The superoxide dismutase and catalase play an important role in protecting worker bees from reactive oxygen species, and their enzyme activities and expression levels are associated with honeybee longevity and disease resistance which affect the performance of the colony. This study was to observe the developmental regulation of CuZnSOD, MnSOD and CAT genes in Apis mellifera ligustica worker bees at mRNA level by real-time quantitative PCR, so as to provide a theoretical basis for the study of aging and disease resistance of honeybees. During the larval, pupal and adult stages, we collected the samples of larva (5 d after egg laying by the queen), pupa (14 d after egg laying by the queen) and adult (1 d, 12 d, 32 d after emergence), respectively. The results showed that these three antioxidant enzyme genes were expressed in all developmental stages. Both CuZnSOD and MnSOD mRNA expression levels decreased during larval to pupal stages and the adult stage at 12-32 d after emergence, with the highest level at 1 d adult after emergence and the lowest level in pupa. However, no significant decrease was found in the expression of CuZnSOD gene at the adult stage, while the expression of MnSOD gene decreased obviously. The lower mRNA level of the CAT gene was detected in pupa. However, the CAT gene expression steadily increased during the development of honeybees, and the expression at 12-32 d adult after emergence was significantly higher than those at the other stages (P<0.05). The study illustrates the function of antioxidant enzymes at the molecular level during different developmental stages of A. mellifera ligustica worker bees.

For the expression and purification of Tenebrio molitor antifreeze proteins, RT-PCR method was adopted to obtain the cDNA of antifreeze protein gene afp84a which was then cloned into prokaryotic plasmid pMAL-p2X and expressed in Escherichia coli TBI strain. The recombination protein was purified through an amylose affinity column and assayed for antifreeze activity by observing bacterial survival rate in the presence of purified antifreeze protein following incubation for various intervals at 0℃. The results showed that the content of fusion protein was 40% of total dissolved proteins. SDS-PAGE analysis indicated that fusion protein could be released from cells treated with MgSO₄ or sonicate. A single band of target protein was acquired after the fusion protein was purified through amylose affinity column and incised by Factor Xa. Furthermore, the antifreeze protein AFP84a could increase the low temperature resistance of bacteria as shown in the biological activity analysis. The cloning and prokaryotic expression of afp84a from T. molitor could provide useful experimental materials for the further study of the nature and function of antifreeze protein AFP84a.

 In order to clarify the effects of body size and fat content on cold tolerance in adults of Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), two developmental temperatures (18℃ and 25℃) were used to induce a phenotypic variation in insect size and the relationships were analyzed among the adult size, fat reserves, the supercooling point (SCP) and the survival rate exposed to constant low temperatures (CLTs) or fluctuating thermal regimes (FTRs) in the laboratory condition. The results showed that the adults grown at the lower temperature (18℃) were significantly larger than those reared at 25℃ (P<0.01). The adult fat content was positively related to dry mass of body, indicating that larger individuals contained a higher proportion of fat. The SCP frequency distribution showed that the lower SCPs (-10℃- -9℃) were found in adults bred at the higher temperature (25℃), whereas the higher SCPs (-8℃- -6℃) in adults bred at the lower temperature (18℃). The negative linear relation was observed between SCP and adult body size. The survival rate, especially under CLT, decreased significantly with duration of coldexposure. The type of exposure (FTR versus CLT) had a dramatic impact on the survival rate, which was distinctively higher under FTR than under CLT as in the following order: FTR18℃＞FTR25℃＞CLT18℃＞CLT25℃. The results indicate that energy storage is an important factor, but the vulnerability to chill-injury is supposed to be the primary factor regulating survival at low temperature.

The ovariole structure and oogenesis have significance in exploring the phylogenetic relationships in insects. Detailed investigation on ovariole structure and oogenesis of Panorpidae could provide the evidence of phylogenetic position of Mecoptera in Holometabola. The ovariole structure and oogenesis of the scorpionfly Panorpa liui Hua were investigated using light microscopy and scanning and transmission electron microscopy. The results showed that each of the two ovaries consists of 12 polytrophic ovarioles, which can be differentiated into the terminal filament, a germarium, a vitellarium and a pedicel. Based on the variations of the nurse cells, oocyte, and follicle cells, the process of oogenesis can be divided into five stages: early, middle and late previtellogenesis, vitellogenesis and choriogenesis. During the vitellogenesis, the nurse cells provided the nutrition for the oocyte and then degenerated gradually. When the nurse cells disappear completely, the nutrition of the oocyte might be supported by the hemolymph through the interspace between follicle cells. Three types of follicle cells surrounding the oocyte are responsible for the eggshell formation in different regions, resulting in the distinct meshwork on the eggshell. Through comparison with the nurse cell number in other orders, we speculate that three nurse cells in each egg chamber might be a plesiomorphic character, suggesting that Mecoptera represent a basal lineage in Holometabola.

The development of new cell lines from unusual and non-pest species may provide important source for basic research and biotechnology application. In this study, three new insect cell lines designated as QB-Ms1-8, QB-Ms2-2, and QB-Ms2-7, respectively, were established from eggs of Manduca sexta. These cells were subcultured approximately 50 passages in TNMFH medium at 28℃. The majority of cells of the lines generally are spindle-shaped. The population doubling time of QB-Ms1-8, QB-Ms2-2 and QB-Ms2-7 cells was 51, 31 and 49 h, respectively. Although these lines were insusceptible to Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) with the infection rate of 33%-40% at 96 h post infection, QB-Ms2-2 cell line showed higher level of secreted SEAP production in comparison with BTI-Tn5B1-4 cell line. In this study, a SEAP high expression type cell line, QB-M2-2 was isolated from the three new cell lines established from M. sexta, which may provide a source for searching novel clones with higher production of recombinant proteins.

 F3-28 is an insecticidal fraction which was isolated from the extract of Periploca sepium bark.  In order to understand the action mechanism of the insecticidal fraction F3-28, the effects of F3-28 on the activities of proteases, trypsin-like protease, chymotrypsin-like protease, lipase and amylase in the midgut of the 5th instar larvae of Mythimna separata and Agrotis ypsilon were studied. The results showed that after A. ypsilon larvae which are insensitive to F3-28 ingested this fraction, the activities of proteases, lipase and amylase in the midgut changed indistinctly. However, after M. separata larvae which are sensitive to F3-28 ingested this fraction, at 2, 4, 8, 10, 12, 24 and 48 h post treatment their midgut protease activity was 0.76, 2.53, 1.45, 1.88, 1.54, 1.46 and 1.70 times as high as that of the control, respectively, and their midgut trysin-like protease activity was 1.60, 1.75, 1.60, 1.12, 1.39, 1.16 and 1.15 times （BAPNA as the substrate） and 1.68, 1.95, 1.53, 1.26, 1.15, 1.13 and 1.14 times (TAME as the substrate ) as high as that of the control, respectively. Similarly, after M. separata larvae ingested F3-28, at 2, 4, 8, 10, 12, 24 and 48 h post treatment their midgut chymotrypsin-like protease activity was 0.50, 1.66, 1.44, 1.18, 1.54, 1.08 and 1.03 times as high as that of the control, respectively, and their midgut amylase activity was 1.60, 1.35, 1.27, 1.31, 1.23 and 1.20 times as high as the control, respectively. The increase in activities of proteases and trysin-like protease in the midgut was dependent on the concentration of F3-28, but that of chymotrypsin-like protease and amylase in the midgut was not. After M. separata larvae ingested F3-28, their midgut lipase activity changed inconspicuously. These results imply that the insecticidal mechanism of F3-28 may be related with the activation of proteases, especially trypsin-like protease, in the larval midgut.

The migration process of the rice leaffolder, Cnaphalocrocis medinalis (Guenée), is composed of several successive long-distance nocturnal flights. It is the unpredictable re-migration ability that makes the outbreak forecasting of rice leaffolder extremely difficult. So it is important to understand the parameters of the flight behaviour of rice leaffolder for its prediction. In this study, the flight and re-migration capacity of the rice leaffolder moth were examined by tethered flight on a computerized flight mill. The results indicated that there were no significant differences in flight capacity of moths among different ages and sex, and the mean flight duration of males and females were 156.94±103.19 min and 147.71±111.38 min, respectively. Based on the accumulative flight duration (AFD) of the individuals on flight mill, the moths could be divided into three types: the residence type (AFD<40 min), the migratory type (40 min ≤AFD≤130 min) and the strong migratory type (AFD>130 min). The proportions of individuals and mean flight duration of the three types were 46.1% and 11 min, 27.3% and 82 min, and 26.6% and 232 min, respectively. Strong re-migration capacity of C. medinalis had been observed during the experiment. In all of the tested moths, the proportion of the one-off migration was higher than 90%, the twice re-migration above 70%, about half of the moths could fly for 4-5 successive nights, and the maximum re-migration was nine nights. There were no significant effects on re-migration capacity of the moths by foods, but feeding honey could enhance the flight capacity. These results will provide useful behavioral parameters for trajectory analysis and immigration forecasting of the rice leaffolder.

 To establish the basis for identifying and synthesizing the sex pheromone of Holcocerus vicarius (Walker), the eclosion, mating and oviposition of H. vicarius adults were studied by means of observations and experiments both in the fields and laboratory. The circadian rhythm of sex pheromone production and release of H. vicarius was studied by means of electroantennogram (EAG) response. The results showed that the eclosion of H. vicarius happened throughout the day, especially from 15:00 to 19:00. Mating behavior started on the 1st night after the eclosion and the nuptial flight and mating took place from 21:30 to 0:30. The peak of mating occurred at 23:00 or so. The mating behavior was not observed during the photophase. Calling varied with the age of female, with older (2 to 3-day-old) virgin females calling earlier and for longer duration than younger (1-day-old) females. They had shorter time of mating when they were young. Females became sex mature one day later. The percentage of mating of 2-day-old was the highest (41.8%). The females searched the gap with ovipositor, and then began to oviposit. They oviposited at various places. The maximum of oviposition was up to 720 eggs, and the minimum was 105 eggs. The egg stage lasted 12-22 d. The hatch rate was from 72% to 88%. The amount of pheromone in the gland was relatively low on the first night after the eclosion, with a peak during the second night, and then decreased gradually when they aged. The amount of pheromone of 2-day-old virgin females began to increase gradually post 17:00, with the peak between 21:30 and 22:00, and then decreased gradually post 22:00. EAG response was the greatest at 22:30. The peak of sex pheromone production and release of H. vicarius occurred between 21:30 and 23:00. There was a circadian rhythm in the eclosion and mating of adults. The sex pheromone release and mating behavior of H. vicarius were synchronized, which was confirmed by tests of using virgin females to trap males in the fields.

The attractive activities of the volatiles from frass of both sexes of Apriona germari feeding on different plants (Morus alba, Cudrania tricuspidata and Broussonetia papyrifera) towards the parasitoid Aprostocetus prolixus was compared by olfactory bioassays, and the components of those volatiles were analyzed by TCT-GC-MS, aiming at deeply understanding of the role of volatiles from frass of A. germari in host location of A. prolixus. The results indicated that the volatiles from frass of female or male A. germari feeding on the different host plants attracted the parasitoid significantly. But the attractive activities towards the parasitoid were not different significantly between different frass, suggesting that the host plant species or the sex of A. germari does not affect the attractiveness of frass to A. prolixus. The volatiles from A. germari frass included mainly alkynes, ketones, aldehydes, esters and terpenes. The components of the frass volatiles of A. germari feeding on C. tricuspidata were the same as on B. papyrifera. However, more volatile components were found in frass of A. germari feeding on M. alba than feeding on the other two species of plants. The components of frass volatiles of both sexes of A. germari feeding on the same species of host plant were roughly the same. The results suggest that the species difference of host plants affects the volatile components of A. germari frass significantly, while the sexuality of A. germari does not.

 In 2009 and 2010, the nest and cell structures and behavior of the Colletes gigas were studied in Guangdong province, China. We excavated some nests of the species aggregating in one place, and had a detailed description of two nests’ structure. We also observed and described the external morphology of the egg, larva and pupa of the species using scanning electron microscopy and light microscope. In addition, through using molecular and morphology methods, we identified another species of the genus Colletes, whose nest are found beside the nests of C. gigas. We found that C. gigas likes to construct nests in soil mixed with sand, and the nests often distribute in an aggregation. The nests consist of a main burrow and some lateral ones, each of which ends with a single cell. The abandoned nests of C. gigas can be reused by the female of the next generation, which constructs its new lateral burrows or expands the old ones. C. gigas feeds obligately on some species of the family Theaceae in Southern China, especially Camellia oleifera. The food mass of C. gigas contains a high proportion of nectar, liquid and even water. In addition, both the COI and 28S D2 data confirmed that the C. gigas can share nest habitat with another Colletes species.

Up to now, the higher level taxonomic and phylogenetic studies about the butterflies are still limited and remain many controversial issues. In this study, the complete mitochondrial genome of Pieris rapae Linnaeus was sequenced by using long-PCR and conserved primer walking techniques, aiming at supporting more mitogenomic data to further clarify these problems. The results of genome characterization analysis showed that the mitogenome is 15 157 bp in size, containing 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA, and an A + T-rich region, with the length of 11 196 bp, 1 474 bp, 2 093 bp and 393 bp, respectively; all the 37 genes are arranged in the same orientations as those of other butterflies determined; there are 10 overlapping sequences totalling 59 bp (1-35 bp for each sequence) and 13 intergenic spacer sequences totalling 120 bp (1-46 bp for each sequence), scattered throughout the genome, respectively, and the largest (46 bp) spacer region is located between the tRNAIle and tRNAGln genes. The neighbor-joining (NJ) and the maximum parsimony (MP) phylogenetic trees of the eleven representative butterfly species were reconstructed based on the complete 13 protein coding gene sequence data, and the results showed that Papilionidae (including papilionids and parnassiids) is a monophyletic group, and Pieridae is closely related with Lycaenidae and Nymphalidae (including nymphalids and acraeids in this study).

Cantharidin is a kind of defensive substance which is synthesized by blister beetles, and it has been proven that cantharidin has special efficacy to cure several cancers, such as liver cancer and lung cancer, and some other diseases. The cantharidin can exist in free cantharidin, bound cantharidin and derivants in blister beetles, such as magnesium cantharidate, calcium cantharidate, cantharidinimide, dimethyl anhydride, hydroxyl-cantharidin, demethyl-cantharidin and so on. The volume of the chemical may vary depending on the sex, age, mating records and feeding conditions. The cantharidin measure can reach as high as 10% in some sexually matured male beetles. Though there are some research reports, no final conclusions about biosynthesis pathway of cantharidin and its distribution in the body of meloid have been achieved yet. In this article, we make a review of the cantharidin on its content, distribution, biosynthesis, metabolism and biological function in blister beetles. This review may be helpful for understanding the biosynthesis pathway of cantharidin, full excavation of meloid insect resource, guiding artificial breeding of the insects and rational use of the resources, and artificial synthesis of cantharidin.

In order to understand the physiological and biochemical mechanism that Beauveria bassiana successfully infected the red imported fire ant (RIFA), Solenopsis invicta, the protein content in infected adults, larvae and pupae of RIFA was determined in the laboratory. Following fungal infections, the protein content varied in different developmental stages of infected RIFA. At 12-72 h after inoculation, the protein content in worker body reached as high as 47.12 mg/g at 24 h, showing significant difference from control group (56.40 mg/g)(P＜0.05), and then dropped to 25.16 mg/g at 72 h. The highest protein content of 24.13 mg/g was observed in larval body at 36 h after inoculation. After that, the protein content in larval body decreased, and was 8.95 mg/g at 72 h after inoculation, which was dramatically lower than that of the control (24.80 mg/g). Protein content in pupae infected by B. bassiana dropped from 36.57 mg/g at 12 h to 10.42 mg/g at 72 h after inoculation, which was significantly lower than that of the control (38.61 mg/g)(P＜0.05). These results showed that the protein content in different developmental stages of RIFA treated with B. bassiana declined dramatically.