Hormetic effects of thiamethoxam on Schizaphis graminum: demographics and feeding behavior.

In agroecosystems, insects contend with chemical insecticides often encountered at sublethal concentrations. Insects' exposure to these mild stresses may induce hormetic effects, which has consequences for managing insect pests. In this study, we used an electrical penetration graph (EPG) technique to investigate the feeding behavior and an age-stage, two-sex life table approach to estimate the sublethal effects of thiamethoxam on greenbug, Schizaphis graminum. The LC5 and LC10 of thiamethoxam significantly decreased longevity and fecundity of directly exposed adult aphids (F0). However, the adult longevity, fecundity, and reproductive days (RPd)-indicating the number of days in which the females produce offspring - in the progeny generation (F1) exhibited significant increase when parental aphids (F0) were treated with LC5 of the active ingredient. Subsequently, key demographic parameters such as intrinsic rate of increase (r) and net reproductive rate (R0) significantly increased at LC5 treatment. EPG recordings showed that total durations of non-probing (Np), intercellular stylet pathway (C), and salivary secretion into the sieve element (E1) were significantly increased, while mean duration of probing (Pr) and total duration of phloem sap ingestion and concurrent salivation (E2) were decreased in F0 adults exposed to LC5 and LC10. Interestingly, in the F1 generation, total duration of Np was significantly decreased while total duration of E2 was increased in LC5 treatment. Taken together, our results showed that an LC5 of thiamethoxam induces intergenerational hormetic effects on the demographic parameters and feeding behavior of F1 individuals of S. graminum. These findings have important implications on chemical control against S. graminum and highlight the need for a deeper understanding of the ecological consequences of such exposures within pest management strategies across the agricultural landscapes.

In Silico Characterisation of the Aedes aegypti Gustatory Receptors.

Aedes aegypti, also known as the dengue mosquito or the yellow fewer mosquito, is the vector of dengue, chikungunya, Zika, Mayaro and yellow fever viruses. The A. aegypti genome contains an array of gustatory receptor (GR) proteins that are related to the recognition of taste. In this study, we performed in silico molecular characterization of all 72 A. aegypti GRs reported in the latest version of A. aegypti genome AaegL5. Phylogenetic analysis classified the receptors into three major clads. Multiple GRs were found to encode multiple transcripts. Physicochemical attributes such as the aliphatic index, hydropathicity index and isoelectric point indicated that A. aegypti gustatory receptors are highly stable and are tailored to perform under a variety of cellular environments. Analysis for subcellular localization indicated that all the GRs are located either in the extracellular matrix or the plasma membrane. Results also indicated that the GRs are distributed mainly on chromosomes 2 and 3, which house 22 and 49 GRs, respectively, whereas chromosome 1 houses only one GR. NCBI-CDD analysis showed the presence of a highly conserved 7tm_7 chemosensory receptor protein superfamily that includes gustatory and odorant receptors from insect species Anopheles gambiae and Drosophila melanogaster. Further, three significantly enriched ungapped motifs in the protein sequence of all 72 A. aegypti gustatory receptors were found. High-quality 3D models for the tertiary structures were predicted with significantly higher confidence, along with ligand-binding residues. Prediction of S-nitrosylation sites indicated the presence of target cysteines in all the GRs with close proximity to the ligand-bindings sites within the 3D structure of the receptors. In addition, two highly conserved motifs inside the GR proteins were discovered that house a tyrosine (Y) and a cysteine (C) residue which may serve as targets for NO-mediated tyrosine nitration and S-nitrosylation, respectively. This study will help devise strategies for functional genomic studies of these important receptor molecules in A. aegypti and other mosquito species through in vitro and in vivo studies.

Impact of sublethal concentrations of flonicamid on key demographic parameters and feeding behavior of Schizaphis graminum.

Flonicamid is a novel systemic insecticide that efficiently controls sap-sucking insect pests. However, the impact of sublethal concentrations of flonicamid on key demographic parameters and the feeding behavior of greenbug, Schizaphis graminum has not yet been studied. In this study, we used the age stage, two-sex life table approach, and electrical penetration graphs (EPGs) to investigate the sublethal effects of flonicamid on the biological traits and feeding behavior of S. graminum. Bioassays showed that flonicamid possesses high toxicity to adult S. graminum with LC50 of 5.111 mg L-1 following 48 h exposure. Sublethal concentrations of flonicamid (LC5 and LC10) significantly decreased the longevity and fecundity of directly exposed parental aphids (F0), while the reproductive days were reduced only at LC10. The pre-adult stage and total pre-reproductive period (TPRP) increased in F1 individuals after exposure of F0 aphids to the sublethal concentrations of flonicamid. Furthermore, the adult longevity, fecundity and key demographic parameters (R0, r, and λ) were significantly reduced in progeny generation (F1). EPG recordings showed that the total duration of phloem sap ingestion and concurrent salivation (E2) decreased substantially in F0 and F1 aphids after exposure to LC5 and LC10 of flonicamid. Taken together, our results showed that the sublethal concentrations of flonicamid affect the demographic parameters and feeding behavior that ultimately suppress the population growth of S. graminum. This study provides in-depth information about the overall effects of flonicamid on S. graminum that might help to manage this key pest.

Sequence and expression analysis of the spermatogenesis-specific gene cognates, wampa and Prosα6T, in Drosophila suzukii.

The sterile insect technique (SIT) is a highly effective biologically-based method for the population suppression of highly invasive insect pests of medical and agricultural importance. The efficacy of SIT could be significantly enhanced, however, by improved methods of male sterilization that avoid the fitness costs of irradiation. An alternative sterilization method is possible by gene-editing that targets genes essential for sperm maturation and motility, rendering them nonfunctional, similar to the CRISPR-Cas9 targeting of ß2-tubulin in the genetic model system, Drosophila melanogaster. However, since genetic strategies for sterility are susceptible to breakdown or resistance in mass-reared populations, alternative targets for sterility are important for redundancy or strain replacement. Here we have identified and characterized the sequence and transcriptional expression of two genes in a Florida strain of Drosophila suzukii, that are cognates of the D. melanogaster spermatocyte-specific genes wampa and Prosalpha6T. Wampa encodes a coiled-coil dynein subunit required for axonemal assembly, and the proteasome subunit gene, Prosalpha6T, is required for spermatid individualization and nuclear maturation. The reading frames of these genes differed from their NCBI database entries derived from a D. suzukii California strain by 44 and 8 nucleotide substitutions/polymorphisms, respectively, though all substitutions were synonymous resulting in identical peptide sequences. Expression of both genes is predominant in the male testis, and they share similar transcriptional profiles in adult males with ß2-tubulin. Their amino acid sequences are highly conserved in dipteran species, including pest species subject to SIT control, supporting their potential use in targeted male sterilization strategies.

Flubendiamide Resistance and Its Mode of Inheritance in Tomato Pinworm Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae).

Tomato pinworm, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is the major pest of tomato crops in Pakistan. Insecticides are commonly used for the management of this insect-pest. To develop a better insecticide resistance management strategy and evaluate the risk of resistance evolution, a field collected population of the tomato pinworm was selected with flubendiamide in the laboratory. We investigated the genetics of flubendiamide resistance and concentration-mortality response to other insecticides by selecting a field strain of tomato pinworm with commercial flubendiamide formulation. Tuta absoluta was reciprocally crossed with resistant strain (Fluben-R) and was selected up to 13 generations, while F1 progeny was back-crossed with resistant parent (Fluben-R). The results of LC50 and Resistance Ratio (RR) demonstrated a higher resistance developed in field and laboratory-selected strains (G2 and G13, respectively). Field-collected and laboratory-selected (Fluben-R) strains demonstrated higher intensity of concentration-mortality response against chlorantraniliprole, thiamethoxam, permethrin, abamectin and tebufenozide compared to susceptible ones. Based on the overlapping of 95% FL, it demonstrated significant differences, revealing that it was not sex linked (autosomal) with no maternal effects. The backcross analysis of the F1´ resistant parent resulting in significant differences at all concentrations suggests that resistance is controlled by more than one factor; the null hypothesis was rejected and inheritance was under polygenic control. Resistance progression from 38 to 550 folds demonstrated that T. absoluta can develop a higher level of resistance to flubendiamide. Concentration-mortality response experiments demonstrated that the LC50 of some tested insecticides was higher for field-collected and laboratory-selected strains, suggesting that resistance mechanisms should be studied at a molecular level for better understanding. These results could be helpful to design resistance management strategies against the tomato pinworm.

Influence of Bacterial Secondary Symbionts in Sitobion avenae on Its Survival Fitness against Entomopathogenic Fungi, Beauveria bassiana and Metarhizium brunneum.

The research was focused on the ability of wheat aphids Sitobion avenae, harboring bacterial secondary symbionts (BSS) Hamiltonella defensa or Regiella insecticola, to withstand exposure to fungal isolates of Beauveria bassiana and Metarhizium brunneum. In comparison to aphids lacking bacterial secondary symbionts, BSS considerably increased the lifespan of wheat aphids exposed to B. bassiana strains (Bb1022, EABb04/01-Tip) and M. brunneum strains (ART 2825 and BIPESCO 5) and also reduced the aphids' mortality. The wheat aphid clones lacking bacterial secondary symbionts were shown to be particularly vulnerable to M. brunneum strain BIPESCO 5. As opposed to wheat aphids carrying bacterial symbionts, fungal pathogens infected the wheat aphids lacking H. defensa and R. insecticola more quickly. When treated with fungal pathogens, bacterial endosymbionts had a favorable effect on the fecundity of their host aphids compared to the aphids lacking these symbionts, but there was no change in fungal sporulation on the deceased aphids. By defending their insect hosts against natural enemies, BSS increase the population of their host society and may have a significant impact on the development of their hosts.

miR-125-3p and miR-276b-3p Regulate the Spermatogenesis of Bactrocera dorsalis by Targeting the orb2 Gene.

Bactrocera dorsalis is considered a major threat to horticultural crops. It has evolved resistance against insecticides. It is believed that development of new methods is highly desirable to control this destructive agricultural pest. Sterile insect technique is emerging as a potential tool to control this insect pest by reducing their reproductive ability. Here we report that orb2 has high expression in the testis of B. dorsalis which is the target of miR-125-3p and miR-276b-3p and plays a critical role in the spermatogenesis. Dual luciferase reporter assay using HEKT293 cells demonstrates that orb2 gene is downregulated by miR-125-3p and miR-276b-3p and is a common target of these miRNAs. Dietary treatment of adult male flies separately and in combination of agomir-125-3p (Ago-125-3p) and agomir-276b-3p (Ago-276b-3p) significantly downregulated the mRNA of orb2. The combined treatments of agomirs suppressed the level of mRNA of orb2 significantly more than any single treatment. Altered expression of miR-125-3p and miR-276b-3p significantly decreased the total and live spermatozoa in the testis which ultimately caused reduction in male fertility. Furthermore, we demonstrate that miR-125-3p, miR-276b-3p, and orb2 dsRNA are the novel agents that could be used in a genetic-based sterile insect technique (SIT) to control the B. dorsalis.

Trophic transfer and toxicity of heavy metals from dengue mosquito Aedes aegypti to predator dragonfly Tramea cophysa.

Heavy metal pollution in aquatic habitats can be detrimental to both prey and predators in a food web. To investigate the potential for bio-transfer and bioaccumulation of heavy metals between specific trophic levels, 3rd instar larvae of Aedes aegypti were exposed to mercury (Hg), lead (Pb), cadmium (Cd), copper (Cu), and zinc (Zn) for three consecutive generations and fed to dragonfly (Tramea cophysa) nymphs. Exposure to Hg caused the highest mortality in A. aegypti larvae and T. cophysa nymphs. Bioaccumulation and life-history parameters of A. aegypti, including egg hatching time, larval and pupal duration, male and female life span, and fecundity, were also evaluated after metals exposure. All life-history parameters except larval duration were significantly affected by heavy metal treatments. Bioaccumulation of metals in A. aegypti larvae and adults gradually and significantly increased from 1st to 3rd generation. To the best of our knowledge, this is the first study describing the acute toxicity of heavy metals to mosquitoes. Our study shows that heavy metals cause dietary toxicity to an aquatic predator, dragonfly, via trophic transfer, which could have considerable consequences on aquatic ecosystems.

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

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

Toxicological risk assessment of some commonly used insecticides on Cotesia flavipes, a larval parasitoid of the spotted stem borer Chilo partellus.

Cotesia flavipes Cameron is an important larval parasitoid exploited for the control of the spotted stem borer, Chilo partellus (Swinhoe). Several studies have evaluated the toxic effects of insecticides on C. partellus, however, little is known about non-target effects of insecticides on this parasitoid, when used to control C. partellus. This laboratory study evaluated the lethal and sublethal effects of twelve insecticides on C. flavipes. Residual toxicity tests showed that organophosphates (chlorpyrifos, triazophos and profenofos) exhibited highest contact toxicity to C. flavipes adults with a LC50 range from 0.63 to 1.05 mg a.i/l, while neonicotinoids (nitenpyram, acetamiprid and imidacloprid) were less toxic to C. flavipes with a LC50 range from 1.27 to 139.48 mg a.i/l. Sugar-insecticide feeding bioassays showed that organophosphates, pyrethroids (cypermethrin, bifenthrin and lambda-cyhalothrin) and carbamates (thiodicarb, carbaryl and methomyl) were highly toxic to C. flavipes adults and caused 100% mortality at 48 h of exposure, while imidacloprid caused 66% mortality at 48 h of exposure. Risk quotient analysis showed that among all tested insecticides, imidacloprid and acetamiprid were less toxic to C. flavipes adults with a risk quotient value of 0.88 and 1.6, respectively. Furthermore, exposure of immature C. flavipes through their host bodies significantly decreased the parasitism rate at their F1 and F2 generations. Risk quotient analysis of insecticides indicated that imidacloprid and acetamiprid were the least toxic to C. flavipes. This study provides important information that will be used in incorporating the most suitable insecticides in integrated pest management programs with reduced negative impacts on non-target beneficial arthropods.

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

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

Author Correction: Impact of low lethal concentrations of buprofezin on biological traits and expression profile of chitin synthase 1 gene (CHS1) in melon aphid, Aphis gossypii.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Fitness costs in clothianidin-resistant population of the melon aphid, Aphis gossypii.

Clothianidin is a second-generation neonicotinoid insecticide, widely used against sap-sucking insect pest including melon aphid, Aphis gossypii Glover (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 of A. 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 of A. 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 (λ), net reproductive rate (R0), 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 of A. 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 against A. gossypii.

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

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

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

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

Impact of low lethal concentrations of buprofezin on biological traits and expression profile of chitin synthase 1 gene (CHS1) in melon aphid, Aphis gossypii.

Buprofezin, a chitin synthesis inhibitor that can be used for the control of hemipteran pests, especially melon aphid, Aphis gossypii. The impact of low lethal concentrations of buprofezin on the biological parameters and expression profile of CHS1 gene were estimated for two successive generations of A. gossypii. The present result shows that the LC15 and LC30 of buprofezin significantly decreased the fecundity and longevity of both generations. Exposure of F0 individuals to both concentrations delay the developmental period in F1. Furthermore, the survival rate, intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) were reduced significantly in progeny generation at both concentrations. However, the reduction in gross reproductive rate (GRR) was observed only at LC30. Although, the mean generation time (T) prolonged substantially at LC30. Additionally, expression of the CHS1 gene was significantly increased in F0 adults. Significant increase in the relative abundance of CHS1 mRNA transcript was also observed at the juvenile and adult stages of F1 generation following exposure to LC15 and LC30. Therefore, our results show that buprofezin could affect the biological traits by diminishing the chitin contents owing to the inhibition of chitin synthase activity in the succeeding generation of melon aphid.

RNAi-Mediated Knockdown of Tssk1 and Tektin1 Genes Impair Male Fertility in Bactrocera dorsalis.

The genetic-based sterile insect technique (SIT) is an effective and environmentally safe strategy to diminish populations of agricultural and horticultural insect pests. Functional characterization of genes related to male fertility can enhance the genetic-based SIT. Tssk1 has been involved to control male fertility in both mammals and insects. Moreover, Tektin1 has also been revealed to influence male fertility in both human and mammals. These findings suggested that Tssk1 and Tektin1 identified from Bactrocera dorsalis could be required for male fertility in B. dorsalis. In this study, expression profiles of these two genes were studied at different developmental stages and in various tissues of adult males. Remarkably, it was found that Tssk1 and Tektin1 were highly expressed in the testis of mature adult males of B. dorsalis. Furthermore, Tssk1 and Tektin1 genes were downregulated by using the RNA interference (RNAi) method. Fertility assays including egg laying, hatching, and spermatozoa count were also performed to investigate male fertility of B. dorsalis. Results showed that knockdown of Tssk1 and Tektin1 caused male sterility up to 58.99% and 64.49%, respectively. As expected, the total numbers of spermatozoa were also significantly reduced by 65.83% and 73.9%, respectively. These results suggested that male sterility was happened wing to the low number of spermatozoa. In conclusion, we demonstrate that Tssk1 and Tektin1 are the novel agents that could be used to enhance the genetic-based SIT, or their double-stranded RNA (dsRNA) can be used as biopesticides to control the population of B. dorsalis.

RNA interference-mediated knockdown of voltage-gated sodium channel (MpNav) gene causes mortality in peach-potato aphid, Myzus persicae.

Voltage-gated sodium channels (VGSC) are transmembrane proteins that generate an action potential in excitable cells and play an essential role in neuronal signaling. Since VGSCs play a crucial role in nerve transmission they have become primary targets for a broad range of commercial insecticides. RNA interference (RNAi) is a valuable reverse genetics tool used in functional genomics, but recently, it has also shown promise as a novel agent that could be used to control agricultural insect pests. In this study, we targeted the VGSC (MpNav) gene in the peach-potato aphid Myzus persicae, by oral feeding of artificial diets mixed with dsRNAs. Knock-down of MpNav gene expression caused up to 65% mortality in 3rd instar nymphs. Moreover, significantly lower fecundity and longevity was observed in adult aphids that had been fed with dsMpNav solution at the nymphal stage. Analysis of gene expression by qRT-PCR indicated that the aphid mortality rates and the lowered fecundity and longevity were attributable to the down-regulation of MpNav by RNAi. Taken together, our results show that MpNav is a viable candidate target gene for the development of an RNAi-based bio-aphicide.

MicroRNA Let-7 targets the ecdysone signaling pathway E75 gene to control larval-pupal development in Bactrocera dorsalis.

MicroRNAs (miRNAs) regulate various biological processes during insect development; however, their role in larval-pupal development in oriental fruit fly, Bactrocera dorsalis (Hendel) remains unknown. In the current study, we address the biological function of a conserved miRNA, Bdo-Let-7 in the regulation of BdE75 gene, which belongs to the ecdysone signaling pathway and participates in the larval-pupal development in B. dorsalis. Using dual luciferase reporter assay in HEK293T cells we show that Bdo-Let-7 miRNA interacts with the 3' untranslated region of BdE75 gene and suppresses its expression. The Bdo-Let-7 and BdE75 are also co-expressed in the larval-pupal stages and in different tissues of B. dorsalis. In in vivo experiments, the injection of Bdo-Let-7 agomir and antagomir in third instar larvae down- and up-regulated the expression of BdE75, respectively. The 20-hydroxyecdysone (20E) injection assay shows that 20E up-regulated the expression of Bdo-Let-7 on the 5th day of the larvae. Moreover, abnormal pupation and eclosion were observed after larval Bdo-Let-7 antagomir injection. Based on these results, we show that Bdo-Let-7 regulates the ecdysone signaling pathway through the exact dose of BdE75 gene, and is indispensable for normal larval-pupal development in B. dorsalis.

The toxicity of flonicamid to cotton leafhopper, Amrasca biguttula (Ishida), is by disruption of ingestion: an electropenetrography study.

BACKGROUND: The cotton leafhopper, Amrasca biguttula, is one of the most destructive pests of cotton in Asia. This species is thought to cause damage by injecting enzymatic saliva into various, presently unknown, cotton tissues and ingesting the resulting macerate. Flonicamid is a novel systemic insecticide used to control the cotton leafhopper; however, its mode of action is unknown. RESULTS: The mechanism of action of flonicamid on cotton leafhopper was investigated using electropenetrography (EPG). EPG recordings revealed six waveforms, i.e. Np (non-probing), A1 (channel cutting), A2, A3, A4, A5 and A6. Waveforms A2 and A3 probably represent active ingestion with (A2) and without (A3) simultaneous watery salivation. The meanings of A4, A5 and A6 are presently unknown, but minor in duration. Flonicamid significantly increased the mean duration of non-probing events and strongly inhibited ingestion by treated insects, which resulted in the slow death of leafhoppers. Inhibition of ingestion was dose dependent, and near-complete suppression was observed when the flonicamid concentration was increased to 10 000 mg L-1 . CONCLUSIONS: We propose that starvation caused by inhibition of active ingestion is the mechanism of toxicity for flonicamid. This knowledge could aid in applicability and use of this new insecticide for field management of leafhopper populations. © 2016 Society of Chemical Industry.