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  • Monthly, Founded in 1950
    Supervisor:Chinese Academy of Sciences
    Sponsor:Institute of Zoology,Chinese Academy of Sciences
    The Entomological Society of China
    Domestic postal code: 2-153
    Foreign issuance code: Q61
    ISSN 0454-6296
    CN 11-1832/Q
Table of Content
20 January 2021, Volume 64 Issue 1
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  • RESEARCH PAPERS
    Differences in the accumulation and allocation of nutrients in the long-winged and short-winged male adults of Velarifictorus aspersus (Orthoptera: Gryllidae)
    ZENG Yang, ZHANG Bin, HE Yi-Yuan, ZHAO Xin, ZHU Dao-Hong
    2021, 64(1):  1-9.  doi:10.16380/j.kcxb.2021.01.001
    Abstract ( 561 )   PDF (1671KB) ( 130 )   PDF(mobile) (1671KB) ( 47 )     
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     【Aim】 For wing polymorphic insects the energy investment of males in reproduction is different from that of females, and this difference may lead to changes in the physiological mechanism of the trade-off between flight and reproduction. To test this speculation, we investigated the accumulation and allocation of nutrients in wing-dimorphic male adults of Velarifictorus aspersus. 【Methods】 We compared the temporal changes in body weight between the long-winged (LW) and short-winged (SW) male adults of V. aspersus with similar head width within 7 d after emergence. The contents of protein, total lipids and glycogen in the whole body, and flight and reproductive organs in the wing-dimorphic male adults were determined by spectrophotometry. 【Results】 The body weight of the LW male adults of V. aspersus remained unchanged within the first 7 d after emergence, while that of the SW male adults increased significantly. The protein contents in the LW males and SW males on the day of adult emergence were 46.5±2.7 and 42.3±1.9 mg, respectively, showing significant difference between the two wing morphs. In addition, the protein content increased faster in the LW male adults than in the SW male adults thereafter. The contents of total lipids in the LW and SW males showed no significant difference on the day of adult emergence, but the contents of total lipids in the SW males were significantly higher than those in the LW males on the 5th and 7th days after adult emergence. Within 7 d after adult emergence, the contents of proteins and total lipids in flight muscles of the LW males were significantly higher than that of the SW males. On the day of adult emergence, there was no significant difference in the contents of proteins and total lipids between the accessory glands of the LW and SW males. However, the contents of proteins and total lipids in the accessory glands of the SW males were significantly higher than those of the LW males from the 3rd day after adult emergence. 【Conclusion】 The results suggest that the trade-off between flight and reproduction can influence the accumulation and allocation of nutrients in wing-dimorphic male adults of V. aspersus.
    Immunolocalization and ligand binding characteristics of the odorant binding protein AplaOBP3 of Agrilus planipennis(Coleoptera: Buprestidae)
    SONG Xuan, WANG Ze-Hua, SHAN Shuang, ZHANG Yong-Jun, WANG Shan-Ning
    2021, 64(1):  10-18.  doi:10.16380/j.kcxb.2021.01.002
    Abstract ( 355 )   PDF (12948KB) ( 102 )   PDF(mobile) (12948KB) ( 154 )     
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    【Aim】 The aim of this study is to explore the localization of the odorant binding protein AplaOBP3 in antennae of Agrilus planipennis and its ligand binding characteristics, and to compare the function of AplaOBP3 with the reported AplaOBP1 and AplaOBP2. 【Methods】 The recombinant AplaOBP3 of A. planipennis was expressed in prokaryotic expression system. The polyclonal antibody against AplaOBP3 was prepared and detected by Western blot assay. Immunocytochemical technique was used to localize the expression of AplaOBP3 in antennal sensilla of A. planipennis. Its binding characteristics to 58 compounds were determined by fluorescence competitive binding assay, and compared with those of AplaOBP1 and AplaOBP2 previously reported. 【Results】 The recombinant protein AplaOBP3 was successfully expressed in prokaryotic expression system. The immunolocalization results showed that AplaOBP3 was expressed in the antennal sensilla basiconica type I. The results of fluorescence competitive binding assay showed that AplaOBP3 had strong binding capabilities with trans-2-hexenal, benzaldehyde, 4′-ethylacetophenone, β-ionone and ocimene, with the dissociation constant KD values of 6.20, 4.03, 5.24, 1.72 and 4.83 μmol/L, respectively. AplaOBP3 and AplaOBP2 had similar expression and ligand binding properties. 【Conclusion】 AplaOBP3 and AplaOBP2 have similar expression and ligand binding properties, and both may be involved in olfactory perception together and play a role in host localization in A. planipennis.
    Expression, purification and characterization of the cuticular proteins TdCPR12611 and TdCPR7854 from Trypoxylus dichotomus (Coleoptera: Scarabaeidae)
    YE Chang-Qing, BAO Han, LIU Tian, YANG Qing
    2021, 64(1):  19-29.  doi:10.16380/j.kcxb.2021.01.003
    Abstract ( 518 )   PDF (6437KB) ( 94 )   PDF(mobile) (6437KB) ( 12 )     
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    【Aim】 To explore the sequence characteristics and biochemical properties of cuticular proteins from Trypoxylus dichotomus. 【Methods】 RT-PCR was used to clone cuticular protein genes of T. dichotomus, and the structural features and phylogeny of cuticular proteins were analyzed by bioinformatics methods. Recombinant cuticular proteins of T. dichotomus were expressed in Escherichia coli expression system, and purified by metal-chelating affinity chromatography. In vitro binding experiments were performed to detect the binding ability of cuticular proteins of T. dichotomus with chitins including α-chitin, β-chitin, chitosan and colloidal chitin. Liquid-liquid phase separation (LLPS) was observed and determined. 【Results】 Two cuticular protein genes TdCPR12611 (GenBank accession no.: MT813021) and TdCPR7854 (GenBank accession no.: MT813022) of T. dichotomus were cloned and obtained. Phylogenetic analysis results showed that TdCPR12611 is closely related to OtCP-1 from Onthophagus taurus, while TdCPR7854 is closely related to OtCP-acp20-1, OtCP-acp20-2, and OtCP-acp20-3 from O. taurus, all of which belong to the CPR_RR-2 family. Recombinant TdCPR12611 and TdCPR7854 proteins were expressed by prokaryotic expression and purified. The two recombinant proteins had different binding abilities with four types of chitins, among which 41.4% of TdCPR12611 could bind with chitosan, while 62.3% of TdCPR7854 could bind with β-chitin. TdCPR12611 was predicted to have a relatively disordered structure and could spontaneously coacervate at room temperature to form liquid-liquid phase separation, while TdCPR7854 could not. 【Conclusion】 In this study we assayed and analyzed the sequence characteristics and chitin binding properties of two RR-2 cuticular proteins, TdCPR7854 and TdCPR12611 of T. dichotomus. The results not only deepen our understanding of insect cuticle assembly mechanism, but also provide ideas for the development of protein biomimetic materials.
    Analysis of the functions of allatostatin genes in the reproductive diapause preparation of Colaphellus bowringi (Coleoptera: Chrysomelidae)
    TIAN Zhong, LIU Xi, ZHU Li, LIU Wen, ZHU Fen, WANG Xiao-Ping
    2021, 64(1):  30-40.  doi:10.16380/j.kcxb.2021.01.004
    Abstract ( 479 )   PDF (4340KB) ( 95 )   PDF(mobile) (4340KB) ( 10 )     
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    【Aim】 This study aims to clarify the molecular characteristics and expression difference of allatotropin (AT) and allatostatin (AST) genes during the reproduction and diapause of Colaphellus bowringi, so as to further reveal the functions of these genes in the reproductive diapause preparation of C. bowringi. 【Methods】 We identified the sequences of CbAT and CbASTs in C. bowringi based on the previously constructed transcriptome database, cloned their ORFs and carried out the bioinformatics analysis of their sequences. Then, we assayed the expression patterns of CbAT and CbASTs in the heads of the diapause-destined and non-diapause-destined female pupae and female adults via RT-PCR. RNAi experiments of CbAST-B, CbAST-C and CbAST-B+CbAST-C were done in the diapause-destined 2-day-old female pupae, and the expression levels of juvenile hormone (JH) signaling genes and vitellogenin genes in 4-day-old female adults were detected. 【Results】 One allatotropin gene CbAT (GenBank accession no.: MT977128) and two allatostatin genes CbAST-B (GenBank accession no.: MT977126) and CbAST-C (GenBank accession no.: MT977127) of C. bowringi were identified and cloned, with the ORFs of 408, 600 and 303 bp, respectively. By amino acid sequence alignment and phylogenetic analysis, we found that CbAT, CbAST-B and CbAST-C respectively cluster into the same branch with homologous proteins of Coleoptera insects, showing high conservation in the evolution. The qRT-PCR assay indicated that CbAT had no significant expression differences in the female heads between the diapause-destined and non-diapause-destined individuals from the 2-day-old pupal stage to the 4-day-old adult stage. CbAST-B and CbAST-C were significantly highly expressed in the heads of diapause-destined female adult since the 1- and 2-day-old, respectively, and the differential expression state between the diapause-destined and non-diapause-destined individuals lasted until the end of diapause preparation period. After knocking down CbAST-B, CbAST-C and CbAST-B+CbAST-C in the diapause-destined 2-day-old female pupae, the expression of CbAST-B and CbAST-C in the heads of 4-day-old female adults was significantly suppressed with the RNAi efficiency over 70%. Further study revealed that the expression of JH biosynthesis genes AACT, FPPS and JHAMT, in the head without antenna, JH-responsive genes Kr-h1 and JHE1 in the fat body, and vitellogenin genes Vg1 and Vg2 in the fat body was all significantly up-regulated in the treatment groups of dsCbAST-B, dsCbAST-C and dsCbAST-B+dsCbAST-C【Conclusion】 CbAT may be not a key regulator mediating the differences of JH signaling between the preoviposition period and diapause preparation phase in C. bowringi. However, CbAST-B and CbAST-C suppress the synthesis of JH and further down-regulate the expression of Vg genes in C. bowringi during the reproductive diapause preparation phase, thereby promoting the occurrence of diapause. This study further reveals an upstream regulation mechanism for the JH signaling in insects during the reproductive diapause preparation phase, being helpful to understand the seasonal adaptation strategy of insects and providing new guidance and reference for potential target genes for pest control.
    Cloning, prokaryotic expression and tissue expression analysis of the silk gland transcription factor gene AaSGF-1 of Antheraea assama (Lepidoptera: Saturniidae)
    LI Qiong-Yan, CHEN An-Li, XUN Li-Jie, LIU Zeng-Hu, LIAO Peng-Fei, YANG Wei-Ke, DONG Zhan-Peng
    2021, 64(1):  41-50.  doi:10.16380/j.kcxb.2021.01.005
    Abstract ( 420 )   PDF (4602KB) ( 101 )   PDF(mobile) (4602KB) ( 13 )     
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    【Aim】 This study aims to clone the silk gland transcription factor gene AaSGF-1 from Antheraea assama, to analyze its sequence features and expression pattern, and to obtain the polyclonal antibody, so as to lay a basis for further studying the function of this gene. 【Methods】 The cDNA sequence of AaSGF-1 was cloned from the silk gland of A. assama by RT-PCR and RACE techniques and subjected to bioinformatical analysis. qPCR was employed to analyze the expression profile of this gene in different tissues (head, midgut, fat body, silk gland, hemolymph, and cuticle) of the day-4 5th instar larvae of A. assama. Prokaryotic expression plasmid was constructed and the fusion protein AaSGF-1 was expressed in Escherichia coli BL21. The polyclonal antibody with high serum titer was prepared using the purified fusion protein to immunize New Zealand rabbit. The protein level of AaSGF-1 in the silk gland and cuticle of the newly hatched larva and the silk gland of the 4th instar larva of A. assama was detected by immunofluorescence technique. 【Results】 The cDNA sequence of AaSGF-1 (GenBank accession no.: MK889510.1) of A. assama was cloned. The ORF of AaSGF-1 is 1 050 bp in length, encoding a polypeptide of 349 amino acids with the molecular weight of 38.8 kD and the isoelectric point (pI) of 8.74. The qPCR analysis results showed that AaSGF-1 was highly expressed in the silk gland of the 5th instar larvae of A. assama, especially in the posterior silk gland, but hardly expressed in other tissues. Immunofluorescence assay showed that AaSGF-1 was expressed in silk glands of the newly hatched larva and 4th instar larva of A. assama. 【Conclusion】 In this study AaSGF-1 was expressed by prokaryotic expression system, the polyclonal antibody was prepared, and AaSGF-1 was confirmed to be highly expressed in the silk gland of A. assama larva, providing a basis for further studying its roles in silk gland development and silk protein synthesis in A. assama.
    Mdogg1 is involved in maintaining redox balance in Musca domestica
    ZHANG Yu-Ming, SHAO Meng-Hua, LI Ya-Jing, FENG Qin, WEI Li-Ya, LIU Feng-Song
    2021, 64(1):  51-60.  doi:10.16380/j.kcxb.2021.01.006
    Abstract ( 402 )   PDF (10230KB) ( 79 )   PDF(mobile) (10230KB) ( 11 )     
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    【Aim】 The objective of this study is to clone and identify the 8-hydroxyguanine glycosylase 1 gene from Musca domestica (Mdogg1), so as to verify its function in regulating redox balance. 【Methods】 Based on the previous transcriptome data of M. domestica, the cDNA sequence of Mdogg1 was cloned by RT-PCR and then subjected to bioinformatics analysis. The expression levels of Mdogg1 at different developmental stages (egg, 1st-3rd instar larva, pupa and adult), in different tissues (hemocyte, muscle, gut, and fat body) of the 3rd instar larvae and in the 2nd instar larvae of M. domestica after different stresses [exposed to 2.5-100 mmol/L CdCl2 for 24 h, soaked in 0.1 g/L doxorubicin hydrochloride (DOX) solution for 30 min and then recovered for 6, 12 and 24 h, and exposed to 280-315 nm ultraviolet (UV) at the dose of 5 J/cm2 for 5-30 min, respectively] were detected by qRT-PCR. The expression of Mdogg1 in the 2nd instar larvae of M. domestica was knocked down by RNAi, and the changes in the malondialdehyde (MDA) content, reactive oxygen species (ROS) level, superoxide dismutase (SOD) activity, and 8-oxoguanine (8-oxoG) content were determined. 【Results】 The full-length cDNA sequence of Mdogg1 (GenBank accession number: AYK27449.1) was obtained. It is 1 062 bp in length, encoding 353 amino acid residues, with the theoretical molecular weight of 41.08 kD and the isoelectric point of 9.02. The amino acid sequence of MdOGG1 includes a HhH (Helix-hairpin-Helix) motif and the conserved endonuclease III domain ENDO3c domain. The qRT-PCR results showed that Mdogg1 was highly expressed in the egg stage and the fat body of the 3rd instar larvae of M. domestica. When the 2nd instar larvae of M. domestica were exposed to 2.5-100 mmol/L CdCl2, the expression level of Mdogg1 increased first and then decreased, and the 8-oxoG content increased. The expression levels of Mdogg1 in the 2nd instar larvae soaked in 0.1 g/L of DOX solution for 30 min and then recovered for 12 and 24 h, and those exposed to UV for 30 min, were significantly up-regulated as compared to that in the untreated control. After knockdown of Mdogg1 in the 2nd instar larvae of M. domestica by RNAi, the ROS level, MDA content and 8oxoG content in M. domestica larvae increased significantly as compared to the control (dsGFP injection group), while the SOD activity decreased significantly. 【Conclusion】 Mdogg1 is involved in maintaining redox balance in M. domestica, playing an important role in protecting the insect from oxidative damage.
    Expression of the fungal-resistance factor BmSPI39 in Bombyx mori in response to Beauveria bassiana invasion
    LI You-Shan, LU Qing-Jun, YANG Xi, ZHANG Jie, LUO Zhu-Xing, XIA Qing-You, ZHAO Ping
    2021, 64(1):  61-69.  doi:10.16380/j.kcxb.2021.01.007
    Abstract ( 484 )   PDF (8264KB) ( 96 )   PDF(mobile) (8264KB) ( 15 )     
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    【Aim】 To prepare polyclonal antibody of the fungal-resistance factor BmSPI39 in Bombyx mori and to analyze its expression in response to Beauveria bassiana invasion. 【Methods】 Prokaryotic expression and immobilized nickel ion affinity chromatography were used to obtain the highly-purified recombinant BmSPI39 protein. The inhibitory activity of the recombinant BmSPI39 protein to Bacillus subtilis protease A was detected by in-gel activity staining. Polyclonal antibodies to BmSPI39 were prepared by immunizing New Zealand white rabbit with the recombinant BmSPI39 protein. Based on the open database SilkDB 3.0 of B. mori, the spatiotemporal expression characteristics of BmSPI39 in the immune-related tissues, integument, midgut and fat body, of B. mori from the day-3 4th instar larva to 1 day-old pupa were analyzed. And the expression of BmSPI39 in response to B. bassiana invasion in the integument of B. mori larvae and pupae at different time after B. bassiana infection was assayed using the prepared BmSPI39 antibodies by Western blot. 【Results】 The results of in-gel activity staining showed that the recombinant BmSPI39 protein could strongly inhibit the activity of subtilisin A. Enzyme-linked immunosorbent assay (ELISA) showed that the antibody titer of BmSPI39 antiserum reached 1∶128 000. The results of Western blot and in-gel activity staining showed that the purified BmSPI39 antibody could effectively bind to the BmSPI39 antigen protein in different multimerization states. The heat map analysis of the expression data of BmSPI39 showed that BmSPI39 was expressed in the integument of B. mori at the wandering stage and the pre-pupal stage, with a higher expression level in the fat body at the pre-pupal stage, but not in the midgut at all the tested developmental stages. The results of Western blot showed that the expression of BmSPI39 in the integument of B. mori larvae was up-regulated at 84 h and down-regulated at 108 and 120 h after B. bassiana infection. 【Conclusion】 Polyclonal antibody to BmSPI39 was successfully prepared in this study. The expression of BmSPI39 protein can respond to B. bassiana invasion and may be involved in the process of resisting fungal invasion in the pupal stage of B. mori.
    Biological characteristics of Asobara japonica (Hymenoptera: Braconidae) parasitizing Drosophila melanogaster and the effects of its parasitization on the host growth and immune responses
    ZHANG Xian, ZHOU Si-Cong, CHEN Jia-Ni, PANG Lan, ZHANG Qi-Chao, WANG Ying, SHI Min, CHEN Xue-Xin, HUANG Jian-Hua
    2021, 64(1):  70-79.  doi:10.16380/j.kcxb.2021.01.008
    Abstract ( 384 )   PDF (3451KB) ( 96 )   PDF(mobile) (3451KB) ( 36 )     
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    【Aim】 This study aims to investigate the biological characteristics of the parasitoid Asobara japonica parasitizing Drosophila melanogaster and to determine the effects of its parasitization on the host growth and immune responses. 【Methods】The developmental duration and morphological characterastics of A. japonica at different developmental stages, the parasitism rate and emergence rate of A. japonica after parasitization on the 2nd instar larvae of D.melanogaster, the changes of host pupation time and transcriptional levels of 15 important genes of host in different immune pathways (SPE, Toll, Myd88, Dif and Drosomycin involved in Toll pathway, PGRP-LE, PGRP-LC, imd, Relish and Diptericin involved in Imd pathway and Spn27A, MP2, yellow-f2, DoxA2 and PPOinvolved in PO pathway) in the parasitized 2nd instar larvae of D. melanogaster were investigated and analyzed by anatomical imaging and qRT-PCR technology. 【Results】 The developmental duration of egg stage, larval stage and pupal stage of A. japonica under the conditions of 25±1℃, 50%±1% relative humidity and photoperiod of 16L∶8D was 2.38±0.01, 5.36±0.07 and 8.30±0.04 d, respectively. After parasitizing the 2nd instar larvae of D. melanogaster, the parasitism rate and wasp emergence rate of A. japonica were 94.9%±4.0% and 64.3%±7.1%, respectively. Moreover, after being parasitized by A. japonica, the pupation time of D. melanogaster with 50% pupation rate was prolonged by 0.5 d, and the transcriptional levels of the antibacterial peptide genes Drosomycin and Diptericin were up-regulated significantly, whereas that of the prophenoloxidase gene PPOwas down-regulated as compared to the unparasitized control. 【Conclusion】 A. japonica can successfully and effectively parasitize D. melanogaster larvae through triggering host developmental delay and suppressing host melanization immune responses. The results provide some useful information for the massive release programs of the agriculturally important parasitoid species A. japonica. 
    Parasitism and development performance of bisexual and thelytokous lines of Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae) on eggs of different strains of Antheraea pernyi (Lepidoptera: Saturniidae)
    ZHOU Jin-Cheng, HE Yue, ZHAO Qian, DONG Hui
    2021, 64(1):  80-89.  doi:10.16380/j.kcxb.2021.01.009
    Abstract ( 450 )   PDF (1781KB) ( 64 )   PDF(mobile) (1781KB) ( 7 )     
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    【Aim】 To determine the parasitism and development performance of bisexual and thelytokous lines of Trichogramma dendrolimi on eggs of different strains of Antheraea pernyi, so as to provide the basis for the mass-rearing of richogrammatid parasitoids using A. pernyi eggs as the alternative host. 【Methods】 The biological parameters including parasitism rate, brood size of offspring (number of offspring wasps emerged from a single brood), and proportion of female offspring of bisexual and thelytokous lines of T. dendrolimi on eggs of six strains of A. pernyi [Kangda (KD), Dasi (DS), Gaoxin (GX), 988 (NEE), Qingda (QD), and Teda (TD)] were detected, and the quality parameters of eggs of the six strains of A. pernyi (wet weight per egg, protein content, total carbohydrate content and triglyceride content) were determined. The correlation between the quality parameters of A. pernyi eggs and the biological parameters of T. dendrolimi was detected by principal component analysis. 【Results】 The body size of offspring of the bisexual wasps of T. dendrolimi developed on eggs of the NEE strain of A. pernyi were significantly higher than those developed on eggs of the other strains of A. pernyi except for the KD and QD strains. The parasitism rate, brood size of offspring, and proportion of female offspring of the bisexual wasps of T. dendrolimi were not significantly influenced by the strain of A. pernyi. The brood size of offspring of thelytokous wasps of T. dendrolimi developed on eggs of the KD strain of A. pernyi was the highest, being significantly higher than those developed on eggs of the NEE and QD strains of A. pernyi, but without significant difference with those developed on eggs of the DS, GX, and TD strains of A. pernyi. The parasitism rate and brood size of offspring of the thelytokous wasps of T. dendrolimi were not significantly influenced by the strain of A. pernyi. The protein content in eggs of the TD strain of A. pernyi was the highest, significantly higher than those in eggs of the other strains of A. pernyi except for the GX strain. The total carbohydrate content in eggs of the KD strain of A. pernyi and the triglyceride content in eggs of the QD strain of A. pernyi were the highest, significantly higher than those in eggs of the other strains, respectively. The principal component analysis showed that the parasitism rate of bisexual wasps of T. dendrolimi, and the protein content and triglyceride content in A. pernyi eggs were negatively correlated with the total carbohydrate content in A. pernyi eggs. The body size of offspring of the bisexual wasps of T. dendrolimi was negatively correlated with the wet weight per egg of A. pernyi. The parasitism rate of the thelytokous wasps of T. dendrolimi and the triglyceride content in A. pernyi eggs were negatively correlated with the brood size of offspring of T. dendrolimi, the total carbohydrate content, and the wet weight per egg of A. pernyi. The body size of offspring of thelytokous wasps of T. dendrolimi and the total carbohydrate content in A. pernyi eggs were negatively correlated with the protein content in A. pernyi eggs. 【Conclusion】 This study determined the suitable strain of A. pernyi eggs for the mass-rearing procedure of bisexual and thelytokous lines of T. dendrolimi and revealed the relationship between the nutritive indexes of A. pernyi eggs and the biological parameters of T. dendrolimi. The results provide basic data for improving mass-rearing of T. dendrolimi.
    Component analysis of the sugar solution stored in wax cells by Bombus friseanus (Hymenoptera: Apidae)
    SU Rui, QIN Jia-Min, DONG Kun, SONG Wen-Fei, JIA Hong-Wei, LIANG Cheng
    2021, 64(1):  90-98.  doi:10.16380/j.kcxb.2021.01.010
    Abstract ( 264 )   PDF (6009KB) ( 85 )   PDF(mobile) (6009KB) ( 12 )     
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    【Aim】 Bumblebees are important pollinators of many plants. They forage and store floral nectar and pollen as their primary food. This study aims to explore the nutritional requirements of bumblebees and to determine whether they can make sugar solution into honey in the process of feeding food provided by breeders. 【Methods】 B. friseanus populations were fed with the artificial sugar solution of commercial granulated sugar (the sugar concentration 50%), and the sugar solutions were collected from capping cells which had been stored by B. friseanus for 1-7 d as the treatment group samples. Meanwhile, the artificial sugar solutions were placed in sterile centrifuge tubes which bumblebees could not access and used as the control group samples. The pH value, sugar concentration and components, and α-amylase and invertase activities in the treatment and control groups were detected over the storage period. 【Results】 The mean pH value of sugar solution stored in wax cells by B. friseanus for 1-7 d was 3.74±0.13, significantly lower than that in the control group (6.55±0.15). There was a significantly positive correlation between the sugar concentration and storage time of sugar solution stored by B. friseanus. The average sugar concentration in the sugar solution stored in wax cells for 6 d was significantly higher than that stored for 1-3 d. Components of carbohydrates in the sugar solution stored in wax cells by B. friseanus became richer, and sucrose, fructose, glucose, maltose, and trehalose were detected by HPLC analysis. The hexose contents in the sugar solution stored in wax cells for 4-5 d were extremely significantly higher than those stored for 1-3 d and 6-7 d. The hexose content and the maltose content showed highly significantly negative correlation with the sucrose content, respectively. There was an extremely significantly negative correlation between the content of fructose or maltose in the total sugar and the storage time of sugar solution stored by B. friseanus. However, there was an extremely significantly positive correlation between the contents of glucose, sucrose and trehalose and the storage time of sugar solution stored in wax cells, respectively. The α-amylase activity in the sugar solution stored in wax wells for 6 d was significantly higher than those in samples stored for the other storage time, and there was no significant difference in the α-amylase activity in sugar solution in wax wells among the other storage time. The invertase activity in all samples in wax cells varied from 21.17 to 38.05 U/g FW, and showed no significant difference during the storage period. 【Conclusion】 The data proved that bumblebees B. friseanus forage and store artificial sugar solution in wax cells. The sugar solution undergoes physical and biochemical changes after processing by B. friseanus, revealing the honeymaking ability of bumblebees. The results of this study provide a scientific basis for researching bumblebee biology and breeding. 
    Culturing of pupal Wiebesia pumilae (Hymenoptera: Agaonidae) on agar culture medium
    ZHAO Meng, WU Wen-Shan, CHEN You-Ling, WU Ting-Ting
    2021, 64(1):  99-108.  doi:10.16380/j.kcxb.2021.01.011
    Abstract ( 260 )   PDF (4196KB) ( 67 )   PDF(mobile) (4196KB) ( 20 )     
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     【Aim】 The achene of Ficus pumila var. pumila has extremely high nutritional and edible values, and F. pumila depends on the pollination of Wiebesia pumilae so that it can bear fruit. This study aims to explore the possibility of laboratory culture of herbivorous parasitic wasps in pupal stage. 【Methods】 The W. pumilae pupae were cultured on agar culture medium at different temperatures (22, 24, 26, 28, 30 and 32℃) and photoperiods (0L∶24D, 10L∶14D, 12L∶12D, 14L∶10D and 24L∶0D), the developmental morphology and duration of W. pumilae pupae at various stages, the female adult longevity and the water content of galls were determined, and the processes of morphological development of galls were observed. 【Results】 As the temperature increased within the range of 22-28℃, the developmental rate of W. pumilae pupae increased, the emergence time became earlier, but the emergence rate showed no significant difference. The developmental duration of W. pumilae pupae was the shortest at 28℃ in agar culture, and the developmental duration from prepupa to adult were 25.65 d for female and 21.79 d for male, which was 44.00 d shorter than that of female and male fig wasps in the wild. A few W. pumilae pupae could develop to mature pupal stage at 30℃, and W. pumilae prepupae entered into diapause at 32℃, suggesting that 30℃ may be the extreme temperature of heat tolerance for the development of W. pumilae pupae. There were no significant differences in the developmental rate and emergence rate of W. pumilae in the pupal stage under different photoperiods. The water content of galls decreased fast with the temperature increasing within the range of 22-28℃, the longer the culture time, the lower the water content of galls. When W. pumilae wasps cultured on agar culture medium emerged at 28℃, the water content of galls was 41.79%, which was significantly lower than that in the wild (51.63%). The gall shell was very hard due to the low water content, and it is difficult for W. pumilae wasps to dig out the hole for flying out, which may be the main reason for the failure of wasp flying out after emergence in agar culture. 【Conclusion】 The agar culture method of this study provides possibilities for continuous observation of the development of herbivorous parasitic wasps at the pupal stage and acquisition of experimental wasp materials.
    REVIEW ARTICLES
    Molecular mechanisms of expression regulation of insect cytochrome P450 genes involved in insecticide resistance
    ZHU Jiang, QIU Xing-Hui
    2021, 64(1):  109-120.  doi:10.16380/j.kcxb.2021.01.012
    Abstract ( 592 )   PDF (1643KB) ( 204 )   PDF(mobile) (1643KB) ( 39 )     
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    The frequent and extensive use of insecticides leads to the evolution of insecticide resistance. It is well recognized that over-expression of cytochrome P450s involved in insecticide detoxification is one common and major mechanism of resistance to
    various classes of insecticides in insects. However, the molecular mechanisms for overexpression of insecticide resistance-related P450 genes in insects have not been well identified until now. In the recent decade, with the development of biological science and technology, many significant advances have been achieved in this direction. In this article, an overview of the current knowledge with respect to expression regulation of insect cytochrome P450 genes was provided. In addition to gene duplication and amplification, up-regulation of P450s at the transcriptional level can be ascribed to the interaction between the cis-regulatory elements and the trans-acting factors. Transcription factors such as CncC, CREB and AhR directly regulate the expression of P450s. The G-protein-coupled receptors and their downstream effectors play indirect roles in the transcriptional regulation of P450 genes. The universality of the role of CncC:Maf/Keap1 in the expression regulation of P450s is supported by studies on a variety of insect species. Increasing reports confirm the involvement of microRNAs in the regulation of P450 expression. Current findings reveal the diversity of regulatory factors and signal transduction pathways, and the complexity of underlying mechanisms in the regulation of P450 expression.
    Research advances in symbiotic microorganisms in insects and their functions
    WANG Wei-Xia, ZHU Ting-Heng, LAI Feng-Xiang
    2021, 64(1):  121-140.  doi:10.16380/j.kcxb.2021.01.013
    Abstract ( 771 )   PDF (2355KB) ( 317 )   PDF(mobile) (2355KB) ( 70 )     
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    Symbiotic microorganisms can account for 1%-10% of insect biomass, including acteria, fungi, archaea and viruses. Insects and symbiotic microorganisms co-evolve to form holobionts. Symbiotic microorganisms play an important role in the biological characteristics, diversity formation, ecological adaptability and stress resistance of insects. Crop pest insects seriously affect agricultural production. In this article, the research advances in the diversity, research methods, and functional mechanisms of insect symbiotic microorganisms, the interaction between symbiotic microorganisms and their application in pest control since 2000 were reviewed and prospected. With the continuous development and application of advanced research methods such as molecular microbial ecology and metagenomic sequencing, breakthroughs have been made in the research of symbiotic microorganisms of agricultural pest insects. It was found that symbiotic microorganisms mainly affect host insects in the following ways: (1) Synthesis of nutrients or production of digestive enzymes to promote host growth and development and to expand host ecological niche; (2) Production of protective metabolites to directly protect the host against stress or indirectly protect the host by regulating the defense response of host plants; and (3) Production of active substances to regulate host propagation, mating, aggregation and movement. The abundance and community composition of insect symbiotic microorganisms maintain dynamic changes in a certain spatial-temporal range and have an important impact on host phenotype, which is the result of the benefit trade-off among host, environment, and interactive microorganisms. We suggest that future research should focus on clarifying the molecular mechanisms underlying the formation and maintenance of symbionts, ascertaining the complex interactions among symbiotic microorganisms, host insects, plants, natural enemies and the environment in more spatial-temporal dimensions, and designing green and efficient pest control strategies through targeted regulation of pest insect symbionts.
    CONTENTS
    Contents of Vol. 64 Issue 1
    2021, 64(1):  141-141. 
    Abstract ( 286 )   PDF (473KB) ( 33 )   PDF(mobile) (473KB) ( 11 )     
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