In this study, we produced germline transgenic silkworm that spin cocoons containing recombinant phyase in the fibroin layer. Using the piggyback-derived vector pPIGA3GFp and pBac{3×P3-EGFPaf}, we successfully developed stable germline transformation in the silkworm Bombyx mori L. We further constructed a piggyback-based transformation vector that carried a fusion cDNA of phyase with partial fibroin light chain (FibL) and red fluorescent protein (DsRed). The fusion cDNA was driven by FibL gene promoter. Silkworm eggs were injected with the vectors, producing worms displaying red fluorescence in their silkglands and cocoons. Southern blot and inverse-PCR results indicated that the insertion fragment was recombined with the chromosome of silkworm. RT-PCR analysis showed that the phytase gene was expressed especially in the posterior silkgland, and this expression pattern was the same as that of fibroin light chain gene. These results showed that we successfully got phy-transgene silkworm. This study demonstrates the viability of transgenic silkworms as a tool for producing useful proteins in the silkglands.

Members of the Caspase family play a central and evolutionary role in apoptosis, which remove the unwanted, damaged and dangerous cells during development to maintain homeostasis. In this study, by using the total RNA isolated from the whole body of the house fly, Musca domestica, the full-length sequence of Mdom-caspase-1 gene encoding the Caspase-1 was cloned by the methods of reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification cDNA ends (RACE) to explore the function of caspase in insect growth and metamorphism. The sequence analysis showed that Mdom-caspase-1 encodes 289 amino acid residues and the 1 295 bp full-length cDNA contains an 870 bp open reading frame (ORF). The GenBank accession numbers of cDNA and amino acid of Mdom-caspase-1 are EU854472 and ACF71490, respectively. The deduced Mdom-caspase-1 protein (MW 32.83 kDa; pI 8.67) has 5 conserved cysteines QACQG, which is one of the typical characteristics of Caspase. The Mdom-caspase-1 protein is hydrophilic, while 8 regions containing 89 amino acids are hydrophobic. The protein secondary structure mainly consists of 11 α helices, 7 β folders and 17 turns. The phylogenetic tree was created from the alignments of the amino acid sequences, and the analysis revealed that Caspase-1 possessed distinct conservation in evolution and the amino acid sequence of Mdom-caspase-1 is 65%-77% identical to Caspase-1 from Drosophila melanogaster, Aedes aegypti and Culex quinquefasciatus. The results of RT-PCR revealed that Mdom-caspase-1 was expressed in every stage of M. domestica, and in five stages including the egg, 3rd instar larva, pre-pupa, pupa and the 5 d adult female the expression levels were obviously higher than those in other stages. The results suggest that the Caspase-1 gene may have some relationships with the growth and metamorphism of insects. These results would provide some valuable information to explore the molecular function of Caspase-1 in insects and develop the specific Caspase-1 inhibitors.

Olfactory receptors (ORs) play a critical role in insect olfactory system. Research on ORs can help us understand the molecular mechanism of odors discrimination in insect. In this study, 11 Helicoverpa armigera OR genes (GenBank accession numbers: EU599565-EU599568, EU818702-EU818706, FJ393455) were cloned using RT-PCR. The results of sequence analysis showed that HarmOR2 was atypical OR and the rest 10 ORs were typical ORs. Phylogenetic tree indicated that HarmOR10 and DOr46a belonged to a single cluster, while HarmOR4 belonged to a cluster of gustatory receptors (Grs). The other H. armigera ORs belonged to the cluster which had a long distance with Drosophila melanogaster ORs. We also found that HarmOR2 was expressed in antennae and proboscis of H. armigera, while other HarmORs were specially expressed only in the antenna of adult moth, of which HarmOR3，HarmOR13 and HarmOR14 were expressed only in the antenna of male adult. Moreover, HarmOR12 and HarmOR20 were expressed at higher level in antenna of male than that of female, while other HarmORs were expressed at the same level both in female and in male.

Chitin, synthesized by chitin synthase (CHS), is not only the essential component for epidermis and peritrophic membrane of the midgut in insects, but also a very pivotal target for insect pest control. Based on the cDNA sequences and genomic sequences of the two CHS genes in Spodoptera exigua, the 5′flanking sequences were cloned using specific primers. Promoter sequences of SeCHSA and SeCHSB were obtained after initiation sites of transcriptional regulation were verified by means of 5′RACE. These results are important for the research of chitin synthesis and transcriptional regulation in insects.

【Aim】 Bac-to-Bac baculovirus expression system was used to obtain a soluble fusion protein sPDGFRβ/Fc, combining the extracellular domain of PDGFR β chain and the Fc region of human IgG in insect cells Sf9, and the specificity and bioactivity of the recombinant protein are detected. 【Methods】 The gene sPDGFRβ/Fc was cloned into a transfer vector pFastBac to form the recombinant donor plasmid pFastbac-sPDGFRβ/Fc, which was transformed into Escherichia coli DH10Bac. By transposition, sPDGFRβ/Fc gene was integrated into Bacmid, and a recombinant shuttle vector rBacmid-sPDGFRβ/Fc was constructed. Then the rBacmid-sPDGFRβ/Fc recombinant genome DNA was used to transfect Sf9 mediated by lipidbody, and the recombinant baculovirus rBacmid-sPDGFRβ/Fc was obtained, which was further used to infect the serum-free cell Sf9 to express sPDGFRβ/Fc. Then the recombinant protein was purified with Protein A chromatography and analyzed by Western blotting and MTT assay. 【Results】 Western blotting analysis showed a specific band about 97 kDa, consistent with the target protein, 75% pure sPDGFRβ/Fc was obtained by using Protein A chromatography with a yield of 1 μg/mL culture medium, which was confirmed to be able to inhibit PDGF-induced proliferation of Balb/c 3T3 cell. 【Conclusion】 The recombinant protein sPDGFRβ/Fc with activity of inhibiting PDGF-induced cell proliferation can be successfully expressed in insect cell.

The endocrine cells in the digestive tract of Oxya chinensis were identified and localized by Grimelius silver impregnation and immunohistochemical method combining with biostatistical analysis. The results showed that argyrophil cells were detected in the gastric caecum, the midgut and throughout the hindgut, but not found in the foregut by silver impregnation, with the highest density in the midgut and rectum （P<0.05）. 5-hydroxytryptaminc （5-HT）， gastrin （Gas）， glucagon （Glu）and pancreatic polypeptide （PP） cells were identified in gastric caecum, the midgut and the hindgut by immunohistochemical method, but somatostatin （SS） cells were not detected throughout the digestive tract. Both 5-HT and Gas cells are distributed in the gastric caecum, midgut and rectum with the highest density in the rectum（P<0.05）. Glu cells are distributed throughout the midgut and the hindgut, with the highest density in the midgut and rectum （P<0.05）. PP cells are distributed in the midgut, ileum and rectum with the highest density in the midgut（P<0.05）. The results indicate that there are several types of endocrine cells in the digestive tract of O. chinensis; and the distributional pattern of these cells are similar to those of other arthropods, but some unique features of the species were also found, which might be related to its special structure and physiological function of digestive tract.

Using SMART RACE RT-PCR, the complete full-length cDNA encoding a chitinase was cloned from the biocontrol agent Isaria fumosorosea. The gene, designated as Ifuchi1, contains an open reading frame (ORF) with 1 275 bp encoding 424 amino acids, which contains a N-terminal 24 amino acid residues displaying the characteristics of signal peptide. The mature chitinase has a molecular mass of 47.6 kDa and a calculated pI of 4.89. The protein sequence contains two highly conserved regions including a putative enzymatic active site and a potential chitin-binding domain. The chinitase belongs to the class V of family 18 of glycosyl hydrolases. The Ifuchi1 gene from I. fumosorosea was engineered into Beauveria bassiana by blastospore transformation. Compared to the wild-type strain Bb13, the chitinase activity of a selected transformant was increased by 86.2% after induction for 36 h. Bioassayed against the larvae of Masson’s pine caterpillar Dendrolimus punctatus, the median lethal time (LT₅₀) of transgenic isolates was reduced by 29%, and the mortality rate was increased by 52.9%, as compared to the wild-type at a concentration of 1×10⁷ conidia / mL. The median lethal concentration (LC₅₀) values of the wild-type and transgenic isolates were 4.71×10⁶ conidia / mL, and 1.74×10⁶ conidia / mL (approximately 1.7-fold decrease), respectively. The transgenic strain thus showed significantly improved virulence against D. punctatus.

Fluorescent brightener 28 is used as an influencing factor to study the mechanism of M2R on the peritrophic membrane (PM) of Loxostege sticticalis larvae. The environmental scanning electron microscopy and biochemical technique were employed to analyze the morphological structure and the protein profiles of the PM of L. sticticalis larvae treated by M2R, and the disruptive effect of M2R on the PM and its synergistic effect on Bt virulence were studied. The results indicated that there were many types of PM proteins. Separation of PM proteins by SDS-PAGE showed that there were at least 19 polypeptides with molecular weight less than 94 kD. The chitin binding proteins （CBP） content of PM could be influenced when the larvae were fed with 1.0% M2R. The surface of the normal PM was smooth and wrinkled without pores or slits while the surface of the treated PM had pores and slits. Although M2R did not affect the larval growth, it could enhance the sensitivity of larvae to Bt infection, shorten the killing time and reduce the application dose of Bt. These results suggest that the M2R could damage the PM of the larvae, and thus increase the control effect of Bt.

To investigate the relationship of locomotor activity and dopamine levels in the head in fruit fly Drosophila melanogaster affected by rotenone, the effects of rotenone on locomotor activity and gene expression related to dopamine biosynthesis and dopamine levels in heads of fruit flies were determined. The results showed that locomotor activity was significantly reduced in male and female adult flies fed on Drosophila instant food containing 0.2-0.8 mmol/L rotenone for 6 d. The dopamine levels were reduced to 79.3%, 66.8% and 53.2% of that of the male control in heads of male adult flies and 83.2%, 72.3% and 59.8% of that of the female control in heads of female adult flies after feeding on instant food containing 0.8 mmol/L rotenone for continuous 6, 12 and 21 d, respectively. Under the above same conditions the expression levels of pale gene were reduced to 76.3%, 51.4% and 37.3% of that of the control, and Ddc gene to 87.1%, 78.2% and 63.5% of that of the control, respectively. The results suggest that rotenone can significantly interfere with pale and Ddc gene expression and reduce dopamine levels in heads of adult flies, and correspondingly affect the adult fly locomotor activity.

To study the ecological regulation effects of species diversity in wheat fields on Sitobion avenae, field experiments were carried out in Langfang Experimental Station of Institute of Plant Protection, Chinese Academy of Agricultural Sciences from October，2007 to July，2008. The intercropping patterns of wheat and pea, by the proportions of planting row of pea and wheat in 2∶2, 2∶4, 2∶6 and 2∶8 (referred to as 2-2 intercropping, 2-4 intercropping, 2-6 intercropping and 2-8 intercropping, respectively), were plotted, and the field cultivar monoculture of wheat was planted as the control. Population dynamics of apterae and alatae S. avenae, population dynamics, species richness, diversity index and evenness of main natural enemies were systematically investigated and analyzed. The results showed that, compared with monoculture of wheat, the amount of S. avenae apterae per 100 plants (square-root transformed) in aphid peak period were very significantly lower in the intercropping treatments than in the control (P<0.01), and the cascade was as wheat monoculture (77.38) > 2-2 intercropping (68.62) > 2-4 intercropping (68.51) > 2-8 intercropping (65.19) > 2-6 intercropping (64.94). Although population dynamics of main natural enemies showed a similar trend with time, wheat-pea intercropping could preserve and augment natural enemies more than monoculture of wheat, and there were higher population densities of ladybeetles and aphid parasitoids, and higher species richness and diversity index of natural enemies, but lower evenness index in every intercropping field. It is so concluded that wheat-pea intercropping system can not only reduce the population of S. avenae, but also improve the stability and sustainability of controlling wheat pests by natural enemies.

To explore the defensive behavior in firefly Pyrocoelia pectoralis Oliver 1883, morphological and behavioral experiments were conducted. P. pectoralis was discovered to possess complex, multiple and effective defensive behavior. Adults use glowing, death mimic and reflex bleeding to defense, whereas larvae defend predators by glowing, death mimic, and small pairs of eversible glands. Male adults of P. pectoralis discharge fluids from pronotal and elytral margins, but when tactually stimulated, females only discharge fluids from pronotal margins. With regard to its content of haemocytes, the discharged fluid conforms to haemolymph. A series of circular pits about 32 μm in diameter, but irregularly spaced depressions in the cuticle of the firefly along the pronotal and elytral margins, seems to be related to the ‘reflexbleeding’ response. The larvae of P. pectoralis possess 9 pairs of small pale white, mammiform eversible glands. TEM revealed that the protuberances of eversible organs of P. pectoralis larva are hollow at the base. The cytoplasm of the secretory cells of eversible organs is characterized by extensive system of cisternae and tubular endoplasmic reticulum. Behavioral experiments confirmed that the blood of P. pectoralis is very effectively repellent to ants.

Cellular immunity that includes phagocytosis, nodulation and encapsulation mediated by hemocytes is a very important part of the innate immune system in insects. In this article we reviewed the current understanding of cellular immune responses in insects. We summarized the hemocyte types involved in cellular immunity, some cellular receptors that recognize different microbes and multicellular parasites, and some enzymes and chemicals that can affect the immune activity. We also discussed the modes of hemocyte-mediated phagocytosis and the morphological change of hemocytes during the progress of nodulation and encapsulation.

Drosophila melanogaster is one of the most popular animal models for biological research, especially for studies on olfaction. Recent advancement in Drosophila olfactory research uncovered much about the mechanisms underlying odor detection and discrimination, sensory transduction, coding, olfactory memory, etc. It showed that～1 300 Drosophila olfactory receptor neurons (ORNs) express a total of 62 different olfactory receptor proteins (ORs) to detect all chemical molecules a fly senses. The odorant ligands of many Drosophila ORs have been elucidated: General odors, such as fruit odors, are encoded by combinations of different ORs; whereas pheromones that elicit mating and social interactions are encoded by single specific OR. Odor information is transformed into electrical neural signal in ORN, then the signal is relayed to the antennal lobe (AL) along the axon of ORN, and further the signal is relayed to the mushroom body (MB) and lateral horn (LH) by the projection neurons (PNs). Some proteins such as Dock, Ncadherin, and Fruitless play crucial roles in the physiology of Drosophila olfaction; loss of these proteins will lead to olfactory deficiencies. In this article we reviewed the major progress made in recent years.

The morphology and histology of the alimentary canal in the scorpionfly Panorpa obtusa Cheng were investigated using light and scanning electron microscopy. In an antero-posterior direction, the foregut includes a pharynx, an oesophagus, and a proventriculus, but is devoid of crop. The elongate pharynx is subdivided into the anterior pharynx and the posterior pharynx or pharyngeal pump, which is inserted with dilator muscles. The proventriculus possesses numerous uniform brown proventricular acanthae on the sclerotized intima, performing the functions of filtrating, grinding, and temporarily storing food particles. Six cardiac valves protrude into the midgut. The long midgut is distensible with longitudinal muscles outside circular muscles; the epithelium consists of large columnar and small regenerative cells. There are no gastric caeca and peritrophic membrane. Six long brown Malpighian tubules are located at the junction between the midgut and the hindgut, extending freely into the heamocoel. The hindgut is subdivided into a V-shaped ileum, a loop-shaped colon, and a translucent, distensible rectum where six rectal pads are located alternatively. Finally, the structure and function of the alimentary canal and its significances in Panorpidae taxonomy are briefly discussed.

【Aims】 Behavioural responses of adults of Nassophasis sp., a species infesting medicinal herb Dendrobium thyrsiflorum, to different odor sources were studied to explore new control methods towards this pest.【Methods】Behavioural responses of Nassophasis sp. adults to different odor sources including conspecifics, puparium, frass, and damaged stem of host plant, Dendrobium thyrsiflorum, were tested with pitfall olfactometer. 【Results】 The results indicated that odor emitted from females could only attract males but not females. The odor from males was not attractive to both sexes. Both sexes could be significantly attracted to puparium. Neither femaleproduced frass nor maleproduced frass showed significant attraction to both sexes. When females and males aggregated, however, their frass demonstrated significant attraction to both sexes. Female adults only responded to female-damaged stem, while male adults only to male-damaged stem.【Conclusion】 These preliminary results may serve as basis to isolate and identify the attractive compounds from these odor sources.

This article reported the anti-senile effect of water extraction of Martianus dermestoides adults and larvae by testing the activities of SOD, GSH-Px and CAT and MDA content in blood serum of mice. Water extraction was distilled from adults and larvae of M. dermestoides which were separately reared by the advanced forage (consisting of safflower, medlar, Chinese date and Juglans regia nuts）, rice and wheat bran. We use water extraction of M. dermestoides larvae to clystere the mice which were of a senile model established by injecting D-galactose. The results demonstrated that all the water extraction of M. dermestoides adults reared by three kinds of forage increased the activities of SOD， GSH-Px and CAT (P<0.05 or P<0.01) in blood serum of mice, and decreased the MDA content (P<0.05); the effect of the advanced forage group was stronger than the other two groups, but there were no significant differences among the three groups. SOD and CAT activities in blood serum of the mice which had been clystered with water extraction of M. dermestoides larvae were obviously higher than those of the adults, both reared by the advanced forage (P<0.05); however, there were no significant differences between GSH-Px activity and MDA content. The results suggest that water extraction of both adults and larvae of M. dermestoides have antioxidant activity, amd M. dermestoides can be exploited as a kind of health food with antisenile effect.