【Aim】 The aim of this study is to clone and express the venom serine protease homologue (SPH) gene of Sclerotium guani (SgSPH), and to investigate the effect of the venom protein encoded by this gene on the phenoloxidase activity in the host hemolymph. 【Methods】 The open reading frame (ORF) of venom SgSPH gene was cloned from S. guani by RT-PCR. Its sequence features were analyzed using bioinformatic software. The relative expression levels of the venom SgSPH gene at different developmental stages (egg, early instar larva, late instar larva, mature larva, spinning larva, pupa in yellow cocoon, pupa in black cocoon, and 1-5-day-old adults) and in different female adult tissues (head, thorax, abdomen without venom apparatus and venom apparatus) of S. guani were determined by qPCR. This venom gene was expressed with prokaryotic expression vector pSUMO-Mut. The expressed recombinant protein was purified using Ni-chelating affinity chromatography, and examined by SDS-PAGE and Western blot analysis. The inhibitory effect of the recombinant SgSPH on the phenoloxidase activity in the pupal haemolymph of Tenebrio molitor was measured by enzymatic activity assay. 【Results】 The ORF of the venom SgSPH gene (GenBank accession number: MT920663) of S. guani was cloned. It is 798 bp in length, encoding 265 amino acids, with the signal peptide consisting of amino acids 1-20, and the predicted protein molecular mass of 30.53 kD and pI of 9.59. Multiple sequence alignment results showed that SgSPH of S. guani shares low amino acid sequence identity (9%-17%) with the serine proteases and SPHs from venoms of other parasitoid wasps, and lacks a conservative catalytic triad. The qPCR results indicated that SgSPH gene was abundantly expressed at the adult stage and in the venom apparatus of S. guani. SDSPAGE and Western blot analyses showed that the recombinant SgSPH was successfully expressed and the highly purified recombinant SgSPH was obtained. Enzymatic activity assay results showed that the recombinant SgSPH was able to inhibit the phenoloxidase activity in the pupal hemolymph of the host T. molitor. 【Conclusion】 The results suggest that the venom SgSPH of S. guani can interfere with the host phenoloxidase cascade.

 Insect immunity has gradually attracted broad attention in recent years because of its important physiological functions in the process of insect resistance to harmful exogenous stimuli. Although both internal and external immunity are parts of the insect immune defense system, it is generally believed that the internal immunity is an immediate and primary immune response of insects while the external immune defense is only considered as a kind of secondary immune response. Nevertheless, the external immunity using defensive secretions as the core component constitutes the first barrier to foreign substances which invade insects. Accordingly, external immunity and internal immunity are two kinds of immune defense strategies for insects. When insects suffer exotic organism infection or adverse environmental factors, the external immunity is often the priority to be employed to fight against the invasion of natural enemies or adversity. However, the internal immunity and external immunity are both synchronized in the whole process of defense, which will cause a dilemma on energy allocation between the two defense strategies. Moreover, it is essential for insects to employ a series of immune reactions to prevent potential threats, which will seriously impair the effectiveness of pest biological control. This review focuses on insect external immunity and summarizes the current understandings on defensive secretions in insects, as well as the two immune defense strategies and their trade-offs, aiming to provide a synthesis of important updates, potential applications, and novel ideas and technologies for further research in this area. Also, the related research will not only contribute to further analysis about the immune interaction and diversified immunization strategies between insects and exogenous factors to improve the control effect in the field, but also pave the way for the development of novel insecticides targeting the insect immune system.

【Aim】 This study aims to evaluate the influence of feeding different concentrations of exogenous trehalose on the survival and cold hardiness of adult Dastarcus helophoroides, an important and most effective natural enemy of longhorned beetles. 【Methods】 D. helophoroides adults were fed on a semi-artificial diets containing 3%, 6% and 9% trehalose, respectively, in the laboratory, and those fed on the semi-artificial diet without trehalose were used as the control group. After 10 weeks, the survival rate of adults was calculated. In addition, the supercooling point and water content of adults not subjected to low temperature treatment and subjected to low temperature treatment (10℃ for 3 d) were measured. 【Results】 The survival rate of D. helophoroides adults fed on the semi-artificial diets containing 6% trehalose was the highest (86.67%) among all the treatments. Whether subjected to low temperature treatment or not, D. helophoroides adults fed on the semi-artificial diet containing 9% trehalose had the lowest supercooling point as compared to those fed on the semi-artificial diet containing 3% and 6% trehalose and without trehalose (the control): the supercooling point of adults unsubjected to low temperature was -19.30℃, while that of adults subjected to low temperature treatment was -21.60℃. Low temperature treatment had a significant effect on the water content of adults fed on the diet without trehalose but no significant effect on the water content of adults fed on the semi-artificial diet containing trehalose. 【Conclusion】 Exogenous trehalose has significant influence on the survival and cold hardiness of D. helophoroides adults. Thus exogenous trehalose can be utilized to improve the survival rate and cold hardiness of D. helophoroides adults in laboratory rearing.

【Aim】 This study aims to explore the sublethal effects of carbendazim on the growth and development and detoxifying enzyme activities of the larvae of the Italian honeybee, Apis mellifera ligustica. 【Methods】 Larvae in two treatment groups were fed with diets containing 0.25 and 0.75 mg/g a.i. carbendazim (relative mortality rate<5%), respectively, while those in the control group with the normal diets. All larvae were reared with different diets until eclosion, and their growth indexes (pupal weight, pupation rate and eclosion rate), total protein concentration, and T-SOD and detoxifying enzyme activities were assayed. 【Results】 There were no significant differences in the pupal weight, pupation rate and eclosion rate of A. m. ligustica among all groups ( P>0.05). The total protein concentration in pupae decreased in the treatment group fed with diets containing 0.75 mg/g a.i. carbendazim ( P<0.05). The total superoxide dismutase activities in the treatment groups fed with diets containing 0.25 and 0.75 mg/g a.i. carbendazim tended to be significantly higher than that in the control group, being 1.35- and 1.28.fold as high as the control group, respectively. The juvenile hormone titer in the treatment groups fed with diets containing 0.25 and 0.75 mg/g a.i. carbendazim were significantly higher than that in the control group, being 1.57- and 1.75-fold as high as the control group, respectively. Ecdysone titers in the treatment groups fed with diets containing 0.25 and 0.75 mg/g a.i. carbendazim were only 62% and 65% of the control group, respectively ( P<0.05). The cytochrome P450 (CYP450) and carboxylesterase (CarE) activities were increased in the low-dose (0.25 mg/g a.i.) treatment group ( P<0.05), and then returned to the normal level in the high-dose (0.75 mg/g a.i.) treatment group, but no inhibitory effect on the activities of the two enzymes was found ( P>0.05). There was no significant difference in glutathione-S-transferase (GST) activity between treatment groups and the control group ( P>0.05). 【Conclusion】 These results demonstrate that sublethal doses of carbendazim restrain the growth and development of A. m. ligustica larvae, but do not cause their acute death, and may cause potential harm to the stability and development of the colony.

【Aim】 Ubiquitin regulatory X (UBX) domain-containing proteins function as cofactors of p97/CDC48 which is involved in multiple ubiquitin-related processes including protein degradation through the ubiquitin-proteasome system and homotypic membrane fusion. Our study aims to clone the UBX domain-containing protein gene from Locusta migratoria manilensis, and to analyze its expression patterns in different tissues and at different developmental stages, so as to provide the foundation for further function research. 【Methods】 A transcriptome database for L. migratoria manilensis was mined through bioinformatic analysis. Gene expression levels were analyzed in different tissues of L. migratoria manilensis adults and at different developmental stages using real-time PCR. 【Results】 A UBX domain-containing protein gene was identified, and named LmUBX2. The open reading frame (ORF) encodes a protein of 339 amino acids with a predicted molecular weight of 37.8 kDa and a theoretical pI of 6.03. BLAST analysis showed that it shares 37%-64% amino acid sequence identities to other UBX domain-containing proteins. LmUBX2 possesses a conserved N-terminal UBA domain and a C-terminal UBX domain. Sequence comparison and phylogenetic analysis revealed that LmUBX2 is a member of the SAKS1 subfamily. Gene expression analysis results showed that LmUBX2 was expressed during the whole life cycle, and highly expressed in ovary and testis. 【Conclusion】 These findings suggest that LmUBX2  might be involved in multiple physiological processes of L. migratoria manilensis. Especially, LmUBX2 might be related to the reproduction of L. migratoria manilensis, although further research needs to be performed to confirm this correlation.

 【Aim】 Chromosomal characteristics play significant roles in phylogenetic analyses of insects. However, chromosomes in Bittacidae (Mecoptera) have rarely been studied to date. 【Methods】 Larvae, pupae and adults of the hangingfly Bittacus flavidus Huang & Hua were obtained by rearing in the laboratory. Chromosome preparations for the testes of the last (4th) instar larvae, pupae and newly-emerged adults of the hangingfly were stained with 4′,6-diamidino-2-phenylindole (DAPI) to investigate the karyotype, meiotic behavior, and sex determination. 【Results】 The results showed that the chromosome number of B. flavidus is 2n=26+X0, and all the chromosomes are metacentric, showing a symmetric karyotype. The absolute length of bivalents ranged from 3.20±0.07 to 1.53±0.19 μm, and the relative length from 5.31±0.29 to 2.73±0.24, with their size decreasing gradually from pair to pair. The meiosis of B. flavidus is chiasmate with a mean chiasma frequency of 11.5 per nucleus and a mean chiasma frequency of 0.88 per autosomal bivalent. The sex determination mechanism is of the XX/X0 type. Fluorescent bands with DAPI staining revealed a large AT-rich block covering one terminal region on all pachytene bivalents. 【Conclusion】 The chromosomes exhibit marked variations with respect to the diploid number, fundamental number and karyotype formula, suggesting that chromosome rearrangements (especially fusion, fission and inversion) play an important part in the lineage differentiation and chromosome evolution in Bittacidae.

【Aim】 To explore the complete mitochondrial genome structure and molecular phylogenetics of Limenitis helmanni. 【Methods】 The complete mitochondrial genome of L.  helmanni was sequenced and analyzed by using PCR and primer walking. Based on the sequences of 13 protein-coding genes and two rRNA genes of the mitochondrial genome, the phylogenetic tree of 66 lepidopteran species were constructed with Bayesian inference method. 【Results】 The complete mitochondrial genome of L. helmanni is a circular molecule of 15 178 bp （GenBank accession no.: KY290566）, including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a 346 bp A+T-rich region. The mitogenomic gene arrangement is consistent with those of other closely related species. The mitochondrial genome of L. helmanni is biased toward a high A+T content (81.1％). All protein-coding genes start with a typical ATN initiation codon, except that COI starts with the CGA codon and ND5 with the GTT codon. Most of the 13 PCGs have a complete termination codon (TAA), except COII and ND4 genes which have incomplete stop codons (T). All tRNA genes show the classic clover-leaf structure, except that the dihydrouridine (DHU) arm of tRNA ^Ser(AGN) forms a simple loop. The A+T-rich region of L. helmanni contains some conserved structures such as the motif ATAGA, 20 bp poly(T) stretch and some tandem repeat units, which is similar to those of other related lepidopteran species. Bayesian phylogenetic analyses supported that the relationship of Nymphalidae subfamilies is (Calinaginae＋Satyrinae)＋((Nymphalinae＋Apaturinae)＋(Heliconiinae＋Limenitidinae)). 【Conclusion】 Limenitini is closely related with the Euthaliini, and Parthenini may be the earliest diverged lineage in Limenitidinae. The phylogenetic relationship of Limenitidinae species based on the mitochondrial genome is not consistent with the results of the traditional morphology-based taxonomy

【Aim】To explore the role of heat shock protein Hsp70 genes of Myzus persicae in response to high and low temperature stress. 【Methods】 RT-qPCR was used to detect the relative expression levels of eight MpHsp70 genes (MpHsp70-1, MpHsp70-2, MpHsp70A1, MpHsp70B2, MpHsp68a, MpHsp68b, MpHsc70-4 and MpHsp70) in different wingless adult tissues (head, midgut, embryo and cuticle) and wingless adults under high temperature (36 ℃) and low temperature (4 ℃) stress for different duration (0, 30, 60, 90, 120 and 150 min), respectively. RNAi was used to silence two key MpHsp70 genes (MpHsp70A1 and MpHsp68a), and the survival rate and number of nymphs produced of wingless adults were observed and calculated at 120 min under high temperature treatment (36 ℃) and 30 min under low temperature treatment (4 ℃).【Results】 The eight MpHsp70 genes were expressed in different wingless adult tissues of M. persicae, and the expression levels of MpHsp70, MpHsp70A1, MpHsp70B2, MpHsc70-4 and MpHsp68b in the cuticile were significantly higher than those in the other tissues. The expression levels of MpHsp70-1, MpHsp70-2 and MpHsp68a in the embryo of M. persicae were significantly higher than those in other tissues. High and low temperature stress had significant induction effect on the expression of MpHsp70 genes in wingless adult of M. persicae. The expression levels of MpHsp70-1, MpHsp70A1, MpHsp70-2, MpHsp68a and MpHsp68b all increased and then decreased under 4 ℃ stress, and reached the highest at 30 min under 4 ℃ stress, which were significantly higher than those of the control. Under 36 ℃ stress, the expressions levels of MpHsp70-1, MpHsp70A1, MpHsp70-2, MpHsc70-4, MpHsp68a and MpHsp68b increased first and then decreased. The expression level of MpHsp70-1 reached the highest at 60 min after 36 ℃ stress, and those of the other genes reached the highest at 120 min after 36 ℃ stress. After the silence of MpHsp68a, the survival rate and number of nymphs produced of M. persicae under high and low temperature stress were significantly decreased and those after the silence of MpHsp70A1 under high temperature stress were extremely significantly decreased as compared with those of the control group. 【Conclusion】MpHsp70 genes of M. persicae can respond to high and low temperature stress, and play an important role in the molecular mechanism of resistance to temperature stress of M. persicae.
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Monochamus alternatus Hope is the major insect vector transmitting pinewood nematode disease, an important quarantine disease of pine forest. The virulence of a strain of Metarhizium anisopliae, MaYTTR-04, to adult M. alternatus was assayed by using the adult-conidial attaching method (attaching all tarsi of adult with dry conidial). The results showed that M. alternatus began to die at 6 days post inoculation (dpi) day at constant temperature (25℃±1℃, RH 70%±10%, light period 10L∶14D). The peak of death was between 18－21 dpi. The mortality of this beetle was 85% at 18 dpi when applied with the amount of inoculum at 2.3×10 ⁶±0.2×10 ⁶ spores/adult, and the LT ₅₀ was 14.7 d. At room temperature (24℃－33℃, RH 40%－60%, natural light from north), the adult of M. alternatus began to die at 3 dpi. The peak of death was between 15－21 dpi. The mortality was 85% at 15 dpi and 100% at 21 dpi when applied with the amount of inoculum at 2.3×10 ⁶±0.2×10 ⁶ spores/adult, and the LT ₅₀ was 12.9 d. When the adults of M. alternatus were kept in the cage in natural pine forest for 21 days, the average mortality of the adults dealt with conidia suspension was 60%, and the cadaver rate was 48.9%, while the average mortality of the adult deal with non-woven fabric strips of M. anisopliae was 86.7%, and the cadaver rate was 75.6%. It was concluded that this strain of M. anisopliae is high virulent to M. alternatus, and more virulent at higher temperature, which may serve as a productive strain of biocontrol agents and be used in forest for controlling M. alternatus.

【Aim】 Picromerus lewisi is a significant predatory natural enemy insect distributed in multiple countries of Asia, such as China, Korea and Japan, primarily used for controlling lepidopteran pests. Venom plays a crucial role in causing rapid paralysis and death of preys during hunting. The aim of this study is to understand the transcriptomic characteristics of the venom glands of P. lewisi, explore the diversity of toxins in P. lewisi, and establish a foundation for further research on the composition and function of the venom in P. lewisi.【Methods】Transcriptome sequencing was conducted on the venom glands of adults and the 5th instar nymphs of P. lewisi collected from Qianxinan Prefecture, Guizhou Province using the high-throughput Illumina NovaSeq 6000 platform. The resulting data were annotated using the NR, NT, Pfam, KOG/COG, Swiss-Prot, KEGG and GO databases. Gene expression in the venom gland samples of P. lewisi was assessed using the FPKM method, and DESeq was employed for the differential expression analysis of venom gland transcriptomes between adult and the 5th instar nymph. The differentially expressed genes (DEGs) between the adult and the 5th instar nymphal venom gland transcriptomes were screened using the criteria of |log₂(Fold change)|>1 and P<0.05, followed by GO functional enrichment analysis and KEGG pathway enrichment analysis of the identified DEGs. The 33 215 transcripts obtained from the sequenced venom gland transcriptomes of adults and the 5th instar nymphs of P. lewisi were subjected to BLAST comparisons in the UniProt database.【Results】Transcriptome sequencing of the venom glands of adults and the 5th instar nymphs of P. lewisi assembled to 22 242 unigenes with an average length of 949 bp. A total of 15 364 unigenes were annotated to the NR, NT, Pfam, Swiss-Prot, GO, KOG/COG and KEGG databases, corresponding to 10 closely related species including three species of true bugs and two species of spiders. A total of 344 DEGs were screened between the venom gland transcriptomes of adults and the 5th instar nymphs of P. lewisi, with 218 genes up-regulated and 126 genes down-regulated. A total of 443 sequences encoding 33 distinct types of toxin-related proteins were identified.【Conclusion】The transcriptome data from the venom glands of both the 5th instar nymphs and adults of P. lewisi were sequenced and obtained in this study. Differential proteins between the venom gland transcriptomes of adults and the 5th instar nymphs were screened, and sequences associated with toxin proteins were identified. This research lays a theoretical foundation for the identification of components in the venom of P. lewisi and the investigation of their biological functions.

 From the point of view of natural history, both the concepts and characteristics of the monophyletic group, phylogenetic systematics and natural classification system have been clarified. Natural classification system is a taxonomic system and natural evolution process of all members in the monophyletic group and their phylogenetic relationships. Only a monophyletic group including all extinct and extant members can be called a natural taxonomic system. At present, systematic biology mainly uses the cladistic principles and methods to reconstruct an evolutionary tree. During the preparation and application of a variety of system programming languages, researchers often focus on the morphological and molecular data acquisition, and the operating and calculating process, but largely ignore the hypotheses and associated weakness of the parsimony principle and the maximum likelihood or Bayesian inference. There are two common misunderstandings in the process of phylogenetic reconstruction: (1) a monophyletic group is claimed as a natural system due to incomplete selection of ingroup members; (2) a parsimony or likelihood phylogenetic relationship through mathematics and program operations is claimed as a natural evolutionary system. A natural monophyletic group often has a long evolutionary history and cannot be directly observed in nature or repeated in the laboratory. The systematical relationship among a monophyletic group established by cladistics principles is only the maximum parsimony or the likelihood speculation, thus, cannot truly reflect the development process of natural history. Fossils can provide better spatiotemporal framework information, but the number and accuracy of the available features are not sufficient. Living organisms have various abundant characteristic and genetic data. However, due to their long evolutionary history and the lack of a large number of important extinct members, the most deduced phylogenies are paraphyletic group or polyphyletic group and cannot constitute a natural system. It is impossible to construct a natural system of the whole monophyletic group by using only macro and microinformation of extant organisms. With the help of various techniques and research methods, the optimal approach to study a natural system is to combine the morphological and molecular data and integrate the whole ancient and modern members. In this process, six principles can be used to test whether the classification system is consistent with the development of natural history.

 Reactive oxygen species (ROS) is a general term for a class of oxygen-containing free radicals formed due to incomplete oxidation of oxygen molecules, or peroxides that are easy to form oxygen free radicals. When insects are invaded by pathogens, the ROS defense system mediated by dual oxidase (DUOX) will respond quickly to produce a large amount of ROS to resist the invasion of pathogenic bacteria, and then play a role in regulating the immune defense process of insects. However, high level of ROS can damage the biological macromolecules such as proteins, DNA and lipids in cells, causing damage to insect cells and affecting the normal development of insects. In order to avoid damage from excessive oxidative stress, insects have formed a complete antioxidant defense system mainly composed of antioxidant enzymes and small molecule antioxidants to prevent excessive damage from occurring. It is interesting that changes in ROS levels in cells can play completely different roles in regulating insect lifespan: for certain insects the accumulation of a large amount of ROS could lead to a shortened lifespan, while for some other insects the presence of high physiological levels of ROS could also induce diapause and prolong their lifespan. Studies on the regulatory mechanisms of ROS in insect lifespan have achieved a lot of progress in recent years. Therefore, in this article, we comprehensively reviewed the sources and influencing factors of insect ROS, the defense mechanisms mediated by ROS, and for the first time made a summary and outlook on the specific roles of ROS in regulating insect lifespan, so as to provide a reference for subsequent research on ROS-related topics.

 【Aim】 The small brown planthopper Laodelphax striatellus (Fallén), an important rice pest insect, feeds on rice and causes the decrease of yield and quality. Especially the rice stripe virus transmitted by L. striatellus induces more serious loss of rice. The small brown plant hopper is widely distributed and adapts to various environments. This study aims to clarify the molecular adaption mechanisms of L. striatellus under temperature stress preliminarily. 【Methods】 A full-length cDNA encoding Hsp90 from L. striatellus was cloned using RT-PCR and RACE technique. Different bioinformatics methods were used to analyze the characteristics of Hsp90. The expression levels of the Hsp90 gene among different developmental stages and under different temperatures were detected by the real-time PCR. 【Results】 The complete cDNA of LsHsp90 deposited in GenBank accession no. KF660250 is 2 740 bp in length, which encodes a protein of 729 amino acids, with the theoretical isoelectric point of 5.0 and molecular weight of 83.7 kDa. The amino acid sequence contains five signature sequences of Hsp90 family and a C-terminal cytoplasmic character sequence (EEVD). The LsHsp90 carries a classic Hsp90 family structural signature. The phylogenic tree demonstrated that LsHsp90 has high homology with Hsp90 proteins from some other insect species. The real-time PCR analyses exhibited that the expression levels of LsHsp90 varied remarkably in different developmental stages. The mRNA level observed was the highest in the 4th instar nymphs of L. striatellus, the lowest in female adults, and showed significant differences between male and female adults ( P=0.008). The LsHsp90 mRNA level of L. striatellus could be induced by heat and cold temperature stress. The highest LsHsp90 mRNA level was observed at 40℃ and -9℃, respectively. At 40℃, LsHsp90 mRNA level reached a peak at 0.5 h after treatment, and then decreased with treatment time. However, at -4℃, the peak of LsHsp90 mRNA level was observed at 1 h after treatment. 【Conclusion】 The LsHsp90 gene in L. striatellus responds to temperature stress, and it may play an important role in the adaption process of the pest to temperature stress.

As the most diverse group of animals on earth, insects are closely related to human production and activities, making entomological research both theoretically significant and practically valuable. In the past decade, the rapid development of novel methods and technologies has greatly promoted the research progress of entomological research. In this article, we provided a comprehensive overview of the applications of emerging methods and technologies in such fields as entomological morphological identification, molecular mechanism and pest management, focusing on the main contents of this special issue from seven aspects: Micro-computed tomography, RNA interference, gene editing, artificial intelligence-driven intelligent recognition, nanotechnology, regulation of insect-microorganism symbiotes, and olfactory behavior regulation. We also proposed the challenges faced by the large-scale application and sustainable development of these technologies, and prospected the future development trend.

【Aim】 This study aims to investigate the effect of short-time heat shock on the resistance of the silkworm, Bombyx mori to nucleopolyhedrovirus, so as to provide references for further study on the antiviral immune mechanism of the silkworm in response to heat shock. 【Methods】 The 5th instar larvae of B. mori were exposed to 42℃ for 15 min, and then fed with B. mori nucleopolyhedrovirus (BmNPV) (5.4×10 ⁶ PIBs/individual). All the tested larvae were reared as usual. The transcription levels of seven major antimicrobial genes ( BmcecD, Bmmor, Bmglve2, BmdefeB, Bmatta1, Bmenbo2 and Bmlebo3) and six heat shock protein genes ( Bmhsp90, Bmhsp70, Bmhsp40, Bmhsp19.9, Bmhsp21.4 and Bmhsp25.4) in the midgut of the 5th instar larvae after heat-shock treatment were analyzed by quantitative real-time PCR. The amino acid sequences of all BmHSPs were also phylogenetically analyzed using MEGA 5.0. 【Results】 Short-time heat shock significantly increased the survival rate of the BmNPV-infected 5th instar larvae of B. mori, as compared with the nonheat shock group. Quantitative real-time PCR results showed that the expression levels of Bmglve2 and Bmhsp25.4 were significantly up-regulated in the midgut of the BmNPV-infected 5th instar larvae of B. mori exposed to heat shock in comparison with the non-heat shock groups. The phylogenetic analysis showed that BmHSP25.4 was special in evolution, and individually classified into the same cluster with large HSPs (BmHSP90, BmHSP70 and BmHSP40). 【Conclusion】 Short-time heat shock before BmNPV feeding can enhance the resistance of B. mori to BmNPV. Bmhsp25.4 and Bmglve2 may relatively function in the immunity induction of B. mori against BmNPV through heat shock response.

Using two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and image analysis system, the changes of the proteins from later stages (after the shortening stage) of embryo of silkworm Bombyx mori were analyzed to discover the protein changing pattern during embryo development. A total of 209 specific protein spots were found in the 2D-PAGE pattern of embryos at the head thorax differentiation, reverse, tubercle appearance,head pigmentation, body pigmentation and hatch stages. Among them, the specific protein spots expressed in the head pigmentation stage and body pigmentation stage embryo contributed the largest quantity, namely 55 and 77 respectively. Similar to the changing pattern of the specific protein spots of embryo at the earlier stages, most of these specific protein spots disappeared in the embryos shortly afterwards. This suggested that those specific proteins might be related to the corresponding body characteristics in embryo development.

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【Aim】 Riptortus pedestris is a major soybean pest which feeds juice in the stages of adult and nymph, causing serious damage to production. However, R. pedestris has been rarely researched, that leads to the lack of genomic resources. The objective of this study is to obtain the antennal transcriptome data of R. pedestris and to seek new methods for pest control through olfaction. 【Methods】 The adult antennal transcriptome of R. pedestris is sequenced using Illumina HiSeq ^TM 4000 platform and analyzed bioinformatically. 【Results】 In total, 45 802 812 clean reads with 6.87 Gb were obtained (GenBank accession no.: SRR4429103). There were 92 259 unigenes with the mean length of 618 bp and an N50 of 1 013 bp. A total of 21 365 unigenes were annotated based on seven databases. By further analyzing the transcriptome data, we identified 219 chemoreception-related genes including 188 for olfactory receptors (ORs), 6 for gustatory receptors (GRs), 2 for ionotropic receptors (IRs), 4 for sensory neuron membrane proteins (SNMPs), 8 for odorant-binding proteins (OBPs) and 11 for chemosensory proteins (CSPs). The analysis of amino acid sequence indicated that RpedOBP1 and RpedOBP2 have three additional conserved cysteine residues immediately after the sixth cysteine and belong to the plus-C OBP family. 【Conclusion】 This study acquired the antennal transcriptom data of R. pedestris and identified olfaction-related genes. The results provide the important information data of molecular biology for the control of R . pedestris using olfaction-related gene targets.

Insects, the largest group of multicellular organisms on the Earth, are notable for their numerous species and varied morphology, generally harboring abundant microbes in their digestive tract. These microorganisms help the host in digestion, nutrition absorption, pheromone synthesis, as well as playing a role in the defense of harmful invaders, which strengthen the host immune activity. Lepidoptera is the second largest order in Insecta, with many common pest and beneficial insect species. Many methods and techniques have been developed for the research on the gut microbiota, which becomes a hot research topic in recent years. This review outlines the research of gut microbiota in lepidopteran insects ( e.g., Bombyx mori, Spodoptera littoralis and Plutella xylostella), including gut environment, microbial diversity and research methods. Furthermore, the community structure of several lepidopteran model insects and the roles of gut microbes in host’s detoxification and immune system are summarized. The findings we review here should be valuable for the further research on the gut microbiota in lepidopteran insects, and pave the way for developing novel strategies to control pests and protect beneficial insects.

【Aim】 Imidaclorprid is the most widely used pesticide among neonicotinoid pesticides and acts on the brain’s nicotinic acetylcholine receptor of honey bees. In addition, imidaclorprid interferes with the growth and development of honey bees. This study aims to clarify the effects of imidacloprid at field realistic doses on learning and memory of honey bees, so as to provide evidence for the prediction of the widespread death of colonies in some areas and reference for the safe use of neonicotinoid pesticide in the field. 【Methods】 Newly emerged 1-day-old bees ( Apis mellifera ligustica) were marked by paint marker. They were kept in the colony for 8 d, and then were reared within boxes (50 individuals/box) in an incubator with constant temperature and humidity (30±1℃, relative humidity 40%±10%, dark) for 9 d. Meanwhile, the treatment group fed 30% (w/v) syrup containing 0.01 ng/μL of imidacloprid ad libitum, while the control group fed 30% (w/v) syrup containing 0.01 ng/μL of acetone ad libitum. Following being trained with three paired stimulations (lemon odor paired with sucrose stimulation) performed using a customized device, the 18-day-old bees were used for the olfactory associative learning and memory experiments. 【Results】 There was no significant difference in the mortality between the treatment group and the control group during the 9-day feeding period ( P＞0.05). In the three olfactory associative learning experiments, the learning ability of bees of the treatment group in the 2nd and 3rd experiments was significantly reduced ( P＜0.01) compared with the control group, but there was no difference in the 1st experiment ［proboscis extension response (PER)%=0］. After 24 h, the proboscis extension reflex rate was not significantly different between the treatment and control groups ( P＞0.05). 【Conclusion】 The results demonstrate that 0.01 ng/μL imidacloprid does not cause acute death of honey bees. This dose of imidacloprid does not affect the 24 h long-term memory of honey bees, but impairs learning ability of honey bees significantly and may even adversely affect the foraging behavior of honey bees.

【Aim】To elucidate the repellent activities of three plant-derived compounds neochamaejasmine A, thymol and azadirachtin against adult Tribolium confusum and their effects on its olfaction-related genes. 【Methods】 The relative electroantennogram (EAG) response values of T. confusum adults to 0.005, 0.05, 0.5, 5 and 50 μg/μL neochamaejasmine A, thymol and azadirachtin were measured. The olfactory behavior responses of T. confusum adults to neochamaejasmine A, thymol and azadirachtin at the concentrations of 0.5, 5 and 50 μg/μL were studied by Y-tube olfactometer. Impregnated filter paper method was used to determine the repellent rates of 20, 30, 40, 50 and 60 mg/L neochamaejasmine A, thymol and azadirachtin to T. confusum adults. Transcriptomes of the head of T. confusum adults treated with median lethal concentration (LC₅₀)(28.37 mg/L) neochamaejasmine A, LC₅₀(51.05 mg/L) thymol and LC₅₀(42.20 mg/L) azadirachtin were sequenced and comparatively analyzed using RNA-Seq. 【Results】The relative EAG response value of T. confusum adults was proportional to the concentrations of neochamaejasmine A, thymol and azadirachtin. At the maximum concentration of 50 μg/μL, neochamaejasmine A, thymol and azadirachtin resulted in the relative EAG response values of T. confusum adults of 1.61, 2.69 and 2.34 mV, respectively, and the selection rates of 86.48%, 73.89% and 78.33%, respectively. The repellent rates (74.67% and 80.82%, respectively) of 60 mg/L thymol to T. confusum adults were the highest at 2 and 4 h, and the repellent grades reached grades Ⅳ and Ⅴ, respectively. After treatment with thymol, neochamaejasmine A and azadirachtin, there were 569, 597 and 661 differentially expressed genes in the head transcriptomes of T. confusum adults, respectively, 412, 434 and 498 differentially expressed genes were annotated with GO function, respectively, and the differentially expressed genes were identified to 26, 33 and 32 KEGG pathways, respectively, compared with the control group. Thirteen, four and five olfaction-related genes with differential expression were found after treatment with thymol, neochamaejasmine A and azadirachtin, respectively. 【Conclusion】In this study, the repellent activities of the three plant-derived compounds to T. confusum adults were determined and the head transcriptome of T. confusum adults was sequenced. It is speculated that the differentially expressed genes may play an important role in T. confusum repelling the three plant-derived compounds.

【Aim】 With the extensive application of chemical pesticides, the small brown planthopper,  Laodelphax striatellus has increased its resistance to pesticides so that the control effect of pesticides on L. striatellus is weakened. Therefore it is crucial to explore a new method for controlling this pest. In this study, we attempt to inhibit the abundance of symbiotes inside L. striatellus by fungicides to weaken the vitality of this pest, so as to provide a theoretical foundation for controlling L. striatellus through inhibiting yeast-like symbiont (YLS). 【Methods】 The yeast-like symbiont was isolated from L. striatellus via in vitro culture and the susceptibility of the isolated YLS to different concentrations of fungicides was examined. The fungicides with apparently higher ability to inhibit the growth of YLS were applied on wheat seedlings used for feeding L. striatellus. The mortality and body weight of L. striatellus adults were measured and the variation of the amount of YLS inside L. striatellus was examined through real-time florescent quantitative PCR. 【Results】 A strain of YLS was successfully isolated and identified as  Pichia anomala based on the corresponding carbon assimilation experiment and 18S rDNA phylogenic analysis. Denaturing gradient gel electrophoresis (DGGE) result further confirmed the existence of this strain in L. striatellus. The test results of sensitivity to fungicides indicated that 70% propineb and 50% tebuconazole+25% trifloxystrobin had obvious inhibitory effects on the growth of P. anomala while fluopicolide+ propamocarb hydrochloride（687.5 g a.i./L） and 40% pyrimethanil had relatively less inhibitory effects on the growth of P. anomala.When two fungicides (70% propineb and 50% tebuconazole +25% trifloxystrobin with obvious inhibitory effect were applied on wheat seedlings at the concentration of 800 mg/L, the mortality of L. striatellus adults feeding the treated wheat seedlings reached 46.7% and 63.3%, respectively, which was apparently higher than that of the control group, and the body weight of L. striatellus adults in the treatment group was also less than that of the control group. Real-time florescent quantitative-PCR results indicated that the amount of YLS of three strains ( Hypomyces chrysospermus, Pichia guilliermondii, and P. anomala) inside L. striatellus dropped sharply after different fungicides were applied. 【Conclusion】 The results suggest that the fungicides applied in vitro on  L. striatellus  have inhibitory effects on their YLS, thus impacting the survival of L. striatellus, and this validates the feasibility of “reduction in use of pesticides through use of fungicides” strategy in controlling L. striatellus.

 The emerging epidemics of obesity and diabetes have been recognized as major public health problems worldwide, and the primary etiology is an elevation of blood glucose and lipid levels resulting from an imbalance in energy availability and expenditure. Numerous reports have underscored that insects can be used as in vivo model organisms for human metabolic disorders, such as identification of evolutionarily conserved hormones (such as insulin-like peptide and adipokinetic hormone), signaling networks (such as target of rapamycin signaling pathway), and analogous organs or tissues (such as midgut and fat body) that regulate carbohydrate and lipid metabolism in arthropods and mammals. Here, we reviewed the regulatory mechanism of carbohydrate and lipid metabolism in insects, which involves the physiological function of the fat body and oenocytes, the antagonism between insulin-like peptide and adipokinetic hormone on hemolymph glucose regulation, the insulin and insulin-like growth factor signaling pathway (IIS) participated in nutrient metabolism and the cholesterol metabolism associated with steroid hormone synthesis, and also summarized the recent findings on Drosophila genes related with carbohydrate and lipid metabolism. This review will provide reference information for insect physiology and contribute to a better understanding of human metabolic disorders.  

【Aim】 This study aims to screen and verify stably expressed genes under given conditions as reference genes for quantitative real-time PCR (qRT-PCR) in Conogethes punctiferalis, so as to provide the basis for quantitative studies of genes of this moth. 【Methods】 Based on transcriptomics sequencing results in C. punctiferalis and reported reference genes in other insect species, six candidate genes including β-actin gene ( ACT), glyceraldehyde 3-phosphate dehydrogenase gene ( GAPDH), ribosomal protein 49 gene ( RP49), alpha tubulin gene (α-tub), ribosomal protein L13 gene ( RPL13) and vacuolar-type ATPase gene (V-ATPase) were cloned, and their expression levels in different developmental stages and different adult tissues were measured by qRT-PCR. Then the stabilities of these candidate genes were evaluated by a series of programs including ΔCt method, GeNorm, NormFinder, BestKeeper and an online program, RefFinder. Finally, the stabilities of selected reference genes were validated with an olfactory receptor co-receptor gene ( Orco) and pheromone binding protein 1 gene ( PBP1). 【Results】 Analyzed by four programs including ΔCt method, GeNorm, NormFinder and BestKeeper, similar rankings of six candidate genes were obtained, among which RP49, RPL13 and GAPDH were the most stable genes in different developmental stages and different adult tissues, and ACT was ranked as the least stable gene despite of experimental conditions by all programs. Comprehensive analysis with RefFinder further showed that in different adult tissues, RPL13 was the most stable gene, followed by RP49, and in different developmental stages, RP49 was the most stable gene, followed by GAPDH. Additionally, the optimal number of reference genes was calculated by GeNorm as 2. Finally, the stabilities of selected reference genes were validated with Orco and PBP1 as target genes. The results indicated that when two pairs of genes, RPL13 and RP49, and RP49 and GAPDH, were respectively used as reference genes, Orco and PBP1 showed reliable expression patterns, which were consistent with life habits of C. punctiferalis and the results of the previous research. The expression patterns of Orco and PBP1, however, were irregular when ACT was used as the reference gene. 【Conclusion】 In C. punctiferalis, two pairs of genes, RPL13 and RP49, and RP49 and GAPDH, are recommended as reference genes in different adult tissues and different developmental stages, respectively.

【Aim】 In the biological control of Myzus persicae, Aphidius gifuensis is widely used in production practice due to its significant parasitic advantages. However, in the process of large-scale breeding of this parasitoid wasp species, after many generations of breeding, the parasitization ability is weakened and the body size becomes smaller. This study aims to clarify the physiological and biochemical degradation mechanism of the parasitization ability of A. gifuensis by measuring and analyzing the changes in parasitization ability on M. persicae and contents and activities of growth and development-related enzymes of A. gifuensis reared in laboratory for 12 generations. 【Methods】 A. gifuensis collected in the field was randomly selected, and reared for five generations (1st, 3rd, 6th, 9th and 12th generations) in groups indoors. The parasitization ability ［parasitism rate and body size (hind tibia length)］, and the contents and activities of growth and development-related enzymes including carboxylesterase (CarE), phenoloxidase (PO), superoxide dismutase (SOD) and trehalase in A. gifuensis adults of different generations were measured. 【Results】 The parasitism rate and hind tibia length of adult offspring of A. gifuensis were significantly affected by the indoor-rearing generations. The parasitism rate and hind tibia length of the F₁₂ generation of A. gifuensis adults were decreased by 36.70% and 38.54%, respectively, as compared to those in the F₁ generation. Indoor-rearing generations significantly affected the contents of growth and development-related enzymes in adults of A. gifuensis offspring. The contents of CarE, PO, SOD and trehalase in adults of the F₁₂ generation of A. gifuensis were decreased by 54.00%, 43.13%, 35.82% and 45.87%, respectively, meanwhile the activities of PO and SOD in adults of the F₁₂ generation of A. gifuensis showed no significant change, as compared to those in the F₁ generation. Additionally, the contents of CarE, PO, SOD and trehalase in adult offspring of A. gifuensis were significantly positively correlated with the parasitism rate, the CarE content and CarE activity in adult offspring of A. gifuensis were also significantly positively correlated with the hind tibia length of A. gifuensis. 【Conclusion】 With the increase of indoor-rearing generations, the parasitism rate and the body size of the adult offspring of A. gifuensis generally decline, and the contents of the growth and development-related enzymes and CarE activity in the adult offspring of A. gifuensis exhibit significant change.

【Aim】 To understand the bacteria species in saliva of adults of the brown planthopper, Nilaparvata lugens， by identifying the bacterial proteins in their saliva. 【Methods】 Saliva of N. lugens adults was collected by stretching two layers of Parafilm with sucrose diet. Protein solution was subjected to electrophoresis after concentration by ultrafiltration. Protein identification was conducted by liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) analysis, and the spectra were searched against the Uniprot database of bacterial protein to identify the bacterial proteins in saliva. 【Results】 Thirty five proteins of twenty-two species of bacteria, mainly related to energy metabolism, protein folding and synthesis and amino acid metabolism, were identified in saliva of N. lugens adults. These bacteria belong to Proteobacteria, Actinobacteria and Firmicutes, and most are from Proteobacteria. 【Conclusion】 The endosymbiont bacteria in N. lugens, whose proteins were identified, may play an important role in the life history of this insect.

【Aim】To develop an efficient mating disruption technique for Helicoverpa armigera and to explore the changes of the calling and mating behaviors, number of eggs laid and egg hatching rate of H. armigera under high-dose sex pheromone environment. 【Methods】 In the laboratory wind tunnel, an active aerosol dispenser was set up to release high-dose sex pheromones at regular intervals. Four treatments were provided, including the complete sex pheromone blend of H. armigera (Z11-16∶Ald∶Z9-16∶Ald=10∶1) and single sex pheromone components Z11-16∶Ald, Z11-16∶OH and Z9-16∶Ald. The complete sex pheromone blend of H. armigera was sprayed once at the intervals of 20, 10 and 5 min, and the spraying interval of the three single sex pheromone components was 5 min. The total duration of sex pheromone spraying treatment was 12 h (from 16:00 to 4:00 of the next day). The calling and mating behaviors of female H. armigera moths, the number of eggs laid and their hatching rates were observed and compared. 【Results】In the high-dose complete sex pheromone blend environment, the calling onset time of female H. armigera moths was delayed in both day-old age and circadian rhythm, and the proportion of female adults extending ovipositor also decreased, as compared with those in the control (in the environment without sex pheromone). The duration of ovipositor extrusion of female moths significantly differed only at the 5-day-old when the complete sex pheromone blend was sprayed at the intervals of 10 and 5 min, but the onset time of ovipositor extrusion of female moths significantly differed at the 1-day-old when the complete sex pheromone blend was sprayed at the intervals of 20, 10 and 5 min. The average mating rate in the control group without sex pheromones was 84.0%±2.1%, as compared with that (5.0%-17.5%) in the complete sex pheromone blend treatment group. The onset age of mating in the complete sex pheromone blend treatment group was also delayed by 1-5-2-day-old as compared with that in the control group. In the experiment of single-component sex pheromone tests, the mating rate of the control group was 88.0%±3.7%, while the single sex pheromone components Z11-16∶Ald and Z11-16∶OH had significant effects on the mating rates, which were suppressed to only 14.0%±4.0% and 14.0%±2.4%, respectively, while the inhibitory effect of Z9-16∶Ald on the mating rate was minor, with the mating rate of 73.3%±3.3%, and  Z11-16∶Ald delayed the onset age of mating by 1.5-day-old, and the effect of Z11-16∶OH and Z9-16∶Ald on the onset age of mating was not obvious. Both the complete sex pheromone blend and single sex pheromone components significantly inhibited the number of matings of female moths (resulted in reduction by 1.3-1.5 times). In the high-dose sex pheromone environment, the egg-laying time of female moths was significantly delayed by 2 d and the average number of eggs laid by mated female moth decreased, but the egg hatching rate of the mated female moths showed no significant change as compared with those in the control. 【Conclusion】The high-dose sex pheromone environment significantly inhibited the calling and mating behaviors of female moths of H. armigera, which further confirmed the autodetection ability of female moths to sex pheromones. By inhibiting calling and mating, sex pheromones delay the mating and oviposition day-old age of female moths of H. armigera, resulting in decreases in the number of eggs laid and hatching rate. The synthetic sex pheromone released by the active aerosol not only prevents the behavioral orientation response of male moths of H. armigera to female moths, but also affects the calling of female moths, and inhibits and delays the mating with male moths, resulting in a decrease in the number of egg laid. Therefore, it may have the potential for higher control efficacy when used in the field.

【Aim】 To identify the ionotropic receptor genes IR8a and IR25a ， and the representative iGluRs and IRs genes in four vector mosquito species of medical importance， Anopheles sinensis， Anopheles gambiae， Aedes aegypti and Culex quinquefasciatus， and to investigate the characteristics and phylogenetic relationships of these genes， so as to determine the classification position of the IR8a and IR25a genes in the group of ionotropic glutamate receptor genes. 【Methods】 By using BLASTP and BLASTN searching against the genomes of these four mosquito species with the iGluRs and IRs gene sequences of Drosophila melanogaster as query sequences， the sequences of iGluRs and IRs genes of these four mosquito species were identified. The characteristics of the amino acid sequences of IR8a and IR25a and the representative iGluRs and IRs genes identified were predicted and comparatively analyzed by bioinformatics methods. The phylogenetic relationships of these genes in the five species were deduced and discussed by using maximum likelihood， Bayesian and maximum parsimony methods based on amino acid sequences， and the selection pressures of these genes were analyzed with the dN/dS(ω) values calculated by using PAML software. 【Results】 The sequences of IR8a and IR25a， 24 representative traditional iGluRs genes and 30 representative IRs genes in mosquitoes An. sinensis， An. gambiae， An. aegypti and Cx. quinquefasciatus were identified by bioinformatics methods. The comparative analyses of characteristics and phylogenetic relationships of these genes showed that the amino acid sequence lengths of IR8a and IR25a in the four mosquito species and D. melanogaster are closer to those of iGluRs， and their mean values are significantly longer than those of other IRs. IR25a has the aminoterminal domain (ATD) that is the characteristic of traditional iGluRs， and IR8a also has a region similar to ATD， although it is not conservative， while IRs have no ATD. IR8a and IR25 also have the conserved S1 and S2 ligand-binding domain (LBD)， the characteristics of iGluRs， but IRs do not have it. The dN/dS(ω) values of all these genes were far less than 1， suggesting the purifying selection in evolution， while the dN/dS(ω) values of IR8a and IR25a were closer to those of iGluRs. In addition， like non-NMDA， IR8a and IR25a have specific sites which do not exit in IRs. IR8a and IR25a cluster together with non-NMDA to form a sister group， with the genetic distance between the sequences within the group less than that of any other pairs of sequences in this study. Based on these results， IR8a and IR25a were classified into the non-NMDA receptors in iGluRs， and named as Putative receptors. 【Conclusion】 This study developed the information frame of iGluRs and IRs genes of mosquito species and determined the classification position of IR8a and IR25a genes， which is of significance for further functional study of these genes.

【Aim】 This research is designed to conduct transcriptomic analysis of the differentially expressed genes (DEGs) of Ascosphaera apis stressing larval guts of Apis mellifera ligustica via trend analysis. 【Methods】 The purified A. apis spores at a concentration of 1×10 ⁷spores/mL was used to feed 3-day-old larvae of A. m. ligustica， and then the cDNA of stressed larval guts was sequenced at Illumina HiSeq 2500 platform. After filtration， the clean reads were used to mapping the ribosome database and the reference genome of A. m. ligustica， and the unmapped reads were used to mapping the reference transcriptome of A. apis assembled previously. The STEM software was used to analyze the gene expression patterns. Gene ontology (GO) enrichment analysis for DEGs involved in significant expression patterns was performed using WEGOsoftware. KEGG enrichment analysis for DEGs associated with significant expression patterns was carried out by using Blastall. Finally， RT-qPCR analysis of six randomly selected DEGs was performed to validate the RNA-seq data. 【Results】 The RNA-seq of A. apis produced 25 454 076 raw reads， and after filtration， 24 909 820 clean reads with Q30 above 93.46% were obtained. Trend analysis results showed that 19 893 DEGs were grouped into eight gene expression patterns， among which 12 151 DEGs were assigned to three expression patterns with significantly up-regulated expression trend. GO enrichment analysis results indicated that all DEGs within significantly up-regulated expression patterns were enriched in 40 GO terms， and the mostly enriched one was cellular process (2 601 unigenes)， followed by metabolic process (2 553 unigenes) and cell (2 522 unigenes). KEGG enrichment analysis result displayed that all DEGs within significantly up-regulated expression patterns were enriched in 119 metabolism pathways， and the mostly enriched one was ribosome (213 unigenes)， followed by biosynthesis of amino acids (154 unigenes) and protein  processing in endoplasmic reticulum (130 unigenes). Furthermore，it was found that 48 DEGs were enriched in MAPK signaling pathway， and the cluster result suggested that the expression levels of these DEGs increased as the stressing time prolonged. RT-qPCR results demonstrated that the expression patterns of the six DEGs were consistent with those of RNA-seq data， confirming that our transcriptome data are credible.【Conclusion】 The findings in this study not only provide the key information for uncovering the pathogenesis of A. apis at the molecular level， but also lay a foundation for clarifying the pathogen-host interaction in A. m. ligustica under the stress of A. apis.

【Aim】 To investigate the molecular characteristics of the endocuticle structural glycoprotein gene (TmAbd4) and its role in endocuticle formation in Tenebrio molitor, and so as to provide a theoretical basis for screening the key target genes involved in the growth and development of T. molitor. 【Methods】 The full-length cDNA sequence of TmAbd4 was obtained based on the transcriptome database of T. molitor, and the functional domains of TmAbd4 were analyzed using bioinformatics tools. RT-qPCR was employed to detect the expression levels of TmAbd4 in different tissues (integument, fat body, Malpighian tubules, foregut, midgut and hindgut) of the 13th instar mature larvae and in the integument of the 1-15-day-old mature larvae of T. molitor. After the silence of TmAbd4 in the 13th instar mature larvae of T. molitor by RNAi, the microstructure of the integument was observed using hematoxylin and eosin staining (H&E), and the ultrastructure of the integument was observed through transmission electron microscopy (TEM). 【Results】 The full-length cDNA sequence of TmAbd4 of T. molitor was 506 bp, with an open reading frame (ORF) of 396 bp, encoding 131 amino acids. TmAbd4 contained a signal peptide, a chitin-binding domain 4 (ChtBD4), and a conserved RR-1 motif, classifying it within the RR-1 subclass of the CPR family. TmAbd4 was specifically highly expressed in the integument, and was significantly highly expressed in the integument of the first 3 d after the molting of the 13th instar mature larvae. Injection of dsTmAbd4 caused no visible abnormal phenotypes in T. molitor from molt to pupal stage, however, the endocuticle was observed significantly thinner compared to the dsGFP-injected control group. 【Conclusion】TmAbd4 belongs to the RR-1 subclass of the CPR family and is highly expressed in the integument of T. molitor, with significantly higher expression during endocuticle formation. Silencing of TmAbd4 did not impair the ability of the larvae to molt into the pupal stage, however, it led to a significant reduction in endocuticle thickness, indicating that TmAbd4 involves the formation of endocuticle.

【 Aim 】 Circular RNA (circRNA) plays a key role in alternative splicing, transcription regulation and expression regulation of source genes. This study aims to analyze the quantity, variety, structural characteristics and function of circRNAs in the midgut of Apis mellifera ligustica workers, and to explore the regulatory function of circRNAs via constructing and analyzing regulatory networks. 【 Methods 】 A. m. ligustica workers were reared under laboratory conditions, and the midgut samples from 7 - and 10 - day - old workers were subjected to deep sequencing using circRNA - seq technology. CircRNAs were predicted from sequencing data after quality control using find_circ software. Source genes of these circRNAs were annotated to GO and KEGG databases to gain function and pathway annotations via BLAST. The target mRNAs of circRNAs and miRNAs were predicted with TargetFinder software, and the regulatory networks between circRNAs and miRNAs and between circRNAs, miRNAs and mRNAs were constructed and visualized using Cytoscape v.3.2.1software. The predicted circRNAs were validated by RT - PCR with the divergent and convergent primers designed. 【 Results 】 Sequencing of midgut samples from A. m. ligustica workers produced a mean of 136 463 071 clean reads, and for each sample over 136 779 122 anchor reads were obtained after removing rRNA. A total of 10 833 circRNAs were predicted, and their length ranged mainly from 15 to 1 000 nt. The types of these circRNAs were abundant, and the largest one was annotated to be exonic circRNA. The number of circRNAs distributed on chromosome 1 of Apis mellifera was the most and that distributed on chromosome 8 was the second. The source genes of circRNAs could be annotated to 45 GO terms including binding, cellular process, and cell, as well as 121 KEGG metabolic pathways including endocytosis, protein processing in endoplasmic reticulum, and ribosome, suggesting that circRNAs may play key roles in such biological processes as growth, development, metabolism, and cellular activity in the midgut of A. m. ligustica workers. Furthermore, the regulatory networks between circRNA and miRNA and between circRNA, miRNA and mRNA were constructed, and the analysis result demonstrated that partial circRNAs could bind microRNAs as competitive endogenous RNAs (ceRNAs). Finally, the true existence of randomly selected three circRNAs was validated by RT - PCR. 【 Conclusion 】 In this study, we predicted, analyzed, and verified circRNAs in the midgut of A. m. ligustica workers. Our findings provide the data of the quantity, variety, structural characteristics, roles, and regulatory network of circRNAs in the midgut, indicating that circRNAs can play roles in the developmental and immune defense processes in the midgut of A. m. ligustica workers through affecting source genes and acting as ceRNAs. This study lays a foundation for further studying the roles of circRNAs in the development and stress response of the midgut of A. m. ligustica.

The tachinid fly, Nemorilla maculosa Meigen (Diptera: Tachinidae) is an important parasitoid of the beet webworm, Loxostege sticticalis L. (Lepidoptera: Pyralidae) in mainland China. Females of N. maculosa lay heavy-shelled macrotype eggs on the last-stadium host larvae of L. sticticalis and other lepidopterous larvae. Little has been studied and understood so far in the host species and instar preference, and parasitic position of N. maculosa. Studies were therefore designed to investigate and understand the host preference of this parasitoid under laboratory conditions (22℃, L16∶D8). The results indicated that N. maculosa could parasitize the beet webworm L. sticticalis, the beet armyworm Spodoptera exigua Hübner (Lepidoptera:Noctuidae), and the oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae) when the final instar of the three host larvae were presented simultaneously. However, the parasitism rate for L. sticticalis was significantly higher than that for the beet armyworm, which in turn was significantly higher than that for the oriental armyworm. Besides, the number of tachinid eggs per host larva received also followed the trend of parasitism rate in these three species, indicating that N. maculosa was more likely to select L. sticticalis larvae as its host. The parasitoid mainly selected the last instar larvae as its host when 3rd, 4th, and 5th instar larvae of L. sticticalis were mixed together. The parasitism rates for these three instars of larvae were increased as the larval instar increased. The tachinid eggs were mainly located on the thorax (66.4%), followed by on the head (23.4%) and the abdomen (10.9%) of the host larvae. The number of tachinid eggs on the thorax was significantly greater than that on the head and abdomen of the host larvae, while the egg numbers between the head and abdomen were insignificantly different. Mechanisms underlying the host instar, position and species preference of N. maculosa were finally discussed.

【Aim】 Pyridoxal-5′-phosphate (PLP) functions as a coenzyme in many cellular processes including the interconversion and catabolism of amino acids. Pyridoxal kinase (PLK) is a key enzyme related to VB ₆ metabolism. This study aims to reveal the relationship of transcription regulation between the kinase of PLP and the apo-B6 enzymes. 【Methods】 Three siRNAs (siRNA1, siRNA2 and siRNA3) based on PLK gene sequence were designed and synthesized, and injected to the day-3 5th instar larvae of the silkworm, Bombyx mori. The expression levels of phosphoserine aminotransferase (SerB) and asparate aminotransferase (AST) genes were detected by real-time quantitative PCR, to explore their interference effects on PLK replication. 【Results】 The best interference efficiency was achieved at 48 h after injection. The interference efficiency of the three fragments from high to low was siRNA1, siRNA2 and siRNA3. The interference efficiency was different in different tissues, and that in the midgut was the highest with the relative expression level of the PLK gene decreased by 55%. The relative expression levels of SerB and AST genes were also decreased by 90% and 29%, respectively, after RNA interference of PLK gene. 【Conclusion】 This study successfully down-regulated the PLK gene in B. mori  larvae. The results further demonstrate that phosphoserine aminotransferase (SerB) and asparate aminotransferase (AST) genes are related to the pyridoxal kinase gene.

【Aim】 This research aims to evaluate the fumigation activities of the essential oils (EOs) from Cinnamomum caphora fruits, Cinnamomum pedunculatum leaves and Sabina chinensis var. chinensis leaves, and their mixtures against the adults of the housefly, Musca domestica, so as to provide theoretical basis for developing botanical insecticides. 【Methods】 The fumigation and knock-down toxicities of these EOs extracted by steam distillation and their mixtures against the 5-day-old adults of M. domestica were detected with the erlenmeyer flask method. 【Results】 The median lethal concentration (LC ₅₀) values of the three EOs against M. domestica adults were 5.28, 16.86, and 14.54 μg/cm ³, respectively, and the median knock-down time (KT ₅₀) values in the treatment of LC ₉₀ were 12.03, 16.56, and 13.37 min, respectively. The fumigation toxicity of EO from C. pedunculatum leaves against M. domestica adults showed strong synergistic effect when it was mixed with EO from S. chinensis var. chinensis leaves at the LC ₅₀ value ratio of 9∶1, with the co-toxicity coefficient (CTC) as high as 367.95. 【Conclusion】 The EO from C. caphora fruits has strong fumigation and knock-down toxicities against M. domestica adults, followed by EO from S. chinensis var. chinensis leaves and then EO from C. pedunculatum leaves. Mixtures of EO from C. pedunculatum leaves and S. chinensis var. chinensis leaves have synergistic effect on their fumigation toxicities against M. domestica adults. The three EOs have the potential to be developed as environmental insecticides.

【Aim】The aim of this study is to explore the function of the ferritin1HCH (Fer1) gene HvFer1 in the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata, and to provide a theoretical basis for the biological control of H. vigintioctopunctata. 【Methods】The amino acid sequence of HvFer1 was subjected to blastp analysis followed by multiple alignment with Fer1 sequences from different species. The expression levels of HvFer1 in different developmental stages (egg, 1st-4th instar larvae, pupa, and female and male adults) and different tissues (head, midgut, fat body, cuticle and Malpighian tubules) of the 4th instar larvae of H. vigintioctopunctata were detected by RT-qPCR. HvFer1 was silenced by injecting 200 ng/individual of dsHvFer1 into the 3rd instar larvae of H. vigintioctopunctata. Then the larval phenotype, survival rate and duration from the 4th instar larva to pupa, as well as the expression level of dual oxidase (Duox) gene HvDuox, the level of reactive oxygen species (ROS), the content of malondialdehyde (MDA), and bacterial diversity in the midgut were observed and determined. 【Results】The coleopteran Fer1 sequences clustered into one clade, and HvFer1 was the most closely related to Fer1s of Harmonia axyridis and Coccinella septempunctata. The expression level of HvFer1 was the highest in male adults and the lowest in the 4th instar larvae of H. vigintioctopunctata. The expression level of HvFer1 in the midgut of the 4th instar larvae of H. vigintioctopunctata was significantly higher than those in the other tissues. Compared with the control group (dsGFP injection), injection of 200 ng/individual of dsHvFer1 into the 3rd instar larvae of H. vigintioctopunctata significantly prolonged the duration from the 4th instar larva to pupa, and prevented the formation of some spines and spots in pupae. The elyta of the emerged adults of H. vigintioctopunctata were abnormally developed after dsHvFer1 injection, with spots fused or deficient. The expression level of HvFer1 in the 3rd instar larvae of H. vigintioctopunctata on the 2nd day post injection of dsHvFer1 showed no significant difference from that in the control group, but the expression level of HvFer1 on the 4th day post injection of dsHvFer1 was significantly lower than that in the control group. Compared with the control group, HvFer1 silencing caused severe oxidative stress in H. vigintioctopunctata larvae, significantly up-regulated the expression level of HvDuox, ROS level and MDA content in the larval midgut, changed the structural composition of midgut bacteria, reduced the relative abundance of Enterobacter and Chryseobacterium, and also increased the relative abundance of Serratia, Pseudomonas, Acinetobacter and other intestinal bacteria.【Conclusion】 HvFer1 plays a crucial role in regulating the larval development, adult phenotype, and maintaining the larval gut immunity and microbial homeostasis of H. vigintioctopunctata. HvFer1 is expected to be a candidate target gene for controlling H. vigintioctopunctata based on RNAi technology.

Benzoxazinoids which exist in most Gramineae plants, with 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one （DIMBOA） as the representative, show broad-spectrum of biological activities. Researches on benzoxazinoids were initialized in 1950－1960's. Recently their biological functions have been much further elucidated due to application of molecular biology technique and the theoretical development of plant induced resistance. The present paper reviews the most recent advances in researches on benzoxazinoids. Benzoxazinoids are secondary metabolites branching out from the tryptophan synthesis pathway. The concentrations of benzoxazinoids vary with plant tissues and ages; and benzoxazinoid accumulation and hydrolyzation of benzoxazinoid glucosides can be induced by feeding of herbivores and infection of pathogens. Benzoxazinoids have many bioactivities, such as pest resistance, phytopoison, pathogen resistance, allopathic effects, etc.

【Aim】 Calliptamus italicus is a major plague pest in the arid and semi-arid grasslands of Xinjiang, with a lack of information of its reproduction related genes. The objective of this study is to reveal the characteristics of the transcriptome of C. italicus and to obtain the gonada development related genes in C. italicus through the transcriptome sequencing of its testicular and ovarian tissues. 【Methods】 The transcriptome of C. italicus was sequenced using the Illumina HiSeq/MiSeq platform and bioinformatically analyzed. 【Results】 A total of 718 872 unigenes were obtained, with a mean length of 631 bp and an N50 of 1 186 bp. Through a similarity search, 254 597 unigenes were annotated. Most unigenes (28.87%) were annotated to Nr database, and the unigenes of C. italicus had the highest similarity (10.80%) with those of Lasius niger annotated in Nr database. According to GO database, all unigenes were broadly annotated into three categories: 31 392 unigenes to be related to molecular function, 20 586 unigenes related to cellular component, and 57 014 unigenes related to biological processes. In KEGG database, a total of 23 666 unigenes were assigned to 251 metabolic pathways, of which many pathways are involved in the reproductive system development, including oocyte meiosis, insulin signaling pathway, Wnt signaling pathway, GnRH signaling pathway, TGF-β signaling pathway, ubiquitin mediated proteolysis, progesterone-mediated oocyte maturation, and insect hormone biosynthesis. By further screening and identification, gonadal development related genes of C. italicus, including genes of vg, vgr, sox 21b, testis-specific serine/threonine-protein kinase, spermatogenesis-associated protein and vasalike gene, were obtained. 【Conclusion】 This study acquired the transcriptome data and gonadal development related genes of C. italicus, providing a molecular foundation for further studying the mechanism of reproductive regulation of C. italicus.