【Aim】 To identify the juvenile hormone binding protein (JHBP) gene of Ostrinia furnacalis (OfJHBP), and explore its function in growth, development and diapause of O. furnacalis. 【Methods】 Based on the transcriptome database of O. furnacalis larvae at different diapause statuses assembled by our laboratory (NCBI accession number: PRJNA1030200), the open reading frame (ORF) sequence of OfJHBP was cloned and identified by RT-PCR. The molecular weight and isoelectric point of OfJHBP of O. furnacalis were predicted using the Expasy-ProtParam online tool. The OfJHBP-encoded amino acid sequence of O. furnacalis was aligned with those of JHBPs from other insects using DNAMAN 6.0 software. A phylogenetic tree was constructed using the neighbor-joining (NJ) method in MEGA 11 software. The expression levels of OfJHBP in the 5th instar larvae of O. furnacalis at non-diapause, diapause and diapause-terminated stages were determined by qPCR. The expression level of OfJHBP was measured by qPCR at 48 h after OfJHBP interference by injecting dsRNA into the diapausing 5th instar larvae of O. furnacalis. The triglyceride content in larvae was detected at 96 h after OfJHBP interference by injecting dsRNA into the diapausing 5th instar larvae of O. furnacalis. The body weight, diapause rate, mortality rate and phenotypic changes of O. furnacalis were observed and determined daily within 9 d after dsRNA injection. 【Results】 The 726 bp ORF sequence of OfJHBP (NCBI accession number: XM_028317397.1) of O. furnacalis was obtained. OfJHBP encodes 241 amino acids. The predicted molecular weight of OfJHBP was 27.06 kD, and its theoretical isoelectric point was 5.99. OfJHBP shared the highest amino acid sequence identity (99.17%) with OnJHBP from O. nubilalis. The evolutionary relationship between OfJHBP and OnJHBP was the closest. OfJHBP was expressed in the 5th instar larvae at different diapause stages, with the highest expression level observed in diapause stage and lower expression levels in non-diapause and diapause-terminated stages. Compared to the dsGFP-injected control group, dsOfJHBP injection resulted in a 78.32% down-regulation of OfJHBP expression level at 48 after injection, a 1.18-fold increase in the triglyceride content at 96 after injection, as well as a mortality rate of 46.67%， a proportional reduction in the body weight, a 53.33% decrease in the diapause rate, developmental abnormalities, various malformations leading to death at different degrees at 9 d after injection, and surviving individuals predominantly developing into black pupae that failed to complete adult eclosion. 【Conclusion】 OfJHBP is closely associated with the growth and development of O. furnacalis and likely plays a crucial role in diapause process of O. furnacalis.

【Aim】 The gustatory receptor genes SfruGR8 and SfruGR9, expressed in the larval head of the fall armyworm, Spodoptera frugiperda, were cloned and the ligand-binding properties of SfruGR8 and SfruGR9 were characterized, to lay the theoretical foundation for revealing the molecular mechanisms of sugar perception in S. frugiperda larvae. 【Methods】 Based on the transcriptome sequencing data of S. frugiperda larvae and the identification results of gustatory receptor genes, the cDNA sequences of SfruGR8 and SfruGR9 containing full-length open reading frames (ORFs) were cloned by PCR. The structural features of SfruGR8 and SfruGR9 were analyzed through phylogenetic analysis and multiple sequence alignment. Using the Xenopus laevis oocyte heterologous expression system combined with the two-electrode voltage clamp technique, the electrophysiological responses of the recombinant SfruGR8 and SfruGR9 to 11 candidate ligand compounds (inositol and 10 carbohydrates) were measured. The potential binding sites of SfruGR8 and SfruGR9 with glucose and fructose, respectively, were predicted via three-dimensional structural modeling and molecular docking simulations. 【Results】 The full-length cDNA sequences of SfruGR8 (GenBank accession no.: LOC118262948) and SfruGR9 (GenBank accession no.: LOC118274941) were cloned from S. frugiperda. SfruGR8 and SfruGR9 possess 8 and 7 transmembrane domains, respectively, characterized by a long intracellular N-terminus and a short extracellular C-terminus. SfruGR8 and SfruGR9 contain a conserved motif, -hh(G/A/S)(A/S)hhTYhhhhhQF, at the C-terminus and belong to the sugar receptor family. The recombinant SfruGR8 exhibited the greatest response to glucose, while the recombinant SfruGR9 showed an exclusive response to fructose. Both SfruGR8 and SfruGR9 showed dose-dependent responses to glucose and fructose, respectively. The recombinant SfruGR8 interacted with glucose via hydrogen bonds and van der Waals forces, with a binding energy of -25.26 kJ/mol, while the recombinant SfruGR9 bound to fructose through the same interactions, exhibiting a binding energy of -26.67 kJ/mol. 【Conclusion】 Both gustatory receptors SfruGR8 and SfruGR9 are sugar receptors. SfruGR8 shows a strong response to glucose, whereas SfruGR9 specifically binds to fructose. It is inferred that SfruGR8 and SfruGR9 play important roles in the feeding or digestion of sugar compounds in S. frugiperda larvae.

【Aim】 To clone and obtain the Halloween family genes involved in ecdysteroid synthesis in the Chinese oak silkworm, Antheraea pernyi, and to characterize their expression patterns during pupal diapause termination under long photoperiod conditions, so as to lay a foundation for elucidating the features of ecdysteroid synthesis throughout diapause termination in A. pernyi pupae. 【Methods】 Five Halloween family genes (ApCyp307a1, ApCyp306a1, ApCyp302a1, ApCyp315a1 and ApCyp314a1) were cloned from A. pernyi using RT-PCR and subjected to bioinformatic analysis. The expression levels of the above five Halloween genes in the brain, prothoracic gland, hemolymph, midgut, fat body, testis/ovary, trachea and cuticle of pupae of A. pernyi at diapause and developmental stages were examined via semi-quantitative RT-PCR. RT-qPCR was employed to analyze the expression levels of the above five Halloween genes, as well as the ecdysone receptor genes ApEcR and ApUSP, in the prothoracic gland and fat body of A. pernyi pupae stored at both room temperature (19-25 ℃) and low temperature (6 ℃) conditions under a long photoperiod of 17L∶7D. RT-qPCR was used to detect the  expression levels of ApCyp314a1, ApEcR and ApUSP in the fat body at 24 h after RNAi of Cyp314a1 in the 4th instar larvae and prepupae of A. pernyi, and the effects of RNAi on the development of larvae and prepupae were further observed and detected. 【Results】Five Halloween family genes of A. pernyi were cloned, and designated as ApCyp307a1, ApCyp306a1, ApCyp302a1, ApCyp315a1 and ApCyp314a1 (GenBank accession numbers were MW677192, MW677193, MW677194, MW677195 and MW677196, respectively), based on homologous sequence alignment. Phylogenetic tree revealed that Halloween genes are highly conserved throughout evolution. Tissue-specific semi-quantitative PCR results indicated high expression levels of ApCyp307a1, ApCyp306a1, ApCyp302a1 and ApCyp315a1 in the prothoracic gland of A. pernyi pupae, with ApCyp302a1 and ApCyp315a1 showing marked upregulation during developmental stage of A. pernyi pupae. ApCyp314a1 was highly expressed in the midgut, fat body and testis/ovary of A. pernyi pupae at the developmental stage. A. pernyi pupae stored at low temperatures and exposed to the photoperiod of 17L∶7D exhibited accelerated emergence compared to those maintained at room temperature. The expression level of ApCyp315a1 in A. pernyi pupae stored under the photoperiod of 17L∶7D at room temperature peaked at 7 d after treatment, those of ApCyp306a1, ApCyp302a1 and ApUSP were increased at 21 d after treatment, whereas those of ApCyp314a1 and ApEcR peaked near eclosion (at 35 d after treatment). After low-temperature storage, the expression level of ApCyp307a1 in A. pernyi pupae was increased substantially during the pre-eclosion phase at 7 d after treatment under the photoperiod of 17L∶7D, while ApCyp302a1 was highly expressed at 21－35 d after treatment under the photoperiod of 17L∶7D, and the expression level of ApCyp315a1 peaked near eclosion, which showed a different expression pattern from that during the termination of pupae diapause at normal temperatures. The expression level of ApCyp314a1 peaked near eclosion, and ApEcR and ApUSP exhibited the expression profiles similar to those in the normal temperature group. Compared to the control group injected with dseGFP, RNAi of ApCyp314a1 significantly decreased the expression levels of ApCyp314a1 in the fat bodies of the 4th instar larvae and prepupae, and significantly suppressed the larval ecdysis and pupation of A. pernyi. 【Conclusion】These results demonstrate that Halloween genes play crucial roles in 20-hydroxyecdysone (20E) synthesis, and growth and development of A. pernyi. During long photoperiod-induced diapause termination, the expression levels of 20E synthesis-related genes increased during 7－21 d, whereas ApCyp314a1 was highly expressed in the pre-eclosion phase. Low-temperature treatment can accelerate diapause termination in A. pernyi pupae. With the exception of ApCyp306a1, which displayed multiple expression level peaks, the other Halloween genes were predominantly upregulated during the middle and late stages of low-temperature treatment.

【Aim】This study aims to explore the endophytic colonization potential of Lecanicillium lecanii strain JMC-01 in cabbage (Brassica oleracea var. capitata), its physiological impacts on the host plant and the repellent effects of induced cabbage volatile compounds against  Myzus persicae. 【Methods】 The L. lecanii strain JMC-01 spore suspensions at the concentrations of 1×10⁸, 1×10⁷ and 1×10⁶ spores/mL were inoculated to cabbage plants using the leaf spraying method and root drenching method, respectively. The colonization rates were determined using the PDA plate detection method on the 1st, 3rd, 7th, 10th, 14th and 21st day after inoculation. The growth promoting effect of L. lecanii strain JMC-01 on cabbage was investigated by measuring the plant height, root length, fresh weight, dry weight, stem diameter and chlorophyll content of the cabbage on the 21st day after inoculation. The insect Y-tube olfactometer was used to detect the behavioral selection of the wingless adults  and day-1 1st instar nymphs of M. persicae to the cabbage plants at 24 h after inoculation with the spore suspension of L. lecanii strain JMC-01 at the concentration of 1×10⁸ spores/mL. Gas chromatography-mass spectrometry (GC-MS) was used to identify differential volatile compounds from the cabbage inoculated and uninoculated with the spore suspension of L. lecanii strain JMC-01 at the concentration of 1×10⁸ spores/mL. The insect Y-tube olfactometer was used to detect the repellent effects of these differential volatile compounds induced by L. lecanii strain JMC-01 against the wingless adults and day-1 1st instar nymphs of M. persicae. 【Results】 The L. lecanii strain JMC-01 inoculated with leaf spraying method exhibited superior colonization efficiency in cabbage plants to that with root drenching method, achieving 100% colonization rates in cabbage across all the three concentrations on the 1st day post inoculation with leaf spraying method, with detectable colonization signals persisting on the 21st day after treatment with L. lecanii strain JMC-01 spore suspension at the concentration of 1×10⁷ spores/mL. Growth promotion experiments demonstrated that L. lecanii strain JMC-01 spore suspensions at the concentrations of 1×10⁸ and 1×10⁷ spores/mL significantly enhanced the plant height, root length, stem diameter and biomass accumulation, with the L. lecanii strain JMC-01 spore suspension at the concentration of 1×10⁷ spores/mL showing the most pronounced promoting effect on chlorophyll synthesis. The total chlorophyll content in the cabbage treated with L. lecanii strain JMC-01 spore suspension at the concentration of 1×10⁷ spores/mL reached 37 mg/g. The L. lecanii strain JMC-01 spore suspension treatment on the cabbage plants exhibited repellent effects on the wingless adults and day-1 1st instar nymphs of M. persicae. GC-MS analysis identified 12 differential volatile organic compounds induced by L. lecanii strain JMC-01 colonization in cabbage, among which n-hexadecane and 3-methyl-2-oxyindole exhibited repellent effects on the wingless adults and day-1 1st instar nymphs of M. persicae. The wingless adult M. persicae showed stronger olfactory response to the standard substances n-hexadecane and 3-methyl-2-oxyindole than the day-1 1st instar nymphs. The repellent effect of n-hexadecane on the day-1 1st instar nymphs increased with the spore suspension concentration increasing. 【Conclusion】 This study reveals that the L. lecanii strain JMC-01 can endophytically colonize in the cabbage plants, and can achieve a growth-promoting effect on the cabbage through endophytic colonization. At the same time, it can also induce the plant to produce volatile compounds to repel M. persicae, providing a theoretical basis for the application of entomopathogenic fungi in biological control strategies.

【Aim】To evaluate the virulence of biocontrol fungi isolated from vegetable gardens and the toxicity of commonly used insecticides against Megalurothrips usitatus. 【Methods】 Three insect pathogenic fungi isolated from pest samples in vegetable gardens were identified via morphology and ITS sequence analysis. The virulence of the three fungal species (1×10⁵-1×10⁹ cfu/mL) against M. usitatus adults was assessed after infection for 4-9 d. The toxicity of five commonly used insecticides including spinetoram (0.05, 0.1, 0.2, 0.4, 0.8 and 1.6 mg/L), spinosad (0.125, 0.25, 0.5, 1, 2 and 4 mg/L), emamectin benzoate (40, 80, 160, 320, 640 and 1 280 mg/L), thiamethoxam (20, 40, 80, 160, 320 and 640 mg/L) and cyantraniliprole (50, 100, 200, 400, 800 and 1 600 mg/L) against M. usitatus adults at 48 h post-treatment was determined using leaf-tube insecticide film method. 【Results】Three biocontrol fungi Metarhizium anisopliae Ma-g07, Beauveria bassiana Bb-g06 and Akanthomyces attenuatus La-sl04 were isolated and identified. The corrected mortality rates of adult M. usitatus at 9 d after infection by M. anisopliae Ma-g07, B. bassiana Bb-g06 and A. attenuatus La-sl04 at the concentration of 1×10⁹ cfu/mL were 100%, 91.38% and 62.50%, respectively, and the median lethal time (LT₅₀) values of M. anisopliae Ma-g07, B. bassiana Bb-g06 and A. attenuatus La-sl04 at the concentration of 1×10⁹ cfu/mL against M. usitatus adults were 4.39, 5.15 and 6.09 d, respectively. The median lethal concentration (LC₅₀) values of M. anisopliae Ma-g07 against adult M. usitatus at 5－9 d after infection were 1.79×10⁸, 2.99×10⁷, 3.30×10⁶, 1.06×10⁶ and 9.58×10⁵ cfu/mL, respectively, those of B. bassiana Bb-g06 were 7.46×10⁸, 7.79×10⁷, 5.01×10⁶, 6.09×10⁵ and 1.63×10⁵ cfu/mL, respectively, and those of A. attenuatus La-sl04 were 7.01×10⁹, 1.11×10⁹, 4.96×10⁸, 3.51×10⁸ and 2.80×10⁸ cfu/mL, respectively. The LC₅₀ values of spinetoram and spinosad against M. usitatus adults at 48 h after treatment were 0.35 and 0.95 mg/L, respectively, which were much lower than those of emamectin benzoate, thiamethoxam and cyantraniliprole (154.55, 297.88 and 315.14 mg/L, respectively). 【Conclusion】M. anisopliae Ma-g07 and B. bassiana Bb-g06 exhibit high pathogenicity and biocontrol potential against M. usitatus. Spinetoram and spinosad have higher toxicity  to M. usitatus adults among the five insecticides. It is suggested that in the future, the research on the combined application of these two biocontrol agents and two insecticides should be carried out for controlling M. usitatus. This study provides a basis for screening biocontrol resources and guidance for the rational selection of insecticides for M. usitatus and the combined application of biocontrol fungi and insecticides.

【Aim】 The aim of this study is to screen the optimal ratio of artificial diet for Propylaea japonica and evaluate the feeding effect of the optimized artificial diet. 【Methods】 Fresh pork liver, honey, sucrose, yeast powder, olive oil and sorbic acid were used as the basic diet ingredients. An orthogonal experimental design was employed to assess the impacts of artificial diet added with varying ratios of rape pollen, cholesterol and vitamin E on several developmental parameters, including developmental duration, eclosion rate, adult acquisition rate, sex ratio (proportion of females), and female and male adult weight of P. japonica. Additionally, the effects of the optimized diet formula on the developmental duration, number of eggs laid per female and ovarian development of P. japonica, as well as the fat, total sugar and total protein contents in the 6-day-old adults were examined. 【Results】 A 20% inclusion of rape pollen in the artificial diet significantly increased the body weight of both female and male adults of P. japonica, the addition of 0.4% cholesterol yielded the most favorable outcomes, and the addition of 1.6% vitamin E resulted in the optimal proportion of females of P. japonica. The optimized artificial diet formula consisted of 20 g of fresh pig liver, 4 g of honey, 4 g of sucrose, 1 g of yeast powder, 0.06 mL of olive oil, 0.1 g of sorbic acid, 4 g of rape pollen, 0.08 g of cholesterol and 0.32 g of vitamin E. Compared to rearing with A. pisum nymphs, rearing with the optimized artificial diet did not significantly affect the survival rate or eclosion rate of P. japonica. However, when P. japonica was fed with the optimized artificial diet, the larval duration was extended by 40.8%, the pupal duration prolonged by 21.9%, and the female and male weight decreased by 34.1% and 30.7%, respectively, the adult acquisition rate increased significantly by 20.1%, the pre-oviposition period extended by 38.9% and the number of eggs laid per female reduced by 71.1%, as compared with those fed with A. pisum nymphs. However, when P. japonica was fed with the optimized artificial diet at the larval stage and with A. pisum nymphs at the adult stage, there was no significant change in the pre-oviposition period, and the number of eggs laid per female only decreased by 46.3%, as compared with those fed with A. pisum nymphs. Additionally, compared with feeding on A. pisum nymphs, larval feeding on the optimized artificial diet led to delayed ovarian development, significant reduction in ovarian length, increased fat content, decreased total protein content, and no significant change in total sugar content in the 6-day-old female adults of P. japonica. 【Conclusion】 The artificial diet optimized through orthogonal experiments is suitable for rearing P. japonica larvae. Compared to rearing with A. pisum nymphs, rearing with the optimized artificial diet caused no significant changes in larval survival rate and adult eclosion rate of P. japonica, but increased the pupal weight, improved the adult acquisition rate and prolonged the larval duration. Further refinements are necessary to enhance the overall effectiveness of this optimized diet.

【Aim】 This study aims to identify the electrophysiologically active components of the volatiles from corn silk that elicit responses from the antennae of Monolepta signata adults, and to explore the influence of these active compounds on the behavioral preferences of this insect, so as to provide a basis for screening potential behavioral regulators for M. signata. 【Methods】 Dynamic headspace adsorption was employed to collect volatiles from corn (variety Xianyu 335) silks. The electrophysiologically active compounds from corn silk to the antennae of M. signata adults were identified from the volatiles by gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS). Furthermore, electroantennogram (EAG) technique was employed to evaluate the EAG responses of M. signata to the electrophysiologically active volatile compounds from corn silks. Finally, the behavioral responses of M. signata adults to the above-mentioned volatile compounds were assessed using a Y-tube olfactometer. 【Results】 Four active compounds were identified from corn silk volatiles: nonanal, azulene, octanol and decanal. Both female and male adults of M. signata exhibited EAG responses to the four active compounds, showing concentration-dependent characteristics with concentrations increasing. Among them, 100 μg/μL of nonanal elicited the strongest EAG response in both sexes, while 0.1 μg/μL of octanol elicited the weakest EAG response. Behavioral attraction assays demonstrated that octanol at the concentrations of 1, 10 and 100 μg/μL, as well as decanal at the concentrations of 10 and 100 μg/μL, significantly attracted female adults of M. signata. Meanwhile, nonanal and octanol at the concentrations of 10 and 100 μg/μL, along with azulene and decanal at the concentration of 100 μg/μL, showed significant attractive effect on male adults of M. signata. 【Conclusion】 Four compounds of corn silk volatiles, i.e., nonanal, azulene, octanol and decanal, have attractive effects on either female or male adults of M. signata, and can be considered as candidate components for developing attractants against this pest.

The rapid advancement of sequencing technologies has greatly facilitated the application of genomics and multi-omics approaches in studying the mechanisms of adaptive evolution in species. Owing to their short life cycles and high reproductive rates, insects have emerged as ideal model systems for investigating environmental adaptative evolution mechanisms. In this article, we summarized the developmental context of genomics in evolution biology and then discussed the unique advantages of insects in evolutionary mechanism research. We further focused on reviewing the applications and recent advances of comparative genomics, population genomics, pan-genomics and gene-editing technologies in deciphering the molecular basis of insect environmental adaptation, including insecticide resistance, migration and dispersal, and cold tolerance. These aspects are illustrated through the following key examples: the CYP9A and glutathione S-transferase (GST) gene clusters associated with insecticide resistance in Helicoverpa armigera, the resistance mechanisms of Spodoptera frugiperda to the Vip3A insecticidal protein, as well as the global distribution and dispersal pathways of its “rice” and “corn” strains, and the population evolution and adaptive differentiation mechanisms of major agricultural pests in China, including Chilo suppressalis, Ostrinia furnacalis and Agrotis ipsilon. Despite of these advances, current studies remain limited by insufficient genomic resources for non-model insect species and by incomplete understanding of epigenetic regulation. The relatively slow development of telomere-to-telomere (T2T) complete genomes and pan-genomes, together with inadequate integrative analyses across multiple omics layers, hampers the construction of coherent regulatory frameworks that connect genetic variation, transcriptional regulation and adaptive phenotypes. Looking forward, the systematic integration of multi-omics datasets, coupled with the development of advanced functional validation approaches-particularly those targeting non-coding regulation and epigenetic modifications-will be critical for achieving a more comprehensive and in-depth understanding of the complex mechanisms underlying environmental adaptation and evolution in insects.