【Aim】 Epacromius coerulipes (Ivanov) is one of the most widely distributed grassland locusts. However, there is a lack of genomic resources. In order to obtain genetic data of E. coerulipes, this study conducted the transcriptome sequencing and analysis. 【Methods】 De novo assembly of the transcriptome was conducted using Illumina paired-end sequencing technology. 【Results】 In total, 63 033 unigenes with a mean length of 772 bp and an N50 length of 1 589 bp were obtained. Of these unigenes, 25 132(39.87%) unigenes were matched with the known proteins in the National Center for Biotechnology Information (NCBI) database, as determined by BLAST search, with 24 841, 16 490, 11 558 and 8 013 unigenes assigned to Nr database, Swiss-Prot, Gene Ontology (GO) and Clusters of Orthologous Groups (COG), respectively. In KEGG database, a total of 7 218 unigenes were assigned to 218 known pathways. Among these, 189 unigenes were involved in metabolic pathways of xenobiotics or drugs. In-depth analysis of the data showed that 213 unigenes were identified as potentially involved in the detoxification of xenobiotics, and 29 unigenes were identified as encoding insecticide target proteins. In addition, 5 696 simple sequence repeats (SSRs) were detected. 【Conclusion】 This study lays the foundation for further research on gene function analysis and molecular resistance mechanisms to insecticides in E. coerulipes.

【Aim】 This study aims to establish and characterize cell lines developed from the neonate larvae of Papilio demoleus Linnaeus. 【Methods】 Modified Grace’s medium supplemented with 20% fetal bovine serum was used to culture tissues from neonate larvae of P. demoleus. The methods of cell morphology analysis, growth analysis, chromosome analysis, and molecular identification of the cell lines were used to obtain the biological characteristics of new cell lines. In order to study the viral susceptibility of four cell lines derived from P. demoleus, recombinant baculovirus carrying secreted alkaline phosphatase (AcMNPVSEAP) was constructed using Bac-to-Bac baculovirus expression system to infect the cell lines. The median tissue culture infectious dose (TCID₅₀) of each cell line was measured using limiting dilution method by measuring the expression of recombinant SEAP. 【Results】 Four new cell lines developed from the neonate larvae of P. demoleus were established on modified Grace’s medium and named RIRI-PaDe-1, RIRI-PaDe-2, RIRI-PaDe-3 and RIRI-PaDe-4, respectively. These cells were subcultured for approximately 60 passages. There were significant differences in biological characteristics between different cell lines. According to morphology, RIRI-PaDe-1 and RIRI-PaDe-4 were composed of round, spindle and polygonal cells, RIRI-PaDe-2 was mainly composed of round cells, and RIRI-PaDe-4 was mainly composed of epidermal-like and fibrous cells. The cell population doubling time (PDT) of IRI-PaDe-1, RIRI-PaDe-2, RIRI-PaDe-3 and RIRI-PaDe-4 was 69.77, 67.42, 81.48 and 65.43 h, respectively. Karyotype analysis showed that the number of chromosome of each cell line fitted the normal distribution. The chromosome numbers of RIRI-PaDe-2, RIRI-PaDe-3 and RIRI-PaDe-4 varied widely from 45 to 97, while that of RIRI-PaDe-1 varied from 36 to 90. All of the four cell lines could be infected with the AcMNPV-SEAP, but RIRI-PaDe-3 was more susceptible to AcMNPV-SEAP. 【Conclusion】 Although the four new cell lines developed from the neonate larvae of P. demoleus have the same genetic background analyzed by DNA amplification fingerprinting (DAF) using mitochondrial cytochrome c oxidase subunit I (COI) gene, there are still significant differences in the basic biological characteristics among them. It is worthy to be further researched.

【Aim】 The study aims to further explore the mechanism of pathogenicity of entomapathogenic nematodes Heterorhabditis bacteriophora strain Cangzhou infecting white grubs. 【Methods】 Transmission electron microscopy (TEM) was used to observe the morphological changes in fat body and midgut cells of the 2nd instar larvae of Hololtrichia oblita (Faldermann) and H. parallela Motschulsky, respectively. After hemocoel injection, ultrastructural changes of the white grubs were examined by TEM. 【Results】 Changes in the ultrastructure of fat body and midgut cells of the 2nd instar larvae of H. oblita and H. parallela at 24 and 48 h after injection of H. bacteriophora were observed. At 24 h after infection, fat droplets, located at fat body cells, became smaller, and their electron density became lighter in the infected larvae than those in the untreated control. Endoplasmic reticulum and mitochondria in both fat body and midgut cells became distended and mcrovilli of the midgut began to fall off in the infected larvae. At 48 h after infection, the membrane structure containing a lot of fat droplets was found broken in the infected larvae. Chromatin in fat body and midgut cell nucleus dissociated and mitochondria as well as endoplasmic reticulum were disintegrated completely in both cells of the infected larvae. Likewise, microvilli of the midgut were almost completely lost in the infected larvae. 【Conclusion】 After infection by the entomopathogenic nematode H. bacteriophora, the fat body and midgut of the 2nd instar larvae of the two white grub species, H. oblita and H. parallela, show similar pathological changes, which may be associated with the rapid death of white grubs. This study may enrich the theoretical basis for using entomopathogenic nematodes as a biological control strategy in IPM of white grubs.

【Aim】 Silkworm pebrine disease has been affecting the healthy development of the silkworm eggs’ industry. It is of great significance to effectively control the harm of pebrine disease via rapidly and accurately detecting the silkworm eggs parasitized by pebrine. 【Methods】 Loop-mediated isothermal amplification (LAMP) is a method that amplifies the template DNA with rapidity, sensitivity and high specificity under constant temperature in vitro. Based on the LAMP protocols, we selected Nosema bombycis EB1 gene (GenBank accession no. KF421134.1), a gene which is related to the replication of spores, to design the sets of LAMP primers. We investigated the optimal reaction conditions of the LAMP system, including the reaction temperature and the concentration ratio of the internal primer to outer primer, and detected the specificity and sensitivity of the LAMP reaction system. We also set up the LAMP assay for detecting the pebrine disease in silkworm eggs. 【Results】 LAMP assay based on EB1 could effectively detect the silkworm eggs infected with N. bombycis.The detection could be completed within 1.5 h at the constant temperature of 63℃. It was able to detect as low as 5.0×10^-3 ng/μL of the template DNA extracted from N. bombycis spores, and 100 copies of the constructed standard plasmid EB1-pMD^TM 19-T. Furthermore, it could also effectively detect 1/8 of the eggs batch from one infected silkworm moth by pebrine or one infected silkworm egg. The results were simultaneously demonstrated by running the gel electrophoresis, and by observing the fluorescence indicator, and also by visual observation with SYBR Green I, respectively. 【Conclusion】 The LAMP assay based on EB1 gene of N. bombycis for detecting the silkworm eggs infected by pebrine has been successfully established. It is further proved that the LAMP method could be used for the quality inspection in production of silkworm eggs or for quarantine inspection of the commercial eggs on sites.

【Aim】 The objective of this research is to determine the effects of total alkaloids from Tripterygium wilfordii Hook. f. on insect nervous system and to provide a basis for elucidating its insecticidal mechanisms. 【Methods】 The 5th instar larvae of Mythimna separata (Walker) were fed with maize leaves treated with the total alkaloids from T. wilfordii. The activities of acetylcholine esterase (AChE), Na⁺, K⁺-ATPase, Ca²⁺, Mg²⁺ - ATPase, glutamic-pyruvic transaminase (GPT) and glutamic decarboxylase (GAD) in the 5th instar larvae of M. separata treated were determined at three different poisoning stages, and the contents of neurotransmitters including acetylcholine (ACh), γ-aminobutyric acid (GABA) and glutamic acid (Glu) were also assayed. 【Results】 The AChE activity in the 5th instar larvae of M. separata was not affected by the total alkaloids from T. wilfordii. The ACh contents in treated larvae in the weak paralysis stage and the deep paralysis stage were not significantly different from those of the corresponding two controls. The GABA contents in treated larvae increased by 89.86%, 49.28% and 20.29%, while the Glu contents increased by 24.55%, 23.33% and 8.13% in the weak paralysis stage, the deep paralysis stage and the recovery stage, respectively. The GPT activity in treated larvae was remarkably inhibited by the total alkaloids, but the GAD activity was not affected. The activities of Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase in the brain of treated larvae were inhibited, but those in the midgut did not change. 【Conclusion】 The results help us understand the effects of the alkaloids from T. wilfordii on insect nervous system, and provide further scientific basis for revealing their action target.

【Aim】 This study aims to clarify the difference in basic biological characteristics and population parameters like relative fitness and intrinsic rate of increase, between different trichlorfon-resistant strains and susceptible strain of the oriental fruit fly, Bactrocera dorsalis (Hendel). 【Methods】 The high- and medium-level trichlorfon-resistant strains of B. dorsalis were selected in the laboratory. Biological characteristics and parameters were investigated and analyzed between the resistant strains and the susceptible strain. 【Results】 There were no significant differences in the durations of egg, larva and pupa, pupation rate, pre-oviposition period, longevity of male and female adults, average longevity and female rate of B. dorsalis. Egg hatching rate (68.33%) and adult emergence rate (93.73%) of the medium-level trichlorfon resistant strain (Tri-M) was significantly lower than those of the susceptible strain (SS), which were 88.33% and 97.93%, respectively, and those of high-level trichlorfon resistant strain (Tri-H), which were 86.67% and 98.21%, respectively. Compared with the fecundity of SS (864.61 eggs laid per female) and Tri-H (750.70 eggs laid per female), the fecundity of Tri-M decreased to 630.87 eggs laid per female. All strains had coincidently daily fecundity dynamics of females, indicating that the oviposition peak periods come in a short time after sexual maturity of females. The oviposition peaks of the resistant strains were observed earlier than that of the susceptible strain. After the oviposition peak, fecundity decreased in a fluctuation. All the c values of equations to simulate adult survival rate dynamics of different strains were higher than 1, demonstrating that the equations fit to the I-type survival model. Adults of all strains could survive until the average longevity was reached. SS had the highest index of population trend (I =339.41), while Tri-H had a moderate (I =307.82) and Tri-M had the lowest index of population trend (I =175.79), indicating that the ascending of trichlorfon resistance of B. dorsalis populations was suppressed, and for Tri-M the suppression was stronger. The net reproductive rate (R₀) of Tri-M (217.49) was significantly lower than those of SS (327.89) and Tri-H (284.29), respectively. The intrinsic rate of increase (r_m) and finite rate of increase (λ) of the two resistant strains had no significant difference, but were much higher than those of SS, and the average generation period (T ) of the two resistant strains was shorter than that of SS. The relative fitness of the high-and medium-level resistant strains was 0.8670 and 0.6633, respectively, significantly lower than that of SS. 【Conclusion】 Under selection pressure of trichlorfon, there is significant decrease in the average generation period, egg hatching rate and pupation rate of Tri-M, while most of biological characteristics of the resistant strains of B. dorsalis have no significant change. Reproduction and development of populations of the trichlorfon-resistant strains are disadvantaged, while Tri-M is much more disadvantaged. Nevertheless, the resistant strains show better population growth potentials.

【Aim】 The whitefly Bemisia tabaci is the only vector of Tomato yellow leaf curl virus (TYLCV) in the field. In addition to direct acquisition of the virus through feeding, the whitefly can acquire the virus via mating. Although previous studies have shown that both cryptic species Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) of the whitefly complex can achieve horizontal transmission of TYLCV via mating, the results on the efficiency of virus transmission via this means by the two species were not consistent. Moreover, experimental evidence is yet lacking to show whether this type of horizontal transmission of TYLCV may facilitate the spread of the virus in the field. 【Methods】 We first collected the field populations of cryptic species MEAM1 and MED of B. tabaci from four provinces in China, including Zhejiang, Guangdong, Yunnan and Henan, and following molecular identification ofthe species status of each of the populations we established seven pure cultures for experiments. Then, we observed the capacity of TYLCV transmission between viruliferous and non-viruliferous adults in each of the populations. And finally, we used the MEAM1 culture from Guangdong Province to simulate a scenario of group mating and examined the capacity of the adults, which acquired TYLCV via mating, in transmitting the virus to tomato plants. 【Results】 Both MEAM1 and MED populations of B. tabaci collected from the four provinces were able to achieve horizontal transmission of TYLCV via mating, although the efficiency of transmission was in general low, mostly below 10%. No significant differences were detected in the efficiency of transmission between the MED populations from the four provinces, between MEAM1 populations from three of the four provinces, and between the two cryptic species within each of the provinces. In each of the two cryptic species, both viruliferous male and female adults were able to transmit viral DNA to individuals of the opposite sex, and no significant differences in the efficiency of virus transmission between the two sexes were found. The data of the group mating test indicated that the originally nonviruliferous individuals were unable to transmit TYLCV to tomato plants following their acquisition of the virus via mating. 【Conclusion】 These results demonstrate that for the virus TYLCV and whiteflies of the cryptic species MEAM1 and MED of B. tabaci tested here low efficiency of horizontal transmission of TYLCV can be expected. However, the data obtained with the Guangdong MEAM1 population suggest that the low horizontal transmission does not increase the number of individuals capable of transmitting the virus to infect new tomato plants. Consequently, the horizontal mode of transmission is unlikely to have much epidemiological relevance in the field.

【Aim】 Galeruca daurica (Joannis) is a new pest outbreaking seriously in the Inner Mongolian grasslands in recent years and overwinters as eggs under cow pats, stones and grass clusters. Understanding the cold hardiness of the overwintering eggs of G. daurica is helpful to forecast its distribution and population dynamics. 【Methods】 We measured the survival rates and developmental durations of overwintering eggs of G. daurica exposed to different low temperatures (-18--39℃) for 12 and 24 h and exposed to -30℃ for different periods (0-60 d), respectively, in the laboratory. 【Results】 Low temperature intensity and exposure time significantly affected the survival rates of overwintering eggs of G. daurica. The survival rates declined with the decrease of exposure temperature and the extension of exposure time. After the overwintering eggs were exposed to the temperatures ≤-33℃ for 12 h or the temperatures ≤-30℃ for 24 h, their survival rates were significantly lower than the corresponding controls (25℃). The lower half lethal temperatures (LT₅₀) after 12 and 24 h exposure were -33.08 and -32.13℃, respectively, and the half lethal time (Lt₅₀) at -30℃ was 33.33 d. The developmental duration of surviving eggs prolonged significantly after exposure to -36℃ for 12 h or the temperatures ≤-33℃ for 24 h, but had no significant change after exposure to -30℃ for 30 days. 【Conclusion】 The overwintering eggs of G. daurica have strong cold hardiness, and low temperature in winter may usually not cause mass mortality of overwintering eggs.

【Aim】 Agriphila aeneociliella is a new pest of wheat crops, which survives summer as diapausing larvae making cocoon in soil. This study aims to determine the survivorship of A. aeneociliella in sustained high temperature and extreme high temperature in summer, so to provide a basis for population monitoring and management in autumn. 【Methods】 The diapause and non-diapause larvae of A. aeneociliella were subjected to extreme high temperatures (43, 46, 49, 52 and 55℃ ) for 0.5, 1, 2 and 4 h, and to the common high temperature (35, 37, 39 and 41℃) for 1, 2, 4 and 8 d, respectively. 【Results】 Semi-lethal temperatures of diapause larvae with cocoon of A. aeneociliella were 50.04℃ (95% confidence interval: 49.36-50.47℃) and 48.81℃ (95% confidence interval: 47.74-49.37℃), and those with cocoon removed were 48.20℃ (95% confidence interval: 47.66-48.74℃) and 47.36℃(95% confidence interval: 47.08-48.37℃) when subjected to the extreme high temperature for 2 and 4 h, respectively. The median lethal time of diapause larvae with cocoon was 1.29 h (95% confidence interval: 1.17-1.44 h), and that with cocoon removed was 1.02 h (95% confidence interval: 0.93-1.18 h) when subjected to the extreme high temperature of 52℃. The median lethal time of diapause larvae with cocoon was 3.21 d (95% confidence interval: 2.92-3.52 d) and that without cocoon was 2.66 d (95% confidence interval: 2.33-3.02 d) when subjected to the common high temperature at 41℃. 【Conclusion】A. aeneociliella has a strong tolerance to high temperatures, and cocoon has a protective effect on diapausing larvae. Ploughing and stubble cleaning to make cocoons or larvae exposed to the soil surface after wheat harvesting, which takes advantage of high temperature of soil surface to kill diapause larvae, can be used as an efficient agricultural control measure.

【Aim】 This study aims to build up a pest and disease forecast model based on wavelet neural network (WNN), so as to provide a basis for taking measures to prevent pests and diseases, reducing crop damage by pests and diseases and improving quantity and quality of crop yields. 【Methods】 Based on the occurrence degree of wheat aphids from 1980 to 2014 and the meteorological factors in Ruicheng County, Yuncheng City, Shanxi Province, we integrated and created 9 new independent variable input models from 40 fundamental meteorological factors through Principal Component Analysis (PCA) and screened hidden layer nodes by trial and error method, conducted training with data from 1980 to 2009 and retested the occurrence degree of wheat aphids from 2010 to 2014. Finally, the study built up a WNN model by taking wavelet function as transfer function and contrasted itself with BP neural network (BPNN) model which takes Sigmoid function as transfer function. 【Results】 The average fitting accuracy of both models, namely, WNN and BPNN models, were above 80% in at least 10 years. Their MAPE values were 89.83% and 83.07%, and their MSE values were 0.0578 and 0.6192, respectively. 【Conclusion】 Both models can well illustrate the occurrence degree of wheat aphids. As for the forecast accuracy and model stability, however, WNN is better than BPNN.

【Aim】 Among many species of oil-tea camellia pests, the tea tussock moth, Euproctis pseudoconspersa, is one of the most dangerous pests. Aiming to complete the automatic detection of E. pseudoconspersa larvae, the images of the oil-tea camellia pests need to be segmented, and the segmentation effects influence directly the automatic identification of images. 【Methods】 In this paper we proposed an new algorithm based on K-means and region merging to make difference calculation for three components of RGB. The originally segmented images were gotten through combining adjacent pixels if the maximum difference is equal to 0. According to the merging rules, the pixels were further merged to obtain the final segmentation results. 【Results】 The proposed algorithm in this paper could separate image background from pests quickly and effectively. 【Conclusion】 Through comparsion of the image segmentation effects from JSEG argorithm, K-means segmentation argorithm, fast geometry deformable segmentation and the proposed algorithm, we found that the proposed algorithm is better than others because it could get satisfactory results within very short time.

Mandibular gland is an important honeybee exocrine gland. Its secretions are of great importance in maintaining the social organization of honeybee colony. Stearic acid is used as the synthetic precursor in biosynthesis pathways of both queen and worker mandibular gland secretion, but the caste differences in the following fatty acid β-oxidation process lead to the different proportions of components of mandibular gland secretions between castes. Queen mandibular gland secretion mainly consists of 9-oxo-2-decenoic acid (9-ODA) and possesses multiple functions including attracting workers and drones, inhibiting activation of worker ovaries, etc. Worker mandibular gland secretion is dominated by 10-hydroxy-2-decenoic acid (10-HDA) and 10-hydroxydecanoic acid (10-HDAA), and constitutes an important part of royal jelly. This caste specificity in the mandibular gland secretion composition is not only a rigid trait but also shows plasticity within the female phenotype and difference between species. The researches at the transcriptional and protein levels in recent years further revealed the molecular basis of this caste difference. Researches about mandibular glands are of great significance to the studies of such areas as honeybee biology, honeybee behavior and quality control of honeybee products. By summarizing these mandibular gland related researches, we aim to provide insights for further study in the functional mechanisms of mandibular gland secretion, the molecular mechanisms of biosynthesis pathway and other related areas．

【Aim】 The present study aims to clone Caspase family gene in Bombyx mori and to investigate its function preliminarily using RNAi. 【Methods】 A new gene of Caspase family was cloned by rapid amplification of cDNA ends (RACE), and then its functions in apoptosis were analyzed by RNAi and flow cytometry. 【Results】 The cloned Caspase family gene was denominated BmCaspase-X in B.mori, the full length is 2 105 bp, and the open reading frame is 1 494 bp, coding a protein of 497 aa with an estimated molecular mass of 57.8 kD and pI of 5.29. BmCaspase-X contains the typical characteristics of Caspase family. Phylogenetic analysis showed that BmCaspase-X and ICE of the silkworm cluster with Caspase-4 homologs from other insects firstly, and then cluster with Caspase members with long precursor structure domain from other insects. RNAi of BmCaspase-X in BmE-SWU1 cells did not cause significant apoptosis effect. 【Conclusion】 The new BmCaspase-X, with the typical characteristics of Caspase family, should be classified into the Caspase family. The preliminary RNAi data show that BmCaspase-X seems not to cause apoptosis in silkworm cells. This study lays a foundation for further functional investigation on BmCaspase-X.

【Aim】 A large number of genomic transcripts in both prokaryotes and eukaryotes are non-protein coding RNAs (ncRNAs). Accumulated evidences show that ncRNAs play important roles during the development of the silkworm, Bombyx mori. This study aims to explore the molecular mechanisms of ncRNAs in the nervous development of the silkworm. 【Methods】 Real-time PCR was used to study the expression of 22 ncRNAs and three neighboring genes of ncRNAs in the nervous system of the silkworm. 【Results】 Eight ncRNAs including a C/D box snoRNA, four H/ACA box snoRNAs and three unclassified ncRNAs were accumulated much more in the ventral thoracic nerves than in the nerves of head in the 5th instar larvae of the silkworm. Especially, the expression levels of snoRNA Bm-51, Bm-18 and Bm-86 in the ventral thoracic nerves were 23, 5 and 4.7 times as high as those in the nerves of head, respectively. Further studies showed that the expression of host genes was consistent with that of the three intron-origin ncRNAs, with more accumulation in the ventral thoracic nerves than in the nerves of head. 【Conclusion】 ncRNAs with higher expression in the ventral thoracic nerves might play roles in the activity of nerves through participating in the regulation of their neighboring genes. The results provide a molecular basis for the functional study of ncRNAs in the silkworm, as well as new clues for the lepidopterous pest control.