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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 July 2023, Volume 66 Issue 7
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  • RESEARCH PAPERS
    Molecular characteristics and functional study of heat shock protein gene SmHsp60 in Sitodiplosis mosellana (Diptera: Cecidomyidae)
    MA Qian, DONG Jin-Hui, LI Fang-Xiang, ZHU Ke-Yan, CHENG Wei-Ning
    2023, 66(7):  859-869.  doi:10.16380/j.kcxb.2023.07.001
    Abstract ( 241 )   PDF (4468KB) ( 290 )   　　
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    【Aim】The wheat blossom midge, Sitodiplosis mosellana, an important agricultural pest, survives under temperature extremes during summer and winter by diapause. This study aims to explore the relationship between heat shock protein 60 (Hsp60) gene expression and diapause development and temperature tolerance in the diapause process of S. mosellana.【Methods】 The full-length cDNA sequence of Hsp60 of S. mosellana (SmHsp60) was amplified via RACE and RT-PCR technologies, and analyzed via bioinformatics. qPCR was used to detect the expression levels of SmHsp60 in the S. mosellana larvae at different stages from pre-diapause to post-diapause development including pre-diapause, diapause, post-diapause quiescence and post-diapause development, as well as oversummering larvae under extremely high temperature stress ［exposed to water bathes at 34, 40, 45 and 50 ℃ for 1 h, and 35 ℃ for 0 (control), 15, 30, 60 and 120 min］ and overwintering larvae under extremely low temperature stress ［exposed to 0, -5, -10 and -15 ℃ for 1 h, and -5 ℃ for 0 (control), 15, 30, 60 and 120 min］. The recombinant SmHsp60 protein was obtained by prokaryotic expression and affinity column chromatography, and its ability to suppress the thermal aggregation of mitochondrial malate dehydrogenase (MDH) at 43 ℃ was examined by colorimetry and SDS-PAGE.【Results】 The full-length cDNA sequence of SmHsp60 (GenBank accession no: KR733065) of S. mosellana obtained is 2 270 bp in length, which contains an open reading frame (ORF) of 1 722 bp in length encoding a protein of 573 amino acids with the relative molecular weight of 60.7 kD. Amino acid sequence analysis indicated that SmHsp60 contains the classical signature sequences of mitochondrial Hsp60, and displayed the highest amino acid identity and the closest relationship to Hsp60 from Contarinia nasturtii of Cecidomyiidae. qPCR detection result showed that the expression level of SmHsp60 did not change significantly in the diapause stage, but began to increase gradually in the post-diapause quiescence stage with a peak in early-to-mid phase of post-diapause quiescence (i.e. December and next January) and was significantly higher than those in the other developmental stages. Compared to the untreated control, the expression of SmHsp60 in the oversummering larvae under 35 and 40 ℃for 1 h and 35 ℃ for 30-60 min and that in the overwintering larvae under -5 ℃ for 1 h was significantly induced, while the expression level of SmHsp60 did not change significantly above 45 ℃ or below -10 ℃. The highly purified recombinant SmHsp60 was able to effectively suppress the thermal aggregation of MDH, indicating its significant molecular chaperone function.【Conclusion】 SmHsp60 is involved in diapause regulation of S. mosellana and might play a role in diapause termination, as well as heat tolerance and cold tolerance during diapause.
    Gene cloning, expression profiling and ligand binding characteristics of GmolNPC2 from Grapholita molesta (Lepidoptera: Tortricidae)
    LI Chun-Qin, ZHANG Yu-Xi, XU Shi-Cai, LI Bo-Liao, CHEN Xiu-Lin, LUO Kun, LI Guang-Wei
    2023, 66(7):  870-884.  doi:10.16380/j.kcxb.2023.07.002
    Abstract ( 176 )   PDF (8087KB) ( 262 )   　　
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    【Aim】The purpose of this study is to provide a theoretical reference for the further study on the function of the Niemann-Pick type C2 protein (NPC2) of Grapholita molesta GmolNPC2 in olfactory sensing.【Methods】Based on the antennal transcriptome data of G. molesta, the full-length cDNA sequence of GmolNPC2 was amplified by RT-PCR, and the phylogenetic analysis and 3D structure model prediction of GmoNPC2 were performed. The relative expression levels of GmolNPC2 in different developmental stages (egg, 1st-5th instar larvae, pupa, and female and male adults) and different tissues (antenna, head without antenna, thorax, abdomen, leg, and wing) of the 3-day-old male and female adults were detected by RT-qPCR. The inhibitory constant (Ki) values of the recombinant GmolNPC2 to 44 odorant ligands including six sex pheromones and 38 host plant volatile compounds were determined by fluorescence competitive binding assays to analyze the binding ability of GmolNPC2 to odorant ligands. By using molecular docking simulations, the binding energy of GmolNPC2 with different odorant ligands was calculated and the key amino acid residues of protein-ligand interactions were predicted.【Results】The full-length cDNA sequence of GmolNPC2 (GenBank accession no.: OQ054801) of G. molesta was obtained and the open reading frame (ORF) is 438 bp in length, encoding 145 amino acids. Multiple sequence alignment showed that GmolNPC2 has the typical structural characteristics of NPC2 of insects. Phylogenetic analysis showed that the reported NPC2s of lepidopteran insects were clustered into two branches, and the GmolNPC2 sequence was highly identic to the NPC2 sequences of Leguminivora glycinivorella, Pectinophora gossypiella, and Manduca sexta, which were clustered into one branch. RT-qPCR detection result showed that the expression levels of GmolNPC2 in the male adult and egg were significantly higher than those in other developmental stages. The expression level of GmolNPC2 was the highest in the wing of the 3-day-old male and female adults followed by that in the antenna, and the lowest in the thorax, abdomen, and leg. The binding spectrum of the recombinant GmolNPC2 was narrow with only 17 out of 44 tested odorant ligands, among which, the recombinant GmolNPC2 exhibited strong binding abilities to (Z)-8-dodecenyl acetate, benzyl alcohol, and cis-3-hexenyl acetate, with the Ki values of (4.117±0.046), (4.845±0.079) and (3.979±0.167) μmol/L, respectively. The molecular docking simulation result showed that the binding energy between GmolNPC2 and each odorant ligand was different, which was consistent with the results of the above fluorescence binding assays. Hydrophobic amino acid residues and hydrogen bonds play an important role in GmolNPC2 binding with different odorant ligands.【Conclusion】 GmolNPC2 has the conserved structure of the insect NPC2 family and is highly expressed in the egg and adult, and the wing and antenna of adult, speculating that GmolNPC2 participates in different physiological processes in G. molesta. The recombinant GmolNPC2 selectively binds sex pheromones and host plant volatile compounds, indicating the involvement of GmolNPC2 in the detection and transportation of volatile semiochemicals.
    Gene cloning, polyclonal antibody preparation, and expression pattern of m6A methyltransferase in Apis mellifera (Hymenoptera: Apidae)
    WU Ying, LIU Zhi-Tan, ZHAO Hao-Dong, GUO Si-Jia, LIU Xiao-Yu, ZHANG Yi-Qiong, FENG Pei-Lin, ZHAO Hong-Xia, XU Xi-Jian, CHEN Da-Fu, FU Zhong-Min, GUO Rui ,
    2023, 66(7):  885-895.  doi:10.16380/j.kcxb.2023.07.003
    Abstract ( 120 )   PDF (7888KB) ( 108 )   PDF（mobile） (7888KB) ( 5 )   　　
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    【Aim】 This study aims to offer reference and foundation for continuous and further study on RNA m6A methyltransferase 14 (METTL14) gene of Apis mellifera AmMETTL14 through cloning AmMETTL14, analyzing the physicochemical properties and molecular characteristics of AmMETTL14, detecting the expression pattern of AmMETTL14 in different developmental stages and various tissues of workers, and preparing the polyclonal antibody to AmMETTL14. 【Methods】 The CDS of AmMETTL14 in A. mellifera was amplified by PCR followed by Sanger sequencing validation. Bioinformatic analysis and construction of phylogenetic tree of AmMETTL14 were conducted using relevant bioinformatics software. Inducible expression of AmMETTL14 fusion protein was performed through prokaryotic expression system, followed by immunization of New Zealand white rabbits to prepare the polyclonal antibody, and the titer and specificity of the antibody were evaluated by ELISA and Western blot, respectively. RT-qPCR was used to detect the relative expression level of AmMETTL14 in the egg, larva, prepupa, pupa and adult worker, and seven tissues including antenna, venom gland, brain, midgut, hypopharyngeal gland, fat body and cuticle of the newly emerged adult worker.【Results】 The CDS of AmMETTL14 of A. mellifera was successfully cloned. The molecular formula of AmMETTL14 is C1957H3113N567O589S15, the molecular weight is about 44.49 kD, the liposoluble coefficient is 79.97, the theoretical isoelectric point is 8.58, and the average hydrophilic coefficient is -0.59. AmMETTL14 contains no typical transmembrane domain and signal peptide, and is simultaneously located to the nucleus, mitochondria and cytoplasm. METTL14s in A. mellifera, A. cerana cerana, A. laboriosa, A. cerana, A. dorsata, A. florae, Bombus terrestris, Vespa mandarinia, B. impatien and Drosophila melanogaster contain the structural domain MT-A70 and three same conserved motifs. AmMETTL14 and METTL14 of A. laboriosa had the highest amino acid sequence identity (65.60%) and the closest relationship. The prepared polyclonal antibody to AmMETTL14 had high titer (more than 512 K) and strong specificity. The expression levels of AmMETTL14 in the 7- and 8-day-old prepupae and eggs were similar and significantly higher than that in the 3-day-old larva, that in the 12-day-old pupa was significantly higher than those in the egg, larva and prepupa, and those in the 6-, 12- and 15-day-old adult workers were significantly higher than that in the 1-day-old adult worker. Additionally, the expression levels of AmMETTL14 in the brain and hypopharyngeal gland were similar to that in the antenna, and significantly higher than those in the venom gland, midgut, fat body and cuticle of the newly emerged adult worker. 【Conclusion】 This study result indicates that AmMETTL14 may be a hydrophilic, non-transmembrane and intracellular protein, AmMETTL14 plays potential regulatory role in larval growth and development, and the prepared polyclonal antibody to AmMETTL14 has high titer, sensitivity and specificity, laying a foundation for further exploring the function of AmMETTL14 and the involved epigenetic regulation mechanisms.
    Screening and expression profiling of highly expressed genes in the hypopharyngeal glands of Apis mellifera (Hymenoptera: Apidae) foragers
    ZHANG Juan, GAO Yan, WANG Ruo-Hong, LI Qiu-Fang, YANG Shang-Ning, ZHOU Shi-Wen, SHI Dan-Dan, SU Song-Kun, NIE Hong-Yi
    2023, 66(7):  896-908.  doi:10.16380/j.kcxb.2023.07.004
    Abstract ( 162 )   PDF (5484KB) ( 181 )   　　
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    【Aim】 This study aims to screen genes highly expressed in the hypopharyngeal glands (HGs) of Apis mellefera foragers, and to provide a basis for further studying the genes related to honey bee foraging behavior.【Methods】 Differentially expressed genes (DEGs) in the HGs were screened based on the previously sequenced HGs transcriptome data of the 3-day-old workers (3 d_W), 10-day-old nurses (10 d_N), 10-day-old foragers (10 d_F), 21-day-old nurses (21 d_N), 21day-old foragers (21 d_F) and the DEGs highly expressed in the HGs of the 10-day-old foragers and 21-day-old foragers of A. mellefera were subjected to GO and KEGG enrichment analysis. Meanwhile, the expression levels of four highly expressed DEGs (LOC409889, LOC406114, LOC408453 and LOC100578735) in the HGs of foragers in different developmental stages (egg, larva, pupa, newly emerged workers, 10-day-old nurses and 21-day-old foragers) and tissues (abdomen, thorax, antenna, mandibular gland, hypoharyngeal gland, sting, leg, gut, wing and brain) of the 21-day-old foragers of A. mellefera were examined using qPCR.【Results】Comparative transcriptome result showed that the expression levels of 164 up-regulated DEGs in the HGs of the 21-day-old foragers were obviously increased as compared with those of the 3-day-old workers, 10-day-old nurses and 21-day-old nurses, and those of the 10-day-old foragers were also higher than those of the 10-day-old nurses. While the expression trend of the 105 down-regulated DEGs was opposite with that of the above 164 DEGs. GO analysis result showed that the 164 highly expressed DEGs in the HGs of the 10-/21-day-old foragers were enriched significantly in sulfate transmembrane transporter activity, sulfur compound transmembrane transporter activity, iron ion binding and oxidoreductase activity, and the 105 down-regulated DEGs were enriched significantly in ribosome, intracellular ribonucleoprotein complex and ribonucleoprotein complex. KEGG pathway analysis result revealed that the 105 down-regulated DEGs were significantly enriched in two pathways ribosome, and starch and sucrose metabolism. qPCR detection result indicated that the expression levels of LOC409889, LOC406114, LOC408453 and LOC100578735 were the highest in the 21-day-old foragers, and those of LOC406114 and LOC100578735 were the highest in the HGs of the 21-day-old foragers and showed low expression level or no expression in other tissues.【Conclusion】 The highly expressed DEGs identified in the HGs of A. mellifera forager were screened in this study via comparative transcriptome, which provides a valuable source of data for the function study of forager HGs, and also provides a new perspective for further studying the foraging behavior of A. mellefera and the molecular breeding selection of bee species with strong foraging ability.
    Transcriptomic analysis of Acanthoscelides obtectus (Coleoptera: Chrysomelidae) in response to fumigation with ethyl formate and methyl isothiocyanate
    JIANG Zhao-Chun, YANG Lu, ZHANG Yue, YANG Hong, DAI Ren-Huai
    2023, 66(7):  909-917.  doi:10.16380/j.kcxb.2023.07.005
    Abstract ( 121 )   PDF (3593KB) ( 370 )   　　
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    【Aim】This study aims to preliminarily clarify the joint action mechanism of ethyl formate (EF) and methyl isothiocyanate (MITC) to Acanthoscelides obtectus through analyzing the transcriptome data of A. obtectus fumigated with EF, MITC and EF+MITC.【Methods】The A. obtectus adults were treated with fumigation of EF(22.398 μL/L), MITC(0.854 μL/L) and EF+MITC(14.764 μL/L) through wild-mouth bottle fumigation method. The control group (CK) was not fumigated. Transcriptome sequencing of the A. obtectus adults treated with fumigation of EF, MITC and EF+MITC was performed with Illumina Hi SeqTM 4000 sequencing platform. GO classification and KEGG enrichment analysis were performed on differentially expressed genes (DEGs). Six DEGs were selected for RT-qPCR validation.【Results】 There were 171 and 293 DEGs in the CK vs EF and CK vs MITC comparative groups, respectively, and most of them were up-regulated. However, the number of DEGs in the CK vs EF+MITC comparative group was 1 745, and most of them were down-regulated. GO classification of DEGs showed that DEGs in the  CK vs EF and CK vs MITC comparative groups were mainly enriched in cellular anatomical entity, binding, and catalytic activity, while those in the CK vs EF+MITC comparative group were mainly enriched in cellular anatomical entity, binding, and cellular process. KEGG pathway enrichment analysis of DEGs showed that DEGs in the CK vs EF comparative group were mainly enriched in pathways of protein digestion and absorption, lysosome and protein processing in endoplasmic reticulum, those in the CK vs MITC comparative group were mainly enriched in pathways of focal adhesion, ECM-receptor interaction and insect hormone biosynthesis, and those in the CK vs EF+MITC comparative group were mainly enriched in RNA transport, DNA replication and mismatch repair. In addition, the expression levels of two and three detoxification enzyme genes in A. obtectus adults in response to EF and MITC fumigation treatments were significantly down-regulated, respectively, however, the expression levels of five detoxification enzyme genes in A. obtectus adults in response to EF+MITC fumigation treatment were significantly down-regulated as compared with those in the CK. The expression trends of the six selected DEGs were basically consistent with the transcriptome data.【Conclusion】The results of this study indicate that the combination of EF and MITC has a synergistic effect, which can induce cell genotoxic damage in A. obtectus. The inhibition of expression of detoxification enzyme genes seems to be the main reason for its synergistic effects. In this study, the molecular mechanism of fumigation and insecticidal effect of the mixture of EF and MITC on A. obtectus was preliminarily clarified, which provides an important basis for the further study on the combined action mechanism of EF and MITC on A. obtectus.
    Effects of light intensity on the oviposition preference of Grapholita molesta (Lepidoptera: Tortricidae)
    YANG Xiao-Fan, JIAO Hui-Tan, LI Xu-Zhao, LU Zi-Yun, RAN Hong-Fan, MA Ai-Hong, WEI Guo-Shu, LI Jian-Cheng
    2023, 66(7):  918-924.  doi:10.16380/j.kcxb.2023.07.006
    Abstract ( 184 )   PDF (2596KB) ( 308 )   　　
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    【Aim】Grapholita molesta is an important and frequent pest of fruit trees. This study aims to investigate the roles of light intensity on the oviposition preference of G. molesta, and to further reveal the dim-light vision ability in G. molesta. 【Methods】The numbers of eggs of G. molesta laid on leaves on the sunny side and shaded side in the peach tree canopy were investigated in spring, summer and autumn, and the difference in light intensity between the sunny side and shaded side at sunset was measured. Light-dark two-choice experiments were carried out indoor to test the oviposition preference behavior response of G. molesta adults to light intensity. 【Results】 In spring, summer and autumn, G. molesta adults preferred to lay eggs on leaves on the sunny side in the peach tree capony with significantly higher light intensity, the numbers of eggs laid on the sunny side were 4.24, 9.30 and 5.82 times as high as those on the shaded side, respectively. In the light-dark two-choice experiments, G. molesta adults preferred to lay eggs to light than to dark, with the oviposition selection rates of 87.48%, 83.68%, 82.92%, 80.08% and 84.84%, respectively, under 10 000, 100, 1 and 0.01 lx and natural light conditions. The oviposition preferences between to strong light and to weak light revealed that when the light contrast (strong light/weak light) was 10 and 2, respectively, G. molesta adults showed a preference to strong light under 10 000, 100, 1 and 0.01 lx and natural light conditions. Moreover, this preference to strong light did not decrease significantly with the decrease of light intensity, and the oviposition selection rates were higher than 75%.【Conclusion】 Light intensity can affect the oviposition preference of G. molesta adults. Female adults show obvious oviposition preference to strong light, and can still discriminate well the difference in light intensity even under dim light, exhibiting a well dim-light vision ability. The results provide a basis for new physical trapping techniques based on visual behavior regulation.
    Effects of mating on the ovarian development and fecundity of Conopomorpha sinensis (Lepidoptera: Gracillariidae)
    LI Wen-Jing, DONG Yi-Zhi, YAO Qiong, QUAN Lin-Fa, CHI Yan-Yan, CHEN Bing-Xu
    2023, 66(7):  925-933.  doi:10.16380/j.kcxb.2023.07.007
    Abstract ( 119 )   PDF (3337KB) ( 330 )   　　
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    【Aim】To observe the ovarian developmental progress and formulate ovarian developmental grading criteria of Conopomorpha sinensis, explore the influence of mating on the ovarian developmental progress and fecundity. 【Methods】 The newly eclosed female and male adults of C. sinensis were paired in a 1∶1 ratio and reared. The female ovaries were dissected daily, and the mating experience was judged according to the characteristics of the bursa copulatrix. The characteristics of fat body, length of ovarioles, total number of egg chambers and number of mature eggs of mated and unmated females were recorded for grading the ovarian development progress. The newly eclosed female and male adults paired in a 1∶1 ratio, and newly eclosed female adults that were individually reared were used as the mated group and unmated group, respectively, and the pre-oviposition period, oviposition period, number of eggs laid per female and female adult longevity were counted in the two groups.【Results】 The ovarian development of C. sinensis could be divided into five stages, including yolk deposition prophase (stage Ⅰ), yolk deposition phase (stage Ⅱ), egg maturation phase (stage Ⅲ), peak phase of oviposition (stage Ⅳ) and terminal phase of oviposition (stage Ⅴ). Only the ovaries of the mated female adults could completely develop. Otherwise, the ovaries of the unmated female adults only developed into the yolk deposition phase (stage Ⅱ). The preoviposition period, oviposition period, number of eggs laid per female and female adult longevity in the mated group were 3.82 d, 11.23 d, 176.42 grains and 16.35 d, respectively. The female adults in the unmated group could not lay eggs for life, and the female adult longevity in the unmated group was 20.48 d. 【Conclusion】 The ovarian development of C. sinensis adults can be divided into five stages. However, mating is a prerequisite for the completion of ovarian development of the female adult. The ovarian development of the unmated female adults terminates after reaching stage Ⅱ. The fecundity of the mated female adults is significantly higher than that of the unmated females, and the unmated females can not lay any eggs for life.

    Sequencing of the mitochondrial genome of Stephanitis nashi (Hemiptera: Tingidae) and phylogenetic analysis of Cimicomorpha at the family level
    LIN Xing-Yu, ZHAO Te, SONG Nan
    2023, 66(7):  934-945.  doi:10.16380/j.kcxb.2023.07.008
    Abstract ( 99 )   PDF (10642KB) ( 170 )   　　
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    【Aim】 To explore the structure characteristics of the mitochondrial genome of Stephanitis nashi and the phylogenetic relationships within Cimicomorpha at the family level. 【Methods】 The complete mitochondrial genome of S. nashi was sequenced by using high-throughput sequencing technology. The phylogenetic relationships within Cimicomorpha at the family level, with the mitochondrial genomes of 150 species known to Cimicomorpha as ingroups and those of two Pentatomomorpha species as outgroups, were reconstructed using maximum likelihood (ML) and Bayesian inference (BI) methods. 【Results】 The mitochondrial genome of S. nashi (GenBank accession no.: OP650254) is 15 045 bp in full-length, which contains 37 genes including 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes and a non-coding control region. Among the 13 PCGs, except that the start codon of nad5 is ATC, the start codons of the other PCGs are ATT, ATA or ATG. While cox2 and nad4l employ the incomplete T as the stop codon, the other 11 PCGs terminate with complete stop codons of TAA or TAG. All tRNA genes can be folded into typical cloverleaf structure. The lengths of the rRNA genes rrnL and rrnS are 1 237 and 763 bp, with the A+T content of 81.08% and 82.04%, respectively. Both ML and BI produced a similar phylogenetic relationships within Cimicomorpha: the families Tingidae, Cimicidae, Thaumastocoridae, Miridae, Plokiophilidae, Microphysidae, Velocipedidae, Anthocoridae, Nabidae, Pachynomidae and Phymatidae were monophyletic, respectively. However, the family Reduviidae was non-monophyletic. The Tingidae had close relationships to Thaumastocoridae and part of Reduviidae. 【Conclusion】 This study reconstructed the phylogenetic relationships within the Cimicomorpha based on the mitochondrial genome data, revealing the non-monophyly of the family Reduviidae and the monophyly of the other 11 families, and it will contribute to increasing our knowledge about the phylogeny and the mitogenomics of Cimicomorpha.
    Mitochondrial genomes of Caerulea coeligena (Lepidoptera: Lycaenidae) and Dodona maculosa (Lepidoptera: Riodinidae) and phylogenetic relationships in Papilionoidea based on mitochondrial genes
    YAN Zhen-Tian, LUO Si-Te, FAN Zhen-Huai, CHEN Bin
    2023, 66(7):  946-958.  doi:10.16380/j.kcxb.2023.07.009
    Abstract ( 146 )   PDF (6567KB) ( 161 )   　　
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    【Aim】 To sequence and analyze the complete mitochondrial genomes (mtgenomes) of Caerulea coeligena and Dodona maculosa, and to explore the phylogenetic relationships in Papilionoidea based on the known mitochondrial genome data. 【Methods】 The complete mtgenomes of C. coeligena and D. maculosa were sequenced using Illumina NovaSeq 6000 sequencing platform, assembled and annotated, and the secondary structure of tRNA genes was predicted. Based on the nucleotide sequences of 13 protein-coding genes (PCGs) from the mtgenomes of 48 reported butterfly species belonging to six families in Papilionoidea obtained from the NCBI database, the selective pressures on these 13 PCGs in 50 species were analyzed. The phylogenetic relationships among these 50 species, as well as their respective families and subfamilies, were analyzed using maximum likelihood (ML) and Bayesian inference (BI) methods. 【Results】 The mtgenomes of C. coeligena and D. maculosa (GenBank accession numbers are MZ489120 and MZ 489121, respectively) are 15 164 and 15 486 bp in length, respectively. Their genome size, structure and composition, gene arrangement, nucleotide composition, codon usage, and predicted tRNA structure of mitogenomes of C. coeligena and D. maculosa were similar to those of the known closely related butterfly species. The mtgenomes in Papilionoidea exhibited a high level of purifying selection, with the ratios of nonsynonymous substitution rate to synonymous substitution rate (Ka/Ks) for the 13 PCGs obviously lower than 1. The phylogenetic relationships in Papilionoidea were (Papilionidae+Hesperiidae+Pieridae+(Nymphalidae+ (Lycaenidae+Riodinidae))).【Conclusion】 The reconstructed phylogenetic tree based on the mtgenomes of 48 reported butterfly species in Papilionoidea and two newly sequenced species in this study suggests that Riodinidae and Lycaenidae be monophyletic， and sister each other. The sister group would be closely related to Nymphalidae in phylogenetics. Moreover, Papilionidae may represent one of the earliest lineages in Papilionoidea. This study provides new molecular data to support further research on the systematic evolution and refinement of the classification system for butterflies.
    REVIEW ARTICLES
    Hybrid dysgenesis and P-element transposition in Drosophila
    WANG Chun-Ming
    2023, 66(7):  959-968.  doi:10.16380/j.kcxb.2023.07.010
    Abstract ( 107 )   PDF (3859KB) ( 103 )   　　
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    Hybrid dysgenesis in Drosophila refers to the occurrence of some abnormal phenomena in the progeny when crossing certain strains of Drosophila, such as ovarian hypoplasia, abnormal segregation ratio, recombination in male meiosis, high mutation rate, high frequency of chromosomal aberrations, even leading to infertility of the progeny etc. Hybrid dysgenesis in Drosophila is caused by frequent transposition of the transposable element (TE), including P-element, I-element, hobo-element, Penelope-element, Paris-element， Helena-element etc. P-element is the first eukaryotic transposon to have its DNA sequence determined, and it is also one of the most well-studied animal transposons. Developed as a genetic engineering tool, P-element has played an important role in Drosophila transgenic research. Drosophila carrying Pelement in the genome is paternal strain (P strain), while that without Pelement is maternal strain (M strain). In a cross of M female×P male, the progeny shows hybrid dysgenesis, while the progeny of the mating combinations of P female×M male, M female×M male and P female×P male is normal. In the past 20 years, significant progress has been made in the research on the regulation mechanism of P-element. In addition to the original repressor mechanism, a piRNA ［P-element-induced wimpy testis (PIWI)-interacting RNA］ mechanism has been discovered. The repressor mechanism is based on the post-transcriptional splicing of three introns among four exons of P-element transposase. If all the three introns are spliced, the resulted mRNA contains four exons, which are translated into a 87 kD transposase, promoting P-element transposition. If only the first two introns are spliced, the resulted mRNA terminates translation prematurely at the stop codon contained in the 3rd intron segment, producing a 66 kD repressor that prevents P-element transposition. The piRNA mechanism discovered in recent years is a more common TE regulatory mechanism. The mechanism is similar to the CRISPR-derived RNA (crRNA) mechanism found in bacteria, and the genes encoding piRNA are also arranged in cluster, called piRNA cluster. The piRNA cluster transcribes a single-stranded RNA precursor molecule, splices to 23-32 nt piRNA, then binds to the PIWI protein to form a complex, and plays a role through two pathways: One is to degrade the target mRNA complementary to the piRNA sequence and play a post-transcriptional silencing effect; the other is to enter the nucleus, guide the epigenetic modification of the sequence of P-element or piRNA cluster, play a transcriptional inhibition of P-element, and play a transcriptional activation effect on piRNA cluster. It was found that both the repressor mechanism and the piRNA mechanism play a role in the regulation of P-element transposition. This review may help with the teaching and scientific research of the phenomenon of hybrid dysgenesis.
    Advances in epigenetics affecting the caste structure and longevity differentiation behavior in eusocial insects
    MA Qiang, DANG Xiao-Qun, MA Zhen-Gang, ZHOU Ze-Yang
    2023, 66(7):  969-977.  doi:10.16380/j.kcxb.2023.07.011
    Abstract ( 107 )   PDF (972KB) ( 333 )   　　
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    Eusocial insects, such as bees, ants and wasps of the Hymenoptera and termites of the Blattodea, have significant diversity in morphology, behavior and life cycle, although their genetic background and genetic basis are consistent in a population. Most eusocial insects show different caste structure and life span differentiation. In these structure, queens tend to have a longer life span than workers, and their reproductive capacity is only owned by one queen or several queens, while the other members of the group can only act as workers. However, in some species, the caste structure has certain plasticity, and individuals can switch from one caste or behavioral phenotype to another according to specific environmental clues. Due to the common genetic background between different castes, the diversity of eusocial insect populations is largely caused by the gene transcriptional differences between individuals. This means that epigenetic mechanisms characterized by modifying gene expression without changing the gene sequence may play an important role in eusocial insects. Evidence had shown that epigenetic regulatory mechanisms such as DNA methylation or RNA methylation, histone post-translation modification and non-coding RNA had been proven to affect eusocial insects in many aspects, such as caste structure, longevity differentiation and aging. In this article, we reviewed the research progress of these epigenetic regulatory mechanisms and their different roles in insects, so as to deepen the understanding and cognitive degree of the origin and behavior evolution of eusocial insects. In the future, epigenetic regulatory mechanisms have potential application value in the research and development of anti-aging drugs, treatment of aging-related diseases, and slowing down the aging process of organisms.
    Insect odorscape: From odor diffusion to pest control
    JIAO Long, TAN Rong-Rong, CHEN Xun, WANG Hong-Juan, HUANG Dan-Juan, MAO Ying-Xin
    2023, 66(7):  978-991.  doi:10.16380/j.kcxb.2023.07.012
    Abstract ( 248 )   PDF (2423KB) ( 434 )   　　
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    The collective set of odors received by insects is called as insect odorscape. Insects rely on the reception and discrimination of the odorscape to complete life activities, such as the object localization, feeding, mating and oviposition. The behavior of insects can be manipulated by odorscape management for pest control strategy. In this article, we reviewed the research progress in the composition and diffusion of insect odorscape, influences of odorscape on insect behavior, factors affecting odorscape, odorscape reception and discrimination of insects, and the application of odorscape management in pest control. Finally, we analyzed and prospected the development direction and research focus of insect odorscape management in the future. For insects, the odor released by target is dispersed into plumes by air flow and mixed with the background odor carried in the air, which together form the odorscape. Insects search and locate the target along the target odor. The behavior of insects can be affected by the shape, composition and concentration of target odors. The background odor plays a complementary or warning role during the target localization of insects. Different background odors can synergize or interfere with the target localization of insects. The insect odorscape is mainly affected by the temperature, humidity, light, heavy metal elements and plant diseases and insect pests in the environment. Studies showed that the olfactory receptors of insects capture odorscape and transmit those odor signals to olfactory nerve centers such as antennal sensilla lobe along the olfactory nerves. Then, the odorscape is analyzed in the nerve centers by the mode of elemental processing or configural processing. The influences of background odor on insect target localization may be the results of the reciprocal addition, competitive binding or signal crosstalk of different odor molecules during the olfactory sensing and coding. At present, several kinds of green pest control technologies have been developed based on the odorscape management, such as insect behavior regulators, exogenous elicitors, breeding the crop varieties that can release resistant volatiles, “push-pull” technology and plant-mediated support system for natural enemies. In the future, it is necessary to further explore the behavioral, electrophysiological and neurological mechanisms of odorscape discrimination in insects, and optimize and integrate the green control technologies related to odorscape management, so as to build rational and efficient odorscape for insect pest control.
    SHORT COMMUNICATIONS
    Cloning and functional analysis of the E3 ubiquitin ligase gene NlHECTD2 in Nilaparvata lugens (Hemiptera: Delphacidae)
    LUO Yi, GUO Ying-Ying, XIONG Zhen-Ze, SONG Yang, LIU Yi-Peng, YU Xiao-Ping, SHENTU Xu-Ping
    2023, 66(7):  992-998.  doi:10.16380/j.kcxb.2023.07.013
    Abstract ( 126 )   PDF (2459KB) ( 328 )   　　
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    【Aim】 To explore the molecular characteristics, expression patterns and biological function of the E3 ubiquitin ligase gene NlHECTD2 of Nilaparvata lugens.【Methods】 Based on the transcriptome data of N. lugens, the full-length cDNA sequence of NlHECTD2 was cloned by PCR and bioinformatically analyzed. qRT-PCR was used to detect the expression levels of NlHECTD2 in the egg, 1st-5th instar nymphs, female adult newly emerged within 24 h and 1-5-day-old female adults, and the head, throax, abdomen, gut and ovary of the 3-day-old female adult of N. lugens. A single newly emerged female adult of N. lugens was injected with 0.05 μL dsNlHECTD2 (5 000 ng/μL) and the expression level of NlHECTD2 was detected by qRT-PCR at 24, 48, and 72 h after RNAi, and the number of yeast-like symbionts (YLSs) in the hemolymph, survival rate of the female adult of N. lugens, number of eggs laid per female and egg hatching rate were also detected.【Results】 The full-length cDNA sequence of NlHECTD2 (GenBank accession number: XM_039427060.1) was successfully cloned from N. lugens. The open reading frame (ORF) of NlHECTD2 is 2 850 bp in length and encodes 950 amino acids. NlHECTD2 has no signal peptide and contains a C-terminal catalytic domain with a subclass ubiquitin ligase (E3). Phylogenetic analysis indicated that NlHECTD2 was clustered together with other hemipteran HECTD2s, and had the highest homology with HECTD2 of Bemisia tabaci. The qRT-PCR results showed that NlHECTD2 had obvious spatiotemporal expression specificity, with the highest expression level in the abdomen of the 3-day-old female adult of N. lugens. The RNAi results showed that compared with the control (injected with dsGFP), injection of dsNlHECTD2 significantly inhibited the expression level of NlHECTD2 and extremely significantly decreased the number of YLSs in the hemolymph of N. lugens, which led to an extremely significant decrease in the survival rate of female adult, number of eggs laid per female and egg hatching rate of N. lugens.【Conclusion】 NlHECTD2 is closely related to the process of YLS release to hemolymph in N. lugens, plays an important role in the growth, development and reproduction of N. lugens, and can be used as a potential target for the control of N. lugens.
    CONTENTS
    Contents of Vol. 66 Issue 7
    2023, 66(7):  999-999. 
    Abstract ( 55 )   PDF (443KB) ( 163 )   　　
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