Screening of cellulose-degrading bacteria and optimization of cellulase production from Bacillus cereus A49 through response surface methodology.

Cellulose-degrading microorganisms hold immense significance in utilizing cellulose resources efficiently. The screening of natural cellulase bacteria and the optimization of fermentation conditions are the hot spots of research. This study meticulously screened cellulose-degrading bacteria from mixed soil samples adopting a multi-step approach, encompassing preliminary culture medium screening, Congo red medium-based re-screening, and quantification of cellulase activity across various strains. Particularly, three robust cellulase-producing strains were identified: A24 (MT740356.1 Brevibacillus borstelensis), A49 (MT740358.1 Bacillus cereus), and A61 (MT740357.1 Paenibacillus sp.). For subsequent cultivation experiments, the growth curves of the three obtained isolates were monitored diligently. Additionally, optimal CMCase production conditions were determined, keeping CMCase activity as a key metric, through a series of single-factor experiments: agitation speed, cultivation temperature, unit medium concentration, and inoculum volume. Maximum CMCase production was observed at 150 rpm/37 °C, doubling the unit medium addition, and a 5 mL inoculation volume. Further optimization was conducted using the selected isolate A49 employing response surface methodology. The software model recommended a 2.21fold unit medium addition, 36.11 °C temperature, and 4.91 mL inoculant volume for optimal CMCase production. Consequently, three parallel experiments were conducted based on predicted conditions consistently yielding an average CMCase production activity of 15.63 U/mL, closely aligning with the predicted value of 16.41 U/mL. These findings validated the reliability of the model and demonstrated the effectiveness of optimized CMCase production conditions for isolate A49.

Two heat shock cognate 70 genes involved in spermatogenesis regulate the male fertility of Zeugodacus cucurbitae, as potential targets for pest control.

The melon fly Zeugodacus cucurbitae Coquillett (Diptera: Tephritidae) is an agricultural quarantine pest threatening fruit and vegetable production. Heat shock cognate 70 (Hsc70), which is a homolog of the heat shock protein 70 (Hsp70), was first discovered in mice testes and plays an important role in spermatogenesis. In this study, we identified and cloned five Hsc70 genes from melon fly, namely ZcHsc70_1/2/3/4/5. Phylogenetic analysis showed that these proteins are closely related to Hsc70s from other Diptera insects. Spatiotemporal expression analysis showed that ZcHsc70_1 and ZcHsc70_2 are highly expressed in Z. cucurbitae testes. Fluorescence in situ hybridization further demonstrated that ZcHsc70_1 and ZcHsc70_2 are expressed in the transformation and maturation regions of testes, respectively. Moreover, RNA interference-based suppression of ZcHsc70_1 or ZcHsc70_2 resulted in a significant decrease of 74.61% and 63.28% in egg hatchability, respectively. Suppression of ZcHsc70_1 expression delayed the transformation of sperm cells to mature sperms. Meanwhile, suppression of ZcHsc70_2 expression decreased both sperm cells and mature sperms by inhibiting the meiosis of spermatocytes. Our findings show that ZcHsc70_1/2 regulates spermatogenesis and further affects the male fertility in the melon fly, showing potential as targets for pest control in sterile insect technique by genetic manipulation of males.

An Odorant Receptor Expressed in Both Antennae and Ovipositors Regulates Benzothiazole-Induced Oviposition Behavior in Bactrocera dorsalis.

The oriental fruit fly,Bactrocera dorsalis (Hendel), is a notorious pest of fruit crops, causing severe damage to fleshy fruits during oviposition and larval feeding. Gravid females locate suitable oviposition sites by detecting the host volatiles. Here, the oviposition preference of antenna-removed females and the electrophysiological response of ovipositors to benzothiazole indicated that both antennae and ovipositors are involved in perceiving benzothiazole. Subsequently, odorant receptors (ORs) expressed in both antennae and ovipositors were screened, and BdorOR43a-1 was further identified to respond to benzothiazole using voltage-clamp recording. Furthermore, BdorOR43a-1-/- mutants were obtained using the CRISPR/Cas9 system and their oviposition preference to benzothiazole was found to be significantly altered compared to WT females, suggesting that BdorOR43a-1 is one of the important ORs for benzothiazole perception. Our results not only demonstrate the important role of antennae and ovipositors in benzothiazole-induced oviposition but also elucidate on the OR responsible for benzothiazole perception in B. dorsalis.

Plasticity of the olfactory behaviors in Bactrocera dorsalis under various physiological states and environmental conditions.

Insects rely heavily on their olfactory system for various behaviors, including foraging, mating, and oviposition. Numerous studies have demonstrated that insects can adjust their olfactory behaviors in response to different physiological states and environmental conditions. This flexibility allows them to perceive and process odorants according to different conditions. The Oriental fruit fly, Bactrocera dorsalis, is a highly destructive and invasive pest causing significant economic losses to fruit and vegetable crops worldwide. The olfactory behavior of B. dorsalis exhibits strong plasticity, resulting in its successful invasion. To enhance our understanding of B. dorsalis' olfactory behavior and explore potential strategies for behavior control, we have reviewed recent literature on its olfactory plasticity and potential molecular mechanisms.

Mitochondrial coding genes mediate insecticide tolerance in the oriental fruit fly, Bactrocera dorsalis (Hendel).

The oriental fruit fly, Bactrocera dorsalis (Hendel), an invasive insect pest infesting fruits and vegetables, possesses a remarkable capacity for environmental adaptation. The investigation of behind mechanisms of the stress adaptability in B. dorsalis holds significantly practical relevance. Previous studies on the molecular mechanism underlying stress resistance in B. dorsalis have predominantly focused on nuclear-coding genes, with limited exploration on organelle-coding genes. In this study, we assessed alterations in the mitochondrial physiological parameters of B. dorsalis under exposure to malathion, avermectin, and beta-cypermethrin at LD50 dosages. The results showed that all three insecticides were capable of reducing mitochondrial complex IV activity and ATP content. Expression patterns of mitochondrial coding genes across different developmental stages, tissues and insecticide exposures were analyzed by RT-qPCR. The results revealed that these mitochondrial coding genes were expressed in various tissues and at different developmental stages. Particularly noteworthy, atp6, cox2, and cytb exhibited substantial up-regulation in response to malathion and avermectin treatment. Furthermore, RNAi-mediated knockdown of atp6 and cox2 resulted in the increased toxicity of malathion and avermectin against B. dorsalis, and cox2 silencing was also associated with the decreased complex IV activity. These findings suggest that atp6 and cox2 most likely play pivotal roles in mediating tolerance or resistance to malathion and avermectin in B. dorsalis. Our results provide novel insights into the role of mitochondrial coding genes in conferring tolerance to insecticides in B. dorsalis, with practical implications for controlling this pest in the field.

Odorant Binding Protein Expressed in Legs Enhances Malathion Tolerance in Bactrocera dorsalis (Hendel).

Bactrocera dorsalis is a highly invasive species and is one of the most destructive agricultural pests worldwide. Organophosphorus insecticides have been widely and chronically used to control it, leading to the escalating development of resistance. Recently, odorant binding proteins (OBPs) have been found to play a role in reducing insecticide susceptibility. In this study, we used RT-qPCR to measure the expression levels of four highly expressed OBP genes in the legs of B. dorsalis at different developmental stages and observed the effect of malathion exposure on their expression patterns. The results showed that OBP28a-2 had a high expression level in 5 day old adults of B. dorsalis, and its expression increased after exposure to malathion. By CRISPR/Cas9 mutagenesis, we generated OBP28a-2-/- null mutants and found that they were more susceptible to malathion than wild-type adults. Furthermore, in vitro direct affinity assays confirmed that OBP28a-2 has a strong affinity for malathion, suggesting that it plays a role in reducing the susceptibility of B. dorsalis to malathion. Our findings enriched our understanding of the function of OBPs. The results highlighted the potential role of OBPs as buffering proteins that help insects survive exposure to insecticides.

Lethal and sublethal effects of fluralaner on the citrus red mite, Panonychus citri (McGregor).

BACKGROUND: The citrus red mite, Panonychus citri (McGregor) is a globally distributed agricultural pest. Of late, resistance to common acaricides has raised concerns that chemical control of P. citri is an inefficient means of control. Fluralaner, a highly toxic isoxazoline insecticide used to treat various ectoparasites, presents one potential alternative. However, little information has been reported about the effect of fluralaner on the citrus red mite. This study aims to evaluate the toxicity, sublethal and transgenerational effects of fluralaner on P. citri. RESULTS: In both laboratory and field populations of P. citri, we found fluralaner to be more toxic than conventional alternatives, including fenpropathrin, bifenazate, azocyclotin and chlorpyrifos. Interestingly, fluralaner proved more toxic to female adults than to the eggs of P. citri, with median lethal concentrations (LC50 ) of 2.446 and 122.7 mg L-1 , respectively. Exposure to sublethal concentrations of fluralaner (LC10 , LC20 and LC30 ) significantly reduced the fecundity and longevity of female adults P. citri individuals. Although concentrations of fluralaner applied to the parental female adults (F0 ) led to some changes in the developmental parameters, there were no significant changes in most of the life table parameters or population growth of the F1 generation. CONCLUSION: Our results indicate that fluralaner is highly toxic to P. citri, and a significant sublethal effect on F0 could suppress the population growth of P. citri, but not for F1 . Fluralaner may be considered as a pesticide for the future management of the citrus red mite. © 2024 Society of Chemical Industry.

The target sign: a significant CT sign for predicting small-bowel ischemia and necrosis.

OBJECTIVE: To investigate the correlation between changes in the thickness and density of diseased small-bowel wall and small-bowel ischemia and necrosis (SBN) on CT imaging when small-bowel obstruction (SBO) occurs. METHODS: We retrospectively analyzed 186 patients with SBO in our hospital from March 2020 to June 2023. The patients were divided into simple SBO (control group) and SBN (case group) groups. We used logistic regression analysis, the chi-square test, and Fisher's exact test to analyze the correlation between the changes in the thickness and density of the diseased intestinal wall and the SBN. A receiver operating characteristic (ROC) curve was used to calculate the accuracy of the multivariate analysis. RESULTS: Of the 186 patients with SBO, 98 (52.7%) had simple SBO, 88 (47.3%) had SBN, and the rate of SBN was 47.3% (88/186). Multivariate regression analysis revealed that six CT findings were significantly correlated with SBN (p < 0.05), namely, thickening of the diseased intestinal wall with the target sign (OR = 21.615), thinning of the diseased intestinal wall (OR = 48.106), increase in the diseased intestinal wall density (OR = 13.696), mesenteric effusion (OR = 21.635), decrease in the diseased intestinal wall enhancement on enhanced scanning (OR = 41.662), and increase in the diseased intestinal wall enhancement on enhanced scanning (OR = 15.488). The AUC of the multivariate analysis reached 0.987 (95% CI 0.974-0.999). Specifically, the target sign was easily recognizable on CT images and was a significant CT finding for predicting SBN. CONCLUSION: We identified 6 CT findings that were significantly associated with SBN, and may be helpful for clinical treatment.

One-pot domino syntheses of 3-alkyl-3-N-substituted aminobenzofuran-2(3H)-ones based on alkali-promoted Michael addition and lactonization.

In this paper, a novel cascade reaction of caesium carbonate-promoted Michael addition and lactonization for the one-pot synthesis of 3-alkyl-3-N-substituted aminobenzofuran-2(3H)-one derivatives has been established based on the screening of the alkaline reagents and optimization of reaction conditions, in which the N-substituted (ortho-hydroxy)aryl glycine esters were used as the Michael donors to react with different α, ß-unsaturated carbonyl compounds. In the case of using the asymmetric starting material, the epimers could be successfully separated by conventional chromatography. In addition, plausible mechanisms were suggested and the absolute configuration of the epimer was analysed. All the chemical structures of unreported benzofuran-2(3H)-one derivatives were characterized by 1H nuclear magnetic resonance (NMR), 13C NMR, IR and high-resolution mass spectrometry (HRMS).

Characterization of two Bursicon genes and their association with wing development in the brown citrus aphid, Aphis citricidus.

The tanning hormone, Bursicon, is a neuropeptide secreted by the insect nervous system that functions as a heterodimer composed of Burs-α and Burs-ß subunits. It plays a critical role in the processes of cuticle tanning and wing expansion in insects. In this study, we successfully identified the AcBurs-α and AcBurs-ß genes in Aphis citricidus. The open reading frames of AcBurs-α and AcBurs-ß were 480 and 417 bp in length, respectively. Both AcBurs-α and AcBurs-ß exhibited 11 conserved cysteine residues. AcBurs-α and AcBurs-ß were expressed during all developmental stages of A. citricidus and showed high expression levels in the winged aphids. To investigate the potential role of AcBurs-α and AcBurs-ß in wing development, we employed RNA interference (RNAi) techniques. With the efficient silencing of AcBurs-α (44.90%) and AcBurs-ß (52.31%), malformed wings were induced in aphids. The proportions of malformed wings were 22.50%, 25.84%, and 38.34% in dsAcBurs-α-, dsAcBur-ß-, and dsAcBurs-α + dsAcBur-ß-treated groups, respectively. Moreover, feeding protein kinase A inhibitors (H-89) also increased the proportion of malformed wings to 30.00%. Feeding both double-stranded RNA and inhibitors (H-89) significantly downregulated the wing development-related genes nubbin, vestigial, notch and spalt major. Silence of vestigial through RNAi also led to malformed wings. Meanwhile, the exogenous application of 3 hormones that influence wing development did not affect the expression level of AcBursicon genes. These findings indicate that AcBursicon genes plays a crucial role in wing development in A. citricidus; therefore, it represents a potential molecular target for the control of this pest through RNAi-based approaches.

Suppressing the expression of glutathione S-transferase gene GSTd10 increases the sensitivity of Zeugodacus cucurbitae against ß-cypermethrin.

Zeugodacus cucurbitae Coquillett (Diptera: Tephritidae) is an agriculturally and economically important pest worldwide that has developed resistance to ß-cypermethrin. Glutathione S-transferases (GSTs) have been reported to be involved in the detoxification of insecticides in insects. We have found that both ZcGSTd6 and ZcGSTd10 were up-regulated by ß-cypermethrin induction in our previous study, so we aimed to explore their potential relationship with ß-cypermethrin tolerance in this study. The heterologous expression of ZcGSTd6 and ZcGSTd10 in Escherichia coli showed significantly high activities against 1-chloro-2,4-dinitrobenzene (CDNB). The kinetic parameters of ZcGSTd6 and ZcGSTd10 were determined by Lineweaver-Burk. The Vmax and Km of ZcGSTd6 were 0.50 µmol/min·mg and 0.3 mM, respectively. The Vmax and Km of ZcGSTd10 were 1.82 µmol/min·mg and 0.53 mM. The 3D modelling and molecular docking results revealed that ß-cypermethrin exhibited a stronger bounding to the active site SER-9 of ZcGSTd10. The sensitivity to ß-cypermethrin was significantly increased by 18.73% and 27.21%, respectively, after the knockdown of ZcGSTd6 and ZcGSTd10 by using RNA interference. In addition, the inhibition of CDNB at 50% (IC50 ) and the inhibition constants (Ki) of ß-cypermethrin against ZcGSTd10 were determined as 0.41 and 0.33 mM, respectively. The Ki and IC50 of ß-cypermethrin against ZcSGTd6 were not analysed. These results suggested that ZcGSTd10 could be an essential regulator involved in the tolerance of Z. cucurbitae to ß-cypermethrin.

Fusion dsRNA designs incorporating multiple target sequences can enhance the aphid control capacity of an RNAi-based strategy.

BACKGROUND: RNA interference (RNAi) is the sequence-dependent suppression of gene expression by double-stranded RNA (dsRNA). This is a promising strategy for the control of insect pests because dsRNA can be rationally designed to maximize efficacy and biosafety, the latter by using sequences that are found in target pests but are safe for non-target insects. However, this has yet to be optimized in aphids, destructive sap-sucking pests that also transmit plant viruses. We used the green peach aphid (Myzus persicae) as a case study to optimize the efficiency of RNAi by applying a novel fusion dsRNA design. RESULTS: Comparative transcriptomics revealed a number of genes that are induced in feeding aphids, and eight candidate genes were chosen as RNAi targets. To improve RNAi efficiency, our fusion dsRNA design approach combined optimal gene fragments (highly conserved in several aphid species but with less homology in beneficial insects such as the predator ladybeetle Propylea japonica) from three candidate genes. We compared this RNAi-based biological control approach with conventional chemical control using imidacloprid. We found that the fusion dsRNA strategy inhibited the aphid population to a significantly greater extent than single-target RNAi and did not affect ladybeetle fitness, allowing an additive effect between RNAi and natural predation, whereas imidacloprid was harmful to aphids and ladybeetles. CONCLUSION: Our fusion dsRNA design approach enhances the ability of RNAi to control aphids without harming natural predators. © 2024 Society of Chemical Industry.

Overexpression of miR-927-5p suppresses stalky expression and negatively reduces the spermatid production in Zeugodacus cucurbitae.

BACKGROUND: The melon fly, Zeugodacus cucurbitae Coquillett, is one of the major pests attacking Cucurbitaceae crops. Identifying critical genes or proteins regulating fertility is essential for sustainable pest control and a research hotspot in insect physiology. MicroRNAs (miRNAs) are short RNAs that do not directly participate in protein translation, but instead function in post-transcriptional regulation of gene expression involved in male fertility. RESULTS: We found that miR-927-5p is highly expressed in the testes and investigated its function in spermatogenesis in Z. cucurbitae. Fluorescence in situ hybridization (FISH) showed miR-927-5p in the transformation and maturation region of the testis, and overexpression of miR-927-5p reduced the number of sperms by 53%. In continuation, we predicted 12 target genes of miR-927-5p using bioinformatics combined with transcriptome sequencing data, and found that miR-927-5p targets the new gene Stalky in insects, which was validated by quantitative real-time PCR, RNA pull-down and dual luciferase reporter assays. FISH also confirmed the co-localization of miR-927-5p and the transcript Stalky_1 in the testis. Moreover, silencing of Stalky_1 by RNA interference reduced the number of sperms by 32% and reduced sperm viability by 39% in physiologically mature male adults. Meanwhile, the silencing of Stalky_1 also resulted in low hatchability. CONCLUSION: Our work not only presents a new, so far unreported mechanism regulating spermatogenesis by miR-927-5p targeting a new unknown target, Stalky, which is providing new knowledge on the regulatory network of insect spermatogenesis, but also lays a foundation for the development of SIT against important tephritid fly pests. © 2024 Society of Chemical Industry.

An Antenna-Enriched Chemosensory Protein Plays Important Roles in the Perception of Host Plant Volatiles in Bactrocera dorsalis (Diptera: Tephritidae).

Olfaction plays indispensable roles in insect behavior such as host location, foraging, oviposition, and avoiding predators. Chemosensory proteins (CSPs) can discriminate the hydrophobic odorants and transfer them to the odorant receptors. Presently, CSPs have been identified in many insect species. However, their presence and functions remain unknown in Bactrocera dorsalis, a destructive and invasive insect pest in the fruit and vegetable industry. Here, we annotated eight CSP genes in the genome of B. dorsalis. The results of quantitative real-time polymerase chain reaction (RT-qPCR) showed that BdorCSP3 was highly expressed in the antennae. Molecular docking and in vitro binding assays showed that BdorCSP3 had a good binding ability to host volatiles methyl eugenol (ME, male-specific attractant) and ß-caryophyllene (potential female attractant). Subsequently, CRISPR/Cas9 was used to generate BdorCSP3-/- mutants. Electroantennograms (EAGs) and behavioral assays revealed that male mutants significantly reduced the preference for ME, while female mutants lost their oviposition preference to ß-caryophyllene. Our data indicated that BdorCSP3 played important roles in the perception of ME and ß-caryophyllene. The results not only expanded our knowledge of the olfaction perception mechanism of insect CSPs but also provided a potential molecular target for the control of B. dorsalis.

Sublethal and transgenerational effects of lufenuron on the biological traits of Panonychus citri (McGregor) (Acari: Tetranychidae).

The citrus red mite, Panonychus citri (McGregor), is a globally important pest that has developed severe resistance to various pesticides. Lufenuron has been widely used in the control of the related pests in citrus orchard ecosystem. In this study, the susceptibilities of egg, larva, deutonymph and female adult of P. citri to lufenuron was determined, and the LC50 values were 161.354 mg/L, 49.595 mg/L, 81.580 mg/L, and 147.006 mg/L, respectively. Life-table analysis indicated that the fecundities were significantly increased by 11.86% and 26.84% after the mites were treated with LC20 concentrations of lufenuron at the egg or deutonymph stages, respectively. After eggs were treated with lufenuron, the immature stage and longevity were also affected, and resulted in a significant increase in r, R0 and λ. After exposure of female adults to LC20 of lufenuron, the fecundity and longevity of F0 generation significantly decreased by 31.99% and 10.94%, respectively. Furthermore, the expression level of EcR and Vg was significantly inhibited upon mites was treated with lufenuron. However, lufenuron exposure has a positive effect on fecundity and R0 in F1 generation, the expression of all reproduction-related genes was significantly up-regulated. In conclusion, there was a stimulating effect on the offspring population. Our results will contribute to the assessment of the resurgence of P. citri in the field after the application of lufenuron and the development of integrated pest control strategies in citrus orchards.

The endosymbiont Serratia symbiotica improves aphid fitness by disrupting the predation strategy of ladybeetle larvae.

Aphids, the important global agricultural pests, harbor abundant resources of symbionts that can improve the host adaptability to environmental conditions, also control the interactions between host aphid and natural enemy, resulting in a significant decrease in efficiency of biological control. The facultative symbiont Serratia symbiotica has a strong symbiotic association with its aphid hosts, a relationship that is known to interfere with host-parasitoid interactions. We hypothesized that Serratia may also influence other trophic interactions by interfering with the physiology and behavior of major predators to provide host aphid defense. To test this hypothesis, we investigated the effects of Serratia on the host aphid Acyrthosiphon pisum and its predator, the ladybeetle Propylaea japonica. First, the prevalence of Serratia in different A. pisum colonies was confirmed by amplicon sequencing. We then showed that harboring Serratia improved host aphid growth and fecundity but reduced longevity. Finally, our research demonstrated that Serratia defends aphids against P. japonica by impeding the predator's development and predation capacity, and modulating its foraging behavior. Our findings reveal that facultative symbiont Serratia improves aphid fitness by disrupting the predation strategy of ladybeetle larvae, offering new insight into the interactions between aphids and their predators, and providing the basis of a new biological control strategy for aphid pests involving the targeting of endosymbionts.

RNA interference in insects: the link between antiviral defense and pest control.

RNA interference (RNAi) is a form of gene silencing triggered by double-stranded RNA (dsRNA) that operates in all eukaryotic cells. RNAi has been widely investigated in insects to determine the underlying molecular mechanism, to investigate its role in systemic antiviral defense, and to develop strategies for pest control. When insect cells are infected by viruses, viral dsRNA signatures trigger a local RNAi response to block viral replication and generate virus-derived DNA that confers systemic immunity. RNAi-based insect pest control involves the application of exogenous dsRNA targeting genes essential for insect development or survival, but the efficacy of this approach has limited potency in many pests through a combination of rapid dsRNA degradation, inefficient dsRNA uptake/processing, and ineffective RNAi machinery. This could be addressed by dsRNA screening and evaluation, focusing on dsRNA design and off-target management, as well as dsRNA production and delivery. This review summarizes recent progress to determine the role of RNAi in antiviral defense and as a pest control strategy in insects, addressing gaps between our fundamental understanding of the RNAi mechanism and the exploitation of RNAi-based pest control strategies.

Cross-resistance, inheritance and biochemical mechanism of abamectin resistance in a field-derived strain of the citrus red mite, Panonychus citri (Acari: Tetranychidae).

BACKGROUND: The citrus red mite, Panonychus citri (McGregor), a global pest of citrus, has developed different levels of resistance to various acaricides in the field. Abamectin is one of the most important insecticides/acaricides worldwide, targetting a wide number of insect and mite pests. The evolution of abamectin resistance in P. citri is threatening the sustainable use of abamectin for mite control. RESULTS: The abamectin resistant strain (NN-Aba), derived from a field strain NN by consistent selection with abamectin, showed 4279-fold resistance to abamectin compared to a relatively susceptible strain (SS) of P. citri. Cross-resistance of NN-Aba was observed between abamectin and emamectin benzoate, pyridaben, fenpropathrin and cyflumetofen. Inheritance analyses indicated that abamectin resistance in the NN-Aba strain was autosomal, incompletely recessive and polygenic. The synergy experiment showed that abamectin toxicity was synergized by piperonyl butoxide (PBO), diethyl maleate (DEM) and tributyl phosphorotrithiotate (TPP) in the NN-Aba strain, and synergy ratios were 2.72-, 2.48- and 2.13-fold, respectively. The glutathione-S-transferases activity in the NN-Aba strain were significantly increased by 2.08-fold compared with the SS strain. CONCLUSION: The abamectin resistance was autosomal, incompletely recessive and polygenic in P. citri. The NN-Aba strain showed cross-resistance to various acaricides with different modes of action. Metabolic detoxification mechanism participated in abamectin resistance in NN-Aba strain. These findings provide useful information for resistance management of P. citri in the field. © 2023 Society of Chemical Industry.

Bursicon regulates wing expansion via PKA in the oriental fruit fly, Bactrocera dorsalis.

BACKGROUND: Bursicon is a heterodimeric neuropeptide that is involved in many physiological activities such as cuticle tanning, wing expansion, reproduction and immunity in insects. In this study, the role of bursicon in the wing expansion was investigated in Bactrocera dorsalis, an important invasive insect pest in agriculture. RESULTS: The cDNA sequences and deduced amino acids of bursicon genes (named BdBurs-α and BdBurs-ß) were determined, and two proteins typically contained 11 cysteine residues in conserved positions that were highly conserved in other insect species. The spatiotemporal expressions of bursicon genes showed that higher expression occurred at the pupal, early adult stage and ovaries, and lower expression at the late larval stage and in wing tissue (8-day-old pupae). Dysfunction of bursicon genes by dsRNA microinjection into 5-day-old pupae reduced PKA (a downstream component of the bursicon pathway) activity and resulted in malformed adult wings. PKA inhibitor injection into 5-day-old pupae also resulted in similar phenotypes. Hematoxylin & eosin staining of the adult wing showed that RNAi and PKA inhibitor treatment reduced the thickness of the wing cuticle, which wing cuticle thickness were ≈50% thinner than in the control. Furthermore, the expression of hedgehog (Bdhh) (one of 10 tested genes related to wing development) was significantly upregulated after RNAi and PKA inhibitor application. CONCLUSION: The results indicate that bursicon plays a crucial role in the wing expansion of B. dorsalis, suggesting bursicon genes have potential to be the targets for B. dorsalis control. © 2023 Society of Chemical Industry.