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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 2023, Volume 66 Issue 1
For Selected: View Abstracts Toggle Thumbnails
  • RESEARCH PAPERS
    Regulatory role of ame-miR-14 in the developmental process of the larval guts of Apis mellifera ligustica (Hymenoptera: Apidae) workers
    WANG Zi-Xin, XU Ya-Jing, ZHANG Wen-De, ZHANG Kai-Yao, WU Ying, LIU Jia-Mei, ZHU Le-Ran, NIU Qing-Sheng, ZHAO Hong-Xia, CHEN Da-Fu, GUO Rui
    2023, 66(1):  1-10.  doi:10.16380/j.kcxb.2023.01.001
    Abstract ( 280 )   PDF (2778KB) ( 462 )     
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    【Aim】 The objective of this study is to unravel the regulatory role of ame-miR-14 in the developmental process of the larval guts of Apis mellifera ligustica workers.【Methods】 The expression and sequence authenticity of ame-miR-14 in the 6-day-old larval guts of A. m. ligustica workers were proved by Stem-loop RT-PCR and Sanger sequencing, respectively. Overexpression and knockdown of ame-miR-14 were conducted by feeding its corresponding mimic (mimic-ame-miR-14) and inhibitor (inhibitor-ame-miR-14), and their corresponding negative controls mimic-NC and inhibitor-NC, respectively, and then RT-qPCR was used to detect the expression levels of ame-miR-14 in the 4-6-day-old larval guts of A. m. ligustica workers. Bioinformatic software was used to predict and analyze the target genes of ame-miR-14. RT-qPCR was employed to detect the relative expression levels of the target genes FoxO and Hedgehog in the 4-6-day-old larval guts after overexpression and knockdown of ame-miR-14.【Results】 ame-miR-14 truly exists and is expressed in the 6-day-old larval guts of A. m. ligustica workers. The expression levels of ame-miR-14 in the 4-6-day-old larval guts of A. m. ligustica workers fed with mimic-ame-miR-14 were significantly up-regulated as compared to those fed with mimic-NC. The expression levels of ame-miR-14 in the 4-6-day-old larval guts of A. m. ligustica workers fed with inhibitor-ame-miR-14 were significantly down-regulated as compared to those fed with inhibitor-NC. In total, ame-miR-14 can target 309 genes, which could be annotated to 45 KEGG pathways and 36 GO terms. Further analysis showed that ame-miR-14 can target 14 genes associated with growth and development and have potential targeting relationship with target genes FoxO and Hedgehog. After overexpression of ame-miR-14, the expression level of FoxO in the 4dayold larval gut of the mimic-ame-miR-14 group was down-regulated and those in the 5- and 6-day-old larval guts significantly down-regulated as compared to those in the mimic-NC group. After knockdown of ame-miR-14, the expression level of FoxO in the 4-day-old larval gut of the inhibitor-ame-miR-14 group was down-regulated, while that in the 5-day-old larval gut was significantly up-regulated and that in the 6-day-old larval gut was up-regulated as compared to those in the inhibitor-NC group. After overexpression of ame-miR-14, the expression levels of Hedgehog in the 4-6-day-old larval guts of the mimic-ame-miR-14 group were significantly down-regulated in comparison with those in the mimic-NC group, whereas those in the 4-6-day-old larval guts of the inhibitor-ame-miR-14 group were up-regulated in comparison with those in the inhibitor-NC group. 【Conclusion】ame-miR-14 truly exists and is expressed in the larval guts of A. m. ligustica workers. Effective overexpression and knockdown of ame-miR-14 in the larval guts of A. m. ligustica workers can be achieved by feeding mimic and inhibitor, respectively. ame-miR-14 potentially participates in regulation of the larval gut development by negatively regulating the expression of FoxO and Hedgehog.
    Identification and analysis of cytochrome P450 genes and their full-length transcripts in Apis cerana cerana (Hymenoptera: Apidae)#br#
    CAI Zong-Bing, WANG Zi-Xin, WU Ying, WANG Si-Yi, QIAN Jia-Jun, HU Ying, ZHANG Kai-Yao, GU Xiao-Yu, XU Xi-Jian, LUO Qun, CHEN Da-Fu, GUO Rui
    2023, 66(1):  11-18.  doi:10.16380/j.kcxb.2023.01.002
    Abstract ( 255 )   PDF (1867KB) ( 244 )     
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    【Aim】To identify cytochrome P450 (CYP450) genes and their full-length transcripts in the larval guts of Apsi cerana cerana workers using nanopore sequencing technology and offer reference information and basis for further functional study.【Methods】The 4-6-day-old larval gut transcriptomes of A. cerana cerana workers were sequenced by Nanopore PromethION platform. Quality control of raw reads was performed using Guppy software to gain clean reads. Full-length transcript sequences were detected through recognizing primers at both ends. The sequences of the above-mentioned full-length transcripts were aligned to the Nr and GO databases with BLAST tool to identify CYP450 genes and their full-length transcripts. Astalavista software was used to identify alternative splicing (AS) events of genes. RT-PCR was used to validate the reliability of AS events of various types. 【Results】In the 4-6-day-old larval guts of A. cerana cerana workers, 7 338 627, 7 003 419 and 7 434 233 raw reads were obtained, respectively, and after quality control, 7 289 494, 6 959 880 and 7 387 756 clean reads were gained. The total number of identified non-redundant fulllength transcripts was 48 200. Forty-seven CYP450 genes and their 265 full-length transcripts were identified. A total of 90 AS events of CYP450 genes were identified, including 36 exon skipping events, 20 alternative 5′ splicing site events, 17 intron retention events, nine alternative 3′ splicing site events and eight mutually exclusive exon events. The RT-PCR result confirmed the authenticity of randomly selected three types of AS events. 【Conclusion】 The findings provide CYP450 genes and their full-length transcripts in A. cerana cerana, supplement the annotation of A. cerana reference genome, and reveal that CYP450 genes of A. cerana cerana can generate abundant isoforms via multiple AS types.
    Molecular characterization and functional analysis of odorant binding protein gene AtOBP1 in Aethina tumida (Coleoptera: Nitidulidae)
    LI Liang-Bin, WU Li-Xian, XU Ya-Jing, LIU Fang, CHEN Da-Fu, LIU Jia-Li, ZHAO Hong-Xia
    2023, 66(1):  19-26.  doi:10.16380/j.kcxb.2023.01.003
    Abstract ( 231 )   PDF (1614KB) ( 501 )     
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    【Aim】 This study aims to clarify the expression pattern of odorant binding protein 1 (OBP1) in Aethina tumida and analyze the role of AtOBP1 in olfactory recognition of A. tumida.【Methods】 The cDNA full-length sequence of AtOBP1 was cloned based on the transcriptome and genome database of A. tumida and analyzed by bioinformatics. RT-qPCR was used to detect the expression levels of AtOBP1 in different developmental stages (egg, larva, pupa, female adult and male adult), different tissues (head, cuticle, wing, leg, fat body, gut, Malpighian tubules, testis and ovary) of the 7-day-old adult and in the head of different day-old adults of A. tumida after eclosion. The biological function of AtOBP1 in olfactory recognition of A. tumida was studied by RNA interference (RNAi) and Y-tube behavior choice experiment. 【Results】The cDNA full-length sequence of AtOBP1 gene (GenBank accession no.: MT211982.1) has six exons and an open reading frame (ORF) of 447 bp in length. AtOBP1 encodes 148 amino acid residues with PBP_GOBP superfamily conserved domain, and the predicted molecular mass and isoelectric point are 15.9 kD and 4.73, respectively. AtOBP1 protein is a dimer composed of six α-helics with six conserved cysteines that form three disulfide bonds. The phylogenetic tree also showed that AtOBP1 was closely clustered into one branch with TmOBP8 from Tenebrio molitor of Coleoptera. The RT-qPCR results showed that AtOBP1 was highly expressed in the pupal stage and the male adult stage, and was highly specifically expressed in the head and testis of adults. In addition, the expression level of AtOBP1 in adult head increased gradually with the day-old age, reached two peaks in the 5- and 7-day-old adult stages, respectively, and decreased in the 8-day-old adult stage. RNAi in combination with Y-tube behavior choice experiment results revealed that silencing of AtOBP1 resulted in significantly reduced preference of A. tumida adults to the pollen volatile compounds ethyl palmitate and ethyl linolenate.【Conclusion】AtOBP1 belongs to Classical OBPs family. AtOBP1 is mainly expressed in the head and testis of A. tumida adults, and may participate in the recognition process of the pollen volatile compounds ethyl palmitate and ethyl linolenate in A. tumida.
    Key words: Aethina tumida; odorant binding protein; AtOBP1; RNAi; olfactory behavior

    RNAi-mediated silencing of HvCDA1 affects the survival and development of the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata (Coleoptera: Coccinellidae)#br#
    WANG Ya-Jie, LIU Zhuo-Qi, PAN Guang, QIU Bao-Li, PAN Hui-Peng, YANG Chun-Xiao
    2023, 66(1):  27-36.  doi:10.16380/j.kcxb.2023.01.004
    Abstract ( 240 )   PDF (3277KB) ( 289 )     
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    【Aim】The 28-spotted ladybird, Henosepilachna vigintioctopunctata is an important pest of the Solanaceae plants. Chitin deacetylase 1 (CDA1) in insects catalyzes the deacetylation of N-acetylamino-D-glucosamine, facilitating the conversion of chitin to chitosan. Chitosan controls the orderly stacking of chitin fibers in insects and maintains the structural integrity of the cuticle. The silencing of CDA1 gene in insects inhibits the synthesis of chitosan which impairs the formation of cutiular structure and development, and eventually leads to death. 【Methods】The expression patterns of HvCDA1 in different developmental stages (egg, 1st-4th instar larvae and prepupa) and different tissues (cuticle, fat body, midgut and Malpighian tubules) of the 4th instar larvae of H. vigintioctopunctata were analyzed by RT-qPCR. The silencing of HvCDA1 was achieved by the oral feedings of eggplant leaves soaked in different concentrations of dsHvCDA1 solution for 1 min in the 1st instar larvae and direct oral feeding of different concentrations of dsHvCDA1 solution in the 4th instar larvae of H. vigintioctopunctata, and the RNAi-mediated silencing effects on the larval survival and development, and expression level of HvCDA1 were investigated.【Results】 Developmental expression profiling revealed that HvCDA1 was expressed across various developmental stages of H. vigintioctopunctata, with the highest expression level in the late 1st and late 2nd instar larvae. Tissue-specific expression profiling exhibited that the expression level of HvCDA1 in the cuticle of the 4th instar larvae was significantly higher than those in the other tissues. After feeding of eggplant leaves soaked in 50 ng/μL of dsHvCDA1 solution, the 1st instar larvae spines could not stand uprightly after molting, the cuticle could not form normally, and showed hindered feeding until death. The 1st instar larvae at 48 after being fed with the eggplant leaves soaked in dsHvCDA1 solution at the concentrations of 50, 200 and 400 ng/μL had the mortality rates of 84%, 94% and 100%, respectively. The oral feeding of the eggplant leaves soaked in 50 ng/μL of dsHvCDA1 solution reduced the expression levels of HvCDA1 by 38.63% and 79.00% in the 1st instar larvae at 48 and 96 h, respectively, as compared to the control (oral feeding of the eggplant leaves soaked in 50 ng/μL of dsGFP solution). On the other hand, ingestion of 50, 200 and 400 ng/individual of dsHvCDA1 solution by the 4th instar larvae resulted in 29%, 40% and 66% of larvae being deformed and unable to emerge normally after pupation, respectively. Similarly, the expression levels of HvCDA1 in the 4th instar larvae at 48 and 96 h after being fed with 200 ng/individual of dsHvCDA1 solution were significantly reduced by 52.99% and 70.09%, respectively, as compared to those in the control (fed with 200 ng/individual of dsGFP solution). 【Conclusion】 These results indicate that HvCDA1 plays an important role in the survival and development of H. vigintioctopunctata. The silencing of HvCDA1 negatively affects the ecdysis, pupation and emergence of H. vigintioctopunctata, and eventually leads to death. Collectively, the results from this study suggest that HvCDA1 can be a potential target gene for the RNAi-mediated control of H. vigintioctopunctata.
    Cloning, polyclonal antibody preparation and expression profiling of cuticular protein gene AlCP17 from Apolygus lucorum (Hemiptera: Miridae)
    TAN Yong-An, ZHANG Jie-Yu, JIANG Yi-Ping, ZHAO Jing, XIAO Liu-Bin, GE Lin-Quan
    2023, 66(1):  37-44.  doi:10.16380/j.kcxb.2023.01.005
    Abstract ( 219 )   PDF (4216KB) ( 123 )     
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    【Aim】This study aims to lay the foundation for the functional study of cuticular protein (CP) 17 of Apolygus lucorum (AlCP17) and to provide a preliminary basis for further analyzing the signal pathway map of cuticular development of A. lucorum by cloning its gene AlCP17, preparing its polyclonal antibody and analyzing its expression profiles.【Methods】Based on the previous transcriptome data, the full-length cDNA sequence of AlCP17 of A. lucorum was cloned and subjected to bioinformatics analysis. The prokaryotic expression vector pCZN1-AlCP17 was constructed and transformed to Escherichia coli to conduct in vitro expression. The purified recombinant protein was used to prepare polyclonal antibody of AlCP17. qRT-PCR was used to analyze the expression profiles of AlCP17 in different developmental stages (1st-5th instar nymphs and adult) and different tissues (head, thorax, leg, midgut, fat body and cuticle) of the early 3rd instar nymphs of A. lucorum.【Results】The full-length cDNA sequence of AlCP17 (GenBank accession no.: OM302231) was cloned from A. lucorum with an open reading frame of 594 bp in length, encoding a putative protein of 197 amino acids with the predicted molecular weight of 21.69 kD and the theoretical isoelectric point of 6.11. AlCP17 has the typical structure characteristics of cuticular protein, containing the conserved chitin binding domain (non-cysCBD) of arthropod cuticular protein, namely Chitin_bind_4 domain. Amino acid sequence alignment result showed that AlCP17 shows the highest amino acid sequence identity (76.29%) with the CPA2B-like of Cimex lectularius of Cimicidae of Hemiptera. Phylogenetic tree showed that CP is a highly conserved protein with high similarity to CPA2B-like of C. lectularius and CP7-like of Halyomorpha halys. The prokaryotically expressed recombinant AlCP17 was obtained by IPTG induction. After purification, the polyclonal antibody of AlCP17 was obtained and had good purity for subsequent experiments. AlCP17 was expressed in different developmental stages and different tissues of the early 3rd instar nymphs of A. lucorum, and its expression level reached the peak in the newly-hatched 1st instar nymphs and increased significantly at the end of each instar. In addition, the expression level of AlCP17 in the midgut of the early 3rd instar nymphs was the highest.【Conclusion】The full-length cDNA sequence of AlCP17 gene of A. lucorum was cloned. AlCP17 has the typical characteristics of insect cuticular protein. The expression of AlCP17 in A. lucorum shows developmental stage specificity and tissue specificity. These results lay the foundation for the future research on the physiological function of this protein in A. lucorum development.
    Extraction conditions of polysaccharides from Aspongopus chinensis (Hemiptera: Dinidoridae) and their inhibitory effects on human breast cancer HCC1937 cells
    YU Heng-Mei, DENG Yang, ZHAO Shuai, GUO Jian-Jun
    2023, 66(1):  45-54.  doi:10.16380/j.kcxb.2023.01.006
    Abstract ( 172 )   PDF (15141KB) ( 98 )     
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    【Aim】To promote the development and utilization of the traditional Chinese medicine Aspongopus chinensis, and to explore the effects of A. chinensis polysaccharides on human breast cancer HCC1937 cells.【Methods】Single factor experiment and orthogonal test were used to explore the optimal conditions for the extraction of A. chinensis polysaccharides by water extraction and alcohol precipitation. The optimal liquid to material ratio, extraction temperature, extraction time and extraction times were determined based on the extraction rates of A. chinensis polysaccharides. The effects of A. chinensis polysaccharides (0, 2, 4, 6, 8 and 10 mg/mL) on the cell proliferation of human breast cancer HCC1937 cells at 24 and 48 h after treatment were detected by CCK-8 method. At 0, 24 and 48 h after treatment with 0, 4 and 8 mg/mL of A. chinensis polysaccharides, the HCC1937 cell morphology was observed under inverted microscope and at 24 and 48 h the HCC1937 cell apoptosis was detected by Hoechst 33258 staining. The cell cycle and the expression of apoptosis-related proteins of HCC1937 cells at 24 h after treatment with 0, 4 and 8 mg/mL of A. chinensis polysaccharides were detected by flow cytometry and Western blotting, respectively. 【Results】The results of orthogonal experiments indicated that the optimum conditions for the extraction of A. chinensis polysaccharides were 40 mL/g of liquid to material ratio, 90 ℃ of extraction temperature, 3 h of extraction time, and three times of extraction. The extraction rate of A. chinensis polysaccharides was the highest (5.067%±0.071%) under the above conditions. A. chinensis polysaccharides at 4, 6, 8 and 10 mg/mL significantly inhibited the proliferation of HCC1937 cells. A. chinensis polysaccharides at 8 mg/mL blocked the cell cycle of HCC1937 cells at the S phase. A. chinensis polysaccharides at 4 and 8 mg/mL promoted the apoptosis of HCC1937 cells, upregulated the expression of pro-apoptotic proteins CASPASE-3, CASPASE-8 and BAX, and downregulated the expression of anti-apoptotic protein BCL-2. 【Conclusion】The highest extraction rate of A. chinensis polysaccharides was 5.067%±0.071% by water extraction and alcohol precipitation method under the optimum conditions of 40 mL/g of liquid to material ratio, 90 ℃ of extraction temperature, 3 h of extraction time, 3 times of extraction. A. chinensis polysaccharides have the activities of inhibiting human breast cancer HCC1937 cells in vitro and may trigger apoptosis through the death receptor pathway and the mitochondrial pathway.
    Knockout of SeGluCl splicing variants and its effects on the insecticide susceptibility in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    AN Jun-Long, YIN Xu, GAO Xin-Pei, ERNA·Azati, CHEN Li-Yu, PEI Ya-Kun, HU Zhao-Nong, ZUO Ya-Yun
    2023, 66(1):  55-62.  doi:10.16380/j.kcxb.2023.01.007
    Abstract ( 202 )   PDF (1459KB) ( 462 )     
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    【Aim】The purpose of this study is to explore whether glutamate-gated chloride channels (GluCls) of the beet armyworm, Spodoptera exigua (SeGluCls) encoded by different transcription variants of SeGluCl have different susceptibilities to insecticides using CRISPR/Cas9 gene editing technology. 【Methods】 The full-length cDNA sequence of SeGluCl from S. exigua was obtained using RT-PCR and RACE, and analyzed by bioinformatics. Two splicing variants SeGluCl 3a and SeGluCl 3b of SeGluCl were knocked out by CRISPR/Cas9 gene editing technique, and two knockout strains (3a-KO and 3b-KO) of S. exigua were established. The difference in the susceptibilies of the 3rd instar larvae of the susceptible strain WH-S (control strain) and 3a-KO and 3b-KO strains to three insecticides abamectin, emamectin benzoate and fipronil was determined by bioassay. 【Results】 The full-length cDNA sequence of SeGluCl of S. exigua was obtained, and its two splicing variants (SeGluCl 3a and SeGluCl 3b) were found. The open reading frame (ORF) of SeGluCl 3a (GenBank accession no.: OM304353) is 1 362 bp in length encoding 454 amino acids. The ORF of SeGluCl 3b (GenBank accession no.: OM304354) is 1 365 bp in length encoding 455 amino acids. The two SeGluCls encoded by the splicing variants SeGluCl 3a and SeGluCl 3b contain four transmembrane regions and one cysteine loop. Phylogenetic analysis indicated that SeGluCl was most closely related to GluCls of S. litura and S. frugiperda. Compared to the control strain WH-S, the knockout strains (3a-KO and 3b-KO) showed no significant difference in the susceptibilities to abamectin, emamectin benzoate and fipronil. 【Conclusion】 There is no difference in the susceptibility of SeGluCls encoded by the splicing variants SeGluCl 3a and SeGluCl 3b of SeGluCl to the three insecticides abamectin, emamectin benzoate and fipronil in S. exigua.
    Toxicity and field efficacy of metaflumizone against the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)#br#
    YIN Xue, FANG Xiao-Han, NIU Duo-Bang, LIAO Min, SHENG Cheng-Wang, CAO Hai-Qun
    2023, 66(1):  63-70.  doi:10.16380/j.kcxb.2023.01.008
    Abstract ( 290 )   PDF (1245KB) ( 225 )     
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    【Aim】 This study aims to determine the toxicity level and field control efficacy of metaflumizone against the larval fall armyworm, Spodoptera frugiperda, and to provide a reference for scientific use of metaflumizone in the control of S. frugiperda. 【Methods】 The median lethal concentration (LC50) and LC90 values of metaflumizone and four commonly used insecticides including emamectin benzoate, chlorantraniliprole, lufenuron and indoxacarb against the 3rd and 6th instar larvae of S. frugiperda and the medium lethal time (LT50) values of these insecticides at the concentration of LC90 against the 3rd instar larvae were determined in the laboratory with diet-incorporated method. Simultaneously, the control efficacies of 22% metaflumizone suspension concentrate(6.6 g/667 m2), 22% metaflumizone suspension concentrate (17.6 g/667 m2), 5.7% emamectin benzoate water dispersant (1 g/667 m2), and 150 g/L indoxacarb suspension concentrate (2 g/667 m2) against S. frugiperda larvae were determined in corn field by artificial spray approach. 【Results】 Laboratory bioassay results revealed that among the tested five insecticides metaflumizone showed higher toxicity to the 3rd and 6th instar larvae of S. frugiperda, with the LC50 values of 2.64 and 4.36 mg/kg, respectively, and its toxicity to the 6th instar larvae of S. frugiperda was only lower than that of emamectin benzoate. The LT50 value of metaflumizone at the concentration of LC90 against the 3rd instar larvae of S. frugiperda was 11.98 h, comparable to that of indoxacarb (11.50 h) and lower than those of chlorantraniliprole, emamectin benzoate and lufenuron (17.20, 23.40, and 39.24 h, respectively). Field efficacy test showed that 22% metaflumizone suspension concentrate at the application rate of 17.6 g/667 m2 had an excellent control efficacy against S. frugiperda larvae, with the corrected control efficacies of 69.97%, 78.98% and 82.86% at 1, 3 and 7 d after application, respectively, showing no significant difference from that of the control agent 5.7% emamectin benzoate water dispersant at the application rate of 1 g/667 m2. 【Conclusion】 The results of this study suggest that metaflumizone has excellent insecticidal activity and rapid action against S. frugiperda larvae, especially against the 6th instar larvae in the laboratory, and exhibits good control efficacy against S. frugiperda in the field, suggesting that it can be utilized to control S. frugiperda in China.
    Circadian rhythms of the behavioural activities of Bradysia cellarum (Diptera: Sciaridae) adults
    LI Meng-Yao, ZHANG Ya-Nan, GE Ya-Fei, YANG Xiao-Fan, FAN Fan, WEI Guo-Shu
    2023, 66(1):  71-76.  doi:10.16380/j.kcxb.2023.01.009
    Abstract ( 194 )   PDF (3147KB) ( 134 )     
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    【Aim】 This study aims to clarify the circadian rhythms of the behavioural activities of Bradysia cellarum adults, and provide basis for indoor breeding, behavioral research and development of accurate and efficient monitoring and novel control technologies. 【Methods】 The circadian rhythms of courtship, mating, oviposition, movement and resting of B. cellarum adults during the emergence peak were observed and recorded, and the time allocation characteristics of their specific behavior were analyzed. 【Results】 The locomotor and resting behaviors of B. cellarum adults occurred both in the daytime and at night, and the frequencies of reproductive behaviors (courtship, mating and oviposition) occurring at night were higher than those in the daytime, and there were significant differences in the proportion of frequencies among movement, resting and reproductive behaviors at various time points. The female and male adults of B. cellarum showed the same courtship rhythm, with two courtship peaks at 3:00 and 7:00, respectively. The mating rhythms of female and male adults of B. cellarum were similar to the oviposition rhythm of female adults, and the proportion of frequencies of movement reached the peak at 3:00 while that of female adults reached the peak at 19:00. There was no obvious circadian rhythm peak in the proportion of frequencies of resting behavior of female and male adults. As for daily time allocation, the resting behaviors of female and male adults accounted for the largest proportion of time, with the proportions of time of females and males of 89.53% and 89.90%, respectively, and the proportions of time of courtship behaviors of female and male adults were the least, being 0.56% and 0.89%, respectively. There was no significant difference in the daily proportion of time of various behaviors between female and male adults. 【Conclusion】 The courtship, mating, oviposition, movement, and resting behaviors of B. cellarum adults show obvious circadian rhythms, and the frequency of occurrence of different behaviors and the time points of activity peaks are different.
    Effects of larval density on the growth, development and reproduction of the mulberry borer, Diaphania pyloalis (Lepidoptera: Pyralidae)
    CAO Hong-Mei, HU Gui-Ping, SHI Xu-Ping, WANG Jun-Wen, CAI Xiang, HU Li-Chun, WANG Feng
    2023, 66(1):  77-84.  doi:10.16380/j.kcxb.2023.01.010
    Abstract ( 191 )   PDF (1403KB) ( 372 )     
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    【Aim】This study aims to understand the effects of different larval rearing densities of the mulberry borer, Diaphania pyloalis on their growth, development and reproduction.【Methods】 The growth, development and reproduction parameters including the developmental duration, larval survival rate, pupation rate, adult emergence rate, pupal weight, oviposition period and number of eggs laid per female of D. pyloalis reared at five different larval densities (130, 650, 1 300, 1 950 and 2 600 individuals/m2) were determined under the same condition in the laboratory. 【Results】 Larval density had influences on the growth, development and reproduction of D. pyloalis to varying degrees, and low or high larval density was not conducive to its growth, development and reproduction. There was no obvious difference in the growth, development and reproduction parameters of D. pyloalis reared at the larval densities of 1 300 and 1 950 individuals/m2, both with excellent performance. The larval, pupal and adult duration of D. pyloalis reared at the larval density of 1 300 individuals/m2 were 11.32, 6.33 and 5.31 d, respectively, and those reared at the larval density of 1 950 individuals/m2 were 11.50, 6.00 and 5.47 d, respectively. The pupation rate, adult emergence rate, and larval survival rate of D. pyloalis reared at the larval density of 1 300 individuals/m2 were 86.67%, 100% and 86.67%, respectively, and those reared at the larval density of 1 950 individuals/m2 were 84.44%, 94.74%, and 80.00%, respectively. When D. pyloalis larvae were reared at the densities of 1 300 and 1 950 individuals/m2, the oviposition periods of their adults reached more than 2.00 d, the numbers of eggs laid per female reached more than 55.00 grains, and the egg hatching rates were more than 90.00%, showing significant difference from those reared at the other larval densities. In addition, the larval density of D. pyloalis showed an extremely significant correlation with the larval survival rate.【Conclusion】 Larval density is an important factor affecting the growth, development and reproduction of D. pyloalis, and when the larval density is 1 300 individuals/m2 or 1 950 individuals/m2, it is conducive to the efficient establishment of its indoor populations.
    Effects of tobacco varieties, seedling raising methods and sources of Myzus persicae (Hemiptera: Aphididae) on the parasitic ability of Aphidius gifuensis (Hymenoptera: Braconidae)
    MA Fu-Hua, ZHOU Ting, LAI Rong-Quan, GU Gang, HAN Meng
    2023, 66(1):  85-91.  doi:10.16380/j.kcxb.2023.01.011
    Abstract ( 154 )   PDF (1216KB) ( 185 )     
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    【Aim】 This study aims to screen the tobacco varieties, seedling raising methods and sources of Myzus persicae suitable for Aphidius gifuensis parasitization, so as to provide a scientific reference for artificial propagation and rejuvenation of A. gifuensis. 【Methods】We investigated the parasitism rate of A. gifuensis adults on M. persicae nymphs and the emergence rate, life span, body size and proportion of females of A. gifuensis adults when M. persicae nymphs were bred with different varieties of flue-cured tobacco (Yunyan 87, K326, CB-1 and Honghuadajinyuan), from different sources (from tobacco region, from nontobacco region and indoor breeding population), and bred with tobacco with different seedling raising methods (soil seedling raising and moist seedling raising) via pot experiments.【Results】The results showed that different tobacco varieties had significant effects on the parasitism rate of A. gifuensis adults on M. persicae nymphs. The parasitism rates of A. gifuensis adults on M. persicae bred with Honghuadajinyuan and CB-1 were 19.00% and 14.00% higher than that on K326, respectively. However, tobacco varieties had no significant effects on the other indices of A. gifuensis. Sources of M. persicae showed significant effects on the parasitism rate of A. gifuensis. The parasitism rates of A. gifuensis adults on M. persicae nymphs from the non-tobacco region and tobacco region were 20.25% and 16.75% higher than that on M. persicae from indoor breeding population, respectively. However, sources of M. persicae showed no significant effects on the other indices of A. gifuensis adults including emergence rate, life span, body size and proportion of females. Different seedling raising methods of tobacco did not have significant effects on the parasitism rate of A. gifuensis adults. 【Conclusion】 The parasitsm rate of A. gifuensis on M. persicae depends on tobacco varieties and M. persicae sources, but is not impacted by seedling raising methods. Therefore, in practical applications, the parasitic ability of A. gifuensis on M. persicae can be improved by adjustments to tobacco varieties and M. persicae sources, and the application effect of artificial reproduction efficiency of A. gifuensis can be enhanced.
    Identification of Panonychus citri (Acarina: Tetranychidae), an emerging pest mite in the pear orchard in Shandong Province, East China, based on morphological and molecular data
    CHEN Tao, LIU Yong-Jie, LI Bo, MEN Xing-Yuan, YIN Shu-Yan
    2023, 66(1):  92-98.  doi:10.16380/j.kcxb.2023.01.012
    Abstract ( 171 )   PDF (1611KB) ( 337 )     
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    【Aim】 The purpose of this study is to accurately identify an emerging pest mite in the pear orchard in Shandong Province, East China, and lay the foundation for its further research and effective control. 【Methods】 The morphological characteristics of female adult mites were directly observed by stereomicroscope, and the slides of male adult mites were prepared to observe the aedeagal characteristics for morphological identification. The genomic total DNA was extracted from a single female adult mite. The mitochondrial cytochrome oxidase subunitⅠ(COⅠ) gene, and the internal transcribed spacer 1 (ITS1) gene and the internal transcribed spacer 2 (ITS2) gene of nuclear ribosomal DNA were amplified with mitespecific primers and sequenced. The obtained sequences were aligned using Blastn on NCBI. The related sequences with high identity to the obtained sequences were downloaded to construct the phylogenetic trees and calculate the genetic distances for molecular identification. 【Results】 The results showed that the female adult mite on pear trees was born with tubercles on the dorsum, which can be identified as the genus Panonychus pest mite. The characteristics of the aedeagus of the male pest mite were very similar to those of the citrus red mite, Panonychus citri. The COⅠ, ITS1 and ITS2 gene sequences of the pest mite were highly similar to the corresponding sequences of P. citri, with the nucleotide sequence identity of more than 99%. The genetic distances between the pest mite and P. citri calculated based on COⅠ, ITS1 and ITS2 gene sequences were much lower than that between it and the other species of Panonychus. On the phylogenetic trees based on COⅠ, ITS1 and ITS2 gene sequences, the pest mite on pear tree clustered in the same branch with P. citri, with very high bootstrap values (99%, 99% and 100%, respectively). 【Conclusion】 The emerging pest mite that broke out in the pear orchard in Shandong Province was determined to be P. citri based on the morphological characteristics combined with the molecular data, indicating that the distribution range of P. citri in China has moved northward, and great attention needs to be paid to this phenomenon.
    Sequencing and analysis of the mitochondrial genome of Scoliidae 
    YAO Yu-Jiang, CHEN Bin, LI Ting-Jing
    2023, 66(1):  99-107.  doi:10.16380/j.kcxb.2023.01.013
    Abstract ( 232 )   PDF (2989KB) ( 216 )     
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    【Aim】This study aims to understand the monophyletic and phylogenetic positions of the Scoliidae by phylogenetic analysis of the mitochondrial genome of Aculeata insects.【Methods】The mitochondrial genomes of five species of three genera of Scoliidae were sequenced by using Illumina Hiseq2500 next-generation sequencing technology, annotated and analyzed. Maximum likelihood (ML) and Bayesian inference (BI) methods were used to construct the phylogenetic tree of Aculeata insects based on the sequences of 13 protein-coding genes (PCGs) and two rRNA genes of 36 mitochondrial genomes. 【Results】The five newly sequenced mitochondrial genomes of the Scoliidae, including the mitochondrial genomes of Colpa quinquecincta (GenBank accession number: OM103696), Colpa tartara (GenBank accession number: OM103697), Megacampsomeris grossa (GenBank accession number: OM103796), Megacampsomeris formosensis (GenBank accession number: OM142776) and Scolia schrenkii (GenBank accession number: OM103797), are 15 367-20 649 bp in length, containing 37-39 genes including 13 PCGs, 2 rRNA genes (rrnL and rrnS) and 22-24 tRNA genes, with the AT content of over 75%. The start codons of all the 13 PCGs are ATN, and the stop codons are TAA or T--. In contrast to the deduced ancestral sequence, complex gene rearrangement events have occurred in the five newly sequenced mitochondrial genomes of the Scoliidae, which are mainly the changes of some gene positions in the gene region trnS1-trnE-trnF-nad5-trnH, a remote inversion phenomenon of the gene region trnI-trnQ-trnM-nad2-trnW-trnC-trnY-cox1-trnL2-cox2-trnK and the position changes of the genes trnF, trnS1, trnE, trnH and nad5. Both Ka/Ks and nucleotide diversity π of 13 PCGs indicated that cox1 is the most conserved gene. The results of the ML tree and BI tree both showed that the Scoliidae is a monophyletic group, and the Scoliidae and the Pompilidae are sister groups to each other. In the Scoliidae, Colpa and Scolia, and Megacampsomeris and Camposmeris all show close relationship.【Conclusion】This study preliminarily clarified the characteristics of the mitochondrial genomes of the family Scoliidae, laying the foundation for further research. The phylogenetic analysis based on the mitochondrial genome shows that the family Scoliidae is a single lineage.
    REVIEW ARTICLES
    Chemical communication mechanism and strategy for attractant development in Bactrocera dorsalis (Diptera: Tetriphitidae)
    ZHANG Jie, ZHANG Yan, LIU Wei, YAN Shan-Chun, WANG Gui-Rong
    2023, 66(1):  108-120.  doi:10.16380/j.kcxb.2023.01.014
    Abstract ( 324 )   PDF (2173KB) ( 521 )     
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    True fruit flies are important insect pests attacking fruits and vegetables. The total damage caused by them worldwide is estimated to amount to be more than 2 billion US dollars annually. The oriental fruit fly, Bactrocera dorsalis, is one of the representatives of this kind of pests, causing serious losses to China’s citrus industry every year. The techniques based on male attractant and protein bait have been used in environment friendly strategies for pest monitoring and control. However, the performances of those baits in the field are unsatisfied and need to be further improved. With the reduction of the cost of high-throughput sequencing technology and the development of modern molecular biology technology, scientists proposed to discover the key molecular targets for olfaction by resolving the molecular mechanism of pest chemosensory first and develop more stable and efficient attractants with the identified new targets. In order to promote the development of behavioral regulation technology targeting key chemosensory molecules in B. dorsalis, we reviewed the research status of important chemicals regulating the behavior of B. dorsalis and the mechanism of chemosensory perception in this article. The important volatiles regulating the behavior of B. dorsalis include sex pheromones, plant volatiles and food-derived protein odors. Among them, the specific compounds identified by the first two have a clear relationship with the behavior of B. dorsalis adults. For example, pyrazine substances obtained from sex pheromones can attract females, methyl eugenol in plant volatiles can attract males, γ-octalactone can induce females to lay eggs; while the composition of food-derived protein odor is complex, although it has a certain efficacy in the field, there is a lack of function validation of specific compounds in female and male insect behavior. In the olfactory sensory mechanism, there is only a morphological description of the peripheral nerve sensilla and the central antennal lobe, and the function of different types of olfactory neurons is not clear. A large number of chemical sensory proteins have been identified in B. dorsalis, including 49 odorant binding proteins (OBPs), 60 odorant receptors (ORs), 23 ionotropic receptors (IRs) and 17 gustatory receptors (GRs), through bioinformatics analysis at present. However, the number of olfactory genes functionally analyzed is small. In conclusion, although some compounds with behavioral activity on B. dorsalis have been identified, and a large number of olfactory proteins have been used as candidate molecular targets, the corresponding relationship between “chemical substances-olfactory molecular targets and nerve-behavior” is lacking, which greatly limits the role of olfactory molecular targets in attractant development. Therefore, on this basis, we put forward a development strategy for the behavioral regulation technology of B. dorsalis based on olfactory key molecular targets, to provide new ideas for the design and screening of effective behavioral regulators of B. dorsalis.
    CONTENTS
    Contents of Vol. 66 Issue 1
    2023, 66(1):  121-121. 
    Abstract ( 89 )   PDF (531KB) ( 128 )     
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