A method measuring EAG (electroantennogram) response in lamellate antennae of Holotrichia parallela Motschulsky was established in this study. A higher value of EAG was got when lamellae were separated from each other by insect pin. With the concentration of 30 ng·μL^-1, the EAG value of male antennae reached the maximum when the volume ratio of L-isoleucine methyl ester (LIME) to (R)-(-)-linalool was 7∶1, which was almost equal to the EAG value of the crude extracts from 25 sex glands. The male antennae were treated with different concentrations of stimulative reagent with the fixed volume ratio (7∶1) of LIME to (R)-(-)-linalool. The highest EAG value of male antennae was found when the concentration was 30 ng·μL^-1. The results of olfactory response further confirmed that male H. parallela was attracted with the selection efficiency of 93.3% when the volume ratio of  LIME  to (R)-(-)-linalool was 7∶1. So it can be deduced that the sex pheromone of Holotrichia parallela in Shandong province were the mixture of  LIME  and (R)-(-)-linalool with the ratio of 7∶1. The results may serve as basis for developing the techniques of sex pheromone attraction to control H. parallela.

Huwentoxin-III purified from the venom of the Chinese bird spider, Ornithoctonus huwena (Wang et al.), is an insect neurotoxic peptide. The effects of huwentoxin-III on neuronal voltage-gated ion channels were studied by using whole-cell patch clamp technique. Huwentoxin-III specifically inhibited voltage-gated sodium channels in dorsal unpaired median neurons of adult cockroach Periplaneta americana (IC50≈1.106 μmol/L) while having no evident effect on voltage-gated potassium channels. HWTX-III inhibited insect voltage-gated sodium channels through a novel mechanism distinctive from other spider toxins, did not affect the activation and inactivation kinetics, and not evidently shift the steady-state inactivation curve. Thus, its specificity and novel mechanism on insect neuronal voltage-gated sodium channels make it an interesting tool for investigating the multiple molecular forms of voltage-gated sodium channels and exploiting new and safe insecticides.

In order to reveal the accumulation mechanism of trehalose, a main chemical substance to maintain diapause of Sitodiplosis mosellana (Gehin), we measured the activities of four sugar metabolic enzymes including glycogen phosphorylase (Gpase), hexokinase (HK), phosphofructokinase (PFK) and aldolase (ALD) in pre-diapause, diapause and post-diapause larvae of S. mosellana by using testing kits. The results indicated that activities of these sugar metabolic enzymes were obviously different at various diapause stages of S. mosellana larvae. Gpase activity was increased significantly, while activities of glycolysis enzymes (HK, ALK and PFK) were decreased significantly after entering diapause. Gpase activity was decreased, while activities of glycolysis enzymes were increased after diapause termination. Activities of four enzymes were all related with developmental status of diapause larvae during diapause. Gpase and PFK were also influenced by environmental temperature, namely, activities of Gpase and PFK were higher in summer and winter than those in spring and autumn. At the same diapause stage, activities of HK, PFK and ALK of non-cocooned larvae were higher than those of cocooned larvae. On the contrary, Gpase activity of non-cocooned larvae was lower than that of cocooned larvae. No obvious difference in four metabolic enzyme activities was found between the 1st and 2nd year diapause larvae.

We described the identification and characterization of a new cDNA encoding chemosensory protein1 (CSP1) from the antenna of diamondback moth, Plutella xylostella, a species whose CSPs have not been identified to date. We focused our investigation on this olfactory protein family using reverse transcription-polymerase chain reaction strategies. The results showed that the CSP1 gene in diamondback moth, named PxylCSP1 (GenBank accession no. FJ361903), is 405 bp in length and encodes 134 amino acid residues, with six cysteine residues in conserved positions relative to other known CSPs. The predicted N-terminus hydrophobic region contains 19 amino acid residues within the Has-CSP, displaying the characteristic features of a signal peptide. Thus the predicted molecular weight (MW) of the mature protein is 13.56 kD and isoelectric point (pI) is 6.12. The alignment of PxylCSP1 showed high sequence identity (70%-80%) with other full-length sequences of other lepidoteran CSPs from GenBank. RT-PCR analysis revealed that PxylCSP1 was not only expressed in antennae, but also in head, abdomen, wings and legs. The reasult of real-time PCR further indicated that the transcription level of PxylCSP1 depended on the gender, age, mating status and tissues of the tested moths.

The effect of cell membrane perforation caused by myclobutanil on the cell membrane permeability of Spodoptera litura ovary cells (SL cells) that were cultured in vitro was assayed in this study. The results showed that a large number of fluorescent dye PI, which was macromolecular material and could not pass through the SL cell membrane, passed through the cell membrane and entered into cells with the myclobutanil. The increasing rates of cell fluorescence intensity were 25.80% and 11.95% respectively after SL cells were treated with 20 and 40 μg/mL myclobutanil for 12 h, and 27.77% and 57.49% respectively for 24 h. Myclobutanil significantly improved the leakage rate of lactate dehydrogenase which is the macromolecular material in SL cells. The leakage rates were 30.66% and 43.93% respectively after SL cells were treated with 20 and 40 μg/mL myclobutanil for 24 h, and 41.22% and 57.91% respectively for 48 h. Myclobutanil significantly increased the intracellular calcium ion content. Intracellular calcium fluorescence intensity was significantly different from control after SL cells were treated with 20 and 40 μg/mL myclobutanil for 24 and 48 h. After SL cells were treated with myclobutanil for 24 h, the LC50 was 35.84 μg/mL. The activity of myclobutanil was significantly higher than that of rotenone which was a typical cell toxicant. When the SL cells were treated with the mixture of myclobutanil and rotenone at the mass ratios of 1∶1 and 2∶1 for 24 h, the LC50 values were 43.92 and 26.09 μg/mL respectively, and the co-toxicity coefficients were 137.60 and 188.49 respectively. The results showed that the synergism of myclobutanil and rotenone is remarkable and increased with the myclobutanil content increasing. These results suggest that myclobutanil has a good inhibition on the SL cells, can get through the membrane barrier that limits materials to penetrate the cells, and so increases the effective utilization of pesticide active ingredients.

Isolation and identification of the binding proteins of Cry1A toxins in the midgut brush border membrane vesicles (BBMV) of Chilo suppressalis is very important for understanding the mode of action and resistance mechanism of Cry1A toxins. In the present study, insecticidal activity and ligand binding of Bacillus thuringiensis Cry1A toxins to the midgut BBMV of striped rice stem borer C. suppresalis were investigated. The results indicated that Cry1Ab was more toxic than Cry1Ac, and Cry1Ac was more toxic than Cry1Aa against the 3rd instar larvae of the laboratory strain (CN) of C. suppresalis. Ligand blot analysis showed that there were six major proteins (50, 70, 90, 120, 160 and 180 kDa) binding to Cry1Ac in the midgut BBMV of the CN strain. Bands of 180, 160 and 90 kDa binding proteins were much darker than the others, suggesting that these proteins had higher binding concentrations. Results of homologous competition binding indicated that the 180 and 90 kDa protein bands were of low binding affinity, while the other four protein bands (160, 120, 70 and 50 kDa) were of high binding affinity. Heterologous competition binding assays were conducted to study crossreactivity of the binding sites between Cry1Ab and Cry1Ac. Cry1Ab competed for all binding sites recognized by Cry1Ac, with high affinity to 180, 120, 70 and 50 kDa proteins and low affinity to 160 and 90 kDa proteins. These data suggest that Cry1Ac and Cry1Ab share several common binding sites in the midgut BBMV of C. suppressalis, but they have different binding affinity with respect to each binding site. Considering the similarity in binding sites, Cry1Ab and Cry1Ac should not be used simultaneously in the transgenic Bt rice to control the target pest C. suppressalis.

To determine the development and utilization value of Vitex negundo Linn. as grain protectant, this study focused on the component analysis, toxicity, mode of action and action mechanism of essential oil of V. negundo, which was extracted using steam distillation. The GC-MS method was used to analyze the components of essential oil. The results showed that there were thirty-one components whose content was more than 0.3% in the essential oil of V. negundo. The main components were caryophyllene, eucalyptol and α-pinene, with contents of 35.97%，8.21% and 0.69％, respectively. Eucalyptol, α-pinene and V. negundo essential oil all had high overall toxicity to the adults of Sitophilus zeamais in 72 h of treatment and the highest one was eucalyptol, whose LC50 was 0.7171 g·kg^-1. They also could control F₁ population effectively. At the dosage of 2.0 g·kg^-1，the inhibitory rates of eucalyptol, α-pinene and V. negundo essential oil to S. zeamais were 100%, 85.45％ and 89.73％, respectively. The LC₅₀ values of eucalyptol, α-pinene and essential oil of V. negundo to S. zeamais in the contact toxicity were 0.2690, 0.7529 and 0.2969 mg·cm^-2, respectively; and at the dosage of 4 g·kg^-1，they all exhibited distinctly repellent effects and the repellency rates were 96.49%，84.26％ and 90.61%, respectively. The LC₅₀ values to S. zeamais were 14.053, 28.648 and 21.429 μL·L^-1 fumigated by eucalyptol, α-pinene and essential oil of V. negundo for 72 h, respectively. They all showed high fumigation effects, of which eucalyptol had the highest one. The essential oil of V. negundo, and its main components eucalyptol and α-pinene all affected the breath of S. zeamais. The three samples distinctly inhibited the activities of AChE, CAT and CarE of S. zeamais in vitro. All the results suggest that the action mechanisms of essential oil and main components of V. negundo show the characteristics of diversity.

The effects of four podophyllotoxin analogues on spontaneous action potentials of the ventral nerve cord of Periplaneta americana were observed by electrophysiology technique. The results showed that at the concertration of 0.1 mmol/L, the four compounds showed different effects on spontaneous action potentials. Dexoypodophyllotoxin exerted irreversible inhibitory effect. β-apopicropodophyllotoxin first exerted excitation and then showed inhibitory effects. Podophyllotoxin showed inhibitory effect, while 4′-demethylpodophyllotoxin did not. The results suggest that the effects of podophyllotoxin analogues on spontaneous action potentials are different according to their structural varieties.

In order to clarify the control effects of pymetrozine on imidacloprid-resistant population of Aphis gossypii, and bring forward proposals on its management strategies，we used the wingless adult aphids of the imidacloprid-resistant population (RF₂₇), the Xiajin population (XJ) and the susceptible population (SS) to assay the cross-resistance of the RF₂₇ to pymetrozine and other pesticides and the effects of pesticide application on its biological characteristics by using the dipping method, microspot method, biochemical method and systematic observation method, respectively. The results showed that the LD₅₀ of pymetrozine to RF₂₇ was 1.213×10^-5 μg/aphid, to XJ was 8.506×10^-5 μg/aphid, and to SS was 5.140×10^-5 μg/aphid, so RF₂₇ exhibited obviously negative cross-resistance to pymetrozine. The resistance ratio of RF₂₇ to acetamiprid, nitenpyram and buprofezin was 2.35, 2.98 and 1.71, respectively. Activities of carboxylesterases (CarE) and gluathione-S-transferases (GSTs) in RF₂₇ were significantly higher than those in SS, suggesting that the enhancement of activities of CarE and GSTs would be the main metabolic mechanism of the occurrence of resistance. Pymetrozine had an obvious inhibition on the CarE and GSTs activities in RF₂₇, XJ and SS. RF₂₇ treated by pymetrozine showed developmental disadvantages, including the lower nymphal survival rate (only 64.60%), the lower net reproductive rate (10.39) , the longer mean generation time (10.87 d), the lower intrinsic rate of natural increase and the finite rate of increase (0.21 and 1.23 respectively), and the lower relative fitness (only 0.70). These results suggest that pymetrozine has high application value in controlling A. gossypii.

To investigate the geographic adaptation of the grasshopper, Oxya chinensis, populations of this grasshopper were collected in 5 localities ranging from 42.3°N to 20.0°N in China, and incidence of embryonic diapause, diapause intensity and adult body size were compared among the populations using their offsprings. The results indicated that different populations of the grasshopper displayed different patterns of response to photoperiod and temperature. The incidence of diapause was not influenced either by photoperiod or temperature in Tieling, Jining and Sihong populations, and their diapause rate was 100% under any conditions. Partial eggs entered diapause in the southern populations, i.e. Changsha and Haikou populations. A higher proportion of diapause eggs were produced at a short photoperiod than at a long photoperiod and the incubation temperature greatly influenced the induction of diapause in Haikou population, whereas the incidence of diapause in Changsha populations was only regulated by the incubation temperature of eggs. The egg diapause intensity was lowest in the Changsha and Haikou populations, followed by the Tieling population, and highest in the intermediate latitude populations, i.e. Jining and Sihong populations. In addition, diapause intensity increased as the original latitude decreased within the northern univoltine area. Adult body size showed a complicated pattern of variation along the latitudinal gradient. Significantly negative correlations between body size and latitude occurred within the northern populations; the body size of Changsha and Haikou populations, however, was smaller than that of Jining and Sihong populations. These results suggest that O. chinensis geographic populations have developed localityspecific adaptations, and the latitudinal gradient appears to play an important role in shaping their life-cycle and development.

In this study, the phylogenetic relationships of 12 families of the Chinese butterflies were investigated based on mitochondrial ND1 and 16S rRNA gene sequences. Thirty-two Chinese species were included in the analyses, of which 30 species were sequenced for the ND1 and 9 for the 16S rRNA gene. In the alignment, the two combined genes were 869 bp in length, including 373 conserved, 496 variable and 375 parsimonious-informative sites, respectively. The average percentage of A+T is 80.2%, which is much higher than that of C+G (19.8%). With both Zygaena angelicae and Z. transalpina as the outgroup taxa, the phylogenetic trees were reconstructed with the neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian inference (BI) methods. Major results included that: 1) Nymphalidae is not monophyletic; 2) the acraeids, danaids and libytheids are included in Nymphalidae; 3) the Pieridae and Papilionidae have a closest common ancestor; 4) the Parnassiidae should be included in Papilionidae; and 5) the satyrids should be separated from the Nymphalidae and be given a family rank. In addition, our results also suggest that the Riodinidae should be classified into Lycaenidae, and the acraeids should be given a subfamily rank. As to the amathusiidae, its taxonomic level and phylogentic position need further more detailed studies.

Sarcosaphagous flies have very important applications in death time estimation and crime scene reconstruction for homicide cases. DNA barcoding technology can extricate the investigators from the difficulty of culturing eggs and larvae of these flies as well as the reliance on special knowledge to scientifically identify the following imagoes, which is of great help for rapid identification of fly samples collected at the crime scene. We collected 77 samples, which belong to 7 dominant species of sarcosaphagous flies in Beijing, from different districts, and sequenced a 1 120 bp region of the mitochondrial cytochrome oxidase subunit I (COI) gene for each individual. Phylogenetic analysis showed that individual sequences of each species highly clustered together with 99% bootstrap support. Sequence divergences within species were less than 1%, while net divergences between species were all greater than 7.74%, the highest reaching 14.85%. Sliding window analysis demonstrated that variable sites for all species were relatively equally distributed along the entire region. By sequencing part of the COI gene, we established the DNA barcodes for the seven dominant species of sarcosaphagous flies in Beijing, which realized the accurate, rapid, and easy identification of these species and provided basic material for the future selection of variable sites that can be used for direct identification of sarcosaphagous flies.

The honeybee is a group of eusocial insects with high economical and ecological value. The Honeybee Genome Project has provided a molecular basis for further researches on honeybee biological characteristics, molecular mechanism and evolution of social behavior, and thereby equipped the honeybee to be a model organism from an economic insect. We introduced the advantages of the honeybee as a model organism and its model value in some hot research fields based on its biological characteristics, such as learning and memory, labor division, caste differentiation, and immunity, etc. After summarizing the current status and prospecting the future development of biological research on honeybee at home and abroad, we appealed for interdisciplinary research on honeybees at home.

Carboxypeptidase A (CPA) plays an important role in the digestion of dietary proteins in insects. LstiCPA, a full-length cDNA clone encoding carboxylesterase was obtained by screening cDNA expression library of the midgut of Loxostege sticticalis L. using the polyclonal antiserum against Spodoptera exigua (Hübner) invertebrate intestinal mucin (IIM). LstiCPA is 1 380 bp in length (GenBank accession no. EU924506), and the open reading frame (ORF) encodes 434 amino acids, with the predicted MW 49.1 kDa and pI 9.56. LstiCPA possesses a trypsin cleavage site and a catalytic zinc site characterized by CPA. LstiCPA was recombined into pET30 vector. The Western blotting analysis demonstrated that LstiCPA protein was expressed at a low level after IPTG induction. The enzymatic activity of LstiCPA was 0.0005 U/mL with the FAPP as substrate.