【Aim】 In order to investigate the differences of antioxidant system in non-diapause (ND), summer diapause (SD) and winter diapause (WD) pupae of Delia antiqua, the variations of antioxidant enzyme activities and glutathione redox status were determined and compared. 【Methods】 Time-course variations of antioxidant enzyme activities of copper/zinc-SOD (Cu/Zn-SOD), manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR), and contents of reduced glutathione (GSH) and oxidized glutathione (GSSG) and their ratio were measured and compared between different stages of ND, SD and WD pupae of D. antiqua. Canonical discriminate analysis was also conducted to further explore the relationships between the major antioxidant components. 【Results】 The activities of all the five antioxidant enzymes dynamically changed during the whole pupal stage. Compared with ND and SD pupae, WD pupae at the pre-diapause phase had higher Cu/Zn-SOD activities. The mean Cu/Zn-SOD activity in SD or WD pupae at the diapause maintenance phase and post-diapause phase was much lower than that in ND pupae. In the whole pupal stage, ND pupae had higher MnSOD activities than SD and WD pupae, but no difference existed between SD and WD pupae. The average MnSOD activity was distinctly higher than that of Cu/Zn-SOD at the same developmental stage. The CAT activities in SD and WD pupae at the pre-diapause phase were statistically higher than that in ND pupae at the same developmental stage, however, the CAT activities in SD or WD pupae at the diapause maintenance phase and post-diapause phase were significantly lower than those in ND pupae. In either non-diapause or diapause pupae at the same developmental stage, the general GPx activities showed an opposite variation trend with GR activities. Canonical discriminant analysis revealed that antioxidant system in onion fly pupae showed diapause type specificity. 【Conclusion】 Redox status in ND pupae is significantly different from that in SD and WD pupae. Higher antioxidant enzyme activities and lower GSH/GSSG ratio in pupae at the pre-diapause phase and post-diapause phase suggest that the oxidative shift results from a higher respiration rate and development changes.

 【Aim】 Trehalase (Tre), a key enzyme in trehalose metabolism of insects, plays important roles in the energy regulation and development of insects. This study aims to clone a trehalase gene from Galeruca daurica, and to analyze its expression patterns in order to investigate the functions of trehalase in the development of G. daleruca and summer diapause of its adults. 【Methods】 Based on the transcriptome data of G. daleruca, the full-length cDNA of the Tre gene was cloned using the rapid amplification of cDNA ends (RACE)-PCR and analyzed by bioinformatics. Real-time quantitative PCR (RT-qPCR) was performed to detect the expression levels of the Tre gene in different developmental stages (egg, 1st-3rd instar larva, prepupa, pupa, and 3, 7, 10, 15, 25, 40, 60, 80 and 100 day-old adult) and various adult tissues (head, thorax and abdomen) of G. daleruca, and the 3 day-old adults under different stress temperature (15, 20, 25, 30, 35 and 40℃). The Tre activity in different day-old adults was assayed by the 3, 5-dinitrosalicylic acid colorimetry. 【Results】 A soluble trehalase gene was cloned from G. daleruca, and named GdTre1 (GenBank accession no.: KY697913). Its fulllength cDNA is 1 933 bp containing a 1 704 bp open reading frame (ORF) that encodes 567 amino acids with the predicted molecular weight of 66.56 kD and pI of 6.62. The coded protein contains typical functional domains in the trehalase superfamily with a signal peptide but without transmembrane domain. Homology and phylogenetic analyses showed that GdTre1 has the highest amino acid sequence identity (70.25%) with Leptinotarsa decemlineata Tre1b. RT-qPCR results revealed that GdTre1 was expressed throughout various developmental stages of G. daleruca, with higher expression levels in the egg and diapause adult, and lower expression levels in the larva, pre-pupa, pupa and pre-diapause adult. Tissue expression profiles revealed that the expression level of GdTre1 was the highest in the abdomen, second high in the thorax and the lowest in the head of adults. The expression level of GdTre1 in 3 day-old adults increased as the stress temperature increased, with the highest level at 30℃, and afterwards decreased a little with the temperature increasing. The expression levels of GdTre1 and the Tre activity were significantly different among different day-old adults and showed the same change trend. 【Conclusion】 Trehalase has a close relationship with the development and summer diapause in G. daleruca. The results provide a necessary foundation for further investigation on the molecular mechanisms of summer diapause in this insect.

【Aim】 Mfe2, peroxisomal multifunctional enzyme type 2 gene, plays a very important role in the β-oxidation of fatty acids. This study aims to clone and identify the Mfe2 gene from the Japanese sawyer beetle, Monochamus alternatus, to express its encoded protein in prokaryotic expression system, and to explore its expression patterns in different developmental stages and adult tissues of the beetle.【Methods】 The recombinant plasmid was constructed to clone the Mfe2 gene of M. alternatus using the fast cloning method through designing primers with overlapping region between the vector and target fragment, and the cloned sequence was analyzed with various bioinformatics tools. The expressed Mfe2 protein in prokaryotic expression system was inducted with IPTG, and the fusion protein was verified by Western blotting. The expression profiles of this gene in different developmental stages and adult tissues of M. alternatus were investigated using the RT-qPCR technology. 【Results】 Mfe2 was cloned from M. alternatus and named MaMfe2 (GenBank accession no.: MF944109). Its open reading frame is 2 148 bp, encoding a protein of 716 amino acids, with an estimated molecular mass of 77.56 kD. Bioinformatics analysis showed that MaMfe2 has an AICARFT_IMPCHas domain, with 91% amino acid sequence identity with peroxisomal multifunctional enzyme type 2 of Anoplophora glabripennis. The expressed protein in BL21 (DE3) strains of Escherichia coli bound specifically with its antibody in Western blot analysis. Developmental expression profiles showed that the expression level of MaMfe2 was the highest in the pupal stage, and lower in egg and larval stages, and tissue expression profiles revealed that the expression level of MaMfe2 was the highest in the fat body and the lowest in the protothorax of adults. 【Conclusion】 MaMfe2 encodes a peroxisomal multifunctional enzyme in M. alternatus, and its expression level is various in different developmental stages and adult tissues. This study provides the basis for further research on the function of this gene in M. alternatus.

【Aim】 Olfactory receptor Or10 is one of the important receptor proteins for the perception of pheromone in the honeybee, Apis cerana cerana. The aim of this study is to clone the sequence of Or10 gene, and to analyze the expression levels of Or10 in drone antennae and brains across various physiological status, so as to provide the theoretical basis for further studies on the biological function of this gene. 【Methods】 The drone antennae of A. cerana cerana were collected, from which the total RNA was extracted to clone the sequence of Or10. The relative expression levels of Or10 in antennae and brains of drones, which were sexually immature at hive entrance, sexually mature at hive entrance, sexually immature at frames in hive and sexually mature at frames in hive, were detected using quantitative real-time PCR (qPCR). 【Results】 The cDNA sequence of Or10 gene (named as AcOr10) (GenBank accession no.: MF693365) of A. cerana cerana was obtained, with the length of 914 bp and a coding region of 738 bp, encoding 246 amino acids. The encoded protein has the molecular weight of 28.163 kD and isoelectric point of 5.50. The phylogenetic tree showed that AcOr10 clustered with Or10 of A. mellifera and Or4-like of A. florea. The relative expression levels of AcOr10 in antennae of sexually immature drones at hive entrance and sexually mature ones at frames in hive were significantly higher than those of sexually immature drones at hive entrance and sexually immature drones at frames in hive (P<0.05). However there was no significant difference in the relative expression level of AcOr10 in antennae between sexually mature drones at hive entrance and mature ones at frames in hive (P>0.05). There was also no significant difference in the relative expression level of AcOr10 in antennae between sexually immature drones at hive entrance and immature ones at frames in hive (P>0.05). The relative expression level of AcOr10 in brains of sexually mature drones at hive entrance was significantly higher than those of drones sexually immature at hive entrance, sexually immature at frames in hive and sexually mature at frames in hive (P<0.05), while there was no significant difference among the latter three groups (P>0.05). The relative expression levels of AcOr10 in antennae of drones sexually immature at hive entrance, sexually immature at frames in hive and sexually mature at frames in hive were all significantly higher than those in brains of drones with the same physiological status (P<0.05), but there was no significant difference in the expression level of AcOr10 between antennae and brains of drones sexually mature at hive entrance (P>0.05). 【Conclusion】 It is inferred that AcOr10 may be related with sexual maturity of A. cerana cerana drones, and flight activities have little influence on the expression of AcOr10 in antennae but cause the changes of brains, thereby affecting the mating behaviour of drones.

 【Aim】 The objective of this study is to investigate the effects of mixtures of brood pheromone esters on the queen quality of the Chinese honeybee, Apis cerana cerana, reared by queenrearing technology without larvae-grafting, and to find a way to rear high-quality queen. 【Methods】 Queen-rearing technology without larvae-grafting was employed to rear queen of A. cerana cerana. Using a micro sampling syringe injector, 1 μL of mixture of brood pheromone esters with the concentration gradients of 0, 0.1%, 1.0% and 10.0% was added into queen cells, respectively, when the queen larvae were 60-64 h old. The body weight, the weight and width of thorax, the number of ovarioles in one ovary of the newly-emerged queens were tested. And the gene expression levels of vitellogenin (Vg), hexamerin70b (hex70b) and hexamerin110 (hex110) were quantified by qPCR. 【Results】 The results showed that the body weight of the newly-emerged queens of A. cerana cerana was increased significantly (P＜0.05) in the treatments with the mixture of three brood pheromone esters (3E) (1.0% methyl linoleate, 16.0% methyl linolenate, and 35.0% methyl oleate) at all the three concentrations, while the width and weight of thorax of reared queens had no significant change as compared with the control (paraffin oil) (P＞0.05). The number of ovarioles in one ovary was increased significantly (P＜0.05) in 0.1% and 1.0% treatment groups. The expression levels of Vg and hex70b in all 3E treatment groups had no significant change, while that of hex110 in 1.0% and 10.0% 3E treatment groups increased significantly (P＜0.05) as compared with the control. When the queens were reared with the mixture of four brood pheromone esters (4E) (4.5% methyl palmitate, 1.0% methyl linoleate, 16.0% methyl linolenate, and 35.0% methyl oleate), the body weight of the newly-emerged queens and the number of ovarioles in one ovary significantly increased in 10.0% 4E treatment group (P＜0.05), while no significant difference of developmental indexes existed in 0.1% and 1.0% 4E treatment groups (P＞005). The expression levels of Vg in ovaries in 0.1% and 1.0% 4E treatment groups, hex70b in 10.0% 4E treatment group and hex110 in 1.0% and 10.0% 4E treatment groups all significantly increased as compared with the control (P＜0.05). When the queens were reared with the mixture of 10 brood pheromone esters (10E) (4.5% methyl palmitate, 2.5% methyl stearate, 35.0% methyl oleate, 1.0% methyl linoleate, 16.0% methyl linolenate, 3.5% ethyl palmitate, 1.5% ethyl stearate, 18.0% ethyl oleate, 0.5% ethyl linoleate, and 17.5% ethyl linolenate), the body weight of the newly-emerged queens, and the expression levels of Vg and hex110 in ovaries in all treatment groups significantly decreased as compared with the control (P＜0.05), while the number of ovarioles in one ovary and the thorax indexes showed no significant change (P＞0.05). The expression level of hex70b also reduced significantly in 10% 10E treatment group (P＜0.05). 【Conclusion】 To a certain extent, the 1.0% 3E and 10.0% 4E added in queen-rearing process of A. cerana cerana could improve the queen quality.

【Aim】 The objective of this research is to analyze the roles of tubulin and microfilaments in the Chinese bush cricket, Gampsocleis gratiosa, so as to build the foundation for exploring the mechanism of acrosomal complex formation and nucleus shaping during spermiogenesis in insects. 【Methods】 The development of sperms from testes, seminal vesicles and spermathecas in G. gratiosa adults including the localization of microfilaments and tubulin during spermiogenesis was observed by immunofluorescence, PAS-hematoxylin staining and transmission electron microscopy. 【Results】 In the early round spermatid testis, microfilaments gather to an area and tubulin is randomly distributed in cytoplasm during spermiogenesis. In the elongated spermatid, when the acrosomal complex is forming, microfilaments first emerge in subacrosomal space, become round-shaped and rodlet-shaped, and extend to two sides of the anterior region of the nucleus to form inverted ‘Y-shaped’ and then to form arrow-shaped. Tubulin exists around the acrosomal complex and the nucleus, and also in the flagellum. In sperm and spermatodesms of the seminal vesicle and the spermathecae from male and female adults, tubulin is only present in the flagellum, nevertheless, microfilaments and tubulin are absent in other areas. 【Conclusion】 The results suggest that microfilaments and microtubules as ‘scaffold’ participate in acrosomal complex formation and nucleus morphological reconstruction during spermiogenesis in G. gratiosa, and the ‘scaffold’ is removed in sperm maturing into spermatodesms.

【Aim】 To analyze and compare the community structure and diversity of the gut bacteria in adult males of Odontolabis fallaciosa with different mandibular forms. 【Methods】 The total DNA of guts from 31 male adults of O. fallaciosa with three mandibular forms (large, medium and small mandible forms) (including 15 adult males of large mandible form, 10 adult males of medium mandible form and 6 adult males of small mandible form) were extracted. The 16S rDNA gene fragments (V3-V4 region) of gut bacteria were sequenced by Illumina MiSeq, and the number of operational taxonomic units (OTUs), species composition, abundance, and alpha and beta diversity were analyzed. 【Results】 A total of 2 238 637 high quality sequences were obtained and clustered into 3 256 OTUs at a 97% similarity threshold. Analysis showed that they could be annotated into 42 phyla, 328 families and 542 genera. Among them, four phyla (Proteobacteria, Bacteroidetes, Firmicutes and Tenericutes) were prevalent in the gut bacteria communities of male adults, and four genera (Acinetobacter, Dysgonomonas, Bartonella and Chryseobacterium) were dominant. The alpha diversity analysis showed that the gut bacteria were quite abundant in these male samples of O. fallaciosa, and the beta diversity revealed that the composition of OTUs was different among the large, medium and small mandible males. 【Conclusion】 Using Illumina MiSeq sequencing technology, the structure and composition of the gut bacterial community in adult males of O. fallaciosa were analyzed. The microbial community composition in guts of adult males of O. fallaciosa has significant difference between the large and medium mandible adult males, whereas no significant difference exists between the large and small mandible adult males, or between the medium and small mandible adult males.

【Aim】 Plants secondary metabolites are potentially considered as suitable alternatives for chemical pesticides if their extents of impacts were well studied earlier on different groups of pests. This study aims to evaluate the possible impacts of solvent polarity on the insecticidal activity of different plant extracts. 【Methods】 The extraction solvents were selected from a wide range of polarities, n-hexane, ethyl acetate and methanol. The lethal effects of Petroselinum sativum seed extracts as well as Thymus vulgaris and Stachys lavandulifolia shoot extracts on Callosobruchus maculatus adults at 28±2℃, 50%±5% RH and a photoperiod of 16L∶8D were evaluated, and their ovicidal and oviposition deterrence effects were also evaluated. 【Results】 The lowest LC₅₀ value for contact toxicity to C. maculatus adults was associated with the n-hexane extract of T. vulgaris (0.05 g/mL) following with the ethyl acetate and methanol extracts, respectively. The ethyl acetate extract of T. vulgaris had the lowest LT₅₀ value (10.04 h), with the highest relative speed of kill index (42.23%) among all the extracts. All of the extracts at LC₂₀ and LC₅₀ concentrations resulted in 100% ovicidal activity in contact toxicity to C. maculatus adults. In terms of oviposition deterrence, the methanol extract of S. lavandulifolia had the highest deterrence activity (97.54%). 【Conclusion】 Based on the results, it is concluded that T. vulgaris, P. sativum and S. lavandulifolia extracts show significant performances in terms of insecticidal and ovicidal toxicities as well as oviposition deterrence against C. maculatus adults. However, non-polar (n-hexane) extracts of the studied plants show better performances in comparison with other extracts.

【Aim】 Temperature is the most important environmental factor for ectotherms and affects all aspects of lifehistory traits. This study aims to understand how lifehistory traits vary with temperature in the Asian corn borer, Ostrinia furnacalis. 【Methods】 The developmental duration from egg hatching to pupation and from pupation to adult eclosion, and the pupal and adult weight were examined in the Nanchang population of O. furnacalis at 20, 22, 24, 26, 28, 30 and 32℃ under a photoperiod of 16L∶8D in the laboratory. 【Results】 The larval and pupal duration and total developmental duration of O. furnacalis were significantly decreased with the increasing of rearing temperature. The larval duration and total developmental duration were significantly shorter in males than in females, showing the protandry phenomenon. The growth rate was positively correlated with temperature. The growth rates of females were significantly lower than those of males at low temperatures but significantly higher than those of males at high temperatures. The relationship between body weight and rearing temperature in O. furnacalis did not follow the temperature-size rule, and both males and females gained heavier body weight at high temperature. Females were significantly larger in size than males at all temperatures, showing a female biased sex size dimorphism (SSD). Contrary to Rensch’s rule, the SSD index and body weight of O. furnacalis tended to increase with rising temperature. Male pupae lost significantly more weight at metamorphosis compared to females, resulting in higher SSD index in adults than in pupae. 【Conclusion】 High temperature not only significantly shortens developmental duration, but also results in heavier body weight at maturity in O. furnacalis. There are significant differences in life-history traits between females and males of O. furnacalis.

【Aim】 Alien species may interfere with the normal reproduction of the closely related native species through mating competition. Although the European bumblebee, Bombus terrestris, is the most widely used pollinator in the world, it has caused biological invasion in many countries. This study aims to elucidate the possibility of reproductive disturbance to some Chinese bumblebees by B. terrestris. 【Methods】 The components of male cephalic labial gland secretions of B. terrestris and nine native bumblebee species in China were detected using gas chromatography-quadrupole time of flight mass spectrometry (GC-Q-TOF/MS), and the data were subjected to cluster analysis. The effects of male B. terrestris on the mating of the most important native bumblebee species under controlled environment were compared. 【Results】 The results showed that the GC-MS chromatograms of male cephalic labial gland secretions were unanimous within a bumblebee species, but significantly different between different bumblebee species. The components of male cephalic labial gland secretions of both B. lantschouensis and B. ignitus were similar to that of B. terrestris with 49.23% and 58.49% identity, respectively. The male of B. terrestris could mate with the queen of B. lantschouensis and significantly decreased the mating success of the latter (P<0.01). 【Conclusion】 It is concluded that the European bumblebee, B. terrestris, can interfere with the normal mating and has a high risk of reproductive disturbance to some local bumblebee species of China. In order to protect the local bumblebee resources and ecosystem balance from biological invasion, alien bumblebee species should be cautiously used in China.

Promoter is an important component regulating gene expression. It binds to RNA polymerase to form a transcription initiation complex that controls the initiation time and the level of gene expression. Studies on promoters may contribute to elucidating the underlying mechanism of gene expression regulation so as to lay the foundation for the analysis of gene regulatory networks. In this article, we reviewed the research methods of insect promoters, mainly about the methods for the identification of insect promoters and their specific components and the application of insect promoters.

The engineered endonuclease (EEN)-based genome editing technology is an important tool for genome function analysis and genetic improvement of biological varieties. The silkworm, Bombyx mori, is the first economic insect to be domesticated and bred by humans, and also an important model lepidopteran, which has been used for functional gene analysis in the post-genome era. In recent years, along with the rapid development of zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN) and CRISPR/Cas9 system-based novel genome editing technologies, the targeted genome editing technology in the silkworm also has had significant progress. In this review, the compositions and action principles of the three most commonly used EEN systems were introduced, and the establishment process of targeted genome editing technology in the silkworm and the application and research progress of this technology for point mutation of silkworm endogenous or exogenous genes, genome structure variation and integration of the exogenous genes into the silkworm genome were comprehensively and systematically summarized. This review also discussed the main problems and the corresponding strategies for the low-efficiency of the knockout of endogenous genes and integration of exogenous genes, and the off-target effects of the integration of exogenous genes in targeted genome editing techniques in the silkworm. In addition, the potential for multiplex gene editing and the development trend for the combined application and diversified development of different targeted genome editing systems of this technology in the silkworm were prospected. It is expected to provide insights and references for promoting the development of targeted genome editing technology in gene functional analysis, mutation model establishment and other research areas in insects.

As one kind of phosphagen kinases, arginine kinase is widely distributed in various tissues of invertebrates. In the long-term evolutionary process, the spatial structures of arginine kinases of different insects have differentiated. However, their amino acid sequences are highly conserved. Functionally, arginine kinases are primarily involved in energy metabolism and maintain the normal life activity in insects and other invertebrates. In addition, with the development of modern scientific techniques, more and more studies show that arginine kinase also plays an important role in autoimmunity and paralysis of host insects. In this article, the distribution, structure and functions of arginine kinases in insects were reviewed.