【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】 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.

【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】 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】 The objective of this study is to explore the effects of chronic bee paralysis virus (CBPV) infection on the immunity and gut microbiota of Apis cerana cerana. 【Methods】 The newly emerged adult workers of A. c. cerana were infected with CBPV by injecting CBPV RNA, and the survival rate was recorded. qRT-PCR was used to detect the genomic copy number of CBPV and the expression levels of 15 immune-related genes (PGRP-S2, Cactus1, Toll, PGRPLC-like, Relish, Defensin1, Hymenoptaecin, SP14, PPO, Dicer, HSP70, HSP90, Stat, Stam and PISA3) in the CBPV-infected A. c. cerana workers. Metagenomic sequencing was used to detect the change of the gut microbiota structure in the CSBV-infected A. c. cerana workers, and alpha- and beta- diversity analyses of gut microbiota were performed. 【Results】The survival rate of A. c. cerana adult workers infected with CBPV gradually decreased as compared with that of the control group injected with 1×PBS, and the cumulative survival rate decreased to 27% on the 7th day after infection. The overall trend of the genomic copy number of CBPV increased sharply and steadily from the 1st-4th day, with a 10⁶-fold increase of genomic copy number on the 4th day after infection. Meanwhile, CBPV infection significantly activated the Toll immune pathway genes, especially the antimicrobial peptide genes Defensin1 and Hymenoptaecin, with their expression levels significantly up-regulated on the 1st-5th day after CBPV infection and more than 10-fold on the 5th day. Metagenomic sequencing result showed that CBPV infection affected the gut microbiota structure, and significantly changed the gut microbiota diversity of A. c. cerana workers, especially the relative abundance of Lactobacillus apis and Snodgrassella alvi was significantly down-regulated in the CBPV-infected A. c. cerana as compared with that in the control. 【Conclusion】 CBPV infection can cause the death of A. c. cerana workers, activate the expression of antimicrobial peptide genes and significantly reduce the relative abundance of beneficial bacteria in the gut. The results provide an important theoretical basis for developing bees’ new health management strategies and prevention and control measures.

DNA barcoding has been widely used and received much attention in recent years， pushing the development of multidisciplinary researches including taxonomy， and promoting the studies of evolutionary biology and ecology. DNA barcoding is not only used in initially proposed simple species identification， but also in studies of animal diets， food chain， and food web by using short DNA fragments extracted from in feces and intestinal contents of animals. In this article， we systematically reviewed the recent advances in DNA barcoding and its application in the identification of diets of insect herbivores， and elaborated the selection of DNA barcodes in the identification of herbivore diets and the contribution in the diet composition analysis. Finally， the related research of diets of Lepidoptera larvae was reviewed. Since the herbivorous Lepidoptera larvae cause serious damages and losses to forestry and agriculture，  the study of their trophic relationship has great significance for pest control and management.

【Aim】 To explore the toxicity of terpenoid compounds from Tithonia diversifolia against Spodoptera litura larvae and their effects on the activities of acetylcholinesterase (AChE) and detoxification enzymes, and to screen new active ingredients for the control of S. litura. 【Methods】 The stomach toxicity of three terpenoid compounds including squalene, caryophyllene oxide and α-pinene against Spodoptera litura larvae in 10 d was evaluated with laboratory bioassay by feeding the 3rd instar larvae with the diets supplemented with S-methoprene (positive control) and different concentrations of terpenoid compounds, respectively, and the larval weight was determined at 7 d after treatment. Enzyme-linked immunosorbnent assay was used to measure the effects of different concentrations of caryophyllene oxide on the activities of AChE and detoxification enzymes［carboxylesterase (CarE), cytochrome P450 (CYP450), and glutathione S-transferase (GST)］ in S. litura larvae after treatment for 3 d. Finally, molecular docking techniques were employed to simulate the binding affinity and binding sites of squalene, caryophyllene oxide and α-pinene with AChE, CarE, CYP450 and GST.【Results】 Within 3-5 d after the 3rd instar larvae of S. litura were fed with the diets supplemented with different concentrations of terpenoid compounds, 1×10^-1-1×10^-4 mg/mL caryophyllene oxide exhibited obvious toxicity to S. litura larvae, causing the corrected larval mortality rates significantly higher than squalene, α-pinene and the positive control. At 7 d after treatment, caryophyllene oxide significantly inhibited larval weight gain of S. litura, and the high-concentration caryophyllene oxide treatment group (1×10^－1 mg/mL) showed significantly lower larval weight than the positive control group. At 3 d after treatment with 1×10^-5-1×10^-3 mg/mL caryophyllene oxide, the activities of AChE, CarE and GST in S. litura larvae were significantly reduced, as compared with those in the positive control group, and the CYP450 activity was also significantly reduced as compared with that in the vehicle control (95% alcohol). Notably, when the test concentration of caryophyllene oxide was 1×10^－1 mg/mL, the AChE activity in S. litura larvae was negative. Molecular docking results further demonstrated that caryophyllene oxide had the strongest binding affinity to AChE, with the binding free energy lower than its interactions with CarE, CYP450 and GST. 【Conclusion】 Caryophyllene oxide exhibits high toxicity against S. litura larvae. It exerts insecticidal effects on S. litura by inhibiting AChE activity, highlighting its potential as a green control resource. This provides novel insights for the development and utilization of terpenoid compounds from T. diversifolia.

【Aim】 The purpose of this study is to evaluate the effects of flonicamid on the predation ability of the 3rd instar larvae of Harmonia axyridis. 【Methods】 The residual film method was employed to observe and analyze the effects of exposure of the 3rd instar larvae of H. axyridis to 0.146 and 0.292 mg/mL of flonicamid on their predation ability on Aphis gossypii adults, including the predation amount, instantaneous attack rate, handling time and searching efficiency.【Results】 The predatory functional responses of the 3rd instar larvae of H. axyridis treated with 0.146 and 0.292 mg/mL of flonicamid to A. gossypii adults followed the type Ⅱ functional responses, which can be described by Holling’s disc equation. Compared with the control group treated with acetone, exposure of 0.146 and 0.292 mg/mL of flonicamid had negative impacts on the predation functional response parameters and searching efficiency of the 3rd instar larvae of H. axyridis， and the effects on the predation and searching ability became more significant with the increase of the concentration of flonicamid. In the control group, and treatment groups with 0.146 and 0.292 mg/mL of flonicamid, the instantaneous attack rates of the 3rd instar larvae of H. axyridis were 0.9260, 1.4451 and 2.1197, respectively, the maximum daily predation amounts were 392.62, 52.63 and 32.15 individuals, respectively, and the handling time was 0.0025, 0.0190 and 0.0311 h, respectively. 【Conclusion】 Exposure of the 3rd instar larvae of H. axyridis to flonicamid has negative impacts on their predation function on A. gossypii adults. Therefore, in order to better protect and utilize natural enemies, when implementing the prevention and control of A. gossypii using H. axyridis, consideration should be given to reducing or not using chemical pesticides.

【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】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】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 explore the sublethal effects oflambda-cyhalothrin on Conogethes  punctiferalis, so as to provide a theoretical basis for its scientific application in field management of this pest. 【Methods】 The 48-h toxicity of lambda-cyhalothrin to the 3-day-old adults of C. punctiferalis was determined using the residual film method. Sublethal concentrations (LC₁, LC₅ and LC₁₀) of lambda-cyhalothrin were applied to the 3-day-old adults of C. punctiferalis, the adult longevity, fecundity and survival rate of the F₀ generation, and the larval and pupal duration, fecundity, hatching rate, pupation rate and emergence rate of the F₁ generation were observed and recorded. The age-stage, two-sex life table was constructed for the F₁ generation. 【Results】 Within 48-h exposure, the LC₁, LC₅ and LC₁₀ values of lambda-cyhalothrin against the 3-day-old adults of C. punctiferalis were 0.233, 0.601 and 0.995 mg/L, respectively. Exposure of the 3-day-old adults of C. punctiferalis to LC₁, LC₅ and LC₁₀ of lambda-cyhalothrin resulted in reductions in the average number of eggs laid per female by 5.56%, 24.55% and 51.06%, respectively. After the 3-day-old adults of C. punctiferalis were exposed to LC₁₀ of lambda-cyhalothrin, the adult longevity of the F₀ generation was significantly shortened by 1.60 d, and the oviposition period significantly decreased by 1.29 d, as compared to those in the control group exposed to acetone. The results for the F₁ population showed that, in the treatment groups with LC₁, LC₅ and LC₁₀ of lambda-cyhalothrin, the survival rates at the egg stage decreased by 8.70%, 13.75% and 30.38%, respectively, the average number of eggs laid per female decreased by 38.05%, 30.75% and 24.84%, respectively, and the survival rates at the pupal stage decreased by 2.70%, 13.87% and 27.68%, respectively, as compared to those in the control group. In addition, in the treatment groups with LC₁, LC₅ and LC₁₀ of lambda-cyhalothrin, the peak values of the age-specific reproductive rate (f_x) curve in the F₁ generation of C. punctiferalis decreased by 39.76%, 34.68% and 39.56%, respectively, and the pre-oviposition period, the peak of the female age-specific fecundity, intrinsic rate of increase (r), finite rate of increase (λ) and net rate of increase of the F₁ generation (R₀) were all significantly reduced as compared to those in the control group. 【Conclusion】 Sublethal concentrations of lambda-cyhalothrin significantly reduced the longevity and fecundity of the F₀ generation of C. punctiferalis, and also caused a significant decline in the number of eggs laid of the F₁ generation, thereby suppressing population growth. The results of this study provide a theoretical foundation for the scientific application of lambda-cyhalothrin in the management of C. punctiferalis.

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】 Bactrocera is the most economically significant genus of tephritid flies. The present study aimed to make molecular identification and phylogeny analysis of common species of the genus Bactrocera. 【Methods】 Twenty-one species belonging to eight subgenera of the genus Bactrocera that were frequently intercepted from customs were identified using DNA barcoding technology based on mitochondrial cytochrome c coxidase subunit I (COI) gene. The partial fragments (about 650 bp) of the mtDNA COI gene were amplified using DNA barcoding universal primers. The obtained COI gene fragments were sequenced and aligned. The phylogenetic tree was established by a neighbor-joining (NJ) method. The intra-and inter-species genetic distances were calculated with MEGA version 5.0 software using the Kimura 2-parameter model. 【Results】 The identification results based on the phylogenetic tree were consistent with those based on morphological analyses for the 21 fruit fly species. In these species, 11 species of Bactrocera formed monophylies of themselves, the other 10 species shared one monophyly， and all bootstrap values of the original divergence among different haplotypes within the same species were over 99%. The average inter-species genetic distance between the 21 species was 35.8 times higher than the average intraspecies genetic distance of the 10 species (0.1540 vs. 0.0043). There was no overlap between intra- and inter-species genetic distances. 【Conclusion】 The results indicated that the DNA barcoding based on the partial sequence of mtDNA COI gene cauld provide rapid and accurate identification of Bactrocera species. The technology could be used in identifying and monitoring tephritid fruit fly species.

【Aim】The morphology of Apis cerana cerana is influenced by geographical and ecological differences. This study aims to explore the morphological differences of A. c. cerana in different regions of Guizhou Province, Southwest China, so as to provide a reference for understanding its morphological differentiation across regions. 【Methods】A total of 7 524 worker samples of the A. c. cerana Yunnan-Guizhou Plateau ecotype were collected from 33 regions across Guizhou Province. The morphological parameters related to proboscis, legs, tergites, sternites and forewings were measured post-dissection. The random forest (RF), neural network (NN) and support vector machine (SVM) methods were employed for parameter tuning to identify the optimal morphological analysis model and screen the importance of variables. The K-means unsupervised cluster analysis was conducted based on the importance of variables. The Pearson correlation analysis was performed between important variables and temperature, precipitation and vegetation values. 【Results】 The machine learning results indicated that the optimal parameter tuning was achieved with the mtry set at 5 and random tree at 550. Among the three models, SVM outperformed RF and NN in terms of precision, recall and F1 score. The importance values for the wax mirror interval on sternite Ⅲ, wax mirror length on sternite Ⅲ, wax mirror width on sternite Ⅲ, cubital2 and forewing length were notably high. The cluster analysis results revealed that the worker samples of A. c. cerana from 12 regions such as Zhijin, Zhenfeng and Chishui clustered into one group, those from 8 regions such as Nayong, Hezhang and Ceheng clustered into another, while those from 13 regions such as Xifeng, Pingtang and Luodian were distributed across both groups. The correlation analysis result showed a highly significant positive correlation between the wax mirror length on sternite Ⅲ and the wax mirror width on sternite Ⅲ and forewing width, and the wax mirror width on sternite Ⅲ was also highly positively correlated with the forewing width. The wax mirror interval on sternite Ⅲ was significantly negatively correlated with the cumulative spring-winter temperature difference over three years, and cubital2 exhibited a significant negative correlation with the cumulative spring-winter temperature difference over three years. 【Conclusion】 The machine learning results indicate morphological differentiation of A. c. cerana across different regions in Guizhou Province. This SVM method offers new insights into bee classification and aids in understanding the evolutionary trends of A. c. cerana morphology within Guizhou Province. It also has implications for the conservation of local A. c. cerana germplasm resources in Guizhou Province.

【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 oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), is a worldwide fruit pest, causing serious loss to fruit productivity. Our study aims to explore the thermal tolerance and adaptability of G. molesta adults after exposure to short-term high temperature. 【Methods】 The survival rate of G. molesta adults exposed to various temperatures (36, 38, 40, 42, 44 and 46℃) for different time durations (0.5, 1, 2, 4 and 8 h), and the effects of shortterm high temperature exposure (heat hardening at 38 or 40℃ for 1 h) on their thermal tolerance, longevity, fecundity, and egg hatchability were investigated. 【Results】 The results showed that the survival rate of G. molesta adults was reduced with the increase of exposure duration to high temperature. The survival rate of G. molesta adults at 42℃ was significantly improved after adults were subjected to heat hardening at 38℃ or 42℃ for 1 h ( P<0.05). After treatment at 41℃ for 1 h, G. molesta adults were heat shocked, with longer life span and less fecundity. The further study confirmed that the male adults were greatly affected by high temperature, resulting in the reduction of eggs laid by female adults paired. Nevertheless, there was no remarkable difference in egg hatchability in different treatments though their parents underwent thermal stress. 【Conclusion】 These results suggest that high temperature exceeding 41℃ for 1 h causes negative effects to G. molesta adults, but G. molesta adults have a relatively high thermal tolerance and the potential to adapt to high temperatures.

【Aim】 Tenebrionid beetles are a group of ground-dwelling insects with strong adaptability, complex feeding habits and robust reproductive capabilities. The impact of topographic factors on the distribution of tenebrionid beetles in the northern Qilian Mountains, northwestern China was analyzed, aiming to reveal the diversity distribution patterns of tenebrionid beetles under different topographic conditions. 【Methods】 During July-August, 2022, 65 sampling sites were selected across four altitude gradients in the northern Qilian Mountains region: 1 000-2 000, 2 000-3 000, 3 000-4 000, and above 4 000 m. Active trapping method was employed to investigate the species composition and distribution of tenebrionid beetles. Canonical correspondence analysis (CCA) and curve regression fitting were utilized to analyze the relationships between the diversity distribution of tenebrionid beetles and topographic factors in the northern Qilian Mountains. 【Results】 A total of 5 682 adult tenebrionid beetles, belonging to 124 species of 34 genera, were identified and counted. Among them, Anatolica potanini and Scleropatrum horridum horridum were the dominant species, accounting for 14.4% and 24.8% of the total captured individuals, respectively. The Shannon-Wiener diversity index and Simpson dominance index of tenebrionid beetles were the highest at the altitudes between 2 000-3 000 m, the Margalef’s richness index of tenebrionid beetles was the highest at the altitudes between 1 000-2 000 m, and the Pielou’s evenness index of tenebrionid beetles was the highest at the altitudes between 3 000-4 000 m. At the altitudes above 4 000 m, only a few sciophilous and hygrophilous species of Blaptini and Platyscelidini were found. CCA result showed that altitude and slope had significant effects on the distribution pattern of tenebrionid beetles, with altitude having the highest explanatory power. The Margalef’s richness index and Shannon-Wiener diversity index of tenebrionid beetles showed significant linear relationships with altitude. The Margalef’s richness index of tenebrionid beetles was significantly negatively correlated with slope. The individual number of the dominant species A. potanini showed no significant correlation with any of the five topographic factors, and A. potanini was distributed near the origin of the CCA ordination axis, indicating its widespread distribution in the Northern Qilian Mountains. The individual number of the dominant species S. horridum horridum was only significantly negatively correlated with altitude. The plane curvature of tenebrionid beetles showed a significant normal curve distribution relationship with their individual numbers. 【Conclusion】 The distribution pattern of tenebrionid beetles in the northern Qilian Mountains is influenced by a combination of multiple topographic factors, with altitude being the dominant factor.

【Aim】 The objective of this research is to reveal the odour preference and pollinating behavior characteristics of the Italian honeybee, Apis mellifera ligustica for two rapes, Brassica napus cv. Shengli and B. juncea cv. Mawei. 【Methods】 We conducted the field test of foraging preference, the odour preference test in colonies outdoors, and the olfactory response test with “Y” olfactometer and learning-memory test in the laboratory to evaluate the selective preference of  A. mellifera ligustica colonies and foragers for flowers of the two rapes and analyze the memory and learning ability of A. mellifera ligustica foragers by proboscis extension response in training and after training. 【Results】 The number of A. mellifera ligustica bees visiting B. napus cv. Shengli flowers was significantly higher than that visiting B. juncea cv. Mawei during 12:00-13:00 in the field test ( P<0.05), but the odour memory of A. mellifera ligustica bees to B. napus cv. Shengli was significantly higher than that to B. juncea cv. Mawei in the 24 h memory test. 【Conclusion】 This research suggests that A. mellifera ligustica has a preference for visiting B. napus cv. Shengli. The odour of flowers is an important factor affecting foraging preference of A. mellifera ligustica. The test of odour response can be used for the quantitative analysis of olfactory sensitivity in honeybees.

The molting hormone, 20-hydroxyecdysone (20E), is an insect steroid hormone which plays a dominant role in the regulation of insect molting, metamorphosis, reproduction and other physiological processes. The 20E nuclear receptor, EcR-USP, has been identified for 20 years, and several breakthroughs have been recently made in understanding the transcriptional regulation mechanisms of 20E-cR-USP. Recent research indicates that: (1) the 20E receptor is composed of two nuclear receptor molecules, i. e., EcR and USP. (2) The EcR-USP heterodimer obtains DNA binding activity with the assistance of a molecular chaperone-containing heterocomplex. (3) 20E transcriptionally activates the EcR-USP heterodimer by releasing corepressors and recruiting co-activators. (4) The ligand-receptor complex, 20E-EcR-USP, triggers a transcriptional cascade, including transcription of the 20E primaryresponse genes and the 20E secondaryresponse genes induced by transcription factors encoded by the 20E primary-response genes, to regulate physiological and developmental events.

【Aim】 The aim of this study is to reveal the function of peroxiredoxin 2 (ApPrx2) in the pea aphid, Acyrthosiphon pisum during bacterial infection. 【Methods】 The open reading frame of ApPrx2 was cloned and heterologously expressed in Escherichia coli. The antioxidative function of the purified recombinant ApPrx2 was characterized. The H ₂O ₂ concentration and ApPrx2 transcription level were detected in pea aphids after bacterial infections of Pseudomonas aeruginosa and Staphylococcus aureus. RNA interference was used to knockdown the expression of ApPrx2. H ₂O ₂ concentration, bacterial load and the survival rate of aphids were evaluated after the knockdown of ApPrx2. 【Results】 Sequence alignment showed that ApPrx2 is highly similar to 2-Cys Prxs from other insect species. In vitro assays demonstrated that the recombinant ApPrx2 protein degraded H ₂O ₂, and ApPrx2 expression rendered E. coli cells resistant to H ₂O ₂. Infections of the bacteria P. aeruginosa and S. aureus increased the H ₂O ₂ concentration and ApPrx2 expression in the pea aphids. The knockdown of ApPrx2 expression increased the H ₂O ₂ concentration, reduced the bacteria load and increased the mortality rate of aphids infected with S. aureus. 【Conclusion】 ApPrx2 functions as an antioxidative protein protecting pea aphids from stress induced by infection with bacteria, particularly S. aureus.

【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】 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.

【Aim】 The occurrence regularity of Adoretus puberulus and the peak period of daily activity of adults were investigated to provide a reference for the key period of the prevention and control of this pest, and the insecticides with good control efficacy were screened out to provide a reference for the control of A. puberulus using pesticides. 【Methods】 In pear orchard of Pu County, Shanxi Province, North China from April 25 to July 22, 2024, through the investigation of underground grubs, the pupation of overwintering mature larvae, the stinging of adults, and the time of oviposition and egg hatching were investigated. From June 12 to August 6, 2024, the daily number of eggs laid per female was investigated in the laboratory to study the law of oviposition. From May 6 to August 23, 2024, the occurrence law of adults was studied by trapping lamp, and the peak period of daily activity of adults was studied by trapping tent from July 10 to August 4, 2023. One percent matrine soluble concentrate (SL) was used for gradient dilution, and the final concentrations were 50, 100, 500, 1 000 and 5 000 mg/L matrine, respectively. Zero point three percent azadirachtin emulsifiable concentrate (EC) was used for gradient dilution, and the final concentrations were 5, 10, 20, 25 and 50 mg/L azadirachtin, respectively. And 6% rotenone microemulsion (ME) was used for gradient dilution, and the final concentrations were 5, 20, 50, 100 and 1 000 mg/L rotenone, respectively. The toxicity of these botanical insecticides to A. puberulus adults was determined by immersion method. 【Results】 In Pu County, Shanxi Province, the mature larvae of A. puberulus began to pupate in mid-May. Adults began to damage in early June, the adults emerged in mid-June began to lay eggs, and the peak period of adult oviposition was in early and middle July. At the end of June, the eggs began to hatch into larvae, and the egg duration was about 2 weeks. The peak period of adults was from mid-late June to early July. The number of adults trapped was the largest during 20: 00-22: 00, and the main activity time of adults was 20:00-24:00. The median lethal concentration (LC₅₀) values of three botanical insecticides matrine, azadirachtin and rotenone against A. puberulus adults were 515.6, 18.6 and 33.2 mg/L, respectively. 【Conclusion】 By clarifying the occurrence regularity and the peak period of evening activity of A. puberulus adults, it is recommended that light trapping and artificial capture of adults should be carried out during 20:00-24:00 every day from mid-late June to early July. Six percent rotenone in 500-fold dilution (120 mg/L rotenone) can be used for the management of A. puberulus in the pear orchard.

【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】 The objective of this study is to analyze the number and distribution of microsatellites in the whole genome of the German cockroach, Blattella germanica, and get the functional annotation information of genes containing microsatellites in exons. 【Methods】 The microsatellite number, repetition and location information were calculated by using microsatellite search tool. The distribution information of microsatellites in the genome was calculated by custom Python scripts, and all genes containing microsatellites were annotated by using the programs of Blast2Go and KAAS. 【Results】 A total of 604 386 simple sequence repeats (SSRs) with 1-6 bp nucleotide motifs were identified, with a total length of 15 301 255 bp, indicating that about 0.75% of the B. germancia genome (2.04 Gb) is occupied by SSRs and that there is a locus per 3.37 kb. The length of the microsatellite sequences mainly ranges from 12 to 60 bp. Among different types of microsatellites, trinucleotide microsatellites (226 876, 37.54%) are the most abundant SSRs, followed by tetranucleotide microsatellites (150 355, 24.88%), mononucleotide microsatellites (141 167, 23.36%), dinucleotide microsatellites (60 877, 10.07%), pentanucleotide microsatellites (21 570, 3.57%) and hexa-nucleotide microsatellites (3 541, 0.59%). The predominant repeat types are ATT, AAT, A, T, AAAT, ATTT and AT, with a total number of 411 789, accounting for 68.13% of the total SSRs. The number of each of these 7 categories is greater than 30 000. There are 2 372 microsatellites in the exons of 1 481 genes. The results of GO annotation indicated that 434 GO terms are classified as cellular component, 402 GO terms are related to molecular function and 660 GO terms are related to the biological process. Aligned to KEGG database, most of these genes are associated with metabolism, followed by genes related to organismal system, and genes related to genetic information processing are the least. 【Conclusion】 This study lays a foundation for further in-depth analysis of microsatellite function and developing microsatellite markers of B. germanica.

Genetic sex determination includes somatic sex determination, sex differentiation of germ cell and dosage compensation. A common cascade (primary signal>master factor>doublesex gene) exists to determine insect somatic sex. Generally, the primary signals (X-chromosome dosage, M factor on Y, maternal imprinting, piRNA, etc.) are very complex, the master factors ( sxl and tra/ fem) relatively variable, the doublesex gene ( dsx) relatively conserved, and the splicing mode (alternative splicing) of important genes very conserved in the genetic hierarchies of somatic sex determination among different insect species. Somatic sex determination mechanisms in many insects are unclear or short of systematic progress, which limits the clarification about molecular mechanisms of dimorphism. Combined with our research on insect sex-determination mechanisms, we summarized the progress and common rules of the primary signals, master factors and doublesex genes in the sex-determination hierarchies of the representative insects of Diptera, Hymenoptera and Lepidoptera, so providing a theoretical basis for a deeper understanding of insect sex-determination mechanism, an exploitation of sterile insect technique (SIT), and an artificial control of insect sex.

【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】 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】 This study aims to clarify the types of antennal sensilla and electrophysiological responses of the adult Galeruca daurica to volatiles of host plants, so as to provide the necessary basis for further investigation on the mechanism of chemoreception in G. daurica. 【Methods】 The sensilla types on the antennae of G. daurica adults were observed using scanning electron microscopy (SEM). The volatiles were collected from the host plant Allium mongolium by headspace dynamic adsorption collection method, and the main components of volatiles were determined by GC-MS. The electrophysiological responses of G. daurica adults to the main volatile components of this host plant were determined by electroantennogram (EAG). 【Results】 There are five types of sensilla on the antennae of G. daurica adults, including sensilla trichodea (ST), sensilla chaetica (SC), sensilla basiconica (SB), sensilla campaniformia (SCa) and Bhm bristles (BB). Thirty-two volatile components were identified from A. mongolium. Among them, sulfur compounds accounted for 49.3%. EAG results showed that six compounds including diallyl sulfide, diallyl disulphide, ( Z)-2-hexen-1-ol, 2-hexenal, methyl benzoate, and hexanal elicited relatively strong eletrophysiological responses in female adults of G. daurica, while only diallyl disulphide and 2-hexenal elicited relatively strong EAG responses in male adults. Moreover, most of the tested compounds elicited stronger EAG responses in female adults than in males. At the concentration of 1 mol/L, the tested six compounds except 2-hexenal could elicit the strongest EAG response in G. daurica adults. 【Conclusion】 Six compounds of A. mongolium can elicit strong EAG response of G. daurica adults, including diallyl sulfide, diallyl disulphide, ( Z)-2-hexen-1-ol, 2-hexenal, methyl benzoate, and hexanal. The EAG responses of G. daurica adults to all these compounds except 2-hexenal increase with increasing concentrations.

As the most diverse group of animals on earth, insects are closely related to human production and activities, making entomological research both theoretically significant and practically valuable. In the past decade, the rapid development of novel methods and technologies has greatly promoted the research progress of entomological research. In this article, we provided a comprehensive overview of the applications of emerging methods and technologies in such fields as entomological morphological identification, molecular mechanism and pest management, focusing on the main contents of this special issue from seven aspects: Micro-computed tomography, RNA interference, gene editing, artificial intelligence-driven intelligent recognition, nanotechnology, regulation of insect-microorganism symbiotes, and olfactory behavior regulation. We also proposed the challenges faced by the large-scale application and sustainable development of these technologies, and prospected the future development trend.

【Aim】 This study aims to explore the effects of altitude and seasonal changes on the composition and diversity of pollen-source plants of Apis cerana cerana. 【Methods】 Pollen-source plant DNA was extracted for PCR amplification targeting the ITS2 region by collecting the bee bread of A. c. cerana at various altitude gradients (500, 700, 900, 1 100 and 1 300 m) across different months (March, May, July, September and November) in Simian Mountain, Chongqing, southwestern China. Sequencing was performed using Illumina PE250 second-generation high-throughput sequencing technology. Bioinformatics methods were employed to analyze the composition, abundance and diversity of pollen-source plants of A. c. cerana. 【Results】 A total of 162 species belonging to 128 genera of 61 families across 30 orders were identified among the pollen-source plants of A. c. cerana in Simian Mountain, Chongqing, including 101 species of woody plants (accounting for 62.35%), 51 species of herbaceous plants (accounting for 31.48%) and 10 species of lianas (accounting for 6.17%). Among these pollen-source plants, the order Rosales exhibited the highest number of plant species (21), followed by Fagales and Astererales, which comprised 19 and 15 species, respectively. Analysis of the composition of pollen-source plants of A. c. cerana based on the operational taxonomic units (OTUs) indicated that there were 17 species of pollen-source plants accounting for over 1%, with the top five being Castanea seguinii (accounting for 18.9%), Eurya nitida (accounting for 11.3%), Rhus chinensis(accounting for 8.5%), Brassica oleracea (accounting for 6.3%) and Aralia chinensis (accounting for 5.3%). The results of alpha-diversity analysis showed that the diversity of pollen-source plants of A. c. cerana in Simian Mountain, Chongqing increased first and then decreased with the increase of altitude, and was the lowest in May and had no significant difference between the other months. 【Conclusion】 The pollen-source plant resources of A. c. cerana in the Simian Mountain region of Chongqing are abundant. The diversity of these pollen-source plants does not significantly vary by month, however, it exhibits a trend of initially increasing and then decreasing with altitudes. This study elucidates the characteristics of spatiotemporal variations in plant composition and diversity of pollen-source plants of A. c. cerana in Simian Mountain of Chongqing. The findings offer a novel perspective for further research and conservation efforts concerning A. c. cerana populations in this area, and provide valuable insights for local beekeepers in selecting appropriate apiary locations, ensuring a steady supply of pollen resources, enhancing honeybee survival rates, and ultimately increasing the yield of bee products.

Larvae of the beet armyworm, Spodoptera exigua (H ü bner), originally collected from the field of sugar beet, were reared on artificial diet and evaluated for resistance to lambda-cyhalothrin for 43 generations under laboratory condition without exposure to any insecticide (under non-selection). Lambda-cyhalothrin showed LD₅₀ value of 0 . 9672 μ g/larva at F₁ generation and decreased gradually in onward generations,  resulting in LD₅₀ value of 0 . 0325 μ g/larva at F₄₃ generation by using topical application, and the resistance level of the non-selected strain decreased from 4 836 . 0 fold to 162 . 5-fold compared with the susceptible strain. In leaf dipping bioassay, lambda-cyhalothrin exhibited LC₅₀ value of 185 . 6 mg/L at F₁ generation and decreased gradually to onward generations with LC₅₀ value of 9 . 2 mg/L at F₄₃ generation. Correspondingly, the resistance level of the non-selected strain decreased from 964 . 7-fold to 47 . 8-fold compared with the susceptible strain. Though decreased 29 . 8-fold (topical application) or 20 . 2-fold (leaf dipping bioassay) from F₁ to F₄₃ generation, the resistance levels of F₄₃  generation was still quite high,  and this indicated that it was very difficult for the beet armyworm to recover the sensitivity to lambda-cyhalothrin. Activities of four monoxygenases,  i.e., methoxyresorufin O-demethylase (MROD), ethoxyresorufin O-deethylase (EROD),  arylhydrocarbon hydroxylase (AHH) and aldrin epoxidase (AE), of F₂, F₂₀ and F₄₁ generation were compared with that of the susceptible strain, respectively. Compared with the susceptible strain, the activities of MROD and AE in midguts in the 5th instar larvae of F₂ generation was significantly higher, but those of F₂₀ and F₄₁ generations showed no significant difference. Similarly,  the activities of EROD and AHH of F₂,  F₂₀ and F₄₁ generation were also significantly higher compared with the susceptible strain. The results suggested that the resistance of S. exigua to lambda-cyhalothrin was closely correlated with multi-function oxidase which may play different roles at different resistant levels.

The species of the genus Decma Gorochov, 1993 from China are studied, of which a new subgenus Paradecma and a new species Decma (Paradecma) bispinosa sp. nov. are described. A key to the Chinese species of Decma and new description of female Decma (Idiodecma) birmanica (Bey-Bienko, 1971) are provided. The type specimens are deposited in the Institute of Zoology, Chinese Academy of Sciences, Beijing, China.

【Aim】 To identify a novel galectin gene from the silkworm, Bombyx mori, analyze its sequence and structural characteristics, detect its expression after microbial challenge and measure the binding properties of the expressed recombinant protein with microbe and microbial surface polysaccharides. 【Methods】 We used TBlastN to identify a novel galectin gene (named BmGalectin-4) by searching the silkworm genome database, and analyzed its sequence and structural characteristics with bioinformatics tools. The expression of BmGalectin-4 in different tissues in larval silkworm after microbial challenge (inoculation of Pseudomonas aeruginosa, Staphylococcus aureus and Candidiasis albicans, respectively) was detected by RT-PCR. The recombinant protein was produced in Escherichia coli and purified by affinity chromatography. The binding property of BmGalectin-4 with bacteria and fungus was analyzed by ELISA. Western blot was used to detect its binding to polysaccharides. 【Results】 Sequence analysis showed that BmGalectin-4 is a tandem repeat galectin, containing two carbohydrate recognition domains. BmGalectin-4 was expressed in fat body, hemocytes and midgut tissues, and its expression level changed significantly after bacterial and fungal challenge. Purified recombinant BmGalectin-4 showed significant binding capacity to Gram-positive and Gram-negative bacteria and fungus. Its binding to microbial surface polysaccharides was highly specific. 【Conclusion】 BmGalectin-4 is a typical tandem repeat galectin and might be involved in the silkworm immune responses against microbial pathogens. This study provides the foundation for further functional study of the silkworm galectin.

In this article the up-to-date progress of the ecophysiological effects of heavy metals on insects was reviewed, and the pitfalls concerning the ecophysiological studies of heavy metals on insects were also discussed. Short-term exposure to heavy metals may cause acute toxicity to insects, while chronic exposure probably results in adaptive evolution of insects to these inorganic materials. The toxicity of heavy metals is dependent on concentration, time of exposure and feeding patterns. The toxicity of heavy metals could also be transferred through prey-predator interaction. The physiological toxicity of heavy metals to insects includes decreased amount of energy materials in haemolymph and/or haemocytes, imbalance of redox state, suppression of cellular and/or humoral immunocompetence, and dis-integrity of insect cells and/or tissues. However, insects have developed physiological and ecological strategies to cope with the toxicity of heavy metals, e.g.,storage of heavy metals in specific tissues or organs, excretion of heavy metals through defecation and/or molting, induction of metal-detoxifying proteins and even development of local adaptation.