Helicoverpa armigera (Hübner) saliva play important roles in insecthost plant interactions. In this study we constructed a cDNA library of salivary glands of H. armigera larvae where saliva is secreted. We randomly sequenced 1 501 expressed sequence tags (ESTs), and clustering resulted in a total of 821 unigenes. Blast2 GO program was used to do BLASTx, functional annotation and metabolism analysis. Finally, we classified mRNAs in salivary glands of H. armigera larvae. By annotation of these ESTs, genes encoding 17 enzymes for digestion of fat, 5 enzymes for digestion of carbohydrates, and 20 serine proteases (of which 16 are newly reported) were identified, suggesting that the function of salivary glands is secreting saliva for predigestion. The cuticle protein, odorantbinding protein and chemosensory protein genes were identified in salivary glands of H. armigera larvae for the first time. The results will lay a foundation for studying predigestion system in H. armigera.

The western flower thrips (WFT), Frankliniella occidentalis Pergande (Thysanoptera: Thripidae), is an important agricultural pest worldwide. In recent years, it has spread into many provinces since its first detection in China. The research of the population genetic structure based on microsatellite marker (simple sequence repeats, SSRs) will contribute to revealing its invasion pathway. In this study, we analyzed the characteristics of SSRs from expressed sequence tags (ESTs) in F. occidentalis, screened PCR primers for EST-SSRs and tested the diversity of EST-SSR primers with the capillary electrophoresis. The results showed that 2 623 EST-SSRs were distributed in 1 930 uni-EST sequences, with an average of 1 SSR in every 2.21 kb of uni-EST sequence. Among mono- to hexa-nucleotide repeat types, mononucleotide repeats are the dominant type (83.00%), and tetranucleotide repeats are the second domainant type (11.17%). Furthermore, 4 of 22 pairs EST-SSR primers designed produced discernable PCR products. The capillary electrophoresis revealed that 3 of 4 pairs of EST-SSR primers are polymorphic. The average polymorphism information content (PIC) with the 3 polymorphic EST-SSR primers (0.48-0.69) is lower than that with 5 polymorphic Genomic-SSR primers (0.88-0.92). This study may contribute to further research on the analysis of genetic structure of F. occidentalis populations in future.

Some serine proteases, such as trypsin and chymotrypsin, are important digestive enzymes in the digestive system of mirid bugs. In order to better understand the role of serine proteases in the digestive system of the green plant bug, Apolygus lucorum (Meyer-Dür), we cloned the cDNA encoding serine protease of A. lucorum for the first time in the laboratory, which was named as AlSP4 (GenBank accession no. JQ609682). The results of sequence analysis showed that the open reading frame (ORF) of AlSP4 is 999 bp in length, encoding a 332-amino-acid peptide, with the predicted molecular weight (MW) of 36.84 kDa and the theoretical isoelectric point (pI) of 5.35, and the predicted N-terminal hydrophobic region containing 16 amino acid residues displays the typical feature of a signal peptide. Protein signature analysis revealed that the protein encoded by AlSP4 shares typical structural features of serine proteases with other insects, including His, Asp, and Ser residues for the catalytic amino acid triad of active sites of serine proteases. Putative trypsin precursors from the encoded protein of AlSP4 cDNA contain a signal peptide, activation peptide, and conserved N-termini (IVGG). By the Real-time PCR technique, we determined the expression pattern of AlSP4 in A. lucorum fed on different hosts. The expression level of AlSP4 was the highest in female adult A. lucorum fed on Bt cottons, significantly higher than that fed on conventional cotton (P＜0.01). The expression of AlSP4 increased significantly in male adult A. lucorum fed on Bt cottons, its expression level was only lower than that fed on garland chrysanthemum and significantly higher than that fed on conventional cottons (P＜0.01). Therefore, AlSP4 is the important digestive enzyme gene for adult A. lucorum to feed on Bt cottons, and plays an important role in adaption of A. lucorum to survive on Bt cottons.

Trehalose, which is the blood sugar in insects, is synthesized mainly by trehalose-6-phosphate synthase (TPS) in insects. By homologous cloning and rapid-amplification of cDNA ends (RACE), the full cDNA of the TPS gene in Harmonia axyridis was cloned and named HaTPS (GenBank accession number: FJ501960). This gene with 5′ and 3′ non-coding region of 26 bp and 505 bp, respectively, contains an open reading frame of 2 418 nucleotides encoding a protein of 805 amino acids with the predicted molecular weight of 90.58 kDa and pI of 7.01. The encoded protein contains two potential N-glycosylation sites, without signal peptide and transmembrane domain. The homology comparison showed that insect TPS was highly conserved with two conserved domains. Real-time fluorescent quantitative PCR was conducted at different developmental stages under the cold induction to detect the expression of HaTPS. The results showed the expression level of HaTPS was the highest in the pre-pupal stage. Under the condition of shortterm cold induction, the HaTPS expression was dramatically increased with temperature decreasing; and under the condition of warming and cooling, its expression level increased first, and then declined. The results suggest that TPS plays an important role in the regulation of insect resilience and its regulatory capacity is increased significantly under cold-induction.

It is one of important strategies for many insects to produce antifreeze proteins (AFPs) to resist low temperature. In order to study the mechanism of cold resistance, an antifreeze protein gene Tm-afp was cloned from the larvae of yellow- and black-color varieties of Tenebrio molitor, the nucleotide sequence and the encoded amino acid sequence characteristics of Tm-afp were analyzed, and the mRNA levels were examined in the two varieties by reverse transcriptional PCR and rapid amplification of 5′ and 3′ cDNA ends. The results showed that the full cDNA sequences of Tm-afp cloned from the yellow- and black-color varieties of T. molitor were 579 bp and 588 bp in length, respectively. Both contain a 20 bp 5′ non-coding region, a 402 bp open reading frame and a variant 3′ non-coding region, and their nucleotide sequence identity is 95%. Because of two amino acid variation (D35→E35 and T130→S130) existing between the two mature peptides encoded by the two cDNA sequences, they were considered two isoforms of Tm-afp , i.e.,Tm-afp -1 and Tm-afp -2, respectively. Three amino acid variation (G2→A2, S9→T9 and Y27→N27) exists between signal peptides of the antifreeze protein isoforms (Tm-afp -1 and Tm-afp -2) encoded by Tm-afp-1 and Tm-afp -2. The 1st amino acid of the mature peptides of the two antifreeze protein isoforms is glutamine. Both the mature peptide sequences contain eight short-tandem repeats, each composed of twelve highly conservative amino acid residues. Furthermore, both the 2nd and 8th amino acids of each repeat are cysteines. In addition, Tm-afp -1 and Tm-afp -2 both belong to threonine- and cysteine-rich insect antifreeze proteins. Their secondary structure consists of beta sheets and random coils, and their tertiary structure has eight special right-handed beta helixes, composed of twelve amino acid residues per helix. Beta sheet, composed of conservative XCT, arrays on the lateral surface of the beta helixes. Meanwhile, low temperature up-regulated the expression of Tm-afp in yellow- and black-color varieties of T. molitor. But under long time of cold induction, the relative mRNA expression level of Tm-afp in the black-color variety of T. molitor was significantly higher than that in the yellow-color variety. The results suggest that Tm-afp among different color varieties of T. molitor may have several isoforms and Tm-afp in the black-color variety of T. molitor is more sensitive to cold than that in the yellow-color variety. This study may provide a useful reference to the further research on the molecular mechanisms of cold resistance in T. molitor.

In order to clarify the physiological function of chemosensory proteins (CSPs) in the olfactory system of Grapholita molesta, the full-length cDNA encoding a chemosensory protein was isolated from G. molesta by using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends-PCR (RACE-PCR), and named as GmolCSP (GenBank accession no. JQ821389). The results of sequence analysis indicated that the open reading frame (ORF) of GmolCSP is 384 bp in length, encoding 127 amino acid residues, with the deduced molecular weight (MW) of 12.80 kD and isoelectric point (pI) of 8.33. The deduced amino acid sequence of GmolCSP showed a high identity to CPSs of other lepidopteran insects. RT-PCR analysis revealed that GmolCSP was expressed in antennae, head, thorax, abdomen, wings and legs of the test moth. GmolCSP was then constructed into the expression vector pET-32a and expressed in Escherichia coli BL21 (DE3) after induction with IPTG.-SDS-PAGE and Western Blot analysis confirmed the molecular weight of the recombinant GmolCSP is 29 kD, consistent with the predicted result. The results in this study are helpful for further research on molecular structure and function of GmolCSP.

The suboesophageal ganglion (SOG) receives sensory projections from the nerves of the mouthparts and gives rise to motorneurons that supply muscles of the mouthparts. The anatomical organization and postembryonic development of the SOG in the Chinese honeybee, Apis cerana cerana, was comparatively studied by using anatomy and immunohistochemical method (5-bromo-2-deoxyuridine, BrdU incorporation). The results showed that the SOG of the honeybee is composed of mandibular, maxillary and the labial neuromeres. During postembryonic development of the SOG, extensive proliferation in the SOG could be detected only within a narrow time window from prepupa to day 1 of pupal development during metamorphosis. Proliferative nuclei disappeared during day 4 of pupal development. In the SOG of A. cerana cerana, glial cells can be classified into three categories based on the position of their cell bodies, and cell morphology: surface associated glia, cortex associated glia and neuropil associated glia. This study provides an essential foundation for studying development and function of nervous system of honeybees.

Wolbachia and Cardinium, with the ability to induce cytoplasmic incompatibility (CI), are maternally inherited intracellular bacteria known to manipulate the reproduction of their hosts. The exact mechanisms of CI which is induced by these two endosymbionts in the same host are unknown. This study tried to investigate the reproductive manipulation of Wolbachia or/and Cardinium infected spider mite Tetranychus piercei McGregor by crossing experiment and fluorescence in situ hybridization (FISH). The results indicated that Wolbachia-infected males induced weak CI. In Guangzhou population of the spider mite, approximate 17.8%±1.6% of all eggs did not hatch in the incompatible cross U/Iw. Cardinium-infected and Wolbachia and Cardinium doubly infected males caused severe CI. The unhatched eggs in the incompatible cross U/Ic and U/Iwc accounted for 70.3%±1.3% and 72.9%±1.2%, respectively, the number of eggs laid per doubly infected female was 35.2±1.2, significantly higher than that of singly (Wolbachia or Cardinium) infected female. There is a positive correlation between CI levels and infection status of the developing sperm. Wolbachia and Cardinium mainly enter the oocytes with trophic flow from nurse cells, midgut and oviduct. The results might provide foundation for understanding the maternally inherited mechanisms of Wolbachia and Cardinium.

In order to provide the theoretical basis for the management of the diamondback moth, Plutella xylostella (L.) resistant to abamectin, the sublethal effects of metaflumizone on the experimental populations of both abamectin-resistant (AV-R) and susceptible (AV-S) strains of P. xylostella were evaluated by means of the life table technology. The results showed that the estimated LC₅₀ and LC₂₅ values of leaf dip bioassay of metaflumizone on the 3rd instar larvae were 0.24 and 0.09 mg/L for the AV-S strain, and 0.20 and 0.07 mg/L for the AV-R strain, respectively. Metaflumizone at the sublethal concentration (0.09 mg/L) tested decreased the pupation rate and pupal weight, prolonged the pupal duration and decreased the emergence rate, fecundity, oviposition duration and longevity of adults in the treated generation. Such treatment also decreased the hatchability and larval survival rate of the offsprings and their larval duration. Reproduction parameters such as the intrinsic rate of increase (r_m), finite rate of increase (λ), and the net reproductive rate (R₀) in populations treated with sublethal concentrations of metaflumizone were significantly lower than those of the control populations (P<0.0001). The results also showed that the sublethal concentration of metaflumizone had a greater effect on the AV-R strain than on AV-S strain, and had a greater effect on the current generation than on the offsprings. We conclude that the sublethal concentration of metaflumizone might have significant effects on the population dynamics of P. xylostella, especially the AV-R strain, and metaflumizone might be extremely useful for managing pest resistance to insecticides.

To evaluate the resistance risk of Panonychus citri to amitraz and clarify the relationship between expression level of cytochrome P450 gene and resistance of P. citri, resistance selection was conducted in the laboratory, and cross-resistance of the amitraz-resistant strain was studied with slide-dip method and cytochrome P450 gene expression profiles between the resistant strain and the susceptible strain were compared using RPKM (reads per kb per million reads) method. After selection with amitraz continuously for 12 generations, a resistant strain was obtained with the resistance ratio (R/S) of 26.32 compared with the susceptible strain. The realized heritability of resistance to amitraz was 0.148. Bioassay data showed that the amitraz-resistant strain had positive cross-resistance to spirodiclofen, diafenthiuron, propargite and azocyclotin (the R/S ratio was 16.85, 4.98, 2.13, and 2.05, respectively), but had no crossresistance to abamectin, fenbutatin oxide, pyridaben and petroleum oil (the R/S ratio was 1.10, 1.21, 0.67 and 0.99, respectively). Gene expression difference analysis results indicated that 16 cytochrome P450 genes were up-regulated and 27 cytochrome P450 genes were down-regulated in the resistant strain. CYP389A6 ［log₂ratio(RS/SS)=11.526］ and CYP389A2 ［log2 ratio(RS/SS) =-12.683］ were the top up-regulated and down-regulated genes, respectively, which are very likely to be associated with the resistance of P. citri to amitraz.

In order to reveal the development of the carpenter moth Holcocerus vicarius (Walker) larva and forecast its occurrence time, the head capsule width, body length, body width, pronotum width, mandible length and mandible width were measured, and Crosby law and linear regression analysis were used to choose the optimal morphological index for determining the larval instars of H. vicarius. The results showed that the head capsule width provided the best index to divide instars, since its mean coefficient of variation and Crosby ratio were the least, while those of other indexes were greater. H. vicarius was determined to have twenty instars. Different instars conformed to the growth regularity of larval head capsule width proposed in Dyar’s law. The linear regression equation of the head capsule width against the instar number was: y=0.233+1.686x+0.127x²-0.005x³ (R²=0.996). Determination of the larval instar number of this moth could provide the basis for studying its occurrence and biological characteristics, and facilitate to develop comprehensive prevention and control.

Five genera and 13 species are reported here from China that are considered members of the Anaphothrips genus-group (Thripidae: Thripinae), including two new species of this group, namely Anaphothrips beijingensis sp. n. and Chilothrips jiuxiensis sp. n., are described and illustrated. A key is provided for identifying these 5 genera with comments on each genus and its species. The yellow morph of the male (adult) of Anaphothrips sudanensis Trybom is newly recorded from China.

【Aim】 In the case of in-situ recognition of pest larva, larvae often appear in bending posture, which makes the extracted feature vectors distorted and affects the results of recognition. To solve this problem, this paper proposes an eigenvector extraction method of posture invariant based on sector-shaped transform. The extracted eigenvectors of pest larvae have the properties of translational, proportional, rotational and posture invariance, which facilitates the automatic identification of fat and short larvae in bending postures. 【Methods】 Firstly, the bending region and non-bending region of the pest larvae are determined by optimal consistent approximation after image thinning. Then, the bending region is straightened through sector-shaped transform, and rotation and translation operations are performed on the non-bending region which is combined with the straightened bending region to form a complete larva. The eight-neighborhood mean method is used to fill up the pixels of gaps in the combined larva, and the automatic correction of bending larva is finished. The invariant Hu’s moment extracted from the image has the property of posture invariance. The minimum distance classifier was used to realize automatic identification of larvae with multiple postures. Finally, experiments were carried out to verify the efficacy of the method by using larvae of Prodenia litura, Heliocoverpa armigera, Spodoptera exigua and Ostrinia nubilalis moths as the target. 【Results】 The identification of 24 kinds of bending larvae of the moths in different postures was carried out on condition that the recognition threshold is 80%. The identification rates based on the classic Hu’s moment and the invariant Hu’s moment were 25% and 100%, respectively. 【Conclusion】 The experimental results demonstrate that the method can effectively identify the bending postures of fat and short larvae.

Phloem feeding involves special biological interactions between the herbivores and their host plants. This article reviews the types of plant defense responses, the change of plant defensive substances, signaling pathways and transcriptome studies of plant responses induced by phloem-feeding insects. The mechanisms of plant defense responses induced by phloem-feeding insects mainly include: changing plant nutritional status, producing toxic metabolites and increasing defensive proteins. Furthermore, plant defense responses are closely related to salicylic acid, jasmonic acid and ethylene signaling pathways. Plant defense responses induced by phloem-feeding insects mainly activate the salicylic acid signaling pathway, while the molecular mechanisms are not well understood. The growing genomics resources and the development of molecular biology techniques will reveal the signaling networks, the specific factors that dictate innate immunity and gene-for-gene-mediated defense to phloem-feeding insects. Understanding the plant responses induced by phloemfeeding insects will provide a better understanding of plantinsect interactions and new insights into pest management and the cultivation of pest-resistant plants.

Research on the visual processing pathways is of great significance in neuroscience, bionic applications and medical treatments. The western honeybee, Apis mellifera, as an important model organism for neurobiology research, has been extensively applied to the research on visual processing pathways. The visual organs of bees include a pair of compound eyes and three ocelli, in which the compound eyes are most important for vision formation. In bees, processing of visual information is achieved within the optic lobes which are divided into four levels of neuropils, i.e., lamina, medulla, lobula and anterior optic tubercle. Complex visual information is segregated at successive stages within the brain, and processed in many parallel and sequential visual pathways. Then they are converged in higher-order brain centers, with some integrated with information of other modalities, which finally generate effective outputs to regulate and control bee behaviours. In this article, the research on visual processing pathways in bee compound eyes is reviewed according to the sequence of information processing in the optic lobes.

【Aim】 Experiments were conducted to evaluate the effects of ultraviolet treatment of Grapholita molesta (Busck) eggs on parasitization and emergence of Trichogramma pintoi Voegele. The optimal ultraviolet intensity and irradiation time to treat G. molesta eggs were determined in order to provide the treatment and storage means of host eggs in mass rearing of T. pintoi. 【Methods】 T. pintoi adults within 12 h after emergence were supplied to parasitize G. molesta eggs which were irradiated by various ultraviolet intensities for different time, the parasitization performance was observed, and the parasitization rate and emergence rate were calculated and compared with that of T. pintoi parasitized G. molesta eggs not irradiated by ultraviolet. 【Results】 The results showed that the parasitization rate of T. pintoi significantly decreased when T. pintoi parasitized the G. molesta eggs which were irradiated by ultraviolet. With the enhancement of ultraviolet intensity and irradiation time, the parasitization rate significantly decreased, while the emergence rate had little change. When the G. molesta eggs were irradiated for 1-2 h by 15 W ultraviolet light or irradiated for 1 h by 30 W ultraviolet light, the emergence rate increased over 80% when compared to the control. But when G. molesta eggs were irradiated by ultraviolet for 3 h, the emergence rate decreased obviously.【Conclusion】 Parasitism and development of T. pintoi could be affected by ultraviolet intensity and treatment time. Therefore, irradiation with ultraviolet light of 15 W for 1 h is suggested in mass rearing of Trichogramma by small eggs in the laboratory.