Pyridine Derivatives as Insecticides: Part 6. Design, Synthesis, Molecular Docking, and Insecticidal Activity of 3-(Substituted)methylthio-5,6,7,8-tetrahydroisoquinoline-4-carbonitriles Toward Aphis gossypii (Glover, 1887).

Three new series of 3-(substituted)methylthio-4-cyano-5,6,7,8-tetrahydroisoquinolines were designed and synthesized starting from readily available materials, 7-acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-pyridyl, 3-pyridyl, phenyl, 4-methoxyphenyl, or 4-chlorophenyl)-5,6,7,8-tetrahydrosoquinoline-3(2H)-thiones 2a-e in high yields and very pure states. Thus, compounds 2a-e were reacted with some chloro reagents, namely, N-aryl-2-chloroacetamides 3a-f and N-(naphthalen-2-yl)-2-chloroacetamide (3g) under mild basic conditions to give the first two series of the target compounds, 3-(N-aryl)carbamoylmethylthio-5,6,7,8-tetrahydroisoquinoline-4-carbonitriles 4a-l and 5a-e, respectively. Reaction of compounds 2d,e with ethyl chloroacetate under the same conditions gave the other series, 3-ethoxycarbonyl-methylthio-5,6,7,8-tetrahydroisoquinoline-4-carbonitriles 6d,e. Structural formulas of all of the new compounds were elucidated and confirmed by elemental and spectral analyses. The insecticidal activity of all synthesized 5,6,7,8-tetrahydrosoquinolines toward the nymphs and adults of Aphis gossypii were screened. The results revealed the promising insecticidal activity of some tested compounds. Moreover, the structure-activity relationships as well as molecular docking of some representative compounds were evaluated.

Nanomaterial-Encapsulated dsRNA-Targeting Chitin Pathway─A Potential Efficient and Eco-Friendly Strategy against Cotton Aphid, Aphis gossypii (Hemiptera: Aphididae).

The r-strategy pests are very challenging to effectively control because of their rapid population growth and strong resurgence potential and are more prone to developing pesticide resistance. As a typical r-strategy pest, the cosmopolitan cotton aphid, Aphis gossypii Glover, seriously impacts the growth and production of cucurbits and cotton. The present study developed a SPc/double-stranded RNA (dsRNA)/botanical strategy to enhance the control efficacy of A. gossypii. The results demonstrated that the expression of two chitin pathway genes AgCHS2 and AgHK2 notably changed in A. gossypii after treated by three botanical pesticides, 1% azadirachtin, 1% matrine, and 5% eucalyptol. SPc nanocarrier could significantly enhance the environmental stability, cuticle penetration, and interference efficiency of dsRNA products. The SPc/dsRNA/botanical complex could obviously increase the mortality of A. gossypii in both laboratory and greenhouse conditions. This study provides an eco-friendly control technique for enhanced mortality of A. gossypii and lower application of chemical pesticides. Given the conservative feature of chitin pathway genes, this strategy would also shed light on the promotion of management strategies against other r-strategy pests using dsRNA/botanical complex nanopesticides.

Evolution, identification, evaluation, and characterization of a stable salinity tolerant sugarcane variety CoG 7.

From the fluff generated during 2005, after the preliminary experiments (2005-2007), a promising clone G2005047 has been identified. It showed moderate resistance to red rot (3.6 on a 9-scale scoring system), less susceptibility to shoot borer (13.25%) and internode borers (25.35%), and resistance to woolly aphid (0%). In the Advanced Yield Trials (2008-2011), it showed advantages over check for cane yield (CY) (11.79%), commercial cane sugar percent (CCSP) (0.35%), and sugar yield (SY) (20.33%). To ascertain its large-scale cultivation suitability, it has experimented under adaptive research trials (2012-2014) at farmers' fields. It exhibited 18.04%, 1.27%, and 19.55% supremacy over the check Co 86032 for CY, CCSP, and SY respectively. The stability of G2005047 under salinity was ascertained through a multi-environment-based experiment (2015-2017). AMMI (Additive Main-effects and Multiplicative Interactions) and GGE (Genotype × Genotype-Environment interaction) biplots were utilized. ANOVA revealed that the genotypic variation exerted the most significant effect followed by genotype × environment interaction and environment. G2005047 had the highest mean values for yield and quality traits with minimal ASV (AMMI stability value) (2.38:CY; 0.57: CCSP; & 0.58:SY) indicating its good-yielding ability and stability. AMMI I, AMMI II, and GGE biplots confirmed the stability of G2005047. In the jaggery quality assessment trials (2018 and 2019), it yielded 37.1% increased jaggery over the check. Also, the clone G2005047, exhibited moderate resistance to red rot disease, less susceptibility to shoot borer (13.25%) and internode borer (25.35%), and resistance against sugarcane woolly aphid (SWA). Due to supremacy for yield, quality, better performance under salinized situations, and tolerance to disease and pests, the clone G2005047 was released as a variety CoG 7 in 2022.

The role of OR5, which is highly expressed in the winged grain aphid Sitobion miscanthi, in specific recognition of EBF.

Winged parthenogenetic aphids are mainly responsible for migration and dispersal. Aphid alarm pheromone (E)-ß-Farnesene (EBF) has dual effects on repelling and stimulating wing differentiation in aphids. Previous studies have shown that the odorant coreceptor SmisOrco is involved in the perception of EBF by S. miscanthi; however, its EBF-specific odorant receptor (OR) and the difference between winged and wingless aphids remain unclear. In this study, the Xenopus oocyte expression system and RNAi technology were used to detect the transmission of EBF signals, and it was found that the olfactory receptor SmisOR5 is an EBF-specific OR in S. miscanthi and is specifically highly expressed in the antennae of winged aphids. Furthermore, when OR5 was silenced with dsRNA, the repellent effect of EBF was weakened, and aphids showed more active aimless movements. Therefore, as a specific OR for EBF, the high expression level of SmisOR5 in winged aphids suggests a molecular basis for its high sensitivity to EBF. This study advances our understanding of the molecular mechanisms of aphid EBF perception and provides novel ideas for effective management and prevention of the migration of winged aphids.

Overexpression of Two Odorant Binding Proteins Confers Chlorpyrifos Resistance in the Green Peach Aphid Myzus persicae.

The green peach aphid, Myzus persicae, is a worldwide agricultural pest. Chlorpyrifos has been widely used to control M. persicae for decades, thus leading to a high resistance to chlorpyrifos. Recent studies have found that insect odorant binding proteins (OBPs) play essential roles in insecticide resistance. However, the potential resistance mechanism underlying the cross-link between aphid OBPs and chlorpyrifos remains unclear. In this study, two OBPs (MperOBP3 and MperOBP7) were found overexpressed in M. persicae chlorpyrifos-resistant strains (CRR) compared to chlorpyrifos-sensitive strains (CSS); furthermore, chlorpyrifos can significantly induce the expression of both OBPs. An in vitro binding assay indicated that both OBPs strongly bind with chlorpyrifos; an in vivo RNAi and toxicity bioassay confirmed silencing either of the two OBPs can increase the susceptibility of aphids to chlorpyrifos, suggesting that overexpression of MperOBP3 and MperOBP7 contributes to the development of resistance of M. persicae to chlorpyrifos. Our findings provide novel insights into insect OBPs-mediated resistance mechanisms.

Hormetic dose response induced by sublethal-dose sulfoxaflor leads to reproductive stimulation of Aphis gossypii.

Aphis gossypii Glover is one of the most agriculturally important phloem-feeding economic pests, causing tremendous loss in crop yield annually. The hormesis is an important cause of A. gossypii resistance formation, population resurgence, and re-outbreak. However, whether the hormesises induced by different insecticides interact mutually remain largely unclear. In the study, four-generation A. gossypii experiment found that the 24-h sublethal-dose (LC20) sulfoxaflor treatment on G0 significantly increased the net reproductive rate (R0) and fecundity of G1 and G2 generation A. gossypii, but it did not significantly affect the fecundity of G3 and G4 individuals. Transcriptomic analyses revealed that the insecticide-induced significant up-regulation of pathways ribosome, energy metabolism, and the DNA replication and reparation might be responsible for the enhancement of fecundity in G1 and G2 A. gossypii. Notably, G0 exposure to LC20 sulfoxaflor followed by G1 exposure to LC30 deltamethrin resulted in a stronger reproductive stimulation than sulfoxaflor or deltamethrin exposure alone. Our findings provide valuable reference for optimizing sulfoxaflor application in integrated pest management strategies.

Secondary metabolites in Cordyceps javanica with insecticidal potential.

Cordyceps javanica has been registered as a fungal insecticide in several countries. However, little is known about whether metabolic toxins are involved in the insecticidal process. In this research, we assessed the insecticidal activity of the fermentation broth of C. javanica. Myzus persicae mortality differed when exposed to the metabolized C. javanica broths at 3 days post fermentation (DPF) and 5 DPF. Comparison of the metabolic fluid at 3 DPF and 5 DPF revealed a key alkaloid, heteratisine, which was found to have insecticidal activity and acetylcholinesterase (AChE) inhibitory activity. Heteratisine has high insecticidal activity against adult M. persicae, the absolute 50% lethal concentration (LC50) was only 0.2272 mg/L. Heteratisine showed high inhibitory activity on AChE with the 50% maximal inhibitory concentration (IC50) of 76.69 µM. Molecular docking and dynamic simulations showed that heteratisine conjugation occurs at the peripheral anionic site (PAS) of the AChE of M. persicae, leading to suppression of enzyme activity. Heteratisine was rarely found in fungal metabolites, which helps us to understand the complex and elaborate insecticidal mechanism of C. javanica.

Elucidating ß-1,3-Glucanase and Peroxidase Physicochemical Properties of Wheat Cell Wall Defense Mechanism against Diuraphis noxia Infestation.

Wheat plants infested by Russian wheat aphids (RWA) induce a cascade of defense responses, including the hypersensitive responses (HR) and induction of pathogenesis-related (PR) proteins, such as ß-1,3-glucanase and peroxidase (POD). This study aims to characterize the physicochemical properties of cell wall-associated POD and ß-1,3-glucanase and determine their synergism on the cell wall modification during RWASA2-wheat interaction. The susceptible Tugela, moderately resistant Tugela-Dn1, and resistant Tugela-Dn5 cultivars were pregerminated and planted under greenhouse conditions, fertilized 14 days after planting, and irrigated every 3 days. The plants were infested with 20 parthenogenetic individuals of the same RWASA2 clone at the 3-leaf stage, and leaves were harvested at 1 to 14 days post-infestation. The Intercellular wash fluid (IWF) was extracted using vacuum filtration and stored at -20 °C. Leaf residues were crushed into powder and used for cell wall components. POD activity and characterization were determined using 5 mM guaiacol substrate and H2O2, monitoring change in absorbance at 470 nm. ß-1,3-glucanase activity, pH, and temperature optimum conditions were demonstrated by measuring the total reducing sugars in the hydrolysate with DNS reagent using ß-1,3-glucan and ß-1,3-1,4-glucan substrates, measuring the absorbance at 540 nm, and using glucose standard curve. The pH optimum was determined between pH 4 to 9, temperature optimum between 25 and 50 °C, and thermal stability between 30 °C and 70 °C. ß-1,3-glucanase substrate specificity was determined at 25 °C and 40 °C using curdlan and barley ß-1,3-1,4-glucan substrates. Additionally, the ß-1,3-glucanase mode of action was determined using laminaribiose to laminaripentaose. The oligosaccharide hydrolysis product patterns were qualitatively analyzed with thin-layer chromatography (TLC) and quantitatively analyzed with HPLC. The method presented in this study demonstrates a robust approach for infesting wheat with RWA, extracting peroxidase and ß-1,3-glucanase from the cell wall region and their comprehensive biochemical characterization.

Efficacy of novel insecticides against piercing sucking insects and their natural enemies on sweet pepper plants under field conditions.

Piercing sucking pests attacking sweet pepper plants cause significant losses to its yield. Considering the undesirable effects of synthetic pesticides, field studies were conducted to evaluate the impact of new pesticides against piercing sucking insect pests of sweet pepper, as well as, their effects on some predators and pepper yield along two seasons of 2021-2022. The obtained results indicated that all tested pesticides effectively suppressed the sucking insect populations (aphids, white fly, thrips) 1,7,14 and 21 days after treatment along two sprays during two seasons. Imidacloprid proved to be the superior one over all other treatments where it recorded mean reduction% (98.91 and 97.27%) & (94.8 and 95.19%), (86.23 and 76.64%) & (80.92 and 88.55%) and (77.68 and 78.44%) & (90.70 and 68.57%) in white fly, aphids and thrips, respectively at 1st and 2nd sprays at 2021 and 2022 seasons, respectively. As for side effects of tested insecticides on natural enemies, Dimethoate induced the highest decrease (60.85 and 69.33%) & (54.02 and 63.41%), (65.52 and 64.74%) & (59.23 and 58.38%) and (64.24 and 59.48%) & (61.66 and 60.8%) on Chrysoperla carnea, Paederus alfierii and Coccinella spp at 1st and 2nd sprays at 2021 and 2022 seasons, respectively. On contrary, Spintoram induced the lowest effects on Chrysoperla carnea, Paederus alfierii and Coccinella spp, recording decrease percent (25.41 and 19.84%) & (15.02 and 12.50%), (11.94 and 11.24%) (16.99 and 18.02%) and (18.73 and15.07%) & (18.35 and18.38%) at1st and 2nd sprays at 2021 and 2022 seasons, respectively. With respect to the effect of tested insecticides on pepper yield, all tested insecticides increased the yield of green pepper fruits compared with control. Imidacloprid achieved the highest fruit yields along two seasons 6.43 and 6.52 (ton / fed.4200 m2) with increase percent 34.53 and 36.04% in yield over control at 2021 and 2022 seasons, respectively.

A tiny sample rapid visual detection technology for imidacloprid resistance in Aphis gossypii by CRISPR/Cas12a.

Insecticide resistance monitoring is essential for guiding chemical pest control and resistance management policies. Currently, rapid and effective technology for monitoring the resistance of tiny insects in the field is absent. Aphis gossypii Glover is a typical tiny insect, and one of the most frequently reported insecticide-resistant pests. In this study, we established a novel CRISPR/Cas12a-based rapid visual detection approach for detecting the V62I and R81T mutations in the ß1 subunit of the nAChR in A. gossypii, to reflect target-site resistance to imidacloprid. Based on the nAChR ß1 subunit gene in A. gossypii, the V62I/R81T-specific RPA primers and crRNAs were designed, and the ratio of 10 µM/2 µM/10 µM for ssDNA/Cas12a/crRNA was selected as the optimal dosage for the CRISPR reaction, ensuring that Cas12a only accurately recognizes imidacloprid-resistance templates. Our data show that the field populations of resistant insects possessing V62I and R81T mutations to imidacloprid can be accurately identified within one hour using the RPA-CRISPR/Cas12a detection approach under visible blue light at 440-460 nm. The protocol for RPA-CRISPR detection necessitates a single less than 2 mm specimen of A. gossypii tissues to perform RPA-CRISPR detection, and the process only requires a container at 37 °C and a portable blue light at 440-460 nm. Our research represents the first application of RPA-CRISPR technology in insecticide resistance detection, offers a new method for the resistance monitoring of A. gossypii or other tiny insects, helps delay the development of resistance to imidacloprid, improves the sustainability of chemical control, and provides theoretical guidance for managing pest resistance.

The ATP-binding cassette transporter subfamily G member 4 mediates cuticular hydrocarbon transport to regulate drought tolerance in Acyrthosiphon pisum.

ABC transporters are a highly conserved membrane protein class that promote the transport of substances across membranes. Under drought conditions, insects primarily regulate the content of cuticular hydrocarbon (CHC) to retain water and prevent evaporative loss. Involvement of ABC transporter protein G (ABCG) subfamily genes in insect CHC transport has been relatively understudied. In this study, we demonstrated that ABCG4 gene in Acyrthosiphon pisum (ApABCG4) is involved in CHC transport and affects drought tolerance by regulating CHC accumulation. ApABCG4 is strongly expressed in the abdominal cuticle and embryonic stages of A. pisum. Effective silencing of ApABCG4 was achieved using RNAi, and the silencing duration was analyzed. ApABCG4 silencing resulted in a significant decrease in the total and component contents of the CHC and cuticular waxy coatings of A. pisum. Nevertheless, the internal hydrocarbon content remained unchanged. The lack of cuticular hydrocarbons significantly reduced the drought tolerance of A. pisum, shortening its survival time under drought stress. Drought stress caused significant upregulation of ApABCG4. Molecular docking showed that ApABCG4 has a high binding affinity for nine n-alkanes of CHC through electrostatic interactions. These results indicate that ApABCG4 is a novel RNAi target with key applications in aphid biological control.

microRNA maintains nutrient homeostasis in the symbiont-host interaction.

Endosymbionts provide essential nutrients for hosts, promoting growth, development, and reproduction. However, the molecular regulation of nutrient transport from endosymbiont to host is not well understood. Here, we used bioinformatic analysis, RNA-Sequencing, luciferase assays, RNA immunoprecipitation, and in situ hybridization to show that a bacteriocyte-distributed MRP4 gene (multidrug resistance-associated protein 4) is negatively regulated by a host (aphid)-specific microRNA (miR-3024). Targeted metabolomics, microbiome analysis, vitamin B6 (VB6) supplements, 3D modeling/molecular docking, in vitro binding assays (voltage clamp recording and microscale thermophoresis), and functional complementation of Escherichia coli were jointly used to show that the miR-3024/MRP4 axis controls endosymbiont (Serratia)-produced VB6 transport to the host. The supplementation of miR-3024 increased the mortality of aphids, but partial rescue was achieved by providing an external source of VB6. The use of miR-3024 as part of a sustainable aphid pest-control strategy was evaluated by safety assessments in nontarget organisms (pollinators, predators, and entomopathogenic fungi) using virus-induced gene silencing assays and the expression of miR-3024 in transgenic tobacco. The supplementation of miR-3024 suppresses MRP4 expression, restricting the number of membrane channels, inhibiting VB6 transport, and ultimately killing the host. Under aphids facing stress conditions, the endosymbiont titer is decreased, and the VB6 production is also down-regulated, while the aphid's autonomous inhibition of miR-3024 enhances the expression of MRP4 and then increases the VB6 transport which finally ensures the VB6 homeostasis. The results confirm that miR-3024 regulates nutrient transport in the endosymbiont-host system and is a suitable target for sustainable pest control.

Chiral Chemopolymorphism in the Monoterpenes of Pistacia palaestina Leaves and Galls.

Pistacia palaestina Boiss. is a common tree in the Mediterranean maquis. The leaves of this plant accumulate defensive monoterpenes, whose levels greatly increase in galls induced by the aphid Baizongia pistaciae. We previously found a significant chemopolymorphism in monoterpene content among individual trees, but the chirality of these monoterpenes was unknown. Although most plant species specifically accumulate one enantiomeric form of a given compound, P. palaestina individuals display chemopolymorphism in the chirality of the key monoterpenes accumulated. We report here a marked enantiomeric variation for the limonene, α- and ß-pinene, camphene, sabinene, δ-3-carene, and terpene-4-ol content in leaves and galls of nine different naturally growing P. palaestina trees. Interestingly, insect-induced gall monoterpene composition is an augmentation of the specific enantiopolymorphism originally displayed by each individual tree.

Study of Altenia wagneriella (rebel) as a predator associated with Forda marginata from Iran.

BACKGROUND: The larvae of Altenia spp. and gall aphids are known to feed on plants related to Anacardiaceae. This study documents the aphidophagous habit of Altenia wagneriella, which was verified by molecular techniques, subsequently by the gut dissection test, and direct observation. MATERIALS AND METHODS: To identify the moth larvae and adult aphids, two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (Cytb), were amplified by polymerase chain reaction (PCR). Nested PCR with the aphid-specific primer pairs AphidF and AphidR was used to detect aphids in the body of moth larvae. The specificity of the primers was verified by PCR analysis of DNA from moth larvae and adult aphids. RESULTS: The method detected aphids in moth larvae, and a band of approximately 200 bp was amplified from moth larvae feeding on aphids. No cross reactions with moth larvae were observed. In the laboratory, all moth larvae feeding on aphids (Forda marginata) were also PCR positive for aphids. CONCLUSIONS: Gall-inducing insects are microhabitat engineers that manipulate their host to obtain a better nutrient supply, as well as protection from natural enemies and abiotic factors. This is the first recorded instance worldwide of the carnivorous larva of the moth A. wagneriella acting as an aphid predator, as well as the first record of a host insect for this species. Additionally, it is the first effort to molecularly analyze the predator-prey relationship between the moth larvae and the aphids inside the wild pistachio gall.

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.

Two chromosome-level genome assemblies of galling aphids Slavum lentiscoides and Chaetogeoica ovagalla.

Slavum lentiscoides and Chaetogeoica ovagalla are two aphid species from the subtribe Fordina of Fordini within the subfamily Eriosomatinae, and they produce galls on their primary host plants Pistacia. We assembled chromosome-level genomes of these two species using Nanopore long-read sequencing and Hi-C technology. A 332 Mb genome assembly of S. lentiscoides with a scaffold N50 of 19.77 Mb, including 11,747 genes, and a 289 Mb genome assembly of C. ovagalla with a scaffold N50 of 11.85 Mb, containing 14,492 genes, were obtained. The Benchmarking Universal Single-Copy Orthologs (BUSCO) benchmark of the two genome assemblies reached 93.7% (91.9% single-copy) and 97.0% (95.3% single-copy), respectively. The high-quality genome assemblies in our study provide valuable resources for future genomic research of galling aphids.

Dynamic response of essential amino acid biosynthesis in Buchnera aphidicola to supplement sub-optimal host nutrition.

The endosymbiotic bacterium Buchnera aphidicola allows its host Acyrthosiphon pisum to utilise a nutritionally limited phloem sap diet without significant mortality by providing essential amino acids (EAAs), which it biosynthesises de novo via complex pathways consisting of multiple enzymes. Previous studies have reported how non-essential amino acids (NEAAs) provided by the host are utilised by B. aphidicola, along with how genes within the biosynthetic pathways respond to amino acid deficiency. Although the effect on B. aphidicola gene expression upon the removal of a single EAA and multiple NEAAs from the A. pisum diet has been reported, little is known about the effects of the complete simultaneous removal of multiple EAAs, especially branched-chain amino acids (BCAAs). To investigate this, A. pisum was provided with amino acid deficient diets ilv- (lacking isoleucine, leucine, valine) or thra- (lacking threonine, methionine, lysine). Due to their involvement in the production of several amino acids, the expression of genes ilvC, ilvD (both involved in isoleucine, leucine and valine biosynthesis) and thrA (involved in threonine, methionine and lysine biosynthesis) was analysed and the expression of trpC (involved in tryptophan biosynthesis) was used as a control. Survival was reduced significantly when A. pisum was reared on ilv- or thra- (P < 0.001 and P = 0.000 respectively) compared to optimal artificial diet and was significantly lower on ilv- (P < 0.001) than thra-. This is likely attributed to the EAAs absent from ilv- being required at higher concentrations for aphid growth, than those EAAs absent from thra-. Expression of ilvC and ilvD were upregulated 2.49- and 2.08-fold (respectively) and thrA expression increased 2.35- and 2.12-fold when A. pisum was reared on ilv- and thra- (respectively). The surprisingly large upregulation of thrA when reared on ilv- is likely due to threonine being an intermediate in isoleucine biosynthesis. Expression of trpC was not affected by rearing on either of the two amino acid deficient diets. To our knowledge this study has shown, for the first time, how genes within the biosynthetic pathways of an endosymbiont respond to the simultaneous complete omission of multiple EAAs as well as all three BCAAs (leucine, isoleucine, valine), from the host diet.

Sgabd-2 plays specific role in immune response against biopesticide Metarhizium anisopliae in Aphis citricola.

Metarhizium anisopliae is an effective biopesticide for controlling Aphis citricola, which has developed resistance to many chemical pesticides. However, the powerful immune system of A. citricola has limited the insecticidal efficacy of M. anisopliae. The co-evolution between insects and entomogenous fungi has led to emergence of new antifungal immune genes, which remain incompletely understood. In this study, an important immune gene Sgabd-2 was identified from A. citricola through transcriptome analysis. Sgabd-2 gene showed high expression in the 4th instar nymph and adult stages, and was mainly distributed in the abdominal region of A. citricola. The recombinant protein (rSgabd-2) exhibited no antifungal activity but demonstrated clear agglutination activity towards the conidia of M. anisopliae. RNA interference of Sgabd-2 by dsRNA feeding resulted in decreased phenoloxidase (PO) activity and weakened defense for A. citricola against M. anisopliae. Simultaneous silence of GNBP-1 and Sgabd-2 effectively reduced the immunity of A. citricola against M. anisopliae more than the individual RNAi of GNBP-1 or Sgabd-2. Furthermore, a genetically engineered M. anisopliae expressing double-stranded RNA (dsSgabd-2) targeting Sgabd-2 in A. citricola successfully suppressed the expression of Sgabd-2 and demonstrated increased virulence against A. citricola. Our findings elucidated Sgabd-2 as a critical new antifungal immune gene and proposed a genetic engineering strategy to enhance the insecticidal virulence of entomogenous fungi through RNAi-mediated inhibition of pest immune genes.

Composition and abundance of midgut plasma membrane proteins in two major hemipteran vectors of plant viruses, Bemisia tabaci and Myzus persicae.

Multiple species within the order Hemiptera cause severe agricultural losses on a global scale. Aphids and whiteflies are of particular importance due to their role as vectors for hundreds of plant viruses, many of which enter the insect via the gut. To facilitate the identification of novel targets for disruption of plant virus transmission, we compared the relative abundance and composition of the gut plasma membrane proteomes of adult Bemisia tabaci (Hemiptera: Aleyrodidae) and Myzus persicae (Hemiptera: Aphididae), representing the first study comparing the gut plasma membrane proteomes of two different insect species. Brush border membrane vesicles were prepared from dissected guts, and proteins extracted, identified and quantified from triplicate samples via timsTOF mass spectrometry. A total of 1699 B. tabaci and 1175 M. persicae proteins were identified. Following bioinformatics analysis and manual curation, 151 B. tabaci and 115 M. persicae proteins were predicted to localize to the plasma membrane of the gut microvilli. These proteins were further categorized based on molecular function and biological process according to Gene Ontology terms. The most abundant gut plasma membrane proteins were identified. The ten plasma membrane proteins that differed in abundance between the two insect species were associated with the terms "protein binding" and "viral processes." In addition to providing insight into the gut physiology of hemipteran insects, these gut plasma membrane proteomes provide context for appropriate identification of plant virus receptors based on a combination of bioinformatic prediction and protein localization on the surface of the insect gut.

Densovirus infection facilitates plant-virus transmission by an aphid.

The interactions among plant viruses, insect vectors, and host plants have been well studied; however, the roles of insect viruses in this system have largely been neglected. We investigated the effects of MpnDV infection on aphid and PVY transmission using bioassays, RNA interference (RNAi), and GC-MS methods and green peach aphid (Myzus persicae (Sulzer)), potato virus Y (PVY), and densovirus (Myzus persicae nicotianae densovirus, MpnDV) as model systems. MpnDV increased the activities of its host, promoting population dispersal and leading to significant proliferation in tobacco plants by significantly enhancing the titer of the sesquiterpene (E)-ß-farnesene (EßF) via up-regulation of expression levels of the MpFPPS1 gene. The proliferation and dispersal of MpnDV-positive individuals were faster than that of MpnDV-negative individuals in PVY-infected tobacco plants, which promoted the transmission of PVY. These results combined showed that an insect virus may facilitate the transmission of a plant virus by enhancing the locomotor activity and population proliferation of insect vectors. These findings provide novel opportunities for controlling insect vectors and plant viruses, which can be used in the development of novel management strategies.