【Aim】 The theoretical mole ratio of glucose (Glc) to pyruvate acid (PA), acetyl-CoA (AC), reduced nicotinamide adenine dinucleotide (NADH) and adenosine nucleoside triphosphate (ATP) is 1:2:2:10:30~32, while the theoretical activity ratio of hexokinase (HK) to phosphofructokinase 1 (PFK1), pyruvate kinase (PK), pyruvate dehydrogenase (PDH), citrate synthase (CS), mitochondria isocitrate dehydrogenase (ICDHm), α-ketoglutarate dehydrogenase (α-KGDH), NADH CoQ reductase (NCR), succinate CoQ reductase (SCR), CoQ cytochrome C reductase (CCR), cytochrome C oxidase (CCO) and ATP synthase (AS) is 1:1:2:2:2:2:2:10:2:12:12:26~28 in aerobic oxidation. This study aims to characterize aerobic oxidation in the silkgland of the silkworm,Bombyx mori, during the synthesis and secretion of silk proteins. 【Methods】 The contents of Glc, PA, AC, NADH and ATP, and the enzymatic activities of PFK1, PK, PDH, CS, ICDHm, α-KGDH, NCR, SCR, CCR, CCO and AS were determined using spectrophotometric and high performance liquid chromatographic methods. 【Results】 In the silkgland of B. mori, Glc was undetectable, and the molar concentration increased in the order of 1/30 ATP, 1/10 NADH, 1/2 AC and 1/2 PA during the synthesis and secretion of silk proteins. Among the related enzymes of glycolysis, the PFK1 activity was the lowest; the activities of the related enzymes of tricarboxylic acid cycle increased in the order of 1/2 ICDHm, 1/2 α-KGDH and 1/2 CS; the activities of the related enzymes of oxidative phosphorylation increased in the order of 1/26 AS, 1/10 NCR, 1/2 SCR, 1/12 CCR and 1/12 CCO, with the 1/26 AS activity the lowest. During the synthesis and secretion of silk proteins, the activities of 1/26 AS, 1/2 ICDHm, 1/2 PDH and PFK1 increased sequentially. The concentrations of NADH and ATP and the activities of PFK1, PDH and NCR generally increased during the synthesis of silk proteins but decreased during the secretion of silk proteins. 【Conclusion】 It is so inferred that PFK1, ICDHm and AS are the ratelimiting enzymes of glycolysis, tricarboxylic acid cycle and oxidative phosphorylation, respectively, the rates of glycolysis, pyruvate dehydrogenation, tricarboxylic acid cycle and oxidative phosphorylation decrease progressively in the silkgland during the synthesis and secretion of silk proteins, and the rates of aerobic oxidation increase during the synthesis but decrease during the secretion of silk proteins.

【Aim】 To explore the optimal prokaryotic expression conditions and study the antimicrobial activity of a novel glycine-rich antimicrobial peptide ApAMP1015 from Anatolica polita borealis.【Methods】 The cDNA and deduced amino acid sequences of ApAMP1015 were analyzed by bioinformatics methods. Prokaryotic expression technology was used to express fusion protein TrxA-ApAMP1015. The fusion protein TrxA-ApAMP1015 was verified by Western blot, and purified with affinity chromatography. Inhibition zone experiment was made to assay the antibacterial activity of ApAMP1015. 【Results】 An antimicrobial peptide gene ApAMP1015 was cloned from A. polita borealis. ApAMP1015 has the opening reading frame of 387 bp and encodes a 128-amino-acid peptide consisting of a signal peptide of 19 amino acids and a mature peptide of 75 amino acids. Homology analysis showed that ApAMP1015 is a member of coleoptericins. The prokaryotic expression vector pET-32a-ApAMP1015  was successfully constructed and the recombinant protein expressed in Escherichia coli BL21(DE3) was verified by Western blot. The0.1 mmol/L IPTG induced a higher level of expression of recombinant protein in the supernatant in the culture condition of 25℃ and 150 r/min. The inhibition zone experiment showed that ApAMP1015 exhibited the antibacterial activity against E. coli. 【Conclusion】 The full-length cDNA of ApAMP1015 was cloned from A. polita borealis. The optimal prokaryotic expression conditions were determined. ApAMP1015 can inhibit the growth of E. coli. This study lays the foundation for promoting the application and further study of ApAMP1015 from A. polita borealis.

【Aim】 This study aims to identify and characterize a new C-type lectin gene from the housefly Musca domestica, and to analyze its immune function. 【Methods】 Based on the transcriptome database, a new lectin gene, designated as  Mdlectin-C1, was cloned from M. domestica. Recombinant Mdlectin-C1 was expressed in Escherichia coli,and purified rMdlectin-C1 was used to prepare polyclonal antibody. Quantitative real-time PCR (qRT-PCR) and Western blot techniques were employed to investigate the gene expression profiles in different developmental stages and larvae post bacterial infection. The survival rate of the 1st instar larvae subjected to Mdlectin-C1 RNAi was detected after bacterial challenge. 【Results】 The cDNA sequence of Mdlectin-C1 contains a 546 bp open reading frame encoding 181-amino-acid residues. A putative signal peptide of 21-amino-acid residues is present at the N-terminus. A conservative carbohydrate recognition domain (CRD) is observed in the mature peptide. The results of qRT-PCR and Western blot revealed that the expression level of Mdlectin-C1 was gradually enhanced fromthe 2nd instar larval stage to the pupal stage and reached the maximum value at the pupal stage. The expression level of Mdlectin-C1 was up-regulated significantly in larvae at 36 and 3 h after challenge with Gram-negative bacterium E. coli and Gram-positive bacterium Staphylococcus aureus, respectively. qRT-PCR analysis showed that the expression level of Mdlectin-C1 was knocked down successfully by RNAi. The survival rate of the 1st instar larvae subjected to Mdlectin-C1 RNAi was significantly lower than the normal control post bacterial challenge. 【Conclusion】 Mdlectin-C1 is involved in antimicrobial immune response in the housefly and may play important roles in this process.

【Aim】 Odorant-binding proteins (OBPs) play an important role in olfactory system of Chinese bee, Apis cerana cerana. In this study, we aim to research the binding characteristics of pheromones and some plant volatiles with a novel pheromone-binding protein OBP10 of A. cerana cerana. 【Methods】 We successfully cloned the gene  AcerOBP10 from A. cerana cerana (GenBank accession no. KP717060.1) by RT-PCR, constructed its prokaryotic expression vector pET30-AcerOBP10, and conducted its expression in the optimized conditions in vitro. The recombinant protein with biochemical activities was purified by the method of Ni²⁺-agarose affinity chromatograph. Using 1-NPN as a fluorescence probe, the binding capability of AcerOBP10 with diverse bee pheromones and plant volatiles was measured by competitive fluorescence assay. 【Results】 By analyzing the alignment of the deduced homologous proteins of other insect species, we speculated that AcerOBP10 is a novel PBP of  A. cerana cerana. The results of competitive fluorescence assay showed that eight out of 14 ligands were able to decrease the relative fluorescence intensity of 1-NPN by more than 50%. Among them, methyl 4-hydroxylbenzoate (HOB), one of queen pheromone components, showed the strongest capability to compete with 1-NPN, causing 93.94% reduction in the relative fluorescence intensity with the K_D value of 11.04 μmol/L. AcerOBP10 was also found to have the capability of binding with worker bee pheromones (geraniol and nerol) and alarm pheromones (heptanone and isoamyl acetate), suggesting that it plays a significant role in recognizing pheromones in the bee olfactory system. AcerOBP10 also had strong binding force with some plant volatiles, like β-ionone. 【Conclusion】 As a pheromone-binding protein, AcerOBP10 has a strong binding capability with diverse bee pheromones. Compared with another PBP (AcerASP1), AcerOBP10 seems to have more extensive pheromone binding profiles. The results provide a theoretical basis to explore the molecular mechanism of pheromone recognition and transduction in A. cerana cerana.

【Aim】 The aim of this study is to investigate the anatomy of the brain of Helicoverpa assulta (Lepidotpera: Noctuidae) adults and to create a digital three-dimensional brain model.【Methods】 Immunohistochemical staining with synaptic protein antibody was used to label the neuropil structure of the brain. By using a confocal laser scanning microscope we obtained digital images of the brain, which were analyzed by using the three-dimensional image software, AMIRA. 【Results】 The immunostaining with synaptic protein antibody visualized the neuropil regions of the brain and the gnathal ganglion of H. assulta. In adults, the brain and gnathal are fused, but with a hole in the middle, which results from the bypassing esophagus. The brain is composed of protoecerebrum, deutocerebrum and tritocerebrum. Based on the synapsin staining, such neuropils of the brain as the optic lobes, the anterior optic tubercles, the mushroom bodies, the central complex, the antennal lobes and their sub-regions were identified and reconstructed. In addition, the large part of the protocerebrum surrounding these neuropils, plus the gnathal ganglion, was reconstructed and categorized as the whole midbrain. This structure includes about 55.05% of the total brain neuropil. 【Conclusion】 These results provide knowledge that is essential for understanding the basic neuroanatomical principles underlying information processing, integration of multimodal input, and behavioral regulation. And the findings are important for future projects intending to study the development, variation, and plasticity of insect brain structures.

【Aim】 This study aims to clarify the bioactivities of rosemary and zanthoxylum oils against the carmine spider mite, Tetranychus cinnabarinus (Boisduval), in the laboratory. 【Methods】 The fumigation activities, oviposition inhibition activities and repellency activities of rosemary and zanthoxylum oils against the 5-day-old female adults of T. cinnabarinus were detected by the methods of fumigation and repellency. By using EthoVision XT6, the effects of the two essential oils on the behavior of T. cinnabarinus were analyzed at the sublethal concentrations. 【Results】 The mortalities of female adults of T. cinnabarinus fumigated by rosemary oil were significantly higher than those by zanthoxylum oil at 12, 24 and 48 h after exposure (P <0.05). Moreover, as the fumigation time or the concentration of the two essential oils increased, the fumigation activities were enhanced. At 24 h after treatment, the oviposition inhibition rates of female adults of T. cinnabarinus exposed to the rosemary oil were significantly higher than those exposed to the zanthoxylum oil at different concentrations ( P <0.05). After 24 h of exposure, the LC₃₀ values of rosemary and zanthoxylum oil against female adults of T. cinnabarinus were determined to be 6.731 and 120.142 μL/L, respectively. The repellency activities of the rosemary oil against female adults of T. cinnabarinus were higher than those of zanthoxylum oil. When the female adults of T. cinnabarinus were treated by the two essential oils at the sublethal concentrations, their spontaneous movement tracks changed significantly and the mobility frequencies were the highest at the LC₁₀. Moreover, the spontaneous movement abilities of female adults of T. cinnabarinus were more obviously affected by rosemary oil than by zanthoxylum oil. 【Conclusion】 The bioactivities of T. cinnabarinus induced by the rosemary oil were significantly higher than those induced by the zanthoxylum oil. Therefore, rosemary oil has higher application value in the biological control of T. cinnabarinus. This study provides a scientific basis for the development of botanical acaricides for green vegetables.

【Aim】 To explore the effects of the dual stress of UV-B and cadmium on the growth, development and fecundity of  Sitobion avenae (Fabricius), and to determine whether there is the interaction between UV-B and cadmium. 【Methods】 The age-specific life table was employed to explore the effects of the co-stress of different intensities of UV-B (0, 0.1, 0.2 and 0.7 mW/cm²) and concentrations of cadmium (0, 20, 40, 80 and 160 mg/kg) on the growth, development and fecundity of S. avenae in the laboratory. 【Results】 The life table parameters of S. avenae were adversely influenced by the dual stress of UV-B and cadmium, and they were affected more severely by UV-B than by cadmium stress. In addition, there are significant interactions between the stress of UV-B and cadmium. In particular,under the same concentration of cadmium treatments, the effects of UV-B on the intrinsic rate of increase (r), the finite rate of increase (λ), the net rate of increase (R₀), fecundity (F), oviposition duration and adult longevity of S. avenae showed a trend of first decreasing and then increasing, and fell to the minimum value under the intensity of 0.2 mW/cm² of UV-B. However, the mean length of a generation (T) revealed an adverse trend, namely showing a trend of first increasing and then decreasing, and reached the first peak at the intensity of 0.1 mW/cm² of UV-B. Under the same intensity of UV-B radiation, with the increase of the concentrations of cadmium, all life table parameters of S. avenae displayed fluctuation. Moreover, the parameters of r, λ, R₀, F, oviposition duration and adult longevity of S. avenae in the combined treatments were significantly lower than those in the treatments only exposed to different concentrations of cadmium. However, the parameter  T revealed a reverse result. Every peak point or minimum value of all above life table parameters of S. avenae exposed to the dual stress of UV-B and cadmium was somewhat earlier than that only exposed to different concentrations of cadmium. 【Conclusion】 The growth, development and fecundity of S. avenae were significantly affected by the stress of UV-B and cadmium. Moreover, the influences will be intensified when the two factors act in combination, suggesting that these two factors have significant interactions.

【Aim】 In agriculture, climatic changes affect crop yield, dynamics of aphid pests and their regulation by ladybird predators. While some ladybirds are ubiquitous and others have limited geographical ranges; but how climatic cues influence distribution of ladybirds is poorly explored.【Methods】Present study, therefore, assesses the effects of different temperatures (15℃, 20℃, 25℃, 30℃ and 35℃), and photoperiods (8L:16D, 12L:12D and 16L:8D) on consumption and utilization of the pea aphid, Acyrthosiphon pisum (Harris) by larvae of a ubiquitous ladybird ［Coccinella septempunctata(Linnaeus)］ and an Oriental/south-Asian ladybird (Coccinella transversalis Fabricius). 【Results】Although predation attributes and developmental rates of both ladybirds were the highest at 25℃ and attributes further increased with increase in day length (from 8L:16D to 16L:8D photoperiod), larvae of C. septempunctata had higher consumption, growth and developmental rates, and their newly emerged adults were heavier than those of C. transversalis at all temperatures and photoperiods. Non-linear regression model extrapolated~11℃ and~10.5℃ as lower development thresholds and ~41℃ and ~43℃ as upper development thresholds for C. transversalis and C. septempunctata, respectively. Dependency of larval predation and developmental rates to climatic cues were higher in C. transversalis than C. septempunctata. 【Conclusion】Present study, therefore, reveals that a ubiquitous ladybird is less susceptible to climatic cues than an indigenous ladybird and hence is widely distributed to different geographical regions. Thus, the former may behave as a better biocontrol agent than the latter.

【Aim】 To explore the phylogenetic relationships within and between three major subfamilies (Polyommatinae, Theclinae and Lycaeninae) of the Chinese lycaenids based on the mitochondrial gene sequence data. 【Methods】 The partial mitochondrial  COI and Cytb genes of 53 species were amplified and determined with PCR method, the sequence variation of these species were also analyzed, and the phylogenetic trees of these lycaenid species were reconstructed with maximum-likelihood (ML) and Bayesian-inference (BI) methods. 【Results】 The results of the sequence analysis showed that the two combined genes are 1 431 bp in length, including 855 conserved sites, 576 variable sites and 488 parsimonious-informative sites, and the average percentage of A+T is 74.5%, which is much higher than that of G+C (25.5%). The results of phylogenetic analyses indicated that the Polyommatinae and Lycaeninae were monophyletic groups, whereas the Theclinae was a paraphyletic group. All the taxa in this study contained three main clades: the first was the subfamily Lycaeninae, the second was the subfamily Polyommatinae plus the tribes Hypolycaenini and Aphnaeini of the subfamily Theclinae, and the third was the other four tribes of the subfamily Theclinae, with their relationship as (((Theclini+Arhopalini)+Deudorigini)+Eumaeini). 【Conclusion】 The subfamily Lycaeninae is a monophyletic group, while the Polyommatinae and Theclinae are more closely to each other, with the relationship of their main groups awaiting further investigation.

【Aim】 To determine the ultrastructural characteristics for diagnosis of dominant Dermacentor species, tick-borne vectors, in Xinjiang, northwestern China. 【Methods】 The ultrastructure and morphology of four dominant Dermacentor species, i.e.D. niveus, D. silvarum, D. nuttalli and D. marginatus, were observed by scanning electronic microscope on the basis of observation and indentification with stereo microscope. 【Results】 Differences existed in the idiosomal size and the enamel color of scutum among the four species. There are three odontoid processes on the ventral podomer Ⅳ (femur, tibia and metatarsus) of D. niveus and D. marginatus, and the odontoid processes are present on both tibia and metatarsus of D. nuttalli, but only present on the metatarsus of D. silvarum. Notable differences were also found in the seta arrangement of ventral palpus’s medial margin, the length of claws, and the claw-pulvillus length ratios of the four dominant Dermacentor species. 【Conclusion】 The number and location of the odontoid processes on ventral podomer Ⅳ and the morphology of spiracular plate are diagnostic characteristics for the four Dermacentor species, and a rapid species identification could be made by combining these characteristics with the seta arrangement of ventral palpus, the claw length and the claw-pulvillus length ratio.

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.

Aging is a complex process that involves the accumulation of poisonous materials inducing the decline of life function, leading to the decrease in physiological functions and ultimately leading to disease and death. Drosophila melanogaster represents one of the most important genetical tools and is widely used in the study of aging. In this article, we summarized the relationship of aging with juvenile hormone, insulin/IGF signaling pathways, target of rapamycin pathway, AMPK pathway, caloric restriction, oxidative stress, microRNA and sphingolipid metabolism. In addition, genome wide association studies may be able to find genomic region, which has potential relationship with aging. The studies based on the model organism D. melanogaster are very useful for studies in aging process of other insects. These will result in further understanding the similar mechanisms in Insecta, the most diverse animal class, so as to develop manipulation techniques for extending longevity of natural enemies and other beneficial insects and reducing fecundity and lifespan of harmful insects in agricultural and urban ecosystems.

The fall webworm, Hyphantria cunea (Drury) (Lepidoptera: Arctiidae), a destructiveinvasive pest and native to North America, has spread from Dandong city, Liaoning province of northeastern China to Wuhu city, Anhui province of southeastern China. Effective control measures are urgently needed. In this study, Beauveria bassiana isolates from H. cunea were examined for biological characteristics and virulence, to assess their potential as a bio-control agent for H. cunea. 【Methods】 The biological characteristics of seven isolates (Bb01-1, Bb08-5, Bb08-6, Bb08-11, Bb22-2, Bb22-3 and Bb22-5), including colony morphology, growth rate, conidial yield, conidial germination and tolerance to UV light, were examined. Then, three isolates with better performance were selected for bioassays against the 4th instar larvae of H. cunea. 【Results】 There were significant differences in colony growth rate, conidial yield, conidial germination and tolerance to UV light among the tested  B. bassiana  isolates. The isolates Bb22-2, Bb22-3 and Bb22-5 exhibited better characteristics with faster hyphal growth rate, higher spore production, faSter spore germination and higher UV tolerance. The results of pathogenicity test of B. bassiana to the H. cunea larvae showed that the three isolates of B. bassiana demonstrated a higher efficacy. The three isolates caused the corrected mortality rates of 80%-90% in H. cunea at 11 d post inoculation. And the infection rates of B. bassiana against H. cunea were between 53% and 87% at 11 d post inoculation. The isolate Bb22-2 caused higher infection rates of B. bassiana against H. cunea than the other two isolates. 【Conclusion】 This study demonstrated that B. bassiana is a highly effective entomopathogen against H. cunea. The isolate Bb22-2 has a great potential for sustainable control of H. cunea in forests. Further field experiments are needed to investigate its efficacy under natural conditions.