Cytochrome oxidase subunits 1(CO1) and 2 (CO2), and the entire sequence of the gene for tRNA leucine from twelve species of seven genera of the ant subfamily Myrmicinae were sequenced. The DNA sequences were analyzed, and the secondary structure of tRNA leucine also gene was studied. The molecular phylogenetic analyses of the subfamily Myrmicinae with Forelius chalybaeus, a species of the subfamily Dolichoderinae as the outgroup were performed by using four methods: the maximum likelihood (ML), maximum parsimony (MP), neighbor-joined (NJ), and unweighted pairgroup method with arithmetic means (UPGMA) based on the deoxyribonucleic acid data and the amino acid data. The taxonomic scheme for these genera and species based on molecular phylogeny of above genes consisted with that of the traditional taxonomy and the consistency for genera was better than that for species.

Insecticidal activities and active ingredients of Derris fordii var. lucida were studied for the first time. Insecticidal activities and action mode were determined by bioassay. The active ingredients were isolated from the roots by activity-directed fractionation with column chromatography, thin layer chromatography (TLC) and recrystallization, and identified predominantly on the basisof MS and NMR data. The results showed among methanol extracts from various parts of D. fordii var. lucida only that from the roots had insecticidal activity. The methanol extract from the roots of D. fordii var. lucida had toxicity against the 4th instar larva of Aedes albopictus, Aphis gossypii Glover, Aphis craccivora ， Myzus persicae , the 2nd instar larva of Herse convolvuli (L.), the neonate larva of Scirpophaga incertulas (Walker), the 2nd instar larva of Pieris rapae (L.) and the adult of Phyllotreta striolata (Fabricius) ， with LC₅₀ value 24 h after treatment being 260 . 3 mg/L, 234 . 6 mg/L, 141 . 3 mg/L, 16 . 4 mg/L, 233 . 4 mg/L, 20 . 8 mg/L, 11 . 7 mg/L and 148 . 4 mg/L, respectively. The methanol extract from the roots had high contact toxicity and stomach toxicity to the 3rd instar larva of H. convolvuli at 24 h after treatment, with the values being 101 . 6 mg/L and 234 . 9 mg/L, respectively. Moreover, the sublethal dosage of the extract had sublethal effect of antifeeding and growth inhibition. Three compounds were isolated from the roots and identified: rotenone, β -sitosterol and 6a , 12a -dehydrodegulin. The two compounds, rotenone and 6a , 12a -dehydrodeguelin, possessed poison effect on the neonate larva of S. incertulas, with the LC₅₀ value at 24 h after treatment being 2 . 6 mg/L and 5 . 3 mg/L, respectively. It is so concluded that only the roots of D. fordii var. lucida had insecticidal activities, contact toxicity and stomach toxicity were the important action mode, and rotenone and 6a , 12a -dehydrodeguelin were the main active ingredients in the roots.

【Aim】 This study aims to explore the physiological responses of silicon-treated rice plants to feeding stress by the white-backed planthopper, Sogatella furcifera, so as to provide a theoretical foundation for the rational use of silicon fertilizer in controlling S. furcifera. 【Methods】 The plants of the susceptible rice variety TN1 were treated with two silicon application levels ［grown in nutrient solution added with sodium silicate (Na₂SiO₃·9H₂O) at the concentration of 112 mg/L(Si+treatment group), and without addition of silicon (control group)］, then exposed to the feeding of the 3rd instar nymphs of S. furcifera. The contents of superoxide anion (O^-·₂), hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), the activities of antioxidant enzymes ［superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT)］ and defense enzymes ［phenylanlanine ammonialyase (PAL), polyphenol oxidase (PPO) and lipoxygenase (LOX)］, and lignin content in rice leaf sheaths were measured at 0, 12, 24, 48, 72 and 96 h after feeding on the rice plants in the Si+ treatment group or the control group. 【Results】 Compared to the control group, the Si+ treatment group exhibited significant reductions in O^-·₂ content by 7.8%-17.9% in rice leaf sheaths during 12-72 h after S. furcifera feeding, in H2O2 content by 15.5%-43.1% in rice leaf sheaths during 12-96 h (except at 24 h) after S. furcifera feeding, and in MDA content by 15.1%-35.3% in rice leaf sheaths during 24-96 h after S. furcifera feeding. During 12-96 h (except at 72 h) after S. furcifera feeding, the CAT and SOD activities in rice leaf sheaths in the Si+ treatment group were significantly increased by 25.8%-44.8% and 21.4%-47.6%, respectively, as compared to those in the control group. During 12-72 h (except at 24 h) after S. furcifera feeding, the POD activities in rice leaf sheaths in the Si+ treatment group significantly increased by 19.1%-54.5%, as compared to those in the control group. During 12-72 h after S. furcifera feeding, the PAL and PPO activities in rice leaf sheaths in the Si+ treatment group were significantly elevated by 17.6%-70.8% and 16.7%-38.3%, respectively, as compared to those in the control group. During 12-96 h after S. furcifera feeding, the LOX activities in rice leaf sheaths in the Si+ treatment group significantly increased by 9.9%-105.4%, as compared to those in the control group. During 12-96 h (except at 48 h) after S. furcifera feeding, the lignin contents in rice leaf sheaths in the Si+ treatment group significantly raised by 12.5%-59.3%, as compared to those in the control group. 【Conclusion】 Silicon application enhances rice resistance to S. furcifera by reducing reactive oxygen species and MDA contents, as well as by increasing the activities of antioxidant enzymes and defense enzymes, and the lignin content in rice leaf sheaths infested by S. furcifera.

【Aim】This study aims to explore the effects of exogenous juvenile hormone (JH) on the female ovarian development and transcription levels of the reproduction-key genes of Spodoptera frugiperda.【Methods】Polyclonal antibody was prepared after isolation and purification of vitelline protein of female S. frugiperda. The vitelline protein contents in the 7-8-day-old female pupae and 1-10-day-old female adults of S. frugiperda were determined using indirect enzyme-linked immunosorbent assay (ELISA). The 5-day-old female pupae were treated with 25 μg/individual precocene, and the newly emerged female adults were supplemented with 100 μg/individual exogenous JH analogue methoprene. Subsequently, the preoviposition period, oviposition period, number of eggs laid per female and hatching rate were calculated. The 3-day-old female adults were dissected to observe the ovarian development, measure the ovarian length, and take photographs. Finally, the expression levels of genes of vitellogenin (Vg) and vitellogenin receptor (VgR) in the 3-day-old female adults of each treatment were quantified using qPCR.【Results】The vitellin content of S. frugiperda increased first and then decreased as female adult emerged, reaching its peak in the fat bodies at 1-day-old and ovaries at 4-day-old. Precocene treatment resulted in a downregulation of the endogenous JH in S. frugiperda, leading to obvious impediment of ovarian development compared to the normally reared female adults as the blank control group. Additionally, there was a significant reduction in the amount of ovarian eggs. However, exogenous supplementation of methoprene after the precocene treatment effectively restored normal ovarian development. The average ovarian length in the precocene treatment group was 39.89 mm, which was significantly shorter than that in the blank control group of 49.79 mm. The average ovarian length in the methoprene treatment group was 46.67 mm, and significantly longer than that in the precocene treatment group, but had no significant difference from that in the blank control group. The average number of eggs laid per female in precocene treatment group significantly decreased to 576.33 grains, as compared to that in the blank control group (1 128.37 grains), significantly decreased by 48.91%. The average number of eggs laid per female in the methoprene treatment group was 806.93 grains, which was significantly higher than that in the precocene treatment group. The expression levels of SfVg and SfVgR in female adults treated with precocene were significantly downregulated as compared to those in the blank control group, while methoprene significantly promoted the expression levels of SfVg and SfVgR.【Conclusion】The suppression of endogenous JH through precocene treatment significantly impeded female fecundity, hindered normal ovarian development, and reduced the transcription levels of SfVg and SfVgR in female adults of S. frugiperda. Conversely, exogenous JH supplementation effectively restored the female fecundity. This study further validates the crucial regulatory role of JH in the reproductive process of S. frugiperda and provides a theoretical foundation for future research on the regulatory mechanism of JH of female reproduction.

【Aim】 This study aims to conduct tissue expression profile and RNAi effect analyses of tropomyosin gene of Helicoverpa armigera HaTm-1,  previously identified through transcriptome screening, so as to explore the feasibility and theoretical basis of HaTm-1 as a broad-spectrum RNA biopesticide target for lepidopteran pests. 【Methods】 The cDNA sequence of HaTm-1 of H. armigerawas cloned using PCR and analyzed through bioinformatics. RT-qPCR was used to detect the expression levels of HaTm-1 in different tissues of the 4th instar larvae, including cuticle, fat body, midgut, brain, testis and Malpighian tubules, as well as in the cuticle of the 2nd-5th instar larvae of H. armigera. RNAi of HaTm-1 was performed by spraying dsHaTm-1 on the 1st instar larvae of H. armigera and Spodoptera frugiperda. The expression levels ofHaTm-1 of H. armigera and HaTm-1 of S. frugiperda, and the larval weight and larval survival rate of these species were subsequently measured. dsHaTm-1 was injected into the 4th instar larvae of H. armigera, and the larval weight, pupal weight, relative growth rate, relative metabolic rate and food intake were detected. 【Results】 The full-length cDNA of HaTm-1 of H. armigera was 855 bp (GenBank accession no.: XP021201113.1). HaTm-1 had the amino acid sequence identity of over 90% with homologous proteins in multiple species. Tm-1s of H. armigera and S. frugiperda converged on the same clade, suggesting a close relationship between them. HaTm-1 was specifically and highly expressed in the cuticle of the 4th instar larvae of H. armigera and was also expressed in the cuticle at different larval instars, with peak expression occurring in the cuticle of the 5th instar larva. There was a decrease in the expression level of HaTm-1 by 55.73% and 50.85% at 12 and 24 h, respectively, a significantly reduced larval survival rate by 43.33% at 168 h， and decrease in the larval weight by 17.67%, 14.65% and 18.90% at 72, 120 and 168 h, respectively, after spraying dsHaTm-1 to the 1st instar larvae of H. armigera, as compared with those in the dsEGFP control group. The expression level of SfTm-1 decreased by 44.72% and 39.65% at 12 and 24 h, respectively, after spraying dsHaTm-1 on the 1st instar larvae of S. frugiperda as compared with those of the dsEGFP control group. The larval survival rate was observed to be 60% at 144 h after spraying dsHaTm-1 on the 1st instar larvae of S. frugiperda. The larval weight decreased by 13.00% and 28.11% at 72 and 120 h， respectively, after spraying dsHaTm-1 on the 1st instar larvae of S. frugiperda as compared with those of the dsEGFP control group. The expression level of HaTm-1 was significantly reduced by 55.66% at 24 h, the larval weight was decreased by 50.34% and 54.91%, respectively, at 72 and 120 h， the pupal weight was reduced by 13.14%, the relative growth rate was decreased by 14.00%, the relative metabolic rate was decreased by 15.66% and the food intake was decreased by 14.50%, after injecting the 4th instar larvae of H. armigera with dsHaTm-1 as compared with those of the dsEGFP control group. These alterations in feeding and digestion processes ultimately resulted in larval death. 【Conclusion】 The interference of HaTm-1 expression is vital to the feeding and digestion processes in H. armigera. This study underscores the potential of tropomyosin as a broad-spectrum target for RNA biopesticides, offering valuable insights for its practical application.

【Aim】 This study aims to further understand the symbiotic relationship between auchenorrhynchan insects of the order Hemiptera and endosymbionts by investigating the diversity and functions of obligate and facultative symbiotic bacteria in bacteriomes, fat bodies and other related tissues of adult Tettigetta isshikii at the ultrastructural and genomic levels. 【Methods】 Field-collected female and male adults of T. isshikii were investigated to clarify the distribution of symbiotic bacteria Karelsulcia, Hodgkinia and Wolbachia in the bacteriomes, fat bodies, ovaries, spermathecae, salivary glands, conical segment, filter chamber and gut of female adults, and testes of male adults through transmission electron microscopy and fluorescence in situ hybridization. Metagenomic sequencing, assembly and functions by genome annotation of symbiotic bacteria in the bacteriomes and fat bodies of female adults, and testes of male adults of T. isshikii were conducted. The phylogenetic relationships of Wolbachia in T. isshikii and other insects were determined using maximum-likelihood and Bayesian inference methods to determine the phylogenetic position of Wolbachia. 【Results】 The obligate symbiotic bacteria Karelsulcia and Hodgkinia are harbored in the bacteriomes of female adults of T. isshikii, and the facultative symbiotic bacterium Wolbachia, belonging to the supergroup F, was harbored not only in the cytoplasm and nuclei of the epithelial cells of testicular follicles but also in the nuclei of sperms of male adults and fat bodies of female adults. Genome annotation analysis revealed that genes of Karelsulcia and Hodgkinia were involved in the synthesis of essential amino acids and vitamins for the host cicada, while genes of Wolbachia were involved in riboflavin metabolism, heme pathway, and biosynthesis of purine and pyrimidine. The duplication of Wolbachia occured within the nuclei of sperms, which ultimately may lead to the rupture of nuclei. Phylogenetic relationship revealed that this Wolbachia belongs to the F supergroup and was closely related to the Wolbachia harbored in other arthropods and nematodes. 【Conclusion】 This study clarified the potential nutritional functions of symbionts Karelsulcia and Hodgkinia in the bacteriomes of T. isshikii adults, and elucidated a unique phenomenon that Wolbachia may have both beneficial and detrimental effects for the host insects. The results of this study contribute to a further understanding of the symbiotic relationship and complex co-evolution between Cicadidae and symbiotic bacteria.

The contact toxicity of 11 synthetic polyacetylenes was tested to newly hatched Periplaneta americana larvae with the method of drug film. The results showed these compounds, 1-t-butyl-4-hydroxymethyldiacetylene, 1-benzyl-4-methyldiacetylene and di-ethyl-2-propargylthiophosphate caused high mortality of over 70% to the larvae at 20 μg/cm² The LC₅₀ of 1-benzyl-4-methyl diacetylene and di-ethyl-2-propargyl thiophosphate was 3.91 μg/cm² and 1.50 μg/cm², respectively. The effect on AChE of the insect indicated that 1-t-butyl-4-hydroxymethyldiacetylene, 1-phenyl-4-o-nitrophenyl-diacetylene and di-ethyl-2-propargylthiophosphate inhibited its activity, and the inhibition rate was 12.00%, 27.24% and 62.22%, respectively. To ATPase, 1-t-butyl-4-hydroxymethyl diacetylene and di-ethyl-2-propargyl thiophosphate inhibited the activity of Na⁺K⁺ATPase, and the inhibition rate was 44.55% and 31.44%, respectively; but 1-phenyl-4-(3,4methylenedioxy)-phenyl-diacetylene and 1-phenyl-4-m-nitrophenyl-diacetylene enhanced the activity of the enzyme, and the activation rate was 24.98% and 20.61%; 1-t-butyl-4-hydroxymethyl-diacetylene,1-phenyl-4-p-methoxyphenyldiacetylene and 1-phenyl-4-o-nitrophenyl-diacetylene inhibited the activity of Ca²⁺Mg²⁺ATPase, and the inhibition rate was 49.02%, 38.53% and 35.32%, respectively; but other compounds enhanced activity of the enzyme, among them the highest activity was observed with 1-phenyl-4-(3,4-methylenedioxy)-phenyl-diacetylene, and the activation rate was 81.12%.

【Aim】 To determine the toxicity of plumbagin (an important plant secondary substance from Chinese medicine Plumbago zeylanica) to the 2nd instar larvae of Spodoptera frugiperda and ascertain the sublethal effect of a sublethal concentration (LC₂₅) of plumbagin on the growth, development and detoxifying enzyme activities of the F₀ and F₁ generations of S. frugiperda. 【Methods】 The toxicity of plumbagin against the 2nd instar larvae of S. frugiperda (the 2nd day after ecdysis) was determined using insecticide incorporated artificial diet bioassay. After the 2nd instar larvae were exposed to LC₂₅ (0.343 mg/g) of plumbagin, the larval duration, pupation rate, pupal duration, female and male adult longevity, and number of eggs laid per female adult of the F₀ and F₁ generations of S. frugiperda were recorded, the age-stage, two-sex life table was constructed, and the activities of three detoxifying enzymes including carboxylesterase (CarE), glutathione S-transferase (GST) and cytochrome P450 monooxygenase (CYP450) were determined at 24 h after treatment. 【Results】 The median lethal concentration (LC₅₀) and LC₂₅ values of plumbagin against the 2nd instar larvae of S. frugiperda in 7 d were 0.607 and 0.343 mg/g, respectively. For the parental generation (F₀) of S. frugiperda, LC₂₅ of plumbagin significantly decreased the pupation rate by 25.13%, significantly shortened the female and male pupal duration by 1.29 and 1.08 d, respectively, significantly decreased the number of eggs laid per female by 47%, and significantly shortened the oviposition period and adult longevity by 1.75 and 1.19 d, respectively, compared to the vehicle control (0.1% acetone). For the offspring generation (F₁) of S. frugiperda, LC₂₅ of plumbagin only significantly shortened the mean generation time (T) by 0.91 d, compared to the vehicle control. LC₂₅ of plumbagin significantly induced the activities of the three detoxifying enzymes CarE, GST and CYP450 in the 2nd instar larvae of S. frugiperda after treatment for 24 h, which were increased to 1.28-, 1.30- and 1.42-fold as high as those in the vehicle control. 【Conclusion】 LC₂₅of plumbagin had obvious adverse effects on the growth, development and fecundity of the F₀ generation of S. frugiperda, and significantly increased the activities of the three detoxifying enzymes CarE, GST and CYP450 in the 2nd instar larvae after treatment for 24 h, which will be helpful for using the plant secondary substance plumbagin as one of potent biocontrol strategies for this pest.

【Aim】 This study aims to enrich the basic information of 14-3-3ζ gene of Apis cerana cerana, so as to provide a reference and basis for its further functional study. 【Methods】 The coding sequence (CDS) of 14-3-3ζ gene was amplified by RT-PCR, followed by TA cloning and Sanger sequencing. The physicochemical properties and molecular features of 14-3-3ζ were predicted using the relevant software, and the phylogenetic analysis of 14-3-3ζ was performed. RT-qPCR was used to detect the expression levels of 14-3-3ζ gene in different developmental stages (egg, larva, pre-pupa, pupa and adult), and different tissues (antennae, midgut, fat body, hypopharyngeal gland, brain, cuticle and venom gland) of the newly emerged adult workers of Ap. cerana cerana, as well as in the guts of the 4-, 5- and 6-day-old larvae of Ap. cerana cerana after inoculating the 3-day-old larval workers with Ascosphaera apis.【Results】 The CDS of 14-3-3ζ gene of Ap. cerana cerana was successfully cloned, including 744 nucleotides and encoding 247 amino acids. 14-3-3ζ of Ap. cerana cerana had the molecular weight of about 28.0 kD, included 26 phosphorylation sites, four structural domains and one conserved motif, but had no transmembrane domains and signal peptides. The 14-3-3ζ proteins of Ap. cerana cerana, Ap. mellifera, Ap. laboriosa, Ap. florea, Ceratina calcarata, Bombus pyrosoma, B. terrestris, Megachile rotundata, Osmia lignaria and Habropoda laboriosa all contained four identical conserved motifs and one same structural domain (14-3-3_1). The 14-3-3ζ proteins of Ap. cerana cerana and Ap. mellifera clustered into a single clade on the phylogenetic tree. The expression level of 14-3-3ζ gene in Ap. cerana cerana eggs was significantly higher than those in the 3-day-old larvae, 1-day-old prepupae, 2-day-old prepupae and 4-day-old pupae. The differences in the expression level of 14-3-3ζ gene in various day-old adult workers of Ap. cerana cerana were non-significant. The expression level of 14-3-3ζ gene in the venom gland of the newly emerged adult workers of Ap. cerana cerana was the highest, significantly higher than those in the antennae, midgut, hypopharyngeal gland, brain, cuticle and fat body. Following inoculation of the 3-day-old larval workers of Ap. cerana cerana with As. apis, the expression levels of 14-3-3ζ gene in the 4-, 5- and 6-day-old larval worker guts of Ap. cerana cerana were significantly down-regulated as compared with those in the control group.【Conclusion】 Ap. cerana cerana 14-3-3ζ gene is specifically and highly expressed in the venom gland and egg of worker, and the expression of 14-3-3ζ gene in the larval guts is activated in the process of As. apis infection. 14-3-3ζ is a putative hydrophilic, non-transmembrane and intracellular protein, and highly conserved in Ap. cerana cerana and the above other ten bee species. There is the closest genetic relationship between 14-3-3ζ of Ap. cerana cerana and Ap. mellifera.
Key words: Apis cerana cerana; 14-3-3; molecular features; expression pattern; Ascospaera apis

【Aim】To explore the safety of commonly used insecticides to the natural enemy Picromerus lewisi and their impact on predation function.【Methods】The spraying method was applied to detect the median lethal concentration (LC₅₀) values of 14 insecticides of 6 categories ［broflanilide, cyantraniliprole, chlorantraniliprole and flubendiamide (diamides), lambda-cyhalothrin (pyrethroids), imidacloprid, flupyradifurone and sulfoxaflor (neonicotinoids and analogues), triflumezopyrim (mesoionics), emamectin benzoate, spinosad and abamectin (macrolides), and bioinsecticides (Bacillus thuringiensis and Metarhizium anisopliae CQMa421)］ against the 3rd instar nymphs of P. lewisi, and the safety factor was used to evaluate their safety to P. lewisi. In addition, the predatory efficiency of the 3rd instar nymphs of P. lewisi treated with four insecticides mentioned above (lambda-cyhalothrin, cyantraniliprole, imidacloprid and abamectin) on the 2nd and 3rd instar larvae of Mythimna seperata was analyzed using the disc equation. 【Results】Chlorantraniliprole, cyantraniliprole and abamectin exhibited relatively high safety levels to the 3rd instar nymphs of P. lewisi, and flubendiamide demonstrated the highest safety to the 3rd instar nymphs of P. lewisi, with the LC₅₀ value of 369.71 mg/L in 24 h and the safety factor of 7.39. In contrast, lambda-cyhalothrin, imidacloprid and broflanilide exhibited lower safety to the 3rd instar nymphs of P. lewisi, and broflanilide showed the lowest safety to the 3rd instar nymphs of P. lewisi, with the LC₅₀ value of 0.01 mg/L in 24 h and the safety factor of below 0.01. After 48-h treatment, sulfoxaflor, chlorantraniliprole, flubendiamide and cyantraniliprole maintained relatively high safety to the 3rd instar nymphs of P. lewisi, and flubendiamide was recorded to have the highest safety to the 3rd instar nymphs of P. lewisi, with the LC₅₀ value of 299.28 mg/L and the safety factor of 5.99. Conversely, broflanilide, imidacloprid and lambda-cyhalothrin exhibited lower safety to the 3rd instar nymphs of P. lewisi, and broflanilide had the lowest safety to the 3rd instar nymphs of P. lewisi, with the LC₅₀ value of 0.04 mg/L in 48 h and the safety factor of below 0.01. The bioinsecticides M. anisopliae CQMa421 and B. thuringiensis were found to be relatively safe to the 3rd instar nymphs of P. lewisi. Compared to the control treated with 0.1% Triton X-100, the stress from the four insecticides (lambda-cyhalothrin, cyantraniliprole, imidacloprid and abamectin) reduced the predation efficiency of the 3rd instar nymphs of P. lewisi, although had no significant effect on the instantaneous attack rate. Notably, under the stress of 20 mg/L cyantraniliprole, the 3rd instar nymphs of P. lewisi had the highest instantaneous attack rates (0.623 and 0.586, respectively) on the 2nd and 3rd instar larvae of M. seperata.  Under the stress of 2 mg/L abamectin, the 3rd instar nymphs of P. lewisi nymphs had the shortest handling time for prey (0.091 and 0.076 d, respectively), the highest maximum daily predation amount (11.00 and 13.12 individuals, respectively) and the highest predatory efficiency (4.72 and 5.34, respectively) on the 2nd and 3rd instar larvae of M. seperata, while under the stress of 2 mg/L imidacloprid, the 3rd instar nymphs of P. lewisi had the lowest predation efficiency (2.46 and 2.08, respectively) on the 2nd and 3rd instar larvae of M. seperata. 【Conclusion】Chlorantraniliprole, flubendiamide and M. anisopliae CQMa421 demonstrated higher safety to P. lewisi nymphs, whereas broflanilide and imidacloprid exhibited lower safety to P. lewisi nymphs. Additionally, treatments with abamectin and cyantraniliprole had minimal impact on the predation capability of P. lewisi nymphs, while imidacloprid treatment significantly affected their predation capability. These findings play a crucial role in providing information for pest management strategies in agricultural environments by minimizing the toxicity of insecticides.

【Aim】The purpose of this study is to detect the expression patterns of ass-milR0037-3p and its key target genes in worker larvae of Apis cerana cerana and A. mellifera ligustica in the process of Ascosphaera apis infection, so as to offer a foundation for further exploring the mechanism of ass-milR0037-3p regulating the As. apis infection. 【Methods】 The target genes of ass-milR0037-3p of As. apis were predicted using related software and then annotated to GO and KEGG databases. Stem-loop RT-PCR and RT-qPCR were employed to verify the expression of ass-milR0037-3p in the guts of the 6-day-old worker larvae of Ap. c. cerana and Ap. m. ligustica and to detect the expression levels of ass-milR0037-3p and its two key target genes (genomic protein acetyltransferase gene ESA1 and flavin containing amine oxidase gene FAO) in the guts of the 4-6-day-old worker larvae after feeding the 3-day-old worker larvae of Ap. c. cerana and Ap. m. ligustica with diets containing 1×10⁷ spores/mL As. apis, respectively.【Results】 ass-milR0037-3p could target 225 genes annotated to 28 GO terms such as reproduction, binding and cells, as well as 105 KEGG pathways such as splicing, biosynthesis of secondary metabolites, and amino sugar and nucleotide sugar metabolism. The target fragments of ass-milR0037-3p were amplified in the guts of the 6-day-old worker larvae of both Ap. c. cerana and Ap. m. ligustica infected with As. apis. The expression levels of ass-milR0037-3p in the guts of the 5- and 6-day-old worker larvae of Ap. c. cerana infected with As. apis were significantly up-regulated and the expression levels of ESA1 and FAO in the guts of the 5- and 6-day-old worker larvae of Ap. c. cerana infected with As. apis were significantly down-regulated, as compared with those of the 4-day-old worker larvae of Ap. c. cerana infected with As. apis. The expression level of ass-milR0037-3p in the gut of the 5-day-old worker larvae of Ap. m. ligustica infected with As. apis was up-regulated as compared with that of the 4-day-old worker larvae of Ap. m. ligustica infected with As. apis and that of the 6-day-old worker larvae of Ap. c. cerana infected with As. apis was significantly up-regulated as compared with that of the 4-day-old worker larvae of Ap. m. ligustica infected with As. apis. The expression levels of the target genes ESA1 and FAO in the gut of the 5-day-old worker larvae of Ap. m. ligustica infected with As. apis were up-regulated and those of the 6-day-old worker larvae of Ap. m. ligustica infected with As. apis were significantly up-regulated as compared with those of the 4-day-old worker larvae of Ap. m. ligustica infected with As. apis.【Conclusion】 ass-milR0037-3p potentially modulates the process of As. apis infecting worker larvae of Ap. c. cerana by negatively regulating the FAO expression, while potentially affects the process of As. apis infecting worker larvae of Ap. m. ligustica through positively regulating the ESA1 expression. The results provide a basis for elucidating the mechanism by which milRNAs respond to the As. apis infection of honeybee larval guts via regulating the expression of target genes.

【Aim】The parasitic beetle, Dastarcus helophoroides (Fairmaire) is the dominant species among the natural enemies against the wood-boring insects of trees. The research of the circadian rhythm of male and female adult behaviors of D. helophoroides can help us to understand their biological characteristics and illustrate their living habits. 【Methods】 The behaviors of D. helophoroides adults were observed and recorded at a 30 min interval by single rearing separately under the temperature 27±1℃ and relative humidity 65%±10% in the laboratory from July 10th to 15th, 2014. The observed behaviors of males and females were divided into five types, i.e., moving, foraging, drinking, resting with contacting wood and resting without contacting wood. 【Results】 The moving and resting with contacting wood behaviors of D. helophoroides adults showed obvious circadian rhythm. The moving behavior mostly took place during the dark period, and the peak of moving was recorded at 20:30-22:30 and 2:00-4:00 of the dark period, while the lowest was recorded at 6:00-16:30 of the light period. The resting with contacting wood occurred mainly at 9:30-16:30 of the light period and 0:00-1:30 of the dark period, while the lowest was recorded at 20:30-23:00 of the dark period. The occurrence of foraging and drinking behavior was very low within a day and happened mainly after 0:00 and before 14:00 o’clock. The peak of resting without contacting wood behavior was recorded at 0:30-3:30 and 20:00-22:00 of the dark period. The percentages and time points of occurrence of various behaviors showed no significant difference between females and males. 【Conclusion】 The behaviors of D. helophoroides are influenced significantly by light and dark conditions. The moving behavior mostly takes place during the dark period; however, there is no significant difference in the percentages of various behaviors between females and males.

【Aim】 To identify genes of the CYP6Y subfamily in Anopheles sinensis, analyze their structure and characteristics, and deduce their possible functions. 【Method】 The CYP6Ys cDNA transcripts were retrieved and identified by two ways of Blasts using An. gambiae CYP6Y1 as query sequence against the transcriptome data of An. sinensis. The structure, characteristics and possible functions of the CYP6Y genes identified were analyzed using bioinformatics methods. 【Results】 Two genes of the CYP6Y subfamily were identified from An. sinensis transcriptome sequencing data, and named as AsCYP6Y1 (GenBank accession number: KF709397) and AsCYP6Y2 (GenBank accession number: KF709398) , respectively. Sequence analysis showed that AsCYP6Y1 and AsCYP6Y2 are 1 713 bp and 1 815 bp in length, encoding 502 and 526 amino acids, respectively. Gene structure analysis showed that the genes of this subfamily only contain one phase “1” intron and form a conserved synteny with other P450 genes. Protein structure prediction showed that the encoded proteins of these two genes contain five P450 characteristic sequences and six specific substrate binding sites, but have no signal peptide sequence, and are localized in cytoplasm. The 3D structural prediction showed that AsCYP6Y1 has 18 α-helix and 13 anti-parallel β-strands, while AsCYP6Y2 has 19 αhelix and 11 antiparallel βstrands. Two CYP6Y genes were also identified in An. darlingi using the same method. Phylogenetic analysis showed that AsCYP6Y1 and AsCYP6Y2 are grouped with CYP6Y1 and CYP6Y2 of other Anopheles species, respectively, with the bootstrap values greater than 90%. The Ka/Ks ratio analysis showed that the Ka/Ks ratio of AsCYP6Y1 and AsCYP6Y2 compared with CYP6Y1 and CYP6Y2 of other Anopheles species were all less than 1. The relative evolutionary rate test showed that the rates of subfamilies CYP6Y and CYP6M were significantly higher than those of the subfamily CYP6P in An. sinensis, while no significant difference between subfamilies CYP6Y and CYP6M was observed. 【Conclusion】 Two genes of the CYP6Y subfamily have been identified and characterized in An. sinensis and An. darlingi, which had earlier been found in two other Anopheles species ( An. gambiae and An. funestus), suggesting the wide and specific existence of the genes of the CYP6Y subfamily in Anopheles.
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【Aim】Prediction of the adult emergence dynamics of the overwintering generation of Chilo suppressalis is crucial for the accurate prediction and control of its offspring generations. This study aims to develop a model simulation for predicting the adult emergence dynamics of the overwintering generation of C. suppressalis.【Methods】To obtain model parameters, the overwintering larval populations of C. suppressalis were collected from the paddy fields in Xing′an, Guangxi, South China in February, March and April 2021 and the developmental duration of immature stages and adult emergence rate were determined at four temperatures (14, 18, 22 and 26 ℃). The developmental rates of immature stages (the reciprocal of developmental duration) were fitted with linear and nonlinear models, and the adult emergence rate was fitted using a three-parameter Weibull equation. Model parameters were calculated using Origin 2022.【Results】From the perspective of model fitting, the model established based on the developmental duration of immature stages and adult emergence rate data of the overwintering larval population of C. suppressalis collected in March performed better (R_adj²_March=0.9445, R_adj²_February=0.9083, R_adj²_April=0.8380). However, the field observation data showed that the nonlinear larval development Schoolfield model {V(T)=0.64×T/298.15×exp［47.11/1.99×(1/298.15-1/T)］} and the adult emergence Weibull equation {F∑(V(T))=1-exp［-(∑V(T)+0.04)/1.05)×5.95］} based on the overwintering larval population collected in February gave the least deviation of adult emergence dynamics between the predicted value and the field-observed value, which was 1.0-5.3 d. 【Conclusion】In practical application, local air temperature data are input into the above models to obtain the predicted adult emergence dynamic values of the overwintering generation of C. suppressalis, which, when corrected by the deviation values obtained in this study, will provide accurate prediction of adult emergence dynamics and aid in the decision-making for the accurate control of C. suppressalis.

Bactrocera fruit flies are an enormous threat to fruit and vegetable production throughout the world, causing great economic loss. Although chemical control based on conventional insecticides and biotechnical tools including sterile insect technique (SIT) and male annihilation technique (MAT) have been the main weapons used in most control programs, they still have many restrictions, and ecofriendly control tools against Bactrocera spp. are urgently needed. Thus, current knowledge about sexual communication and related behaviours in Bactrocera spp. was reviewed in this article to help build behaviour-based control strategies. Two different polygynous mating systems in Tephritidae and behavioural sequences of males before courtship in lekking sites were summarized, and some mated female behaviours in oviposition sites, including oviposition marking behaviour and fighting behaviour for single oviposition sites, were elaborated. Future perspectives were also outlined. The knowledge about sexual communication is expected to provide new insights and references for integrated pest management programs for tephritid pests.

【Aim】This study aims to make clear the sublethal effects of the botanical insecticides azadirachtin and celangulin on Ostrinia furnacalis larvae and their effects on the activities of detoxification enzymes, so as to explore the detoxification mechanism of O. furnacalis on these two pesticides. 【Methods】The toxicity of azadirachtin and celangulin to the 3rd instar larvae of O. furnacalis in 72 h was analyzed by dipping method. The 3rd instar larvae of O. furnacalis were exposed to sublethal concentrations (LC₁₀, LC₂₀ and LC₄₀) of azadirachtin and celangulin for 6, 12, 24, 48 and 72 h, and the activities of three main detoxification enzymes including glutathione S-transferase (GST), carboxylesterase(CarE) and cytochrome P450(CYP450) were measured by enzyme-linked immunosorbent assay (ELISA). 【Results】 TheLC₁₀, LC₂₀ and LC₄₀ values of azadirachtin against the 3rd instar larvae of O. furnacalis in 72 h were 0.352, 0.614 and 1.290 mg/L, respectively, and those of celangulin against the 3rd instar larvae of O. furnacalis in 72 h were 0.032, 0.086 and 0.321 mg/L, respectively. After the 3rd instar larvae of O. furnacalis were exposed to LC₁₀, LC₂₀ and LC₄₀ of azadirachtin, the GST activity was firstly inhibited, then induced, and finally inhibited, and 12-h exposure to LC₁₀ of azadirachtin had an induction effect on the GST activity. LC₁₀, LC₂₀ and LC₄₀ of azadirachtin mainly inhibited the activities of CarE and CYP450. After the 3rd instar larvae of O. furnacalis were exposed to LC₁₀, LC₂₀ and LC₄₀ of celangulin, the activities of GST and CarE were mainly inhibited, and the CYP450 activity was firstly inhibited and then induced, while 72-h exposure to LC₁₀ of celangulin had an induction effect on the CYP450 activity. 【Conclusion】Azadirachtin and celangulin can affect the detoxification metabolism of O. furnacalis, and the inhibition of azadirachtin and celangulin on the activities of GST, CarE and CYP450 in O. furnacalis is one of the reasons for their insecticidal activity, GST only plays a certain role in metabolism on low concentration (LC₁₀)  of azadirachtin and CYP450 only plays a certain role in metabolism on low concentration (LC₁₀)  of celangulin.

【Aim】 To elucidate the biological function of serine protease inhibitor Kazal-type 7 (SPINK7) and its immunorecognition mechanism of Bombyx mori against bacterial infestation. 【Methods】 Bioinformatics analysis of SPINK7 of B. mori was made followed by prokaryotic expression, and the secondary structure and heat resistance of SPINK7 were analyzed by circular dichroism. The tendency of SPINK7 to form aggregates was detected by non-reducing electrophoresis and Western blot. The 5th instar larvae of B. mori were infected by Escherichia coli and Staphylococcus aureus, respectively, and the expression levels of SPINK7 gene in hemocytes and fat bodies were detected by qPCR, and the SPINK7 content in serum was detected by Western blot. The inhibitory activities of SPINK7 against E. coli, S. aureus and Micrococcus luteus were analyzed by bacterial inhibition curves. The binding of SPINK7 to pathogen-associated molecular patterns (PAMPs) such as peptidoglycan and lipopolysaccharide, and the binding of SPINK7 to E. coli, S. aureus and M. luteus were detected by immunofluorescence localization and enzyme-linked immunosorbent assay (ELISA).【Results】 SPINK7 consists of three conserved Kazal structural domains, each of which contains six cysteines, forming three pairs of disulfide bonds. The secondary structure of SPINK7 is mainly α-helix, which is consistent with the typical secondary structure of Kazal structural domains, it is heat-resisting and remains structurally stable at 80 ℃. After induction by E. coli and S. aureus, SPINK7 formed a multimeric structure, and the expression levels of SPINK7 gene in hemocytes and fat bodies and the SPINK7 content in the serum of the 5th instar larvae of B. mori were significantly up-regulated, as compared with those of the phosphate buffer solution (PBS) control group. SPINK7 did not directly inhibit bacterial activity, and promoted bacterial aggregation by binding to bacterial PAMPs. 【Conclusion】SPINK7 is a Kazal-type immune-associated protein, whose expression is up-regulated and multimerized after bacterial infestation in B. mori, and helps hemocytes to aggregate bacteria and exert immune effect by recognizing and binding to bacterial PAMPs.

 【Aim】This study aims to investigate the chemotaxis and thermotaxis of Bursaphelenchus xylophilus, by analyzing the physicochemical properties, structure and functions of the members of TAX family and the expression of TAX family gene Bx-taxs after drug and low-temperature stresses. 【Methods】Based on the NCBI database, the TAX family gene sequences of Caenorhabditis elegans were retrieved and compared with B. xylophilus genome for tBlastn to search candidate TAX family genes in B. xylophilus. The candidate sequences were screened and cleaned by software and online tools such as ExPASy, WoLF PSORT, SOPMA, Alphafold 3, MEME, CD-Search and TBtools, and Bx-taxs of B. xylophilus were identified and the gene structure and chromosomal localization, and the subcellular localization, sequence features, physicochemical properties, secondary structure and tertiary structure of their proteins were analyzed. A phylogenetic tree of TAX protein sequences was constructed by IQtree software to infer phylogenetic relationships. The expression profiles of six Bx-taxs were analyzed to determine the roles of Bx-taxs by subjecting B. xylophilus to emamectin benzoate drug stress and low-temperature stress. 【Results】A total of six members of the TAX family, Bx-TAX-1-6, were identified from B. xylophilus. These proteins had 623-1 115 amino acid residues, relative molecular masses of 71.49-129.27 kD, and theoretical isoelectric points of 5.68-9.04, most of them being hydrophilic proteins. All six TAX family members were located on the cell membrane, especially the Bx-TAX-1 protein was also distributed in the cytoplasm. The secondary structures of the TAX family proteins were mainly composed of α-helixes and random coils, accounting for 42.10%-53.07% and 32.38%-45.80%, respectively. Chromosomal localization and phylogenetic analyses showed that TAX family genes of B. xylophilus were distributed on four chromosomes of which genes were closer to the TAX-4 genes of other nematodes, except for the Bx-TAX-1 gene, which was a separate branch. Under emamectin benzoate stress, all genes of the members of TAX family showed different degrees of down-regulation of expression within 12 h, and up-regulation of expression at 24 h. Among them, the expression levels of Bx-tax-3, Bx-tax-5 and Bx-tax-6 were up- or down-regulated by more than 1-fold. Only Bx-tax-5 was up-regulated under low-temperature stress, while the others were down-regulated. 【Conclusion】 TAX family genes of B. xylophilus show varying degrees of response to emamectin benzoate and low temperature stresses. The results of this study provide a theoretical basis for chemical and temperature tropism studies based on the TAX family, and provide a research basis for B. xylophilus response to chemical stress and its northward migration mechanism.

【Aim】 The camellia weevil,  Curculio chinensis Chevrolat, is an important pest attacking fruits of the oil tea  Camellia, an endemic genus to China, and causes tremendous fruit drop of its host plants. To reveal the oviposition strategy of  C. chinensis, we explored the relationships between oviposition activity of C.chinensis  and the traits of its host fruit. 【Methods】 During the most active oviposition season, 960 fruits of  C. meiocarpa  were collected randomly from an oil tea ( Camellia meiocarpa) farm. The number of punctures and clutch size of C. chinensis, as well as the weight, length and diameter of each fruit were measured, and the difference of fruit size among oviposition-punctured fruits, feeding-punctured fruits and non-damaged fruits was analyzed through one-way ANOVA (LSD), and the effect of fruit traits on feeding and oviposition activity was analyzed through linear regression model. A choice test of the pest on fruits size was carried out, and the difference of fruit size between selected and non-selected fruits was analyzed by paired-samples t test. 【Results】 Oviposition-punctured fruits of C. meiocarpa were significantly larger than feeding-punctured fruits, while the latters were significantly larger than non-damaged fruits in terms of weight, length, diameter, and volume ( P＜0.01). The number of punctures, clutch size and weevil parasitism rate were positively correlated with fruit size ( P＜0.01), suggesting that parental weevils prefer larger fruits to feed and oviposite. The female adults of C. chinensis showed obvious preference to larger fruits when the difference of fruit size between the two fruits tested was significant, and heavily damaged fruits were significantly larger than lightly damaged fruits in terms of length and diameter after infection by female adults of C. chinensis for 12 h. Independent of host fruit traits, C. chinensis laid one egg in each puncture hole. 【Conclusion】 C. chinensis adults prefer larger fruits to feed, which supports the optimal foraging theory. C. chinensis adults also prefer larger fruits to oviposite. The small clutch size would reduce intraspecific competition C. chinensis between offsprings and increase the probability of larval development, and thus they would be more likely to successfully exit from host fruits. We infer that the single egg laying behavior in C. chinensis is a riskspreading strategy, an adaptation toward limited food resource.

5-Hydroxytryptamine (5-HT) is an important biogenic amine in insects, which is synthesized in both neuronal and peripheral tissues and can be reuptaken by serotonin transporter into presynaptic neuron. 5-HT plays various important physiological roles in insects through specific G protein-coupled receptors, such as feeding, circadian behavior, aggregation, learning and memory. There are five types of 5-HT receptors in insects, i.e., 5-HT1A, 5-HT1B, 5-HT2A, 5-HT2B and 5-HT7. 5-HT1A and 5-HT1B inhibit intracellular cAMP production, 5-HT2A and 5-HT2B  increase Ca ²⁺ level, and 5-HT7 induces cAMP production. In recent years, great progress has been made in the research of 5-HT in insects, especially their receptors. More and more 5-HT receptor genes have been cloned, and their functions and pharmacological properties have been analyzed. The pharmacological differences of 5-HT receptors from different insects will provide fundamental basis for designing and developing new specific insecticides for pest management.  

 【Aim】To screen cellulose-degrading bacterial strains from the gut of Allomyrina dichotomus for constructing composite bacterial consortia and to investigate their degradation capacity for spent mushroom substrate (SMS), thereby providing theoretical and practical foundations for SMS waste management.【Methods】Composite bacterial consortia were constructed based on non-antagonistic relationships among six cellulose-degrading bacteria with high enzyme activity (Bacillus velezensis M24, Bacillus subtilis H12, H11, H4 and M33, and Bacillus siamensis M32) isolated and purified from the larval gut of A. dichotomus. The activities of four cellulases (filter paper enzyme, endoglucanase, exoglucanase and β-glucosidase) were determined, the optimal composite bacterial consortia was selected according to its enzymatic activity profile, and the culture time, inoculation amount and incubation temperature for the selected composite bacterial consortia were optimized. The structural changes in the spent mushroom substrate before and after degradation were observed through scanning electron microscope by differential weight method.【Results】The composite bacterial consortia M24∶H11 was constructed based on the enzymatic activities of filter paper enzyme, endoglucanase, exoglucanase and β-glucosidase (18.08, 69.37, 19.09 and 17.95 U/mL, respectively). The optimal culture time, inoculation amount and culture temperature of M24∶H11 were 2 d, 1% and 40 ℃, respectively. Following optimization, the activities of the four cellulases increased significantly by 1.98-2.16-fold. At 25 ℃ within 30 d, M24∶H11 showed a significantly higher SMS degradation rate (up to 38.04%) than individual strains, with the degradation rate increasing over time. The degradation by M24∶H11 effectively disrupted the SMS surface structure, increasing its contact area.【Conclusion】The larval gut-derived composite bacterial consortia M24∶H11 of A. dichotomus was constructed and optimized for enzyme production, with high efficiency in SMS degradation. This study offers novel technical insights and theoretical support for the efficient control of the edible fungi waste.

【Aim】To explore the feasibility and theoretical basis of using sex pheromone and floral scent trapping as monitoring methods for Loxostege sticticalis adults. 【Methods】The developmental status of female ovaries and male testes of L. sticticalis moths trapped by sex pheromone, floral scent and net catching were investigated, dissected and analyzed in the field, and the relationships between the egg quantity, number of trapped moths and the number of field survey populations, and the migration status were analyzed. 【Results】The testes of male moths of L. sticticalis were closely related to the day-old age, and the regression equation was y=1.289-0.1288x+0.003516x2. Field experiments conducted in Kangbao, Hebei during 2020-2023, revealed that when sex pheromone traps successfully captured male moths, the mating rates of female moths were 66.7%-100%, and the proportions of level Ⅳ ovaries were 44.1%-95.4%. When the male moths could not be trapped, the mating rates of female moths were 0%-30.3%, and the level Ⅳ ovaries accounted for 0%-3.6%. From June 12 to June 22, 2024, the average mating rate of female moths trapped by floral scent (94.3%±2.3%) was significantly higher than that of female moths trapped by net catching (76.2%±5.3%) in Dashimen Town, Hexigten Banner, Inner Mongolia Autonomous Region. There was no difference in the average number of matings between female moths trapped by floral scent and by net catching. The number of egg grains remaining in the ovaries of female moths trapped by floral scent was (169.8±15.4), which was significantly lower than the that of female moths trapped by net catching (267.9±20.7). The average testicular volume of the male moths caught by sex pheromone trapping was the largest ［(0.21±0.01) mm³］, followed by that trapped by net catching ［(0.19±0.02) mm³］, and that trapped by floral scent was the smallest ［(0.15±0.01) mm³］. The number of L. sticticalis adults trapped by sex pheromone and floral scent was closely related to their physiological state. Adults attracted by sex pheromone were male moths searching for mates, while those caught by floral scent trapping were female moths which have mated and laid part of eggs, as well as male moths which have mated. Therefore, male moths of the immigrant population and local breeding population of L. sticticalis were sensitive and had strong responses to sex pheromones, while the ovarian development level of female moths of the emigrant population was low, and the corresponding male moths were immature, and had no olfactory behavioral response to sex pheromones and floral scent. This relationship was verified by multiple field experiments in different seasons in Kangbao, Hebei and Hexigten Banner, Inner Mongolia from 2020 to 2024. 【Conclusion】 The combination of sex pheromone trapping and floral scent trapping can help determine the physiological state of L. sticticalis and estimate its migratory status and population dynamics of the next generation in the field.

【Aim】Superparasitism often occurs in parasitoids. In this study, we aimed to investigate the effects of superparasitism on the offspring growth and development of Cotesia ruficrus and to determine the factors that contribute to superparasitism. 【Methods】 The effects of oviposition times of C. ruficrus on the survival rate of the host （the 3rd instar larvae of Cnaphalocrocis medinalis） and the growth and development of the parasitoid offsprings, and the effects of exposure time and parasitoid density on superparasitism were detected in this study. 【Results】Laboratory experiments demonstrated that the superparasitism occurred in C. ruficrus when it parasitized C. medinalis larvae. Female wasps, irrespective of the experience of oviposition, attacked hosts that had been parasitized by themselves or other individuals. The number of parasitoid cocoons increased with oviposition times, while the number of the dead parasitoid larvae increased when the host was parasitized 3 to 5 times. The mortality of the host before emergence of parasitoid adults increased with the oviposition times, and reached to 50% when the host was parasitized 5 times. Superparasitism prolonged the developmental duration, lowered the emergence rate and the female/male ratio of the parasitoid offsprings. The female body size of the parasitoid offsprings significantly decreased as the oviposition times increased, and the superparasitism rate increased with the parasitoid density and exposure time. 【Conclusion】Ovipositing twice on host larvae shows to be the most beneficial to the growth and development of C. ruficrus offsprings, and ovipositing thrice causes superparasitism. Although superparasitism increases the number of parasitoid offsprings, it is detrimental to the progeny development. The superparasitism, therefore, should be avoided in mass rearing of C. ruficrus in laboratory, by means of controlling exposure time and density of the parasitoids.

【Aim】To explore the effects of artificial diet-based rearing technology on the growth, development and reproduction of Anoplophora glabripennis, to clarify the duration and reproductive parameters of A. glabripennis at different developmental stages, and to establish a system of indoor pass-on technology of A. glabripennis based on artificial diet, so as to provide basic data and technical support for the in-depth basic research on A. glabripennis and the development of new preventive and control technologies in the future. 【Methods】 The duration and mortality rates of different developmental stages of A. glabripennis reared on artificial diet were observed and counted. The effects of feeding different host plants (Salix babylonica and Acer negundo branches) on the adult longevity and number of eggs laid by female adult of A. glabripennis were observed and compared to clarify the feeding preference of A. glabripennis adults for different host plants, and to explore the host plant species suitable for indoor nutrient supplementation of A. glabripennis adults. The differences in the adult longevity and number of eggs laid per female of A. glabripennis of different sources (reared on artificial diet indoors and collected in the field) to evaluate the effects of different growing environments on the development and breeding biology of adult A. glabripennis. The pupal weight of A. glabripennis reared on artificial diet indoors and collected in the field was determined to clarify the effects of different growing environments on the pupal weight of A. glabripennis. 【Results】 Under the condition of the temperature 25 ℃, relative humidity of 60% and photoperiod of 16L∶8D, the egg duration of A. glabripennis was (10.39±0.09) d, and the entire larval duration was (153.78±5.93) d. The larval stage of A. glabripennis reared on artificial diet indoors was divided into five instars, and the 1st-5th instar larval duration was (18.28±0.23), (24.75±0.20), (33.30±0.27), (37.67±0.27) and (39.85±0.31) d, respectively. The pupal duration of A. glabripennis was (15.05±0.06) d, and the survival rates of various developmental stages were maintained at more than 90.0%. Nutrient supplementation with A. negundo branches significantly increased the number of eggs laid per female and adult longevity of A. glabripennis, with the average values of (112.97±3.64) grains, (39.90±4.69) d and (30.93±6.75) d, respectively, which were significantly higher than those of females with nutrient supplemention with S. babylonica branches ［(26.33±0.97) grains, (15.23±3.41) d and (15.50±4.35) d, respectively］. In comparison, there were no significant differences in the number of eggs laid per female and female and male adult longevity of A. glabripennis reared on artificial diet indoors ［(101.50±34.17) grains, (41.00±3.82) d, and (30.50±8.23) d, respectively］ and those collected in the field ［(94.25±18.59) grains, (38.63±6.12) d and (25.88±6.60) d, respectively］. In addition, the pupal weight of A. glabripennis reared on artificial diet indoors was (0.95±0.20) g, which was not significantly different from that collected in the field ［(0.93±0.12) g］. 【Conclusion】 The artificial diet rearing method in this study can not only greatly improve the survival rate of artificially bred A. glabripennis, but also achieve the indoor 2 generations/year pass.on rearing, which greatly shortens the breeding cycle of A. glabripennis, and can provide a stable source of test worms for the basic research of A. glabripennis, and lays a foundation for further development of new prevention and control technologies.

【Aim】 This study aims to explore the immune-related genes and immune recognition regulatory network of Plutella xylostella against the entomopathogenic fungus Isaria fumosorosea, so to further investigate the innate immune mechanisms of P. xylostella to I. fumosorosea. 【Methods】 We analyzed the transcriptome of I. fumosorosea infected and uninfected (the control) 4th instar larvae of P. xylostella at 12 h after treatment using high-throughput single-end RNA-sequencing (RNA-seq). The differentially expressed genes and their functions, classifications and signaling pathways were analyzed using bioinformatic tools. 【Results】 The numbers of 12 346 987 and 12 315 210 clean reads were obtained for the infected and uninfected libraries, respectively. Among the two RNA-seq libraries, 60.93% and 61.26% of reads could be mapped to reference genome with a perfect match proportion of 32.15% and 32.73%, respectively. A total of 351 differentially expressed unigenes (DEUs) were identified between infected and uninfected larvae. Among them, 275 DEUs were up-regulated while 76 DEUs down-regulated. In total, there were 156 DEUs putatively involved in immune responses. A Blast2GO analysis indicated that 102 differentially expressed transcripts were able to be assigned to 46 Gene Ontology (GO) terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis mapped 132 DEUs to 13 pathways ( P<0.05).【Conclusion】 Most of the DEUs are putatively linked to innate immune recognition and regulation. These infection-responsive DEUs are enriched in various immune processes, such as energy metabolism, disease and defense response. The findings generated from this study provide a bioinformatic database for searching immune-related genes, and also benefit the future investigations of the innate immune mechanisms of P. xylostella against the entomopathogenic fungus I. fumosorosea.

【Aim】 α-Galactosidase A (GLA), an enzyme crucial for growth and development, nutrient absorption and stress response of organisms, has been extensively studied in both animals and plants. However, the function of GLA in mites has not been reported. This study aims to explore the role of GLA gene in the feeding process of Tetranychus urticae on its host plants, providing important theoretical insights for the development of novel pest mite control strategies. 【Methods】 Based on the genome and salivary proteome of T. urticae, the full-length open reading frame (ORF) sequence of TuGLA was obtained, and bioinformatic analysis was performed using Expasy, Clustal, Jalview and MEGA. RT-qPCR was used to detect the expression levels of TuGLA at different developmental stages (egg, larva, protonymph, deunymph and female adult) of T. urticae, and in female adults of T. urticae at different feeding time points (1, 3, 6, 9, 12 and 24 h) on kidney bean leaves and at 12 h after feeding on different host plant leaves (kidney bean, tomato, cucumber, corn, cotton and tobacco). RNAi was employed to silence TuGLA by feeding female adults of T. urticae with dsTuGLA, and subsequently the mortality within 5 d and number of eggs laid within 5 d, and the feeding damaged area of kidney bean leaves were calculated. 【Results】 The full-length coding sequence (CDS) of TuGLA (GenBank accession number: XP_015794636.1) of T. urticae is 1 275 bp, encoding 424 amino acids, with a predicted protein molecular weight of 48 kD and a theoretical isoelectric point of 5.01. The 1st-18th amino acids at the N-terminus are signal peptide sequences of TuGLA without transmembrane domains. TuGLA was most closely related to GLAs in Tetranychus genus. TuGLA was expressed in various developmental stages of T. urticae, with the highest expression level in the adult stage. The expression level of TuGLA in female adults of T. urticae at 9 h after feeding on kidney bean leaves was the highest and those in female adults of T. urticae at 12 h after feeding on leaves of tomato, cucumber, corn, cotton and tobacco were significantly up-regulated as compared with those in female adults of T. urticae at 12 h after feeding on kidney bean leaves of the control group. After silencing TuGLA, the mortality within 5 d and the number of eggs laid within 5 d of female adults of T. urticae were significantly increased and reduced, respectively, and the feeding damaged area of kidney bean leaves within 24 h significantly decreased after silencing TuGLA as compared to the those of the dsGFP control group. After silencing TuGLA, the mortality of T. urticae fed on the leaves of kidney bean within 3 d significantly increased as compared with that of the dsGFP control group.【Conclusion】 The feeding on different host plants and feeding duration by T. urticae can induce the expression of TuGLA. Silencing TuGLA by RNAi can significantly affect the survival rate and number of eggs laid of T. urticae, and its damage to host plants. The results of this study provide a theoretical basis for further exploring the feeding mechanism of T. urticae.

Tachinids are among the important endoparasitoids for the meadow moth, Loxostege sticticalis  (Lepidoptera: Pyralidae). In most cases, the parasitism rate of tachinids is far higher than that of hymenopterous parasitoids, which exhibits a great potential to regulate the population of meadow moth. In this paper, the tachinid species parasitizing on the meadow moth, types of parasitism, the dominant species and their roles in controlling the host population as well as their protection and utilization were reviewed by combining the results derived from field investigations and literatures. The species of tachinid parasitoids for the meadow moth were as many as 22 species, which were widely distributed along with the meadow moth and most of them were with a broad host range of other phytophagous species. Four dominant species were recorded: Exorista civilis Rondani,  Nemorilla maculosa Meigen, Clemelis pullata Meigen, and Exorista pratensis  Robincau-Uesvoidy. It was found that the controlling effect of these tachinid species on the current generation of meadow moth is limited; however, they could play an important role in controlling the next generation of meadow moth. The conservation and utilization of the tachinid parasitoids were also discussed based on these prospects.

【Aim】 To identify the morphological and molecular characteristics of microsporidium Nosema sp. CP isolated from Catopsilia pyranthe， and then to assess the infectivity and transmissibility of this Nosema sp. CP in the domestic silkworm， Bombyx mori. 【Methods】 After purification of Nosema sp. CP spores isolated from wild C. pyranthe butterflies， the shape， sizes and axial ratios of the spores were measured under the microscope， respectively. The DNA fragments of 16S rDNA from this microsporidian species were amplified by PCR and then sequenced to confirm its classification status. The Nosema sp. CP isolated and N. bombycis was fed respectively to B. mori larvae of day-1 2nd instar and day-1 4th instar to survey the infectivity and transmissibility via embryo during different developmental stages of B. mori. 【Results】 Oblong shape and diplokaryotic nuclei of the isolated microsporidia in this study were visualized under the microscope. Molecular analysis of 16S rDNA sequence showed more than 99% identity to that of the previously reported microsporidian spores isolated from C. pyranthe. The total infection rates of Nosema sp. CP and N. bombycis to B. mori were 68.8% and 98.3%， respectively. In the second generation of B. mori after vaccination， the detection rates of sporozoans of Nosema sp. CP and N. bombycis in newly-hatched larvae were 100% and 100%， and those of their eggshells were 92.9% and 100%， respectively. The embryo transmissibilities of Nosema sp. CP and N. bombycis to B. mori were 9.6% and 23.2%， respectively. 【Conclusion】 The microsporidia isolated in this study are Nosema sp. CP and have typical characteristics of Nosema diplokaryotic nuclei. The Nosema sp. CP isolated can infect B. mori and has transmissibility via embryo in this insect. However， in B. mori both the infection rate and transmission rate of Nosema sp. CP isolated in this study are lower than those of N. bombycis. Thus， it is necessary to prevent and control the prevalence of Nosema sp. CP in sericultural industry.

【Aim】 Heavy metal pollution has become one of important environmental issues globally. This study aims to evaluate the effects of cadmium exposure on the flight performance of the beet armyworm, Spodoptera exigua, a migratory insect. 【Methods】 Insect flight information system was used to study the flight performance of female and male adults of S. exigua at different ages (1, 3, 5 and 7 day-old), which were fed with the artificial diets containing different concentrations of cadmium (0, 0.20, 0.80, 3.20 and 12.80 mg/kg), respectively. 【Results】 There were significant differences in the flight performance of S. exigua adults exposed to different concentrations of cadmium. The 3 day-old female adults stressed by low concentration of cadmium (0.20 mg/kg) were the best fliers with the longest average flight distance (36.96 km). The flight performance of S. exigua adults exposed to 0.20 mg/kg cadmium was not significantly different from that of the control. With the increase of cadmium concentration, the flight performance of both male and female adults decreased significantly. The negative effects of heavy metal cadmium stress on the performance of male adults were more obvious than on that of female adults. 【Conclusion】 Different concentrations of cadmium stress have significant effects on the flight performance of S. exigua adults of different ages and different sexes.  

【Aim】 The camphor sawfly , Mesoneura rufonota, is an important leaf-feeding pest of the camphor tree, Cinnamonum campora. In this study the effects of temperature on the development and reproduction of M. rufonota were investigated so as to provide a foundation for the forecasting and integrated management of this pest. 【Methods】 The duration of different developmental stages, survival rate and reproduction of M. rufonota reared at different constant temperatures (19, 22, 25, 28 and 30℃) were measured and analyzed, the model fitting between the developmental rate and temperature was conducted, and the developmental threshold temperature and effective accumulated temperature were calculated by using the method of least squares. 【Results】 Within the constant temperatures ranging from 19 to 30℃, the average duration of various developmental stages of M. rufonota shortened as temperature rose gradually. This insect needed 34.62 d to complete a life cycle at 19℃, while needed 18.97 d at 30℃. The developmental rates in different stages were positively correlated with temperature, and the relationships between developmental rates and temperature all fitted the quadratic regression model. The developmental thresholds of egg, larva, pupa, adult and whole generation were 5.26, 3.22, 7.66, 8.24 and 5.11℃, respectively, while the effective accumulated temperatures were 65.58, 204.15, 121.94, 65.01 and 460.29 d·℃. The accumulative survival rates in different stages were decreased and the adult longevity shortened with increasing temperature. The fecundity was the highest at the temperatures ranged from 22 to 25℃ and the lowest at 30℃, indicating that higher or lower temperature inhibits oviposition of M. rufonota. 【Conclusion】 Temperature is a key factor affecting the development and reproduction of M. rufonota, and the optimum temperature range for its development and reproduction is from 22 to 25℃. These findings provide a scientific basis for the monitoring and integrated management of this pest.

-【Aim】 Intestinal microbes play important roles in metabolism, growth and development, and immunity in host insects. This study aims to explore the community structure of the intestinal microbes of Picromerus lewisi adults and their ability to metabolize carbon sources. 【Methods】The intestinal microbes were isolated and purified from P. lewisi adults by in vitro culture and identified by molecular biology technologies. The bacterial 16S rDNA and fungal ITS genes were sequenced by Illumina high-throughput sequencing technology to analyze the structure and diversity of the intestinal bacterial and fungal communities of P. lewisi adults. PICRUSt and FUNGuild were used to predict the function of bacteria and fungi and their genes. Biolog ECO technique was used to analyze the carbon source metabolic function of the intestinal bacteria and fungi of P. lewisi adults. 【Results】A total of 10 strains of Enterococcus sp. were isolated from the intestinal culturable dominant bacteria of P. lewisi, and the dominant bacterium was E. faecalis. The results of high-throughput sequencing showed that the dominant phyla in the intestine of P. lewisi adults were Proteobacteria (relative abundance: 58.51%) and Firmicutes (relative abundance: 38.92%), and the most dominant fungal phylum was Ascomycota (relative abundance: 56.53%), followed by Basidiomycota (relative abundance: 11.34%). The most dominant bacterial genus in the intestine of P. lewisi adults was Enterococcus (relative abundance: 25.05%), followed by Lactococcus (relative abundance: 12.23%), Serratia (relative abundance: 11.48%) and Providencia (relative abundance: 2.38%), and the most dominant fungal genus was Biappendiculispora (relative abundance: 18.30%), followed by Cladosporium (relative abundance: 11.83%), Vishniacozyma (relative abundance: 517%), and Phallus (relative abundance: 3.62%). The community structure of the intestinal bacteria and fungi of P. lewisi adults showed high species richness and diversity, and strong metabolic ability on carbon sources, being able to efficiently metabolize 27 carbon sources including α.cyclodextrin, L.serine, sutrescine, D.malic acid etc. Functional predictions showed that the intestinal bacterial taxa of P. lewisi adults were mainly distributed in metabolism, environmental information processing, genetic information processing etc., and the fungal taxa were mainly distributed in unassigned taxa, plant saprotroph, undefined saprotroph, endophyte.plant pathogen, fungal parasite.undefined saprotroph, leaf saprotroph, and animal pathogen.endophyte.plant pathogen.wood saprotroph. 【Conclusion】 P. lewisi adults have a high variety and diversity of intestinal bacteria and fungi. The dominant bacterial genera were Enterococcus, Lactococcus, Serratia and Providencia, and the dominant fungal genera were Biappendiculispora, Cladosporium, Vishniacozyma and Phallus, with a strong metabolic ability on carbon sources..

【Aim】 To analyze the molecular features, phylogeny and gene expression profiles of DNA methyltransferase 3 (DNMT3) from Apis cerana (AcDNMT3), so as to provide a reference and foundation for further functional analysis of AcDNMT3. 【Methods】 Bioinformatics tools were used to predict and analyze the physicochemical properties, molecular features, structural domains, conserved motifs and phylogeny of AcDNMT3 in Ap. cerana. RT-qPCR was used to measure the relative expression levels of AcDNMT3 in different developmental stages (egg, 3-day-old larva, 1-day-old prepupa, 2-day-old prepupa, 4-day-old pupa, and 1-, 2-,6-,12-,15- and 18-day-old adults) of Ap. cerana workers, and various adult tissues (antennae, hypopharyngeal gland, brain, cuticle, midgut, fat body and venom gland) of Ap. cerana workers, as well as in the guts of the 4-6-day-old larval workers of Ap. cerana after inoculating the 3-day-old larval workers with Ascosphaera apis and the midguts of the 2-5-day-old adults after inoculating the 1-day-old adult workers with Nosema ceranae. 【Results】 AcDNMT3 contains 758 amino acids with the approximate molecular weight of 88.24 kD, the molecular formula of C₃₉₄₅H₆₁₈₂N₁₀₇₆O₁₁₃₄S₄₄, the theoretical isoelectric point (pI) of 8.32, and the average hydrophilic index of -0.454. AcDNMT3 was predicted to interact with ten proteins, including adenosylhomocysteinase (A0A2A3ECJ8). DNMT3 proteins from Ap. cerana, Ap. mellifera, Ap. dorsata, Ap. laboriosa, Bombus terrestris, Frieseomelitta varia, Osmia lignaria, Colletes gigas, Eciton burchellii, Nylanderia fulva, Linepithema humile, Solenopsis invicta, Monomorium pharaonis and Vollenhovia emeryi contained four common structural domains (PWWP_DNMT3, ADDz_Dnmt3, Dcm domain and PWWP domain) and five common conserved motifs (Motif1, Motif2, Motif3, Motif4 and Motif5). AcDNMT3 was clustered with Ap. mellifera DNMT3 on the phylogenetic tree. AcDNMT3 was differentially expressed in the eggs, larvae, prepupae and pupae of Ap. cerana workers, with the highest expression level in the 2-day-old prepupae which was significantly higher than those in eggs, the 3-day-old larvae, 1-day-old prepupae and 4-day-old pupae. AcDNMT3 was differentially expressed in adult workers of Ap. cerana at the 1-, 2-, 6-, 12-, 15- and 18-day-old, with the highest expression level in the 1-day-old adults which was significantly higher than those in the 2-, 6-, 12-, 15- and 18-day-old adults. Differential expression of AcDNMT3 was observed across various adult worker tissues including antennae, venom gland, brain, midgut, fat body, cuticle and hypopharyngeal gland of Ap. cerana, with the highest expression level in the antennae and the lowest expression level in the midgut. After infecting the 3--day-old larval workers of Ap. cerana with As. apis, the expression level of AcDNMT3 in the gut of the 4-day-old larval workers was significantly down-regulated as compared with that of the control. After inoculation of the 1-day-old adult workers of Ap. cerana with N. ceranae, the expression levels of AcDNMT3 in the midguts of the 2-, 4- and 5-day-old adult workers were significantly up-regulated and that in the 3-day-old adult workers was extremely significantly down-regulated as compared with those of the control. 【Conclusion】 AcDNMT3 is a putative hydrophilic, intracellular, non-transmembrane protein that is highly conserved across Ap. cerana and the above other insect species. AcDNMT3 shows the highest homology with DNMT3 of Ap. mellifera.AcDNMT3 may play a role in the development of Ap. cerana workers, in the larval responses to As. apis infection, and in the adult responses to N. ceranae infection.

【Aim】 To address the issue of low transfering rate of sperm after artificial insemination of Apis cerana cerana and to extend the time during which inseminated queens lay fertilized eggs within the colony.【Methods】 A. c. cerana gynes at 7 d after eclosion were inseminated with 1, 3 and 5 μL of semen. For each dosage, there were treatment groups and control groups, with 5 gynes in each group. After insemination, a plastic device was immediately fitted onto the end of abdomen of queen in the treatment group, while no plastic device was fitted onto the end of abdomen of queen in the control group. After 24 h, the number of sperms in the spermatheca of each queen was counted. 【Results】Physical isolation via the fitted plastic device significantly improved the transferring rate of sperm of A. c. cerana to spermatheca after artificial insemination. It demonstrated a particularly notable advantage under low semen dosage, with the transferring rate of sperm increasing from approximately 11% under conventional artificial insemination to 43%. This effectively extended the time during which the queen laid fertilized eggs within the colony. The application of this method successfully elucidated the inheritance pattern of body color in A. c. cerana, demonstrating that body color is controlled by a pair of alleles, with yellow being dominant and black recessive. The three castes in the offspring resulting from the backcrossing of the heterozygous queen (F ₁) with the black drone exhibited a stable 1∶1 segregation ratio of body color, consistent with Mendelian inheritance. 【Conclusion】 The optimized sperm transfer method developed in this study significantly enhances the transferring rate of sperm of A. c. cerana from lateral oviduct to spermatheca in artificially inseminated queens, particularly under low semen dosage, such as single-drone insemination. This method can be successfully applied in the study of the inheritance pattern of body color, providing both a theoretical foundation and technical support for the selective breeding and germplasm conservation of A. c. cerana.

【Aim】 Ultraspiracle protein (USP) belongs to an essential component of a heterodimeric receptor complex with the ecdysone receptor (EcR) which binds ecdysteroids. This study aimed to explore the distribution of USP mRNA in heads of different castes of Polyrhachis vicina Roger adults for inferring the influence of PvUSP on the function or behavior of cranial nerve of P. vicina, and to assess the influence of PvUSP on physiological functions in the body and the relationship with EcR gene by feeding PvUSP dsRNA as RNAi to different castes of P. vicina adults. 【Methods】 We used real-time fluorescent quantitative PCR technique and fluorescence in situ hybridization to test the distribution of PvUSP mRNA expression level in the head of different castes of P. vicina adults, conducted RNAi on different castes of P. vicina adults by feeding PvUSP dsRNA, and used the real-time fluorescent quantitative PCR technique to detect the changes in PvUSP and PvEcR mRNA levels. 【Results】 PvUSP was widely expressed in the head of different castes of P. vicina adults, and especially concentrated in the mushroom body. However, its expression level was quite different in the three castes. The highest expression level was found in the head of workers, the moderate expression level was in the head of male adults, and the lowest was in the head of female adults. After using RNAi technique to silence PvUSP, the expression level of PvUSP mRNA decreased obviously in bodies of different castes of P. vicina adults as compared to the control, and an extremely significant difference was observed ( P<0.01), while the expression levels of PvEcR mRNA all increased in different castes. The changes in PvEcR mRNA level in workers and male adults were not distinct （ P>0.05), while those in the female body showed a dramatic increase ( P<0.05). 【Conclusion】 PvUSP may be related to the structure and functions of nervous system of P. vicina. PvUSP and PvEcR may have relevance in the formation of heterogeneous dimers or function complementarily. PvUSP may influence the reproductive functions of female adults of P. vicina.

The insect brain is composed of protocerebrum, deutocerebrum and tritocerebrum. The protocerebrum contains advanced sensory centers, such as mushroom body and central complex, controlling the advanced neural activities of insects, such as learning, memory and mobility. The deutocerebrum includes antennal lobe, which is the center of olfactory systems, while the tritocerebrum, which usually is not developed well, includes endocrine and mobile neurons. Unlike other organisms, the auditory and visual systems of insects are relatively degenerated because of their special biological characteristics. They prey, communicate and call mainly depending on the olfactory system, and thus their olfactory systems have been delicately developed. We here reviewed the research progress in the neuronal structure and function of insect brain (central complex, mushroom body and antennal lobe) and the genetic variation of the brain structures (sex dimorphism, differences between developmental stages, insects, and insects and other animals), and summarized the revealed mechanisms that insect brains process and identify odor signals.

【Aim】 This study aims to provide data for elucidating the application value of  the body temperature of the diamondback moth, Plutella xylostella, in its management.  【Methods】 At different temperatures in the artificial climate incubators (ambient  temperatures), the body temperature of the 2nd, 3rd, and 4th instar larvae of P. xylostella  was measured, and the relation equations between the body temperature of various instar  larvae (y) and the ambient temperature (x) were established. Meanwhile, the body  temperatures of the 3rd instar larvae of P. xylostella at different time after treatment  with different concentrations of avermectin, chlorpyrifos, fipronil and cypermethrin,  respectively, at different ambient temperatures were measured. 【Results】 The relation  equations between the ambient temperature (x) and the body temperature (y) of the 2nd, 3rd  and 4th instar larvae of P. xylostella were y＝0.95x+1.19 (r=0.9463), y＝0.95x+1.18  (r=0.9988), and y＝0.93x+1.45 (r=0.9989) with the corresponding isothermal points of 22.16 ℃, 21.40℃ and 21.41℃, respectively. When the ambient temperature was set at 15℃ or 40 ℃, none of the four pesticides changed the body temperature of the 3rd instar larvae of P.  xylostella. However, at the other ambient temperatures, the body temperature of the 3rd  instar larvae of P. xylostella could be changed by pesticide treatment. For avermectin, at  25℃, the body temperatures of the 3rd instar larvae in the 2, 4 and 8 mg/L treatment  groups at 12 h, 2 and 4 mg/L treatment groups at 24 h, 0.5, 2, 4 and 8 mg/L treatment  groups at 36 h and 0.5, 1, 2 and 8 mg/L treatment groups at 48 h were significantly  increased, while that in the 8 mg/L treatment group at 24 h was significantly decreased; at  30℃, those in the 0.5 mg/L treatment group at 24 h and 1 mg/L treatment group at 36 h were  significantly decreased and those in the 1 mg/L treatment group at 48 h and the treatment  groups at various concentrations at 60 h were significantly increased; and at 35℃, only  those in the 1 and 8 mg/L treatment groups at 48 h were significantly decreased as compared  to that in the control. For chlorpyrifos, at 20℃, the body temperatures of the 3rd instar  larvae in the 50, 200 and 800 mg/L treatment groups at 24 h and 100, 400 and 800 mg/L  treatment groups at 36 h were significantly decreased; at 25℃, those in the 100 and 200  mg/L treatment groups at 12 h, 800 mg/L treatment group at 24 h and 100, 200 and 800 mg/L  treatment groups at 60 h were significantly decreased, but those in the 50, 100, 200 and  400 mg/L treatment groups at 24 h, 100 and 200 mg/L treatment groups at 36 h and 100 and  400 mg/L treatment groups at 48 h were significantly increased; and at 30℃, only that in  the 800 mg/L treatment group at 24 h was significantly decreased and those in the 50, 100,  200 and 800 mg/L treatment groups at 60 h were significantly increased as compared to that  in the control. For fipronil, at 20℃, only the body temperature of the 3rd instar larvae  in the 0.5 mg/L treatment group at 36 h was significantly decreased; at 25℃, that in the 4 
mg/L treatment group at 12 h and those in the treatment groups at various concentrations at  60 h were significantly decreased, and that in the 0.5 mg/L treatment group at 24 h and  those in the 0.25, 1 and 2 mg/L treatment groups at 48 h were significantly increased; at  30℃, those in the 0.25 and 0.5 mg/L treatment groups at 12 h, 0.25 and 2 mg/L treatment  groups at 24 h, 4 mg/L treatment group at 48 h and 2 mg/L treatment group at 60 h were  significantly decreased; and at 35℃, only those in the 0.25 and 0.5 mg/L treatment groups  at 60 h were significantly increased as compared to that in the control. For cypermethrin,  at 20℃, the body temperatures of the 3rd instar larvae in the 2 and 8 g/L treatment groups  at 36 h and 4 and 8 g/L treatment groups at 48 h were significantly increased; at 25℃,  those in the 2, 4 and 8 g/L treatment groups at 12 h were significantly decreased and those  in the 0.5, 4, and 8 g/L treatment groups at 24 h, 1, 4 and 8 g/L treatment groups at 36 h,  and 1, 2 and 4 g/L treatment groups at 60 h were significantly increased; and at 30℃,  those in the 0.5 and 1 g/L treatment groups at 12 h, 0.5, 1, 4 and 8 g/L treatment groups  at 24 h and 1, 2 and 8 g/L treatment groups at 60 h were significantly decreased as  compared to that in the control. 【Conclusion】 The autonomic thermoregulation ability of  P. xylostella larvae is comparatively low. Avermectin, chlorpyrifos, fipronil or  cypermethrin treatment can affect the body temperature of the 3rd instar larvae of P.  xylostella, but the effect varies with the pesticide type and concentration, ambient  temperature and treatment time. The results expand the studies on pesticide toxicology and  pest control.

【Aim】 Junonia coenia densovirus (JcDV) encodes four viral structural proteins, namely VP1, VP2, VP3 and VP4 via leaky scanning. This study aims to explore the expression characteristics of four viral structural protein genes (vp1-4) of JcDV in Spodoptera frugiperda larvae, so as to provide the basis for further studying on the function of the virus structural proteins VP1-4 and the mechanism of their assembly. 【Methods】 The subcellular localization of VP1-4 within Hi5 cells transfected with pJcDV plasmid was detected by immunofluorescence. The expression levels of vp1-4 in the 2nd instar larvae of S. frugiperda at 0, 3, 6, 12, 24, 48, 72, 96 and 120 h after infection by JcDV were analyzed through RT-PCR. The expression levels of VP1-4 in the 2nd instar larvae of S. frugiperda at 0, 24, 48, 72, 96 and 120 h after infection by JcDV were detected by Western blot. Immunohistochemistry was adopted to detect the tissue expression characteristics of VP1-4 in the 2nd instar larvae of S. frugiperda at 0, 24 and 96 h after infection by JcDV. 【Results】 VP1-4 were mainly located in the cytoplasm of Hi5 cells and only distributed in small amounts in the nucleus in the form of polymers. The transcription of vp1-4 could be detected in the 2nd instar larvae of S. frugiperda at 3 h after infection by JcDV, and stably extended to at least 120 h. VP1-4 were expressed at 24－120 h after infection by JcDV. The expression of VP1-4 was detected in the cuticle and trachea of the 2nd instar larvae of S. frugiperda at 96 h after infection by JcDV, however, there were no obvious expression signals in the muscle and fat body. 【Conclusion】 The transcription and translation of the viral structural protein genes vp1-4 could be detected in the 2nd instar larvae of S. frugiperda at 3 and 24 h, respectively, after infection by JcDV, indicating that VP1-4 might play an important role in the early stages of infection. JcDV could infect the cuticle and trachea of S. frugiperda larvae, which might be one of the causes accounting for their death.