate.

The western flower thrips, Frankliniella occidentalis (Pergande), is difficult to control because of high insecticide resistance. In this study, susceptibility to major insecticides was examined in two Japanese strains (H-1 and KC) and a Chinese strain (BJ) using a leaf-dipping method. All three strains were resistant to permethrin and acetamiprid at agriculturally recommended doses. The median lethal concentration (LC50) for acetamiprid was 1720 ppm in strain H-1, 4780 ppm in strain KC and >6680 ppm in strain BJ. In the presence of piperonyl butoxide, an inhibitor of cytochrome P450 monooxygenases, the LC50 for acetamiprid was 312 ppm in strain H-1, 837 ppm in strain KC and 1250 ppm in strain BJ. These results suggested that metabolism by cytochrome P450 monooxygenases is involved in acetamiprid resistance in these strains, though other factors also seem to play a role. Furthermore, cDNA cloning of the nicotinic acetylcholine receptor (nAChR) subunits was performed using degenerate primers, and the presence or absence of a point mutation in nAChR β1 was confirmed. The R81T mutation that had been reported in Myzus persicae (Sulzer) nAChR β1 was not found in F. occidentalis strains tested in this study.

The gene expression and activity of (+)-δ-cadinene synthase during cotton development and in response to stress, as well as the spatial and temporal pattern of sesquiterpene biosynthesis, constitute one of chemical defense mechanisms in cotton plants. In order to explore the effects of omethoate on the cotton defense in relation to (+)-δ-cadinene synthase and gossypol, effects of omethoate treatments on activity of (+)-δ-cadinene synthase and gossypol content in cotton seedlings were investigated. Cotton seedlings treated with 400 mg L−1 omethoate exhibited a significant decrease in the specific activity of (+)-δ-cadinene synthase from 12 to 120 h after treating when compared to the untreated control; significantly lower (+)-δ-cadinene synthase activity was also observed in cotton seedlings treated with 200 mg L−1 omethoate from 72 to 120 h after treating; but for cotton seedlings treated with 100 mg L−1 omethoate, from 12 to 120 h after treating, no significant changes were observed for activity of (+)-δ-cadinene synthase. The gossypol content in cotton seedlings treated with 100, 200 or 400 mg L−1 omethoate for different time periods showed no significant changes compared to that of the control. These results indicated that the activity of (+)-δ-cadinene synthase in cotton seedlings in responses to exposure of omethoate at three concentrations for different time periods followed dose- and time-dependent responses to omethoate exposure. With omethoate as a chemical stress factor for cotton seedlings, the cotton defense in relation to the activity of (+)-δ-cadinene synthase in cotton seedlings may be affected by omethoate application.