S431F mutation on AChE1 and overexpression of P450 genes confer high pirimicarb resistance in Sitobion miscanthi.

Sitobion miscanthi is a destructive wheat pest responsible for significant wheat yield losses. Pirimicarb, one of the most important representatives of N, N-dimethylcarbamate insecticides, is widely used to control wheat aphids. In present work, heterozygous S431F mutation of acetylcholinesterase 1 (AChE1) was identified and verified in three pirimicarb-resistant S. miscanthi populations (two field populations (HA and HS, >955.8-fold) and one lab-selected population (PirR, 486.1-fold)), which has not been reported in S. miscanthi yet. The molecular docking results revealed that AChE1 containing the S431F mutation of S. miscanthi (SmAChE1S431F) showed higher free binding energy to three insecticides (pirimicarb, omethoate, and methomyl) than wild-type AChE1 of S. miscanthi (SmAChE1). Enzyme kinetic and inhibition experiments showed that the recombinant SmAChE1S431F was more insensitive to pirimicarb and omethoate than the recombinant SmAChE1. Furthermore, two overexpression P450 genes (CYP6K1 and CYP6A14) associated with pirimicarb resistance of S. miscanthi were verified by RNAi. These results suggested both target alteration and enhanced metabolism contributed to high pirimicarb resistance of S. miscanthi in the field and laboratory. These findings lay a foundation for further elucidating the mechanism of pirimicarb resistance in S. miscanthi, and have important implications for the resistance management of S. miscanthi control.

microRNA-3037 targeting CYP6CY2 confers imidacloprid resistance to Sitobion miscanthi (Takahashi).

The wheat aphid Sitobion miscanthi is a dominant and destructive pest in agricultural production. Insecticides are the main substances used for effective control of wheat aphids. However, their extensive application has caused severe resistance of wheat aphids to some insecticides; therefore, exploring resistance mechanisms is essential for wheat aphid management. In the present study, CYP6CY2, a new P450 gene, was isolated and overexpressed in the imidacloprid-resistant strain (SM-R) compared to the imidacloprid-susceptible strain (SM-S). The increased sensitivity of S. miscanthi to imidacloprid after knockdown of CYP6CY2 indicates that it could be associated with imidacloprid resistance. Subsequently, the posttranscriptional regulation of CYP6CY2 in the 3' UTR by miR-3037 was confirmed, and CYP6CY2 participated in imidacloprid resistance. This finding is critical for determining the role of P450 in relation to the resistance of S. miscanthi to imidacloprid. It is of great significance to understand this regulatory mechanism of P450 expression in the resistance of S. miscanthi to neonicotinoids.

Hybridisation has shaped a recent radiation of grass-feeding aphids.

BACKGROUND: Aphids are common crop pests. These insects reproduce by facultative parthenogenesis involving several rounds of clonal reproduction interspersed with an occasional sexual cycle. Furthermore, clonal aphids give birth to live young that are already pregnant. These qualities enable rapid population growth and have facilitated the colonisation of crops globally. In several cases, so-called "super clones" have come to dominate agricultural systems. However, the extent to which the sexual stage of the aphid life cycle has shaped global pest populations has remained unclear, as have the origins of successful lineages. Here, we used chromosome-scale genome assemblies to disentangle the evolution of two global pests of cereals-the English (Sitobion avenae) and Indian (Sitobion miscanthi) grain aphids. RESULTS: Genome-wide divergence between S. avenae and S. miscanthi is low. Moreover, comparison of haplotype-resolved assemblies revealed that the S. miscanthi isolate used for genome sequencing is likely a hybrid, with one of its diploid genome copies closely related to S. avenae (~ 0.5% divergence) and the other substantially more divergent (> 1%). Population genomics analyses of UK and China grain aphids showed that S. avenae and S. miscanthi are part of a cryptic species complex with many highly differentiated lineages that predate the origins of agriculture. The complex consists of hybrid lineages that display a tangled history of hybridisation and genetic introgression. CONCLUSIONS: Our analyses reveal that hybridisation has substantially contributed to grain aphid diversity, and hence, to the evolutionary potential of this important pest species. Furthermore, we propose that aphids are particularly well placed to exploit hybridisation events via the rapid propagation of live-born "frozen hybrids" via asexual reproduction, increasing the likelihood of hybrid lineage formation.

CYP4CJ6-mediated resistance to two neonicotinoid insecticides in Sitobion miscanthi (Takahashi).

The wheat aphid Sitobion miscanthi (CWA) is an important harmful pest in wheat fields. Insecticide application is the main method to effectively control wheat aphids. However, CWA has developed resistance to some insecticides due to its extensive application, and understanding resistance mechanisms is crucial for the management of CWA. In our study, a new P450 gene, CYP4CJ6, was identified from CWA and showed a positive response to imidacloprid and thiamethoxam. Transcription of CYP4CJ6 was significantly induced by both imidacloprid and thiamethoxam, and overexpression of CYP4CJ6 in the imidacloprid-resistant strain was also observed. The sensitivity of CWA to these two insecticides was increased after the knockdown of CYP4CJ6. These results indicated that CYP4CJ6 could be associated with CWA resistance to imidacloprid and thiamethoxam. Subsequently, the posttranscriptional regulatory mechanism was assessed, and miR-316 was confirmed to participate in the posttranscriptional regulation of CYP4CJ6. These results are crucial for clarifying the roles of P450 in the resistance of CWA to insecticides.

MicroRNA-263b confers imidacloprid resistance in Sitobion miscanthi (Takahashi) by regulating the expression of the nAChRß1 subunit.

The Chinese wheat aphid Sitobion miscanthi (CWA) is an important harmful pest in wheat fields. Imidacloprid plays a critical role in controlling pests with sucking mouthparts. However, imidacloprid-resistant pests have been observed after insecticide overuse. Point mutations and low expression levels of the nicotinic acetylcholine receptor ß1 (nAchRß1) subunit are the main imidacloprid-resistant mechanisms. However, the regulatory mechanism underlying nAChRß1 subunit expression is poorly understood. In this study, a target of miR-263b was isolated from the 5'UTR of the nAchRß1 subunit in the CWA. Low expression levels were found in the imidacloprid-resistant strain CWA. Luciferase reporter assays showed that miR-263b could combine with the 5'UTR of the nAChRß1 subunit and suppress its expression by binding to a site in the CWA. Aphids treated with the miR-263b agomir exhibited a significantly reduced abundance of the nAchRß1 subunit and increased imidacloprid resistance. In contrast, aphids treated with the miR-263b antagomir exhibited significantly increased nAchRß1 subunit abundance and decreased imidacloprid resistance. These results provide a basis for an improved understanding of the posttranscriptional regulatory mechanism of the nAChRß1 subunit and further elucidate the function of miRNAs in regulating susceptibility to imidacloprid in the CWA. These results provide a better understanding of the mechanisms of posttranscriptional regulation of nAChRß1 and will be helpful for further studies on the role of miRNAs in the regulation of nAChRß1 subunit resistance in homopteran pests.

Identification of differentially expressed microRNAs under imidacloprid exposure in Sitobion miscanthi.

Imidacloprid is a neonicotinoid that targets sucking pests, such as aphids and the green leaf bug and has been widely applied in wheat fields to control wheat aphids in China. To investigate the involvement of miRNAs in imidacloprid resistance, we sequenced small RNA libraries of Sitobion miscanthi Fabricius, across two different treatments using Illumina short-read sequencing technology. As a result, 265 microRNAs (miRNAs), of which 242 were known and 23 were novel, were identified. Quantitative analysis of miRNA levels showed that 23 miRNAs were significantly up-regulated, and 54 miRNAs were significantly down-regulated in the nymphs of S. miscanthi treated with imidacloprid in comparison with those of the control. Modulation of the abundances of differentially expressed miRNAs, smi-miR-316, smi-miR-1000, and smi-miR-iab-4 by the addition of the corresponding antagomir/inhibitor to the artificial diet significantly changed the susceptibility of S. miscanthi to imidacloprid. Subsequently, the post-transcriptional regulatory mechanism was conducted, smi-miR-278 and smi-miR-316 were confirmed to be participated in the post-transcriptional regulation of nAChRα1A and CYP4CJ6, respectively. The results suggested that miRNAs differentially expressed in response to imidacloprid could play a critical regulatory role in the metabolism of S. miscanthi to imidacloprid.

High Spatial Genetic Structure and Genetic Diversity in Chinese Populations of Sitobion miscanthi (Hemiptera: Aphididae).

The wheat aphid, Sitobion miscanthi Takahashi, a serious wheat pest, was previously considered to be highly migratory and anholocyclic in China. We recorded 69 alleles and 346 multilocus genotypes among 708 aphid individuals from 12 populations in China using 5 microsatellite loci. This genotypic diversity indicates that at least some holocyclic lineages exist. Bayesian clustering analysis revealed that there are two differentiated genetic groups of S. misanthi, one northern and one southern, in China. Principal coordinates analysis of population genetic distance, pairwise F(ST)'s, and network analysis of individual minimum spanning distance also supported the division. Low levels of migration were detected between the northern and southern sampling sites, but the high genetic differentiation does not support the hypothesis S. miscanthi overwinters in the south and migrates to the north in the spring annually.

Overexpression of SmUGGT1 Confers Imidacloprid Resistance to Sitobion miscanthi (Takahashi).

Sitobion miscanthi, the main species of wheat aphids, is one kind of harmful pest. Chemical insecticides are the important agrochemical products to effectively control wheat aphids. However, the broad application has led to serious resistance of pests to several insecticides, and understanding insecticide resistance mechanisms is critical for integrated pest management. In this study, SmUGGT1, a new uridine diphosphate (UDP)-glycosyltransferase (UGT) gene, was cloned and more strongly expressed in the SM-R (the resistant strain to imidacloprid) than in the SM-S (the susceptible strain to imidacloprid). The increased susceptibility to imidacloprid was observed after silencing SmUGGT1, indicating that it can be related to the resistance to imidacloprid. Subsequently, SmUGGT1 regulated post-transcriptionally in the coding sequences (CDs) by miR-81 was verified and involved in the resistance to imidacloprid in S. miscanthi. This finding is crucial in the roles of UGT involved in insecticide resistance management in pests.

Seed Coating with Thiamethoxam-Induced Plant Volatiles Mediates the Olfactory Behavior of Sitobion miscanthi.

Pesticides can induce target plants to release odors that are attractive or repellent to their herbivore insects. But, to date, the activity of volatile organic compounds (VOCs), singly or as mixtures, which play a crucial role in the olfactory behavior of herbivore insects, remains unclear. The objective of our research was to investigate the impact of thiamethoxam (TMX), a pesticide, on the emission of odors by wheat plants, and how these odors influence the behavior of grain aphids (Sitobion miscanthi). S. miscanthi showed a greater repellent response to the volatiles emitted by Thx-induced plants compared to those emitted by uncoated plants. Using gas chromatography-mass spectrometry (GCMS), we discovered that TMX greatly induced the release of VOCs in wheat plants. For instance, the levels of Bornyl acetate, 2-Oxepanone, Methyl acrylate, Cyclohexene, α-Pinene, and 1-Nonanol in coated wheat plants were significantly higher as compared to uncoated wheat plants. Moreover, varying concentrations also had an impact on the olfactory behavior of S. miscanthi. For instance, Cyclohexene exhibited clear attractiveness to aphids at concentrations of 100 µL/mL, whereas it displayed evident repellent properties at concentrations of 1 µL/mL and 10 µL/mL. These new findings demonstrate how TMX-induced VOCs affect the behavior of S. miscanthi and could help in developing innovative approaches to manage aphids by manipulating the emission of plant volatiles. Furthermore, these findings can also be utilized to evaluate substances that either attract or repel aphids, with the aim of implementing early monitoring and environmentally friendly methods to manage aphids, while simultaneously impeding the spread of viruses.

Fitness Cost of the Field-Evolved Resistance to Sulfoxaflor and Multi-Insecticide Resistance of the Wheat Aphid Sitobion miscanthi (Takahashi).

Sulfoxaflor belongs to a new class of insecticides that is effective against many sap-feeding pests. In this study on Sitobion miscanthi (Takahashi) (i.e., the predominant wheat pest), a highly sulfoxaflor-resistant (SulR) population was obtained from a field. Its resistance to the other seven insecticides and its biological fitness were analyzed using a leaf-dip method and a two-sex life table approach, respectively. Compared with the relatively susceptible (SS) population, the SulR population was highly resistant to sulfoxaflor, with a relative insecticide resistance ratio (RR) of 199.8 and was moderately resistant to beta-cypermethrin (RR = 14.5) and bifenthrin (RR = 42.1) but exhibited low resistance to chlorpyrifos (RR = 5.7). Additionally, the SulR population had a relative fitness of 0.73, with a significantly prolonged developmental period as well as a lower survival rate and poorer reproductive performance than the SS population. In conclusion, our results suggest that S. miscanthi populations that are highly resistant to sulfoxaflor exist in the field. The possibility that insects may develop multi-resistance between sulfoxaflor and pyrethroids is a concern. Furthermore, the high sulfoxaflor resistance of S. miscanthi was accompanied by a considerable fitness cost. The study data may be useful for improving the rational use of insecticides and for exploring novel insecticide resistance mechanisms.

Population dynamics of migrant wheat aphids in China's main wheat production region and their interactions with bacterial symbionts.

Sitobion miscanthi, Rhopalosiphum padi, and Schizaphis graminum are the three main pests in Chinese wheat-producing regions. In 2020, they are classified into the Chinese Class I list of agricultural diseases and pests, due to their severe harm to wheat plantings. S. miscanthi, R. padi, and S. graminum are migrant pests, and understanding their migration patterns and simulating their migration trajectories would improve forecasting and controlling them. Furthermore, the bacterial community of the migrant wheat aphid is also less known. In this study, we employed a suction trap to uncover the migration patterns of the three wheat aphid species in Yuanyang county, Henan province, during 2018 to 2020. And then the migration trajectories of S. miscanthi and R. padi were simulated using the NOAA HYSPLIT model. The interactions between wheat aphids and bacteria were further revealed by specific PCR and 16S rRNA amplicon sequencing. The results showed that the population dynamics of migrant wheat aphids was varied. Most of the trapped samples were identified to be R. padi, and S. graminum was the least collected sample. Typically, R. padi had two migration peaks in the 3 years, whereas S. miscanthi and S. graminum only exhibited one migration peak in 2018 and 2019. Moreover, the aphid migration trajectories varied over the years. Generally, the aphids originated from the south and migrated to the north. Herein, the infections of three main aphid facultative bacterial symbionts, Serratia symbiotica, Hamiltonella defensa, and Regiella insercticola, were detected in S. miscanthi and R. padi with specific PCR. Rickettsiella, Arsenophonus, Rickettsia, and Wolbachia were further identified with 16S rRNA amplicon sequencing. Biomarker searching indicated that Arsenophonus was significantly enriched in R. padi. Furthermore, diversity analyses showed that the bacterial community of R. padi had a higher richness and evenness than that of S. miscanthi. In conclusion, this study expands our knowledge about the migration patterns of aphids in the main wheat plant region of China and reveals the interactions between bacterial symbionts and migrant aphids.

Sitobion miscanthi L type symbiont enhances the fitness and feeding behavior of the host grain aphid.

BACKGROUND: Symbiotic bacteria affect physiology and ecology of insect hosts. The Sitobion miscanthi L type symbiont (SMLS) is a recently discovered and widely distributed secondary symbiont in the grain aphid Sitobion miscanthi Takahashi in China. RESULTS: In this study, SMLS-infected (SI) and SMLS-uninfected (SU) aphid strains were obtained from field population. The artificially SMLS-re-infected (SRI) strain was established by injecting SU aphids with the SI strain hemolymph containing SMLS. The SRI and SU strains had identical genetic backgrounds and similar microbial community structures. Compared with the SU strain, adult longevity, survival rate, and fecundity were significantly greater in the SRI strain (biological fitness of 1.48). Moreover, the SRI strain spent more time ingesting phloem than the SU strain. A comparative transcriptome analysis indicated that reproduction- and longevity-related genes were more highly expressed in the SRI strain than in the SU strain. CONCLUSION: The findings indicated that the infection with SMLS enhanced the Sitobion miscanthi fitness and feeding behavior. The beneficial effect of the SMLS on hosts could explain why it frequently infects the field populations in the grain aphid Sitobion miscanthi Takahashi in China. © 2022 Society of Chemical Industry.

Two salivary proteins Sm10 and SmC002 from grain aphid Sitobion miscanthi modulate wheat defense and enhance aphid performance.

The grain aphid Sitobion miscanthi is a serious pest of wheat that causes severe economic damage by sucking phloem sap and transmitting plant viruses. Here, two putative salivary effector homologs from S. miscanthi (Sm10 and SmC002) were selected based on sequence similarity to other characterized aphid candidate effectors. These effectors were then delivered into wheat cells separately via the type III secretion system of Pseudomonas fluorescens to elucidate their functions in the regulation of plant defenses and host fitness. The results showed that the delivery of either Sm10 or SmC002 into wheat plants significantly suppressed callose deposition and affected the transcript levels of callose synthase genes. The expression levels of salicylic acid (SA)-associated defense genes were upregulated significantly in wheat leaves carrying either Sm10 or SmC002. Moreover, LC-MS/MS analysis revealed that wheat SA levels significantly increased after the delivery of the two effectors. The results of aphid bioassays conducted on the wheat plants carrying Sm10 or SmC002 showed significant increases in the survival and fecundity of S. miscanthi. This study demonstrated that the Sm10 and SmC002 salivary effectors of S. miscanthi enhanced host plant susceptibility and benefited S. miscanthi performance by regulating wheat defense signaling pathways.

Activation of Wheat Defense Response by Buchnera aphidicola-Derived Small Chaperone Protein GroES in Wheat Aphid Saliva.

Salivary proteins secreted by aphids during feeding play an important role in regulating the plant defense response. We used mass spectrometry to identify 155 proteins from the wheat aphid, Sitobion miscanthi, among which 44 proteins were derived from the primary symbiont, Buchnera aphidicola. GroES, which is a highly abundant molecular chaperone that binds to GroEL, was detected in saliva. In vitro injection of purified GroES protein and overexpression of GroES in wheat leaves verified that GroES induced hydrogen peroxide accumulation and callose deposition in wheat and further activated the plant salic acid and jasmonic acid defense pathways. Our findings indicate that plants may have evolved new strategies to detect aphid attack and trigger defense responses by recognizing proteins derived from B. aphidicola, which is present in almost all aphid species.

Proteomic and Transcriptomic Analysis for Identification of Endosymbiotic Bacteria Associated with BYDV Transmission Efficiency by Sitobion miscanthi.

Sitobion miscanthi, an important viral vector of barley yellow dwarf virus (BYDV), is also symbiotically associated with endosymbionts, but little is known about the interactions between endosymbionts, aphid and BYDV. Therefore, two aphids' geographic populations, differing in their BYDV transmission efficiency, after characterizing their endosymbionts, were treated with antibiotics to investigate how changes in the composition of their endosymbiont population affected BYDV transmission efficiency. After antibiotic treatment, Rickettsia was eliminated from two geographic populations. BYDV transmission efficiency by STY geographic population dropped significantly, by -44.2% with ampicillin and -25.01% with rifampicin, but HDZ geographic population decreased by only 14.19% with ampicillin and 23.88% with rifampicin. Transcriptomic analysis showed that the number of DEGs related to the immune system, carbohydrate metabolism and lipid metabolism did increase in the STY rifampicin treatment, while replication and repair, glycan biosynthesis and metabolism increased in the STY ampicillin treatment. Proteomic analysis showed that the abundance of symbionin symL, nascent polypeptide-associated complex subunit alpha and proteasome differed significantly between the two geographic populations. We found that the endosymbionts can mediate vector viral transmission. They should therefore be included in investigations into aphid-virus interactions and plant disease epidemiology. Our findings should also help with the development of strategies to prevent virus transmission.

Identification of a Novel Densovirus in Aphid, and Uncovering the Possible Antiviral Process During Its Infection.

Densoviruses (DVs) are single-stranded DNA viruses and exclusively happen in invertebrates. Most of DVs reported in insects are pathogenic to their native hosts, however, no pathogenic effect of them has been examined in vertebrates. Hence, DVs are the potential agents used in pest managements. Aphids are the primary vectors of plant viruses. In this study, we identified a novel DV in Chinese Sitobion miscanthi population, provisionally named "Sitobion miscanthi densovirus" (SmDV). Taxonomically, SmDV belongs to genus Hemiambidensovirus. In S. miscanthi, SmDV is hosted in diverse cells and can be horizontally transmitted via wheat feeding. Subject to SmDV, aphids activate their intrinsic antiviral autophagy pathway. Grouped with ascorbate and aldarate metabolism, chlorophyll metabolism, p450 related drug metabolism, and retinoid metabolism, aphids form a complex immune network response to the infection of SmDV. Obviously, it works as elder aphids still alive even they contain the highest examined concentration of SmDV. This study provides a foundation for the identifications of novel DVs, and further improves the understanding of the molecular interactions between insects and DVs.

The Reciprocal Effect of Elevated CO2 and Drought on Wheat-Aphid Interaction System.

Due to the rising concentration of atmospheric CO2, climate change is predicted to intensify episodes of drought. However, our understanding of how combined environmental conditions, such as elevated CO2 and drought together, will influence crop-insect interactions is limited. In the present study, the direct effects of combined elevated CO2 and drought stress on wheat (Triticum aestivum) nutritional quality and insect resistance, and the indirect effects on the grain aphid (Sitobion miscanthi) performance were investigated. The results showed that, in wheat, elevated CO2 alleviated low water content caused by drought stress. Both elevated CO2 and drought promoted soluble sugar accumulation. However, opposite effects were found on amino acid content-it was decreased by elevated CO2 and increased by drought. Further, elevated CO2 down-regulated the jasmonic acid (JA) -dependent defense, but up-regulated the salicylic acid (SA)-dependent defense. Meanwhile, drought enhanced abscisic acid accumulation that promoted the JA-dependent defense. For aphids, their feeding always induced phytohormone resistance in wheat under either elevated CO2 or drought conditions. Similar aphid performance between the control and the combined two factors were observed. We concluded that the aphid damage suffered by wheat in the future under combined elevated CO2 and drier conditions tends to maintain the status quo. We further revealed the mechanism by which it happened from the aspects of wheat water content, nutrition, and resistance to aphids.

Silencing an aphid-specific gene SmDSR33 for aphid control through plant-mediated RNAi in wheat.

Grain aphid (Sitobion miscanthi) is one of the most dominant and devastating insect pests in wheat, which causes substantial losses to wheat production each year. Engineering transgenic plants expressing double strand RNA (dsRNA) targeting an insect-specific gene has been demonstrated to provide an alternative environmentally friendly strategy for aphid management through plant-mediated RNA interference (RNAi). Here we identified and characterized a novel potential RNAi target gene (SmDSR33) which was a gene encoding a putative salivary protein. We then generated stable transgenic wheat lines expressing dsRNA for targeted silencing of SmDSR33 in grain aphids through plant-mediated RNAi. After feeding on transgenic wheat plants expressing SmDSR33-dsRNA, the attenuated expression levels of SmDSR33 in aphids were observed when compared to aphids feeding on wild-type plants. The decreased SmDSR33 expression levels thus resulted in significantly reduced fecundity and survival, and decreased reproduction of aphids. We also observed altered aphid feeding behaviors such as longer duration of intercellular stylet pathway and shorter duration of passive ingestion in electroneurography assays. Furthermore, both the surviving aphids and their offspring exhibited decreased survival rates and fecundity, indicating that the silencing effect could be persistent and transgenerational in grain aphids. The results demonstrated that SmDSR33 can be selected as an effective RNAi target for wheat aphid control. Silencing of an essential salivary protein gene involved in ingestion through plant-mediated RNAi could be exploited as an effective strategy for aphid control in wheat.

Screening and Evaluation for Antixenosis Resistance in Wheat Accessions and Varieties to Grain Aphid, Sitobion miscanthi (Takahashi) (Hemiptera: Aphididae).

The grain aphid, Sitobion miscanthi causes serious damage by removing nutritional content from wheat plants and transmitting viral diseases. The use of resistant wheat cultivars is an effective method of aphid management. To identify S. miscanthi resistant cultivars, preliminary antixenosis resistance screening was conducted on 112 Ethiopian and 21 Chinese wheat accessions and varieties along with bioassay to test for further antixenosis resistance, identification of aphid feeding behavior using electrical penetration graph (EPG), and imaging of leaf trichome densities using a 3D microscope. According to antixenosis resistance screening, one highly-resistant, 25 moderately-resistant, and 38 slightly-resistant wheat cultivars to S. miscanthi were identified. Aphid choice tests showed that Luxuan266, 243726, and 213312 were the least preferred after 12, 24, 48, and 72 h of S. miscanthi release. Longer duration of Np, longer time to first probe, and shorter duration of E2 waveforms were recorded in Lunxuan266, 243726, and 213312 than in Beijing 837. The trichome density on adaxial and abaxial leaf surfaces of Lunxuan266, 243726 and 213312 was significantly higher than on those of Beijing 837. We concluded that Lunxuan266, 243726, and 213312 were antixenosis resistant to S. miscanthi based on the choice test, EPG results, and leaf trichome densities.

Reduced insecticide susceptibility of the wheat aphid Sitobion miscanthi after infection by the secondary bacterial symbiont Hamiltonella defensa.

BACKGROUND: Bacterial symbionts in insects, especially aphids, have a major influence on host adaptation. We previously showed that infection with the secondary symbiont Hamiltonella defensa increases the fitness of the wheat aphid Sitobion miscanthi, yielding increases in fitness parameters such as adult weight and offspring number. However, whether H. defensa affects the sensitivity of host aphids to insecticides remains unknown. RESULTS: We tested the effects of H. defensa on host aphid susceptibility to the insecticides chlorpyrifos methyl, imidacloprid, cyantraniliprole and acetamiprid. Our results showed that compared with Hamiltonella-free aphid clones, Hamiltonella-infected aphid clones exhibited lower sensitivity to most of the tested insecticides at low concentrations. Quantitative polymerase chain reaction showed that the density of H. defensa in the infected clones was slightly decreased at 24 h but then sharply increased until the late stage after treatment with the different insecticides. H. defensa in the host aphids was detected by fluorescence in situ hybridization and was localized to the aphid hindgut. Levels of the detoxification enzymes acetylcholinesterase, glutathione transferase and carboxylesterase were significantly higher in Hamiltonella-infected clones than in Hamiltonella-free clones. CONCLUSIONS: The findings indicated that infection with H. defensa reduced aphid susceptibility to the investigated insecticides at low concentrations, potentially by increasing detoxification enzyme activity in the host. Therefore, symbiont-mediated insecticide resistance should be taken into account when performing resistance-monitoring studies. Studies of symbiont-mediated insecticide resistance may enhance our understanding of the emergence of insecticide resistance in agricultural systems. © 2020 Society of Chemical Industry.