【Aim】 This study aims to investigate the number, distribution and morphology of cell bodies in the antennal lobe of adults of the diamondback moth, Plutella xylostella. 【Methods】 The nuclear staining and laser scanning confocal microscope were used to acquire the confocal image stacks of antennal lobe. The cell digital three-dimensional reconstructions were created by the Imaris software, and the volumes and surface areas of cell bodies were measured by using the statistic tool of the Imaris software. 【Results】 A lot of cell bodies are distributed in the medial cell cluster (MCCL), lateral cell cluster (LCCL) and anterior cell cluster (ACCL) of the antennal lobe of P. xylostella adults, and some irregular giant cells exist in the MCCL and LCCL. There are about 560 cell bodies in the antennal lobe in females, and 810 in males, more in males than in females. The morphology of cell bodies varies significantly, and three-dimensional reconstructions of cell bodies showed that most of them are nearly spherical. The surface area and volume of the smallest cell body are 96.7 μm² and 76.9 μm³, and those of the nuclei are 42.5 μm² and 24.1 μm³, respectively. The surface area and volume of the largest cell body are 389 μm² and 514 μm³, and the surface area and volume of the nuclei are 265 μm² and 261 μm³, respectively. 【Conclusion】 The characteristics of cell bodies in the antennal lobe of P. xylostella have been illustrated in this study, which will provide a basis for further understanding the anatomical structure and physiological function of the central nervous system.

【Aim】 To study the molecular characterization and biological function of the cytochrome P450 gene LmCYP6FD3 from the migratory locust, Locusta migratoria. 【Methods】 The cytochrome P450 gene sequences were searched from the transcriptome database of L. migratoria, the cDNA fragment of the target gene was cloned using RT-PCR, and its expression patterns in various tissues (gastric caeca, foregut, midgut, hindgut, integument, testis, ovary, muscles, hemolymph, fat body and Malpighian tubules) of the 5th instar nymphs and different developmental stages (egg, 1st-5th instar nymph and adult) were analyzed using real-time quantitative PCR (RT-qPCR). The target gene was silenced by RNA interference (RNAi), the silence efficiency was evaluated by injecting dsRNA to the 2nd instar nymphs， and the susceptibility of the nymphs to three insecticides (malathion, carbaryl and deltamethrin) was assessed. 【Results】 The full-length cDNA of cytochrome P450 gene was cloned (GenBank accession no.: KT316378) and named LmCYP6FD3. Its length is 1 563 bp, encoding 521 amino acids. The expression level of LmCYP6FD3 was the highest in the Malpighian tubules of the 5th instar nymphs, moderate in the hindgut and fat body, and lower in other tissues. LmCYP6FD3 was expressed during the whole developmental period and had relatively higher expression level in the nymphal stage. Insecticide bioassay in combination with RNAi showed that the highest silence efficiency of LmCYP6FD3 was achieved at 24 h after RNAi, and the mortality of the 2nd instar nymphs of L. migratoria in the RNAi treatment group (injection with dsLmCYP6FD3) increased by 32% as compared with that of the control group (injection with dsGFP) after exposure to carbaryl by dipping method. 【Conclusion】 In this study, the full-length of cDNA sequence of LmCYP6FD3 was cloned. This gene has the highest expression level in the Malpighian tubules of L. migratoria and might play a significant role in the detoxification of carbaryl in this locust.

【Aim】 Aquaporin (AQP) is one of the transmembrane proteins widely existing in cell biological membrane system, and plays an important role in the balance of water osmotic pressure. This study aims to understand the localization of AQP1 in organelles based on the previously cloned full-length cDNA in Ectropis obliqua Prout (EoAQP1), and to explore its effects on cell morphology and cell proliferation. 【Methods】 The eukaryotic expression vectors of green fluorescent protein (GFP) and its fusion protein with EoAQP1 (EoAQP1-GFP) were constructed by double enzyme digestion. Cell expression characteristics of GFP and EoAQP1-GFP proteins in Drosophila melanogaster embryonic cell line S2 were observed by fluorescence microscope and confocal scanning laser microscope. The localization of EoAQP1 protein in organelles in S2 cells was detected by Lyso-Tracker Red and Golgi-Tracker Red kit. The effects of EoAQP1 on cell granularity, size and proliferation were detected by flow cytometry and microplate reader. 【Results】 The eukaryotic expression vectors of GFP and EoAQP1GFP were constructed successfully, and named pAc5.1-GFP and pAc5.1-EoAQP1-GFP, respectively. Fluorescence microscope and confocal scanning laser microscope observation showed that GFP protein was evenly filled in S2 cells and showed stable expression. The EoAQP1-GFP protein was expressed by two types: one showed scattered expression in the cytoplasm and around the nucleus, in which the EoAQP1-GFP protein was in spherical and spheroidic structure similar to lysosome; the other showed the hemisphere and arched expression in the cytoplasm, in which the EoAQP1-GFP protein was in hemisphere and arch structure similar to the Golgi apparatus. Lyso-Tracker Red and Golgi-Tracker Red kit detection showed that EoAQP1 protein had no expression in lysosome but completely overlapped in the Golgi apparatus. Flow cytometry and microplate reader detection indicated that the overexpression of EoAQP1 protein in S2 cells enhanced cell volume and granularity significantly, but had no significant effect on cell proliferation. 【Conclusion】 The EoAQP1 protein is localized and plays functions in the Golgi apparatus. It can change cell morphology, but not promote cell growth.

【Aim】 To explore the molecular regulation mechanism of grafted larvae on ovarian development of queens in the Western honey bee, Apis mellifera. 【Methods】 The isobaric tags for relative and absolute quantitation (iTRAQ) method was used to quantify the proteins in the ovaries of queens reared from the larvae grafted at different instars and screen the differentially expressed proteins (DEPs). The results were validated by Western blotting. 【Results】 A total of 452 966 spectra were obtained and 3 642 proteins were identified from ovaries of A. mellifera queens. Gene Ontology (GO) analysis revealed that the DEPs in the ovaries of queens reared from the larvae grafted at different instars are associated with cell metabolism, cell division, and protein biosynthesis. The DEPs in the ovaries of queens reared from the larvae grafted between the 1st and 2nd instars are enriched in such pathways as carbohydrate metabolism, lipid metabolism, and exogenous degradation, while those reared from the larvae grafted between the 1st and 3rd instars mainly enriched in developmental pathways, ribosome pathways, and lysosome metabolism. The Western blotting results of the two differentially expressed storage proteins hexamerin 110 and hexamerin 70b showed that their expression levels decreased with the grafted larval instar. 【Conclusion】 The identification of differentially expressed proteins in the ovaries of A. mellifera queens reared from the larvae grafted at different instars provides a foundation for the regulatory mechanism of queen reproductive development and caste differentiation.  

【Aim】 To understand the bacteria species in saliva of adults of the brown planthopper, Nilaparvata lugens， by identifying the bacterial proteins in their saliva. 【Methods】 Saliva of N. lugens adults was collected by stretching two layers of Parafilm with sucrose diet. Protein solution was subjected to electrophoresis after concentration by ultrafiltration. Protein identification was conducted by liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) analysis, and the spectra were searched against the Uniprot database of bacterial protein to identify the bacterial proteins in saliva. 【Results】 Thirty five proteins of twenty-two species of bacteria, mainly related to energy metabolism, protein folding and synthesis and amino acid metabolism, were identified in saliva of N. lugens adults. These bacteria belong to Proteobacteria, Actinobacteria and Firmicutes, and most are from Proteobacteria. 【Conclusion】 The endosymbiont bacteria in N. lugens, whose proteins were identified, may play an important role in the life history of this insect.

【Aim】 This study aims to investigate the effect of wilforgine on muscular structure against adults of the oriental armyworm, Mythimna separata, so as to provide basis for further studying its insecticidal mechanisms. 【Methods】 M. separata adults were fed on acetone-sugar water solution of wilforgine to assay the toxicity and poisoning symptoms. The pathological changes in their muscular structure induced by wilforgine (300 μg/mL) were observed by light and electron microscope. 【Results】 Wilforgine had strong oral toxicity against M. separata adults with the LC₅₀ value of 194.53 μg/mL. The typical toxicity sign was paralysis leading to death. Microstructural examinations demonstrated that the muscular structure was damaged with increasing spaces between muscular fibers. The sarcolemma was digested and band structures disappeared. The ultrastructural examinations showed shed myolemma, swollen mitochondria and dilated sarcoplasmic in the muscle cells. Moreover, the damage to muscle cells increased with poisoning time. 【Conclusion】 Wilforgine could induce pathological changes in the flight muscle of M. separata adults, leading to death. Thus, the action site of wilforgine may be located in the muscle tissues of this insect.

【Aim】 The white-backed planthopper, Sogatella furcifera (Horváth) (Hemiptera: Delphacidae) is an important insect pest of rice. The present study aims to determine the sublethal effects of pymetrozine on the development and reproduction of this insect and its susceptibility to other insecticides. 【Methods】 The life table method was used to evaluate the sublethal effects of pymetrozine (LC₁₀ and LC₂₅) on developmental duration, survival rate, emergence rate and longevity of F₀ and F₁ generations of S. furcifera and rice stem dipping method was adopted to determine its susceptibility to other insecticides after pymetrozine (LC₁₀ and LC₂₅) treatment. 【Results】 The developmental duration of the F₀ generations of S. furcifera treated with sublethal concentrations (LC₁₀ and LC₂₅) of pymetrozine for 48 h was increased by 1.98 d and 4.25 d, the longevities were shortened by 0.49 d and 1.73 d, the survival rates (85.00% and 68.50%, respectively) of the 3rd-5th instar nymphs and the emergence rate (75.89% and 67.78%, respectively) decreased as compared with the control (92.00% and 85.90%, respectively), but significant difference was just observed in generation duration between the LC₂₅ treatment group and the control. Furthermore, the F₁ generation of S. furcifera exposed to sublethal concentrations (LC₁₀ and LC₂₅) of pymetrozine for 48 h had decreased emergence rate (76.97% and 68.94%, respectively), copulation rates (77.79% and 66.44%, respectively), egg hatching rates (73.19% and 68.67%, respectively), fecundity (112.36 and 88.34 eggs laid per female, respectively), and relative fitness (0.41 and 0.20, respectively) as compared with the control (88.22%, 86.67%, 87.26%, 147.80 eggs laid per female and 1.00, respectively). The susceptibility of pymetrozine-treated S. furcifera to thiamethoxam, chlorpyrifos, and buprofezin showed that the LC₅₀ values of the three chemicals were 2.16, 40.87 and 3.12 mg/L in LC₁₀ treatment group, and 4.93, 17.96 and 8.39 mg/L in LC₂₅ treatment group, respectively. The relative toxicity index (RTI) of thiamethoxam, chlorpyrifos and buprofezin to S. furcifera indicated that the susceptibility of pymetrozinetreated S. furcifera to these three insecticides was reduced. 【Conclusion】 Pymetrozine at sublethal concentrations can reduce the fecundity and population growth of S. furcifera and make it less susceptible to thiamethoxam, chlorpyrifos and buprofezin.

【Aim】 This study aims to explore the effects of beta-cypermethrin (β-cypermethrin) on the biological characteristics of Carposina sasakii Matsumura, especially the sublethal effects of beta-cypermethrin on the development, survivorship and reproduction of the F₀ (parental) and F₁ generations of C. sasakii. 【Methods】 The sublethal concentrations (LC₁₀, LC₂₀ and LC₄₀) of β-cypermethrin against the newly hatched larvae of C. sasakii were determined by fruitdipping method. The mortality of C. sasakii was recorded at 24 h after treatment and the LC₁₀, LC₂₀ and LC₄₀ values were estimated using DPS software. Sublethal effects of β-cypermethrin on the development and reproduction of the F₀ and F₁ generations were assessed by establishing a life table. The statistical differences of the survivorship and developmental duration, the adult longevity, the number of eggs laid per female and life table parameters of F₀ and F₁ generations were analyzed with SPSS 21.0. 【Results】 According to the bioassay results, the LC₁₀, LC₂₀ and LC₄₀ values of β-cypermethrin on the newly hatched larvae of C. sasakii at 24 h after treatment were 0.146, 0.267 and 0.579 mg/L, respectively. Sublethal concentrations of β-cypermethrin reduced the survivorship and the proportion of female adults of F₀ and F₁ generations. The values of boring rate, exiting rate, and survival rate of larvae, emergence rate and sex ratio (♀/♂) reduced with the increase of the insecticide concentration. After the newly hatched larvae of F₀ generation were exposed to the sublethal concentrations (LC₁₀, LC₂₀ and LC₄₀) of β-cypermethrin, the average numbers of eggs laid per female per day were 34.09, 33.00 and 30.12, respectively; correspondingly, the numbers of eggs laid per female were 284.90, 276.56 and 252.89, respectively, both were significantly less than those of the control (38.02 and 320.98, respectively). When the newly hatched larvae of F₁ generation were exposed to LC₁₀, LC₂₀ and LC₄₀ of β-cypermethrin, the average numbers of eggs laid per female per day were 34.57, 30.82 and 33.39, respectively, which were significantly higher than that of the control (27.97). Meanwhile, the numbers of eggs laid per female after treatment with β-cypermethrin at LC₂₀ and LC₄₀ were 304.45 and 298.31, respectively, which were significantly higher than that of the control (271.40). However, there was no significant difference in the developmental duration of each stage and the larval weight as compared with the control. Life table analysis showed that the net reproductive rate (R₀) decreased significantly in a dose-dependent manner after treatment. The R₀ values of the F₀ generation decreased significantly after treatment with β-cypermethrin at LC₁₀, LC₂₀ and LC₄₀ with the values of 124.36, 114.33 and 60.16, respectively, as compared with the control (166.54), while the corresponding values of the F₁ generation were 128.84, 112.30 and 85.32, respectively, as compared with the control (128.80). Sublethal concentrations of β-cypermethrin obviously reduced the relative fitness value (R_f) of the F₀ and F₁ generations. The R_f values of the F₀ generation after treatment with β-cypermethrin at LC₁₀, LC₂₀ and LC₄₀ were only 0.75-, 0.68- and 0.37-fold as high as that of the control, while those of the F₁ generation increased to 0.98-, 0.86- and 0.64-fold as high as that of the control. 【Conclusion】 Sublethal concentrations of β-cypermethrin influence the survivorship, growth and reproduction of F₀ and F₁ generations of C. sasakii, consequently affecting its population dynamics. β-cypermethrin applied singly or sequentially would inhibit the population growth rate of C. sasakii, but its reproductive disadvantage would be improved after a secondary or more treatments.

【Aim】 This study aims to clarify the toxicity of sulfoxaflor, spinetoram and their mixture to the buff-tailed bumblebee, Bombus terrestris. 【Methods】 The acute oral and contact toxicities of sulfoxaflor, spinetoram and their mixture (1∶1) to the 1 day-old adult workers of B. terrestris were measured by feeding application and topical application methods, respectively, and meanwhile the acetylcholinesterase (AChE) activity was measured. The risk of insecticides to B. terrestris was assessed based on the hazard quotient (HQ) values of the ecological risk of insecticides to bees. 【Results】 The LD₅₀ values of sulfoxaflor, spinetoram and their mixture (1∶1) against adult workers of B. terrestris at 48 h after oral exposure were 2.50, 4.97 and 4.07 μg a.i. per bee, respectively. Sulfoxaflor showed moderate toxicity to adult B. terrestris with the LD₅₀ value of 8.72 μg a.i. per bee at 48 h by contact, and no bumblebees died after acute contact with spinetoram and its mixture with sulfoxaflor (1∶1) at 120 μg a.i. per bee. The HQ values of the three insecticides to adult workers of B. terrestris were all below 50, suggesting that these three insecticides present low risk to this bumblebee. Sulfoxaflor inhibited AChE activity in adult workers of B. terrestris, and its action strengthened with the increase of treatment dosage. After the adult workers of B. terrestris were treated with spinetoram and the mixture (1∶1) at LD₆₀, the AChE activities were enhanced first and then inhibited. 【Conclusion】 Sulfoxaflor, spinetoram and their mixture (1∶1) show low risk to adult workers of B. terrestris as judged by the HQ value. The poisoning effects of spinetoram and its mixture with sulfoxaflor (1∶1) are slower than sulfoxaflor, and their safety is higher than that of sulfoxaflor in laboratory safety evaluation.

【Aim】 The aim of this study is to clarify the effects of Vip3Aa11 toxin on the Asian corn borer, Ostrinia furnacalis, and its parasitoid Macrocentrus cingulum. 【Methods】 The indirect effects of Vip3Aa11 protein on M. cingulum were assessed by the bioassay with the artificial diet containing 6 μg/g azadirachtin used as the positive control. The mortality and parasitism of O. furnacalis larvae parasitized by M. cingulum were recorded at 10 d after feeding with a diet containing 100 μg/g Vip3Aa11 protein, and the cocoon weight of M. cingulum and the emergence number per O. furnacalis larva after pupation and emergence of M. cingulum were investigated. The direct effects of Vip3Aa11 protein on M. cingulum were assessed by the bioassay with 20% honey solution containing 100 μg/g azadirachtin used as the positive control. The mortality of M. cingulum adults daily after feeding with 20% honey solution containing 100 μg/g Vip3Aa11 protein, and the mortality and parasitism rate of O. furnacalis larvae at 10 d after being parasitized by M. cingulum adults from different treatments were recorded. The cocoon weight of M. cingulum and emergence number per O. furnacalis larva of next generation of M. cingulum were recorded after pupation and emergence. 【Results】 The bioassay results showed that the average mortality and average inhibition rate of body weight of O. furnacalis larvae were 50.7% and 77.1%, respectively, at 7 d after exposure to 100 μg/g Vip3Aa11 protein. In the bioassay of indirect effects, the mortality of O. furnacalis larvae significantly increased when they were fed with the artificial diet containing Vip3Aa11 protein after parasitized by M. cingulum, and the cocoon weight and the number of M. cingulum progenies produced per host decreased; however, the toxin had no effect on the parasitism rate and longevity of M. cingulum adults. In the bioassay of direct effects, the longevity and parasitism rate of M. cingulum adults, the mortality of O. furnacalis larvae, and the cocoon weight, the number of progenies produced per host and the adult longevity of next generation of M. cingulum were not affected when M. cingulum adults were fed with 20% honey solution containing 100 μg/g Vip3Aa11 protein. 【Conclusion】 This study assessed the potential effects of Vip3Aa11 toxin on M. cingulum from the indirect and direct aspects through bioassay system. The results demonstrate that M. cingulum is insensitive to Vip3Aa11 protein at the concentrations exceeding those encountered in Bt crop fields. Vip3Aa11 protein has no direct adverse effects on M. cingulum, and the indirect detrimental effects detected in the bioassay may be due to poor host quality.

【Aim】 In order to better understand the morphology and function of Malpighian tubules of the Cicadomorpha, comparative morphology and ultrastructure of the mute cicada Karenia caelatata Distant and the coleopterous spittlebug Lepyronia coleoptrata (Linnaeus) were investigated. 【Methods】 The Malpighian tubules of adult K. caelatata and L. coleoptrata were observed using both light and transmission electron microscopies. 【Results】 The results show that except the anterior-most segment that takes part in the formation of the filter chamber, each tubule outside the filter chamber of both K. caelatata and L. coleoptrata is morphologically differentiated into four regions, i.e., intermediate duct, proximal segment, distal segment, and terminal segment. Apical parts of the terminal segments in K. caelatata are attached to the ileum, with the posterior-most ends united in pairs and enveloped by a connetcive tissue shortly before arriving at the rectum, whereas the corresponding parts in L. coleoptrata terminate as separated rods and closely attached to the rectum. Ultrastructurally, cells of the tubules in both species possess basal infoldings and apical microvilli. Cells of the proximal segment in K. caelatata possess secretory vesicles, secretory granules, rough endoplasmic reticulum and urospherites. In L. coleoptrata, cells of the proximal segment possess abundant secretory vesicles, and cells of the distal segment contain oval vesicles which appear to fuse with the basal plasma membrane. The microorganisms observed in the proximal and distal segments of K. caelatata are possibly symbiotic bacteria, while those resided in the intermediate duct of L. coleoptrata are likely to be pathogenic. 【Conclusion】 The similarities and differences in morphological characteristics and ultrastructure of Malpighian tubules in the two Cicadomorpha representatives, K. caelatata and L. coleoptrata, not only support the sister relationship of Cicadoidea and Cercopoidea, but also improve our understanding of the functional differentiation of Malpighian tubules in the Cicadomorpha. The discovery of microorganisms in the Malpighian tubules of the two species is informative to the future study of coevolution between the Cicadomorpha and related bacteria and to the biological control of related pests.

【Aim】 To learn how the proboscis of Plutella xylostella is formed by two galeae using the dorsal and ventral linkages. 【Methods】 The mouthpart characteristics of P. xylostella were observed under light microscope. The ultrastructure of the dorsal linkage, ventral linkage and food canal of proboscis were observed under scanning electron microscope. 【Results】 The mouthparts of P. xylostella consist of a pair of labial palpi, and a proboscis is located in the middle. The proboscis is linked by two extended galeae. The dorsal linkage of proboscis is formed by overlapping lance-shaped plates of galea. The dorsal linkage contains two parts, i.e., upper and lower lanceshaped plates. In the apical part of proboscis, the lower lance-shaped plates disappear slowly while the upper lance-shaped plates gradually become larger and a row of triangular process appears in the outside. There are also some pores between upper lance-shaped plates and galea. The dorsal linkage is affected by a series of overlapping distally directed upper and lower lanceshaped plates of cuticle. However, in the subapical part of proboscis, the linkage is formed by the overlapping of the upper lance-shaped plates. The ventral linkage is affected by a series of closely applied, curved, toothed hooks which lock into each other from the two sides. Each toothed hook contains upper branch and lower branch with similar shape, the former being longer than the latter. From the basal part to the apical part of the proboscis, the length of the toothed hook gradually increases. There is a gap between the two branches for the opposite side of the toothed hook inserted. 【Conclusion】 The difference in the structure and linking mode of the dorsal and ventral linkage of the proboscis of P. xylostella may be related to the movement mode of proboscis when it coils or stretches. In the apical part of proboscis, the pores between the dorsal linkage and galea may be the place where food can be sucked in.

Bactericera gobica (Loginova) is an important pest of medlar (Lycium barbarum L.), that can attack buds or tender leaves, causing damage as early leaf drop and fruit drop, and leading to big economic loss. In the investigation of natural enemies conducted in Zhongwei, Ningxia, China, we discovered a species of Psyllaephagus, P. arenarius Trjapitzin, which attacks B. gobica and has high parasitism rate. This is the first record of P. arenarius from China, and B. gobica also a new host record for P. arenarius. In this paper, the species P. arenarius was redescribed and illustrated with photomicrographs.