• 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
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Identification and expression profiling of genes of odorant-binding proteins and  chemosensory proteins in adult antennae of Lytta caraganae (Coleoptera: Meloidae)
LIU Pan-Jing, WEI Hong-Yi, GUO Kun, MA Guang-Yuan, ZHANG Tao
Acta Entomologica Sinica    2022, 65 (8): 927-936.   DOI: 10.16380/j.kcxb.2022.08.001
Abstract332)      PDF(pc) (4785KB)(239)       Save
【Aim】 This study aims to establish the adult antennal transcriptome database of  Lytta caraganae, and explore and identify genes of olfactory-related odorant-binding  proteins (OBPs) and chemosensory proteins (CSPs) in its adult antennae. 【Methods】  Transcriptome analysis of adult antennae of L. caraganae was performed on Illumina HiSeq  platform. Assembled genes were annotated by alignment against public databases NR, NT, KO,  Pfam, Swiss-Prot, GO and KOG. OBP and CSP genes of L. caraganae were screened according to  the annotation results. Structural characteristics and evolutionary relationship of OBP and 
CSP genes were analyzed by ClustalX 1.83 and MEGA 7.0 software, respectively. The  expression levels of OBP and CSP genes in female and male adult antennae of of L. caraganae  were determined by qRTPCR. 【Results】 A total of 51 028 transcripts and 41 998 unigenes  were obtained from the adult antennal transcriptome of L. caraganae. Gene annotation  results showed that L. caraganae genes have the highest match (87.3%) to those of  Tribolium castaneum. Twenty-wo OBP genes and seven CSP genes were screened. Sequence  alignment result showed that 13 LcarOBPs are classified into classic OBPs, which contain  six conserved cysteine residues. Phylogenetic tree showed that LcarOBPs and LcarCSPs show  the highest amino acid sequence identity with OBPs and CSPs of Hycleus cichorii and H.  phaleratus, respectively, indicating the closest evolutionary relationship. qRT-PCR  results showed that two LcarOBP genes and two LcarCSP genes were highly expressed in the  male adult antennae of L. caraganae, while ten LcarOBP genes and two LcarCSP genes highly  expressed in the female adult antennae. 【Conclusion】 The OBP and CSP genes in adult  antennae of L. caraganae have been identified for the first time, providing a theoretical  basis for further study on the mechanism of olfactory recognition in L. caraganae.
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Molecular cloning and functional analysis of  Nl 15 in the brown planthopper,  Nilaparvata lugens  (Hemiptera: Delphacidae)
WANG Fu-Xin, WANG Wei-Xia, WEI Qi, HE Jia-Chun, LAI Feng-Xiang, FU Qiang, WAN Pin-Jun
Acta Entomologica Sinica    2022, 65 (5): 558-567.   DOI: 10.16380/j.kcxb.2022.05.003
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 【Aim】 The phytophagous piercing-sucking insect saliva protein participates in the regulation of plant defense response against insects and affects insect adaptability to host plants. The aim of the present study is to clone the important salivary protein gene Nl15 in the brown planthopper, Nilaparvata lugens, and to investigate its temporal and spatial expression patterns, so as to clarify its roles in virulence o f N. lugens. 【Methods】 Based on the transcriptome data of IR56 population of N. lugens, the cDNA sequence of Nl15 was cloned from N. lugens by RT-PCR, and subjected to bioinformatics analysis. The expression profiles of Nl15 in different developmental stages (egg, 1st-5th instar nymph, and female and male adult) and female adult tissues (head, thorax, abdomen and leg) of TN1 and IR56 populations of N. lugens were determined by qPCR. The RNAi of Nl15 was carried out by dsRNA microinjection into the 4th instar nymphs of TN1 and IR56 populations of N. lugens. The relative expression levels of Nl15 in N. lugens nymphs after RNAi of Nl15 and defense-related genes OsLecRK4, OsMPK10, OsWRKY24, OsLox, OsNPR1, and OsGns5 in rice plants fed by N. lugens nymphs for 3 d following RNAi of Nl15 were detected by qPCR. The survival rate and the honeydew amount and body weight gain of N. lugens adults after RNAi of Nl15 were determined by bioassay. 【Results】 The cDNA sequence of Nl15 (GenBank accession no.: OK181113) of N. lugens was cloned. It has an open reading frame of 1 008 bp in length, encoding 335 amino acids with the predicted isoelectric point of 7.54 and the molecular weight of 38.7 kD. The Nl15 protein contains a signal peptide sequence of 23 aa and a predicted glycosylation modification site, whereas has no transmembrane domain and other known functional domains. Nl15 shares 45% amino acid sequence identity with the homologous protein from Laodelphax striatellus. Developmental expression profile revealed that Nl15 was expressed in various developmental stages of N. lugens, with the highest expression level in the 3rd-4th instar nymphs. Tissue expression profile showed that Nl15 exhibited the highest expression level in the head of female adults of N. lugens, with a higher expression level in the head of IR56 population than in the head of TN1 population. RNAi results showed that the expression level of Nl15 in ds Nl15 injection group was significantly down-regulated by 89.5%, the survival rate and the honeydew amount and body weight gain of adults of N. lugens were significantly decreased, and the expression levels of the above six rice defense-related genes were significantly up-regulated as compared to those in the control group (ds GFP injection group). 【Conclusion】 Nl15 in IR56 population of N. lugens is involved in the interaction of defense and counter defense between N. lugens and rice. This study provides insights into the mechanisms by which N. lugens overcomes resistance genes and the molecular network of interactions between insects and plants.
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Screening and identification of actinomycetes with antimicrobial activity from the gut of  the medicinal insect Blaps rynchopetera(Coleoptera: Tenebrionidae)
WANG Ming-Ming, YIN Peng-Kai, LI Ming-Hui, YANG Zi-Zhong, YANG Da-Song, YANG Yin-He
Acta Entomologica Sinica    2022, 65 (9): 1106-1114.   DOI: 10.16380/j.kcxb.2022.09.004
Abstract305)      PDF(pc) (18899KB)(69)       Save
【Aim】 In order to provide new actinomycete resources for the development of  antimicrobial drugs, actinomycetes with antimicrobial activity from the gut of Blaps  rynchopetera were explored. 【Methods】 Actinomycetes were isolated from the gut of B.  rynchopetera adults by using dilution coating method and selective culture method. With six  pathogenic bacteria methieillin-resistant Staphylococus aureus, S. aureus, Escherichia  coli, Enterococcus faecalis, Salmonella typhimurium and Pseudomonas aeruginosa, and three  pathomycetes Aspergillus niger, Penicillium expansum and Canidia albicans as the indicator  strains,the antimicrobial activities of secondary metabolites of actinomycetes were tested  by Oxford cup method. Subsequently, 16S rRNA sequence analysis was performed to identify  the 18 strains of actinomycetes with significant activity through molecular biological  method and construct a phylogenetic tree. 【Results】 A total of 176 strains of symbiotic  actinomycetes were isolated from the gut of B. rynchopetera adults. Preliminary  antimicrobial screening showed that among them 46 actinomycete strains had different  degrees of antimicrobial activities. Some actinomycetes exhibited broad-spectrum  antimicrobial activity, with their inhibition zone diameters larger than those of the  positive control drugs. Eighteen actinomycete strains with the inhibition zone diameter  larger than 15 mm were selected for molecular identification, and the results showed that  they were Streptomyces spp. 【Conclusion】 There are abundant actinomycete resources with  antimicrobial activity in the gut of B. rynchopetera
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Development and function of halteres in insects 
DONG Wei, WU Wen-Jun, ZHANG Xu-Bo
Acta Entomologica Sinica    2022, 65 (8): 1068-1074.   DOI: 10.16380/j.kcxb.2022.08.015
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 The halteres in dipteran insects evolved from the hindwings and play an important  role in flight. The sensilla at the base of halteres detect the inertial force and provide  feedback to motor neurons that subsequently balance body during flight. The haltere of  insects is developed from imaginal disc and regulated by the HOX gene (Ultrabithorax, Ubx).  Mature haltere is composed of two layers of epithelial cells. The bulb is filled with  vacuolar cells, while the base possesses various sensilla. Interestingly, the halteres  controlled by independent muscles move antiphase relative to ipsilateral wing. However, the  winghaltere coordination is essential for departure and maintaining balance. Recently,  the navigation principles of halteres have been increasingly applied in bionics, and  navigation devices of aircrafts have been developed based on the structure and functions of  halters of flies. In this article we reviewed the progress in the research on the  development, morphological structure, function and bionics application of halteres with the  goal of providing a theoretical basis for further understanding the developmental  meachanisms and biological functions of halteres in insects.
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Research progress on the occurrence, damage and control of Mythimna loreyi (Lepidoptera: Noctuidae)
DUAN Yun, CHEN Qi, GUO Pei, MIAO Jin, XIA Peng-Liang, GONG Zhong-Jun, JIANG Yue-Li, LI Tong, WU Yu-Qing
Acta Entomologica Sinica    2022, 65 (4): 522-532.   DOI: 10.16380/j.kcxb.2022.04.012
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 Mythimna loreyi is a relative species of Mythimna seperata. The two pest species are similar in morphology and have basically the same damage characteristics, but there may be some differences in the occurrence and damage rules in different regions. In recent years, M. loreyi has become a common agricultural pest with frequent outbreaks in many countries and regions. In order to deeply study the outbreak law and to comprehensively prevent and control this pest, we summarized its research status based on the analysis of domestic and foreign research data in this article. At present, M. loreyi has been distributed in nearly 80 countries and regions in Asia, Europe, Africa, Oceania and North America. The larvae of this pest feed on a variety of gramineous crops and weeds, such as corn, rice and wheat. Before the 3rd instar, the larvae eat less, the 5th-6th instar larvae eat more, and enter the gluttony stage. The larvae of M. loreyi are aggregative, omnivorous and gluttonous, and the adults are migratory. The damage caused by this pest is covert, sporadic and fulminant. There are differences in the law of the occurrence and damage of M. loreyi due to different regions, years and seasons. Climatic conditions, food and natural enemies are the main factors that affect its damage. At the same time, it is pointed out that climatic factors, human activities and the biological characteristics of M. loreyi are the important factors that have led to its occurrence expansion and damage aggravation in recent years. Sex attractants, natural enemies and biological pesticides are the main research directions in the biological control of this pest in recent years. Based on the occurrence and research status of M. loreyi in China, the following three aspects are suggested to be carried out in the future work: (1) By learning from the prevention and control strategies and experience of M. seperata, strengthening the prediction and forecast of M. loreyi from two aspects of air forecast and ground forecast; (2) carrying out comprehensive prevention and control work from agricultural control, physical control, biological control and chemical control; and (3) applying modern technologies and methods such as biotechnology and modern agricultural information technology to study the law of outbreak and disaster, integrated prevention and control of M. loreyi, and to establish the comprehensive prevention and control technology systems.
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Molecular mechanisms of reproductive regulation in hematophagous insects
WANG Xue-Li, LI Shan, LYU Xiang-Yang, ZOU Zhen
Acta Entomologica Sinica    2022, 65 (4): 512-521.   DOI: 10.16380/j.kcxb.2022.04.011
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Hematophagous insects are arthropods that can spread insect-borne pathogens, including mosquitoes, sandflies, midges, kissing bugs, fleas and so on. Blood-feeding behavior makes them become the vectors transmitting malaria, dengue fever, filariasis, trypanosomiasis, and other acute infectious diseases. The fast and wide-spreading of vector-borne diseases, in addition to heavy damage to human health, might result in huge economic losses. Due to the scarcity of effective medicines and increasing drug resistance of pathogens, the interruption of reproduction of hematophagous insects is an effective measure to control the spread of insect-borne diseases. Juvenile hormone (JH) and 20-hydroxyecdysone (20E) play important roles in insect reproduction. JH binds to intracellular receptor complex Met/Tai to regulate the expression of JH/Met target genes, and then promotes vitellogenesis, which provides pre-requisite to bloodfeeding and oviposition of insects. Heterodimer EcR/USP is an intracellular receptor of 20E. The combination of 20E with EcR/USP complex can activate downstream gene expression and induce the synthesis of vitellogenin (Vg) to provide nutrition for the developing ovary. Nutrient signaling pathways (insulin signaling pathway and amino acidmediated target of rapamycin signaling pathway) can also activate Vg synthesis and promote insect reproduction. In addition, nutrient signaling pathways can interplay with JH and 20E signaling cascades to regulate the development and reproduction of hematophagous insects. Energy metabolism, such as carbohydrate and lipid metabolism, is the main energy source during insect reproduction, which can meet the extremely high energy requirements in different stages of reproductive development of hematophagous insects. Studies have shown that JH and 20E signaling pathways play important regulatory roles in energy metabolism. MicroRNAs have been proved to be closely related to physiological processes such as gut microbiome homeostasis, blood digestion and lipid metabolism in mosquitoes, further affecting the development mosquitoes. In recent years, with the innovation of molecular biology and sequencing technology, new progress has been made in the study of reproductive regulation mechanisms in hematophagous insects. In this article, we present the research progress and insights into molecular mechanisms of reproductive regulation in hematophagous insects, which will provide important clues for blocking the transmission of vector-borne diseases by regulating the reproduction of hematophagous insects.
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Comparative transcriptome analysis and screening of diapause-associated genes during the  egg development of Gomphocerus sibiricus(Orthoptera: Acrididae)
ZHAO Na, LÜ Xue-Feng, HU Hong-Xia, SONG Yu, JIANG Si-Han, JI Rong, YE Xiao-Fang
Acta Entomologica Sinica    2022, 65 (9): 1075-1089.   DOI: 10.16380/j.kcxb.2022.09.001
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【Aim】This study aims to preliminarily clarify the molecular mechanism of  diapause in Gomphocerus sibiricus eggs through screening diapause genes and metabolic  pathways of the eggs.【Methods】 Transcriptome sequencing was performed on G. sibiricus  eggs at different developmental stages [early developmental stage (ES), diapause stage  (DS), and post-diapause developmental stage (PS)], with the Illumina NovaSeq 6000  sequencing platform. The diapause-associated pathways of G. sibiricus eggs were predicted 
by GO and KEGG enrichment analysis, and analyzed by cluster heat map analysis combined with  literature reports to screen diapause-associated genes. qRT-PCR was used to verify six  important genes of the screened diapause-associated genes. 【Results】 In the DS vs ES and  PS vs DS comparative groups, 12 419 and 4 789 differentially expressed genes (DEGs) were  enriched, respectively, and most of them were up-regulated. A total of 2 206 DEGs of the  two comparative groups were mainly related to glucose metabolism, environmental stress and  growth and development. The most significant enrichment of GO items in the DS vs ES group  was protein binding. The GO items in the PS vs DS group mainly included enzymatic activity,  cytoskeleton construction and protein binding. Diapause-associated genes were mainly  involved in Wnt signaling pathway, insulin signaling pathway, cell cycle pathway and insect  hormone biosynthesis pathway. The expression trends of the six important  diapause-associated genes were consistent with the transcriptome data. 【Conclusion】 In  this study, important metabolic pathways that regulate diapause of G. sibiricus eggs were  preliminarily identified, and a total of 20 diapause-associated genes were screened out,  laying a foundation for further study on the adaptation mechanism of this species.
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Dynamics of ecdysone titer and the interference of methoxyfenozide on diapause during the  development of Carposina sasakii(Lepidoptera: Carposinidae)
LI Yong-Li, YAN Zuo-Bing, YIN Xin-Ming, LEI Zhen-Shan, ZHOU Zhou
Acta Entomologica Sinica    2022, 65 (7): 791-798.   DOI: 10.16380/j.kcxb.2022.07.001
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【Aim】 Under short and long light photoperiod, the mature larvae of Carposina  sasakii undergo diapause and non-diapause development, respectively. This study aims to  interfere with the diapause of C. sasakii by using the ecdysone analog methoxyfenozide so  as to lay a foundation for the new control technologies for C. sasakii. 【Methods】 The 20E  titer dynamics in C. sasakii during larvae diapause and non-diapause developments were  measured by ELISA. The interference effect of methoxyfenozide (0.05, 0.1, 1, 2, 5, 7.5  and 10 mg/mL) and 20E (0.5 and 1 mg/mL) on the cocoon formation of C. sasakii was  determined using the sandblast method with pure water as the control. The effects of 5  mg/mL methoxenozide and 1 mg/mL 20E sandblasting treatments on the 20E titer of C. sasakii  were measured, and the effect of sandblasting with 0.1 and 5 mg/mL methoxenozide on the  diapause of C. sasakii were investigated subsequently. 【Results】 There was no difference  in 20E titer in the developmental process between diapause and non-diapause larvae of C.  sasakii under two photoperiods (15L∶9D and 12L∶12D), and both gradually decreased with  the increase of instar and reched the lowest when they escaped off fruits. However, the 20E  titer in non-diapause larvae escaping off fruits under long photoperiod (0.473 ng/g) was  significantly higher than that in diapause larvae escaping off fruits under short  photoperiod (0.254 ng/g). The non-diapause developmental larvae entered the long cocoon  pupation after defruiting, and the 20E titer increased significantly and maintained a high  level (0.652-1.217 ng/g) in the pupal stage. The diapause developmental larvae entered the 
round cocoon diapause after defruiting, and the 20E titer slowly increased in the first  four days, reaching the first peak (0.656 ng/g) on the 4th day, then decreased and then  increased again, reaching the 2nd peak (0.790 ng/g) on the 8th day. With the entry of  diapause stabilization period, the 20E titer gradually decreased. After 70 d, the diapause  was relieved, and the 20E titer remained at a low level. The lowest titer (0.424 ng/g) of  diapause period was reached on the 90th day, and then the 20E titer began to rise.  Methoxyfenozide at the concentration of 0.05 mg/mL could interfere with the formation of  diapause round cocoons of C. sasakii larvae escaping off fruits under short photoperiod,  and the LD50 value of methoxyfenozide to mature defruiting larvae of C. sasakii was 7.039  μg/individual. Both methoxyhydrazine and 20E at the concentration of more than 0.05 mg/mL  sprayed on the sand surface could effectively interfere with the formation of diapause  round cocoons of C. sasakii, and 20E showed higher interference activity than  methoxyfenozide at the same concentration of 1 mg/mL. Both of them could make the diapause  larvae produce long cocoon, abnormal cocoon and non-cocooning reaction, and the 20E titer  in abnormal cocoon and non-cocooning larvae increased significantly. The percentages of  cocooning failure in 5 mg/mL methoxyfenozide and 1 mg/mL 20E sand spraying treatments were  70.0% and 66.7%, respectively. The 20E titers in abnormal cocooning larvae significantly  increased by 16.3% and 143.0%, respectively. The 20E titers in non-cocooning larvae  significantly increased by 149.3% and 278.6%, respectively. Although some larvae could  form round cocoons after being exposed to 0.1 mg/mL methoxyfenozide, the proportion of  larvae successfully completing diapause and eclosion reduced, and the eclosion rate was  only 20.8%. 【Conclusion】 The 20E titer in C. sasakii keeps a low titer during the  diapause process, and a higher titer is needed in the reproductive developmental stage.  Moreover, methoxyfenozide and 20E can interfere with its diapause state forming round  cocoons. The increase range of 20E titer is consistent with the change of cocooning  phenotype, and 20E shows higher interference activity than methoxyfenozide at the same  concentration. Methoxyfenozide reduces the proportion of larvae successfully completing  diapause.
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Oviposition preference of Serangium japonicum (Coleoptera: Coccinelidae) to different  tomato varieties and its influencing factors
MEI Wen-Juan, LIN Shuo, ZHANG Qian-Rong, DING Xue-Ling, ZHENG Yu, LU Xue-Song, YAO Feng-Luan, HE Yu-Xian, WENG Qi-Yong
Acta Entomologica Sinica    2022, 65 (6): 757-771.   DOI: 10.16380/j.kcxb.2022.06.011
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Aim】 This study aims to explore the oviposition preference of Serangium  japonicum to tomato (Lycopersicon esculentum) varieties with different densities and types  of leaf trichomes. 【Methods】 We selected four tomato varieties including Zhenmu 101,  Minfenying 1, Zhenmu 301 and Yiseliechaojijingang as the host plants of S. japonicum observed the microstructure of the abaxial leaf surface (ALS) under scanning electronic  microscopy, and counted the types and densities of leaf trichomes on the ALS. We determined  the proportions of eggs laid by female adults of S. japonicum on leaf discs and plants of  the four tomato varieties bearing Bemisia tabaci eggs, the offspring and adult performance  such as the development, fecundity and predation capability, and attachment force of S.  japonicum on the leaves of the four tomato varieties, the preference of female adults of S.  japonicum to the odours from healthy leaves and B. tabaci-infected leaves of the four  tomato varieties, and the risk of egg cannibalism on leaf discs and plants of four tomato  varieties. 【Results】There are type Ⅱ, type Ⅲ and type Ⅴ non-glandular trichomes, and  type Ⅰ, type Ⅳ, type Ⅵ and type Ⅶ glandular trichomes on the ALS of four tomato  varieties, and type Ⅴ trichomes have the highest density. Yiseliechaojijingang has the  lowest density of leaf trichomes among the four tested tomato varieties. S. japonicum  preferred to lay eggs on the tomato variety Yiseliechaojijingang. The fecundity of S.  japonicum on the tomato varieties Yiseliechaojijingang, Zhenmu 101 and Minfenying 1 (165- 223.92 eggs per female) was significantly higher than that on the tomato variety Zhenmu  301 (28.09 eggs per female). S. japonicum offspring had the shortest developmental  duration (egg to adult duration) (15.73 d) on Minfenying 1, while had the longest  developmental duration (23.00 d) on Zhenmu 101. However, the proportions of S. japonicum  eggs laid on the four tomato varieties had no significant correlation with the offspring  and adult performance, but were positively correlated with the risk of egg cannibalism. The  proportions of S. japonicum eggs laid on the four tomato varieties had no significant  correlation with the attachment force and the density of leaf trichomes on the ALS. The  densities of the seven types of trichomes were significantly different among the four  tomato varieties. The density of type Ⅰ glandular trichomes was negatively correlated with  the fecundity (number of eggs laid per female in 30 d) of S. japonicum, but the density of  the other six types of trichomes had no correlation with the offspring and adult  performance of S. japonicum. The female adults of S. japonicum had no obvious preference to  odours from the four tomato varieties. The proportions of eggs consumed by cannibalistic S.  japonicum on leaf discs and plants of the four tomato varieties were significantly  different. 【Conclusion】Female adults of S. japonicum prefer to lay eggs on leaves of  tomato varieties with low density of leaf trichomes, and tomato glandular trichomes have  great influence on the life activities of S. japonicum adults.
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Function of vitellogenin receptor gene TaVgR in the regulation of reproductive development  in Tuta absoluta (Lepidoptera: Gelechiidae)
YAN Xin, YANG Hong, SONG Jia-Hui, LI Ren-Gui, ZHANG Yi-Bo, YANG Wen-Jia
Acta Entomologica Sinica    2022, 65 (6): 675-683.   DOI: 10.16380/j.kcxb.2022.06.003
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【Aim】 This study aims to clarify the function of vitellogenin receptor (VgR) in  the reproductive development of the tomato moth, Tuta absoluta, so as to provide a  potential target for the green control of leaf miner pests. 【Methods】 Based on the  transcriptomic data of T. absoluta, the full-length cDNA sequence of TaVgR was amplified  by RT-PCR and analyzed by bioinformatics methods. The expression patterns of TaVgR in  different developmental stages (1st-4th instar larvae, 1-7-day-old female pupae, and  female adults) and female adult tissues (head, integument, foregut, midgut, hindgut, ovary,  fat body, and Malpighian tubules) of T. absoluta were examined by RT-qPCR. RNAi was used  to inhibit the expression of TaVgR in female pupae, and the changes in the ovarian  development and fecundity of T. absoluta were observed after silencing of TaVgR. 【Results 】 The cDNA sequence of TaVgR (GenBank accession no.: MZ682118) of T. absoluta was cloned.  It contains an open reading frame of 5 496 bp in length, encoding 1 831 amino acids with  the predicted molecular weight of 206 kD and isoelectric point of 5.17. The deduced protein  of TaVgR has a signal peptide with 18 amino acid residues at the N-terminus and harbours  the typical conserved functional domains of LDLR family proteins. The RT-qPCR results  showed that the transcriptional level of TaVgR gradually rose with the increase of age of  T. absoluta and reached the peak after female adult emergence. TaVgR exhibited the highest  expression level in the ovaries of female adults of T. absoluta. RNAi of TaVgR inhibited  the expression level of TaVgR in the early female pupae by 62.04%-72.55%, led to disruption  of the deposition of yolk protein in the ovary, shortened the lengths of ovarioles and  eggs, and reduced the total number of eggs laid per female adult in 10 d and the egg  hatching rate of offspring, resulting in a decline in fertility. 【Conclusion】 TaVgR is  ighly expressed in female adults and ovaries of T. absoluta. Silencing of TaVgR severely  blocks the ovarian development of T. absoluta and impairs its fecundity. This study lays a  theoretical foundation for the development of new control strategies of lepidopteran pests  with VgR gene as the target.
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Molecular species definition of common blister beetles (Coleoptera: Meloidae) from northern  China 
CHENG Hai-Yun, DUAN Jia-Chong, ZHANG Chao, PAN Zhao
Acta Entomologica Sinica    2022, 65 (9): 1204-1221.   DOI: 10.16380/j.kcxb.2022.09.014
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【Aim】 This study aims to test the applicability of four analytical methods,  including automatic barcode gap discovery (ABGD), generalized mixed Yule coalescent (GMYC),  Bayesian Poisson tree processes (bPTP), and Bayesian phylogenetics and phylogeography (BPP)  with fragments of two genes, mitochondrial COI and nuclear CAD, in the molecular species  definition of Meloidae insects. 【Methods】 Eighteen meloid morphologic species widely  distributed in northern China and belonging to six genera (Hycleus, Mylabris, Epicauta,  Lytta, Megatrachelus, and Meloe) were sampled. The molecular species definition was carried  out based on COI, CAD, and concatenated COI+CAD sequence data sets by using ABGD, GMYC,  bPTP, and BPP. The molecular species definition results by using different methods were  compared with that by the morphological identification, respectively. 【Results】 The  molecular species definitions based on the COI+CAD concatenated sequence data sets were  consistent with the morphological identification results, those based on COI sequences with  the ABGD and GMYC methods were also consistent with the morphological identification  results, while the number of species identified by using bPTP was more than that by  morphological identification. The definition results based on CAD sequences by using all  three single-gene species definition methods except GMYC were partially different from the  morphological identification results. 【Conclusion】 The molecular species definitions of  Meloidae based on multi-gene concatenated sequences and multiple methods are better than  those based on single gene fragment or single definition method. The results of this study  provide data support and reference for molecular species definition and integrative  taxonomy of the family Meloidae.
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Morphological differentiation in the Asian honey bees ( Apis cerana) in China  ( In English)
HU Xiang-Jie, ZHOU Shu-Jing, XU Xin-Jian, YU Ying-Long, HU Jun-Jun, ZHANG Zhong-Yin, QI Wen-Zhong, WANG Biao, YUAN Chun-Ying, XI Fang-Gui, ZHOU Bing-Feng
Acta Entomologica Sinica    2022, 65 (7): 912-926.   DOI: 10.16380/j.kcxb.2022.07.013
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【Aim】 The genetic differentiation research is an important link to understand the morphological diversity and adaptive evolution of honey bees. It is a prerequisite for the determination of the bioresource management unit and the protection unit and helps to protect the genetic resources of honey bees. This study aims to study the genetic differentiation and genetic resource distribution of the Asian honey bee, Apis cerana across the geographical environment in China by analyzing morphological differentiation. 【Methods】 A total of 6 147 worker bees of A. cerana were collected from 102 sampling sites across the complete distribution area of A. cerana in China. Sixty worker bees of each sampling site from 10-20 colonies were dissected and 33 morphological characteristics associated with the wings, individual size, hind leg, and body color were measured. A multivariate morphometric analysis was conducted and clusters with their morphological traits and distribution patterns were identified. 【Results】 According to the cluster results of discriminant analysis and principal component analysis, A. cerana in China can be divided into 14 morphological clusters. Five clusters with smaller body size were identified. Hainan cluster had the smallest body size, followed by South Yunnan cluster, Taiwan cluster, Southern cluster, and Northern cluster. These five clusters were significantly different in proboscis length, forewing length, the structure of the 3rd submarginal cell in the forewing, body color, and the length of the wax plate. Changbai cluster had the largest cubital index, wax plate size, and width of the stripe of tomentum on tergite 5. However, Bomi cluster of Tibet had the smallest width of the stripe of tomentum on tergite 5 in China. Northwest cluster had the longest hind legs. Five clusters in the West Sichuan Plateau were characterized by larger individuals and black body color. Batang cluster had the smallest cubital index (3.0169) and the largest individual size in China. The cubital index of the Aba cluster was inferior only to that of the Changbai cluster, and the wing lengths and the sizes of sternite 7 were the largest. Derong cluster was the darkest. Yajiang cluster was unique in wing vein angles (A4, N23, E9 and J10 were the smallest and B4 the largest). Chuandian cluster had the smallest body size on the Western Sichuan Plateau. 【Conclusion】 In this study, the morphometric analysis of A.  cerana was conducted based on collection of samples across the complete distribution area of A. cerana in China, especially those from Bomi of Tibet, Taiwan Province, and the Western Sichuan Plateau. Fourteen clusters of A. cerana were obtained in China, including Hainan cluster, southern Yunnan cluster, Changbai cluster, Taiwan cluster, Bomi cluster, Aba cluster, Batang cluster, Derong cluster, Yajiang cluster, Chuandian cluster, Chuangui cluster, Northwest cluster, Southern cluster, and Northern cluster. The results of this study provide a theoretical basis for the protection and exploitation of genetic resources of A. cerana in China.
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Application of the bootstrap technique and the multinomial theorem in the research of age-stage, two-sex life table
LI Jian-Yu, CHEN Yan-Ting, FU Jian-Wei, SHI Meng-Zhu, CHI Hsin, YOU Min-Sheng
Acta Entomologica Sinica    2022, 65 (10): 1389-1400.   DOI: 10.16380/j.kcxb.2022.10.015
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The age-stage, twosex life table,called two-sex life-table for short, is an important theory and an analytical tool that are commonly used in population ecology and pest management. The user-friendly TWOSEX-MSChart program, which had been designed based on the twosex life table theory to help researchers for data analysis in insect population studies, has been more and more widely used by increasing numbers of scientists around the world. There are many statistical techniques and computer simulations embedded in the TWOSEX program, and the bootstrap technique is one of the major procedures included in the program. In this article, we describe the principles, methods, advantages/disadvantages, and the application of the bootstrap technique in the twosex life table analysis, as well as the application of the multinomial theorem in life table research. Compared with the general statistics, the bootstrap technique can be used to estimate and infer the distribution characteristics of data without the assumption of data distribution. In the twosex life table analysis, the bootstrap technique can not only be used to estimate the population parameters or the variances and standard errors of general statistics, but also to be used to assess the differences between treatments by paired bootstrap test to accurately show the population variability. The same bootstrap samples can be used to calculate the hatching rate and the contribution of different reproductive forms to population parameters, and to link the life table and predation rate analysis of natural enemies for an accurate analysis of the reproduction and predation potential of natural enemies. In addition, we also introduce the multinomial theorem, i.e., the mathematical basis of the bootstrap technique. The application of the multinomial theorem demonstrates that stable and reliable estimates can be obtained by using the bootstrap technique. We also elaborate the necessity of considering the ineffective bootstrap samples in the life table research. In recent years, although the twosex life table and the bootstrap technique had been widely adopted in research, few reports discussed the principles and methodology involved. This article will help interested researchers in entomology and ecology understand the basic theories and principles of the bootstrap technique and the multinomial theorem, and their application in twosex life table analysis, so as to better apply them in the related scientific research projects.
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Effects of four pesticides on the body temperature of the diamondback moth, Plutella  xylostella (Lepidoptera: Plutellidae)
TIAN Su-Fen, LI Zhi-Hua, LI Yuan-Tao, LIN Shuo, CHEN Yi-Xin, TIAN Hou-Jun, HU Xiao-Han, WANG Xi-Ying, HUANG Jing-Fei, WEI Hui, GU Xiao-Jun
Acta Entomologica Sinica    2022, 65 (9): 1115-1126.   DOI: 10.16380/j.kcxb.2022.09.005
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【Aim】 This study aims to provide data for elucidating the application value of  the body temperature of the diamondback moth, Plutella xylostella, in its management.  【Methods】 At different temperatures in the artificial climate incubators (ambient  temperatures), the body temperature of the 2nd, 3rd, and 4th instar larvae of P. xylostella  was measured, and the relation equations between the body temperature of various instar  larvae (y) and the ambient temperature (x) were established. Meanwhile, the body  temperatures of the 3rd instar larvae of P. xylostella at different time after treatment  with different concentrations of avermectin, chlorpyrifos, fipronil and cypermethrin,  respectively, at different ambient temperatures were measured. 【Results】 The relation  equations between the ambient temperature (x) and the body temperature (y) of the 2nd, 3rd  and 4th instar larvae of P. xylostella were y=0.95x+1.19 (r=0.9463), y=0.95x+1.18  (r=0.9988), and y=0.93x+1.45 (r=0.9989) with the corresponding isothermal points of 22.16 ℃, 21.40℃ and 21.41℃, respectively. When the ambient temperature was set at 15℃ or 40 ℃, none of the four pesticides changed the body temperature of the 3rd instar larvae of P.  xylostella. However, at the other ambient temperatures, the body temperature of the 3rd  instar larvae of P. xylostella could be changed by pesticide treatment. For avermectin, at  25℃, the body temperatures of the 3rd instar larvae in the 2, 4 and 8 mg/L treatment  groups at 12 h, 2 and 4 mg/L treatment groups at 24 h, 0.5, 2, 4 and 8 mg/L treatment  groups at 36 h and 0.5, 1, 2 and 8 mg/L treatment groups at 48 h were significantly  increased, while that in the 8 mg/L treatment group at 24 h was significantly decreased; at  30℃, those in the 0.5 mg/L treatment group at 24 h and 1 mg/L treatment group at 36 h were  significantly decreased and those in the 1 mg/L treatment group at 48 h and the treatment  groups at various concentrations at 60 h were significantly increased; and at 35℃, only  those in the 1 and 8 mg/L treatment groups at 48 h were significantly decreased as compared  to that in the control. For chlorpyrifos, at 20℃, the body temperatures of the 3rd instar  larvae in the 50, 200 and 800 mg/L treatment groups at 24 h and 100, 400 and 800 mg/L  treatment groups at 36 h were significantly decreased; at 25℃, those in the 100 and 200  mg/L treatment groups at 12 h, 800 mg/L treatment group at 24 h and 100, 200 and 800 mg/L  treatment groups at 60 h were significantly decreased, but those in the 50, 100, 200 and  400 mg/L treatment groups at 24 h, 100 and 200 mg/L treatment groups at 36 h and 100 and  400 mg/L treatment groups at 48 h were significantly increased; and at 30℃, only that in  the 800 mg/L treatment group at 24 h was significantly decreased and those in the 50, 100,  200 and 800 mg/L treatment groups at 60 h were significantly increased as compared to that  in the control. For fipronil, at 20℃, only the body temperature of the 3rd instar larvae  in the 0.5 mg/L treatment group at 36 h was significantly decreased; at 25℃, that in the 4 
mg/L treatment group at 12 h and those in the treatment groups at various concentrations at  60 h were significantly decreased, and that in the 0.5 mg/L treatment group at 24 h and  those in the 0.25, 1 and 2 mg/L treatment groups at 48 h were significantly increased; at  30℃, those in the 0.25 and 0.5 mg/L treatment groups at 12 h, 0.25 and 2 mg/L treatment  groups at 24 h, 4 mg/L treatment group at 48 h and 2 mg/L treatment group at 60 h were  significantly decreased; and at 35℃, only those in the 0.25 and 0.5 mg/L treatment groups  at 60 h were significantly increased as compared to that in the control. For cypermethrin,  at 20℃, the body temperatures of the 3rd instar larvae in the 2 and 8 g/L treatment groups  at 36 h and 4 and 8 g/L treatment groups at 48 h were significantly increased; at 25℃,  those in the 2, 4 and 8 g/L treatment groups at 12 h were significantly decreased and those  in the 0.5, 4, and 8 g/L treatment groups at 24 h, 1, 4 and 8 g/L treatment groups at 36 h,  and 1, 2 and 4 g/L treatment groups at 60 h were significantly increased; and at 30℃,  those in the 0.5 and 1 g/L treatment groups at 12 h, 0.5, 1, 4 and 8 g/L treatment groups  at 24 h and 1, 2 and 8 g/L treatment groups at 60 h were significantly decreased as  compared to that in the control. 【Conclusion】 The autonomic thermoregulation ability of  P. xylostella larvae is comparatively low. Avermectin, chlorpyrifos, fipronil or  cypermethrin treatment can affect the body temperature of the 3rd instar larvae of P.  xylostella, but the effect varies with the pesticide type and concentration, ambient  temperature and treatment time. The results expand the studies on pesticide toxicology and  pest control.
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Behavioral manipulation of the plant secondary metabolites to thrips and their application  in thrips management
LI Xiao-Wei, CHENG Jiang-Hui, HAN Hai-Bin, LÜ Yao-Bin
Acta Entomologica Sinica    2022, 65 (9): 1222-1246.   DOI: 10.16380/j.kcxb.2022.09.015
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Thrips are important pests of agricultural and horticultural crops, causing  enormous economic losses by direct feeding and indirect transmission of the  plant-pathogenic virus. Plant secondary metabolites play a pivotal role in plant-insect  interactions. The manipulation of insect behavior using plant secondary metabolites to  protect crop plants from pest infestation is a promising eco-friendly control tactic. In  this article, plants, plant extracts, essential oils and chemical compounds that have  attractive, repellent, oviposition and feeding deterrent effects, fumigation toxicity and  toxic activities to thrips were reviewed, and the potential of plant secondary metabolites  for thrips management was discussed. Volatiles or essential oils from 54 plant species in  27 families, 29 benzenoids, 17 pyridines and 13 terpenes are attractive to thrips and could  be used as trap plants and attractants. Volatiles or essential oils from 40 plant species  in 17 families, 20 terpenes and 6 benzenoids show repellency against thrips and could be  used as repellent plants and repellents. Extracts or essential oils from 42 plant species  in 20 families, 6 alkaloids, 15 terpenes and 5 benzenoids have oviposition and feeding  deterrent effects, fumigation toxicity and toxic activities to thrips, and could be  developed into botanical pesticides and fumigants. Finally, current problems of plant  secondary metabolites in thrips management, such as unstable effects, lack of field  application technology and unclear muchamisms, were discussed, and potential research  directions were prospected, which are of great significance to thrips management based on  plant secondary metabolites.
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Sequencing and analysis of the complete mitochondrial genome of  Aclerda takahashii  (Hemiptera: Aclerdidae)
DENG Jun, WANG Gang, LU Cong-Cong, ZHANG Jiang-Tao, HUANG Xiao-Lei
Acta Entomologica Sinica    2022, 65 (4): 451-459.   DOI: 10.16380/j.kcxb.2022.04.005
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【Aim】 This study aims to sequence and analyze the complete mitochondrial genome (mitogenome) of Aclerda takahashii, which represents the first complete mitogenome of the family Aclerdidae of Hemiptera, and to investigate the phylogenetic relationships with other Coccoidea groups. 【Methods】 The whole mitogenome of A. takahashii was sequenced based on the Illumina sequencing technique, and analyzed with bioinformatics methods. The phylogenetic trees of hemipteran insects were constructed based on the reported complete mitochondrial genomes of 31 speciec of 15 families of Hemiptera using the maximum likelihood (ML) and Bayesian inference (BI) methods. 【Results】 The mitogenome of A. takahashii is 16 599 bp in length with high A+T content (84.51%). The truncated tRNAs are common in this mitogenome, ten tRNAs lacking of a dihydrouridine (DHU) arm or TΨC (T) arm. The DHU and T arm of tRNAser(S1) and tRNAser(S2) are both missing. The phylogenic trees showed that Aclerdidae has the closest relationship with Coccidae. 【Conclusion】 This study reported the first mitogenome of Aclerdidae, and revealed that truncated tRNAs are prevalent in the mitogenome of A. takahashii, providing data for the further systematic research of mitogenomes of scale insects.
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Effects of residues of pesticides commonly used in corn fields on  Trichogramma ostriniae    (Hymenoptera: Trichogrammatidae) at different time after application
WANG Yu, DAI Xiao-Yan, WANG Rui-Juan, LIU Yan, CHEN Hao, ZHENG Li, DONG Xiao-Lin, ZHAI Yi-Fan
Acta Entomologica Sinica    2022, 65 (7): 852-865.   DOI: 10.16380/j.kcxb.2022.07.007
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【Aim】 This study aims to evaluate the safety of fungicides and insecticides commonly used in corn fields to Trichogramma ostriniae. 【Methods】 Corn leaves were applied with four fungicides (thiophanate-methyl, tebuconazole, bismerthiazol, and pyraclostrobin) and four insecticides (avermectins, chlorantraniliprole, beta-cypermethrin, and imidacloprid) at the field recommended dosages in the laboratory. At 1, 3, 5 and 7 d after pesticide application, the mortality rates, parasitic abilities (numbers of Sitotroga cerealella eggs parasitized) and offspring emergence rates of T.  ostriniae adults exposed to corn leaves containing pesticide residues for 24 h were investigated. 【Results】 Different fungicides and insecticides at different time after application had significant effects on the mortality rate, parasitic ability and emergence rate of T. ostrinia adults. Among the four fungicides, the effects of bismerthiazol and pyraclostrobin on the parasitic ability of T. ostrinia lasted for a long time. After 7 d of application, the number of S. cerealella eggs parasitized by T. ostrinia adults exposed to corn leaves containing bismerthiazol and pyraclostrobin residues for 24 h were 20.25 and 20.80, respectively. Tebuconazole had the greatest effect on T. ostrinia. After T. ostrinia adults were exposed to corn leaves containing tebuconazole residue for 24 h, their emergence rate was 18.48% at 7 d after pesticide application and their mortality was 37.03% at 3 d after pesticide application. Among the four insecticides, avermectins and imidacloprid had a more long-lasting effect on the parasitic ability of T. ostrinia and showed greater toxicity. After T. ostrinia adults were exposed to corn leaves containing avermectins and imidacloprid residues for 24 h, the numbers of S. cerealella eggs parasitized were 3.43 and 9.19, respectively, at 7 d after pesticide application, and their mortality rates were 96.21% and 74.00%, respectively, at 1-3 d after pesticide application. The emergence of T. ostrinia offspring was damaged for a longer time after application of beta-cypermethrin, and the offspring emergence rate was 27.92% at 7 d after application. In general, the four insecticides had a greater impact on T. ostriniae than the four fungicides. 【Conclusion】 It is not recommended to use the three fungicides, tebuconazole, bismerthiazol and pyraclostrobin, and the two insecticides, avermectins and imidacloprid, in the field in 7 d before T. ostriniae are released to control Ostrinia nubilalis, and to use thiophanate-methyl and beta-cypermethrin in 5 d before T. ostriniae release. Chlorantraniliprole should be reasonably used depending on the recommended field dosages.
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Analysis of differentially expressed genes in response to low temperature stress in Apis mellifera worker pupae
LIU Yi-Ming, XU Xin-Jian, ZHOU Shu-Jing, YAO Dan, LI Han, ZHU Chen-Yu, LI Xiang, HE Yu-Chang, ZHOU Bing-Feng, ZHU Xiang-Jie
Acta Entomologica Sinica    2022, 65 (4): 469-479.   DOI: 10.16380/j.kcxb.2022.04.007
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【Aim】 This study aims to screen and analyze the differentially expressed genes (DEGs) across different pupal stages of Apis mellifera workers in response to low temperature stress at the transcriptomic level. 【Methods】 The 3-d-old post-capped prepupae, and 6- and 9-d-old post-capped pupae of A. mellifera workers were exposed to 20℃ for 4 h as the cold treatment groups (T), and those reared at 35℃ (their optimal developmental temperature) for 4 h as the control groups (CK). The DEGs in A. mellifera at the three developmental stages between the cold treatment groups and the control groups were screened by transcriptomic technology, followed by GO functional classification and KEGG pathway analysis. Subsequently, the expression profiles of 8, 6, and 5 DEGs from the 3-d-old post-capped prepupae, and 6- and 9-d-old post-capped pupae, respectively, were validated by RT-qPCR. 【Results】 There were 220, 50 and 26 DEGs between the 3-d-old post-capped prepupae and 6- and 9-d-old post-capped pupae exposed to low temperature and their control groups, respectively. GO functional classification of DEGs showed that the most enriched terms are related to metabolic process, cellular process, catalytic activity and binding. In addition, more DEGs in the 3-d-old post-capped prepupae were enriched in biological process regulation, cell part and organelle. KEGG pathway analysis showed that DEGs in A. mellifera at the three developmental stages between the cold treatment groups and the control groups were enriched in global and overview maps, amino acid metabolism, signal transduction, transport and catabolism. The expression of the shared DEG 3-phosphoinositide-dependent protein kinase 1 gene PDK1 in the 3-d-old post-capped prepupae and 6- and 9-d-old post-capped pupae exposed to low temperature was up-regulated and this gene was enriched in autophagy-animal, and mTOR and FoxO signaling pathways. In the 3-d-old post-capped prepupae in response to low temperature, the expression levels of insulin receptor substrate 1-B gene IRS1-B and Kruppel homolog 1 gene Kr-h1 were up-regulated, while those of the nuclear hormone receptor FTZ-F1 gene Ftz-F1 and ecdysis triggering hormone gene Eth were down-regulated significantly, suggesting that autophagy and ecdysis are inhibited to a greater degree in the 3-d-old post-capped prepupae in response to low temperature. The expression of tyrosine hydroxylase gene TyHyd related to tanning and immunity in the 6-d-old post-capped pupae exposed to low temperature was down-regulated. The number of DEGs identified between the 9-d-old post-capped pupae exposed to low temperature and the control group was the least, suggesting that the 9-d-old post-capped pupae are less affected by low temperature among the three pupal stages. 【Conclusion】 In this study, the DEGs in different developmental stages of A. mellifera pupa in response to low temperature were determined. The results show that most of DEGs are stage-specific, suggesting that different developmental stages of A. mellifera have different response mechanisms to low temperature. The function and mechanism of some shared and stage-specific DEGs are the key content to further study the low temperature response mechanism of honeybee, which provides basic data for exploring the molecular mechanism of the response of suggesting honeybee pupae to low temperature stress.
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Insect development and immunity: An important branch of modern entomology
WANG Mao, ZOU Zhen, XU Wei-Hua
Acta Entomologica Sinica    2022, 65 (12): 1565-1570.   DOI: 10.16380/j.kcxb.2022.12.001
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 As one of the important branches of entomology, insect development and immunity, facing the national demand and scientific frontier, has made great achievements in solving major pest disasters and human health through multidimensional research. Meanwhile, the progress of new biotechnology has greatly promoted the research of insect development and immunity by deepening and widening our understanding of insect development and immune defense. Articles in this special issue of “Insect growth and development and immunity” reflect well the current status and features of research on insect development and immune in China. The growth and development part covers all developmental stages from egg to adult, mainly focusing on the signal transduction, and the immunity part focuses on biological interactions. In the context of big data, more efforts will be made to combine traditional and modern techniques, and strengthen cooperation, thus making the research branch play a greater role in pest control, insect resource utilization, and food security.
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Expression and functional analysis of the autophagy-related gene NlATG13 in the brown  planthopper, Nilaparvata lugens(Hemiptera: Delphacidae)
WU Jian-Gen, JIAO Qi-Qi, YU Fei-Fei, ZHENG Yuan-Yuan, CHEN Tong-Tong, HAO Pei-Ying, YU Xiao-Ping
Acta Entomologica Sinica    2022, 65 (8): 967-976.   DOI: 10.16380/j.kcxb.2022.08.005
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【Aim】 Autophagy participates in various physiological processes of cells. The  autophagy-related protein ATG13 is a component of the Atg1/13 complex and plays an  important role in initiating the autophagy. This study aims  toanalyzethefunctionofATG13inthebrownplanthopper, Nilaparvata  lugens, and to evaluate its potential as a pest control target. 【Methods】 Based on the  transcriptome data of N. lugens, the full-length cDNA sequence of NlATG13 in N. lugens was  cloned via RACE method. The nucleotide and amino acid sequence characteristics of NlATG13  were analyzed with bioinformatics technology. RT-qPCR technology was used to detect the  expression patterns of NlATG13 in different developmental stages (1st-5th instar nymphs,  and female and male adults) and different tissues (the head, thorax, midgut and fat body of  the 5th instar nymphs and the ovary of newly emerged female adults) of N. lugens. The  expression of NlATG13 was knocked down by RNAi through microinjection of dsNlATG13 into the  3rd instar nymphs to explore its effect on the survival, and autophagy in the midgut cells  of N. lugens. RT-qPCR was used to detect the expression levels of NlGSK3, NlGS and NlGP  related to glycogen synthesis and metabolism in the 3rd instar nymphs after RNAi for 4 d.  【Results】 The full-length cDNA sequence of NlATG13 (GenBank accession no.: MF805752) was  cloned. It contains an open reading frame of 1 203 bp in length, encoding a protein of 400  amino acids (GenBank accession no.: AWW05678.1). The phylogenetic analysis showed that  NlATG13 protein was closely related to ATG13 proteins of Cimex lectularius and Halyomorpha  halys among the analyzed species. Developmental expression profiling showed that the  expression levels of NlATG13 in the 3rd and 5th instar nymphs were significantly higher  than those in the 1st-2nd instar nymphs, female adults and male adults of N. lugens. Tissue  expression profiling revealed that the relative expression level of NlATG13 was higher in  the head and fat body, but the lowest in the thorax of the 5th instar nymphs of N. lugens.  RNAi results showed that in the dsNlATG13 treatment group, glycogen granules were  accumulated in midgut cells, the expression levels of NlGS, NlGSK3 and NlGP had no  significant change, the ATP content in tissues was significantly decreased and the survival  rate of N. lugens significantly decreased as compared to those in the dsGFP control group.  The survival rate of N. lugens on day 10 post dsNlATG13 treatment was 41.4%, while that in  the dsGFP control group remained at a higher level of 85.6%. 【Conclusion】 RNA  interference targeting NlATG13 gene has a significant inhibitory effect on the survival and  autophagy of midgut cells in of N. lugens, and NlATG13 gene could be used as a potential  target in controlling N. lugens.
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miR-252-5p affects metamorphosis by targetedly regulating the expression of juvenile  hormone acid methyl transferase gene JHAMTin Drosophila
ZHANG Ze-Lin, JI Jie, XU Wen-Yu, LIU Zhuo, FAN Xiao-Chun, DENG Chang-Xin, HE Qian-Yu
Acta Entomologica Sinica    2022, 65 (7): 799-806.   DOI: 10.16380/j.kcxb.2022.07.002
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【Aim】 Juvenile hormone acid methyl transferase (JHAMT) is the key  rate-limiting enzyme in the juvenile hormone (JH) synthetic pathway. This study aims to  screen and verify the miRNAs targetedly regulating the transcription of JHAMT, and to  eveal the important action mechanism of miRNAs in JH biosynthesis in Drosophila  melanogaster. 【Methods】 MiRNAs targeting JHAMT in D. melanogaster were predicted using  online websites miRanda, TargetScan and microT-CDS, and those predicted by all the three  websites were selected as the miRNA candidates targeting JHAMT. The targeted relationship  between candidate miRNAs and JHAMT were verified using a dual luciferase assay system. The  expression profiles of miRNA and JHAMT during D. melanogaster development were detected by  qRT-PCR. The effects of miRNAs on the JHAMT expression and metamorphosis in D.  melanogaster were tested by overexpressing miRNA in the corpora allata using qRT-PCR and  the fly GAL4-UAS system, respectively. 【Results】 A total of 5, 18 and 16 miRNAs  targeting JHAMT were predicted by miRanda, TargetScan and microT-CDS, respectively, and  four miRNAs including miR-252-5p, miR-277-3p, miR-1002-5p and miR-987-5p were  coincidentally predicted by these three algorithms. Dual luciferase assay results showed  that miR-252-5p mimics significantly decreased the luciferase activities of wild-type  JHAMT 3′UTR luciferase reporter, and the suppression effect was compromised when the miR- 252-5p binding sites within the 3′UTR of JHAMT were mutated. qRT-PCR results showed that  miR-252-5p and JHAMT displayed opposite expression patterns in egg, larval and prepupal  stages of D. melanogaster, and overexpression of miR-252 in the corpora allata  significantly decreased the expression levels of JHAMT and the JH primary response gene Kr -h1. Meanwhile, overexpression of miR-252 resulted in similar phenotypes as JH deletion,  such as delayed pupation, reduced pupal size and increased pupal lethality. 【Conclusion】  miR-252-5p affects Drosophila metamorphosis through targeting JHAMT and regulating JH  biosynthesis.
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Wolbachia regulates the egg hatching of hybrids of Ectropis obliqua and E. grisescens  (Lepidoptera: Geometridae)
WANG Zhi-Bo, LIU Yong-Jian, BAI Jia-He, ZHANG Xin-Xin, XIAO Qiang
Acta Entomologica Sinica    2022, 65 (8): 999-1009.   DOI: 10.16380/j.kcxb.2022.08.008
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【Aim】 The objective of this study is to reveal the impacts of Wolbachia on the  egg hatching of hybrids of two sibling tea geometrids and the underlying mechanism.  【Methods】 A subset of Ectropis grisescens larvae were fed with fresh tea leaves soaked in  2.5 mg/mL tetracycline solution for 1 min to remove Wolbachia. E. grisescens populations  without Wolbachia were established after being fed with tetracycline-treated tea leaves  for three generations. E. obliqua adults were hybridized with E. grisescens adults with and  without Wolbachia to produce hybrids for the investigation of the impacts of Wolbachia on  the egg hatching. The iTRAQ technique was used to detect and analyze the differences in  sperm proteins between E. grisescens with and without Wolbachia. 【Results】 When Wolbachia  was removed from E. grisescens, the egg hatching rates of hybrids between E. grisescens and  E. obliqua were significantly increased from 3.92% to 56.20%, indicating that Wolbachia  induced cytoplasmic incompatibility. Results of differential sperm proteins analysis showed  that a total of 128 proteins were confirmed to have significantly different expression  levels in sperms of E. grisescens with and without Wolbachia. According to the KEGG  database, 45 differential proteins were enriched in 106 pathways, including sphingolipid  metabolism, salivary secretion, lysosome, sphingolipid signaling, sphingolipid  biosynthesis, etc. Among them the expression levels of ceramidases and phosphatases related  to ceramide synthesis in the sphingolipid metabolic pathway were significantly  up-regulated. 【Conclusion】 The symbiotic bacterium, Wolbachia, mediates the prezygotic  reproductive isolation between the two sibling tea geometrids through cytoplasmic  incompatibility, leading to a significant decrease in the egg hatching rate of hybrids. The  observed cytoplasmic incompatibility may be attributable to the modification of the male  sperm proteins caused by the impacts of Wolbachia on the sphingolipid metabolism pathway.
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Impact of overexpression and knockdown of ame-miR-13b on the expression of genes in larval gut of Apis mellifera ligustica
ZHU Zhi-Wei, WANG Jie, LONG Qi, XU Ya-Jing, FENG Rui-Rong, LIU Jia-Mei, ZHAO Hao-Dong, ZHU Le-Ran, HOU Hai-Qing, CHEN Da-Fu, GUO Rui
Acta Entomologica Sinica    2022, 65 (4): 460-468.   DOI: 10.16380/j.kcxb.2022.04.006
Abstract199)      PDF(pc) (3671KB)(69)    PDF(mobile) (3671KB)(14)    Save
【Aim】MicroRNAs (miRNAs) play a pivotal role in important life processes such as reproduction, development and immunity in insects. The aim of this study is to explore the impact of overexpression and knockdown of ame-miR-13b on the expression of target genes in Apis mellifera ligustica larvae, so as to provide theoretical and experimental bases for further investigation of regulation mechanism of ame-miR-13b underlying the development of A. m. ligustica larval gut. 【Methods】 Based on the nucleotide sequence of ame-miR-13b, the sequences of the corresponding mimic-ame-miR-13b and inhibitor-ame-miR-13b and their counterpart controls mimic-NC and inhibitor-NC were designed and synthesized, and then mixed with the diet which was used to feed the 3-day-old larvae for 6 times with the diet changed every 12 h to perform overexpression and knockdown of ame-miR-13b, respectively. RT-qPCR was performed to detect the expression levels of ame-miR-13b and target genes Ecr, Egfr and P450 18a1 in the larval gut of A. m. ligustica after overexpression and knockdown of ame-miR-13b. 【Results】 The expression levels of ame-miR-13b in the guts of the 4- and 6-day-old larvae of A. m. ligustica fed with mimic-ame-miR-13b were significantly up-regulated as compared to that in the group fed with mimic-NC, and those in the guts of the 4-day-old larvae and in the guts of the 5- and 6-dayold larvae fed with inhibitor-ame-miR-13b were down-regulated and significantly down-regulated, respectively, as compared to that in the group fed with inhibitor-NC. After overexpression of ame-miR-13b, the expression level of Egfr in the gut of the 6-day-old larvae was significantly decreased, while those of Ecr and P450 18a1 were significantly increased as compared to that in the group fed with mimic-NC. After knockdown of ame-miR-13b, the expression of Egfr in the gut of the 6 day-old larvae was up-regulated insignificantly, that of Ecr was significantly up-regulated, whereas that of P450 18a1 was significantly down-regulated as compared to that in the group fed with inhibitor-NC. 【Conclusion】 Overexpression and knockdown of miRNA in larval individuals of A. m. ligustica are successfully achieved by feeding method. There is a potential negative regulatory relationship between ame-miR-13b and Egfr.
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Cloning and functional characterization of the peptidoglycan recognition protein gene SePGRP-SA in Spodoptera exigua (Lepidoptera: Noctuidae)
LI Ya-Zi, ZHAO Dan, GUO Xiao-Chang, WU Han, LIU Zhao-Rui, GUO Wei
Acta Entomologica Sinica    2022, 65 (4): 417-426.   DOI: 10.16380/j.kcxb.2022.04.002
Abstract198)      PDF(pc) (5599KB)(133)    PDF(mobile) (5599KB)(30)    Save
【Aim】 This study aims to elucidate the function of peptidoglycan recognition protein (PGRP) in Spodoptera exigua larvae in response to Bacillus thuringiensis (Bt) infection. 【Methods】 The fulllength cDNA of SePGRP-SA from S. exigua larvae was cloned by PCR method. The relative expression levels of SePGRP-SA in different developmental stages (egg, 1st-5th instar larval, prepupal and pupal stages) and different tissues of the 4th instar larvae (midgut, Malpighian tubules, peritrophic membrane, fat body, hemolymph and epidermis) of S. exigua were analyzed by qRT-PCR. At 72 h after silencing SePGRP-SA by RNAi, the silence efficiency of SePGRP-SA, the changes in the expression levels of antimicrobial peptide-related genes (Ceropin, Attacin and Defensin) and the bacterial load in the midgut of the 4th instar larvae were detected by qRT-PCR. At 0, 24, 48, 72, 96 and 120 h after the 4th instar larvae of S. exigua were fed with B. thuringiensis Bt-GS57 strain following silencing SePGRP-SA by RNAi for 24 h, the corrected mortality rates of larvae were calculated. The relative expression levels of SePGRP-SA, Ceropin, Attacin and Defensin in the migdut of the 4th instar larvae fed with Bt-GS57 for 0, 24, 48 and 72 h were detected by qRT-PCR. 【Results】 The full-length cDNA of SePGRP-SA (GenBank accession no.: MW265930) was successfully cloned. Its ORF is 576 bp in length encoding 191 amino acid residues with the molecular weight of 21.59 kD. Sequence analysis results indicated that SePGRP-SA contains typical conserved PGRP and Ami2 domains and a 19-amino-acid signal peptide, being a secretory protein. Phylogenetic analysis indicated that SePGRP-SA is most closely related to SlPGRP of Spodoptera litura, sharing 91.1% amino acid sequence identity. Developmental expression profiles revealed that SePGRP-SA was highly expressed in the 4th and 5th instar larval, pre-pupal and pupal stages of S. exigua. Tissue expression profiles showed that SePGRP-SA was expressed in various tissues of the 4th instar larvae, with the highest expression level in the hemolymph. After the 4th instar larvae of S. exigua were injected with dsSePGRP-SA for 72 h, the expression level of SePGRP-SA in the midgut was down-regulated by 95.26%, while those of Cecropin, Attacin and Defensin were significantly down-regulated, and the microbial load in the midgut was significantly increased as compared to those in the control (infection with dsEGFP). At 72 h after the 4th instar larvae of S. exigua were fed with Bt-GS57 following injection with dsEGFP and dsSePGRP-SA, the corrected mortality rates of larvae were 50.00% and 73.33%, respectively, indicating the significantly increased sensitivity of larvae to Bt-GS57. After the 4th instar larvae of S. exigua were fed with Bt-GS57, the expression levels of SePGRP-SA, Cecropin, Attacin and Defensin in the midgut were increased significantly at 48 h after feeding, but decreased at 72 h after feeding. 【Conclusion】 SePGRP-SA gene of S. exigua can activate the expression of antimicrobial peptide-related genes Cecropin, Attacin and Defensin after infection of Bt.
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Whole-genome resequencing reveals the genetic diversity and adaptive evolution of  Apis  cerana  (Hymenoptera: Apidae) on the eastern and southeastern edges of the Qinghai-Tibet  Plateau
TANG Xiang-You, SONG Hua-Li, SHI Peng, ZHANG Xiao-Yan, TANG Zi-Han, WANG Wen-Feng, ZHA Luo, CHEN Xin-Lan, ZHOU Ze-Yang, XU Jin-Shan
Acta Entomologica Sinica    2022, 65 (5): 638-647.   DOI: 10.16380/j.kcxb.2022.05.012
Abstract198)      PDF(pc) (3213KB)(71)    PDF(mobile) (3213KB)(15)    Save
【Aim】 This study aims to study the population genetic diversity and adaptive evolution of Apis cerana samples from the eastern and southeastern edges of the Qinghai-Tibet Plateau and adjacent areas, so as to provide references for further revealing their genetic resource diversity, population diffusion rules and molecular evolutionary mechanisms of adapting to plateau habitats. 【Methods】 The whole-genomes of 77 colonies of A. cerana samples collected from the eastern and southeastern edges of the Qinghai-Tibet Plateau and adjacent areas were resequenced. The whole-genome reseqeuncing data of the 77 colonies of A. cerana and the reseqeuncing raw data of 90 colonies of A.  cerana downloaded from GenBank database were analyzed using population genetic method based on the population structure, principal component analysis, phylogenetic tree, genetic differentiation index, mitochondrial genome haplotype and selective signal analysis. 【Results】 The 167 colonies of A. cerana were separated into four plateau colonies including colonies from Western Sichuan Plateau, Southern Tibet Plateau, Northern Yunnan Plateau and Northern Sichuan Plateau, which clustered into two clades. The average genetic differentiation index of A. cerana colonies in the plateau region (Fst=0.1178) was higher than that in the non-plateau region ( Fst=0.0411). The analysis of the minimum genetic distance among populations showed that the colonies of Southern Tibet Plateau, Northern Yunnan Plateau and South Yunnan on the southeastern edge had closer genetic relationships. The colonies of Western Sichuan Plateau and Northern Sichuan Plateau on the eastern edge had closer genetic relationship with those of the Western Sichuan Mountain and Qinba, respectively. Combined with the haplotype analysis of the mitochondrial genome, the ancestral haplotype and origin of the plateau populations on the eastern and southeastern edges of the Qinghai-Tibet Plateau were preliminarily inferred. Through selective analysis, potential selected genes involved in signaling pathways of fatty acid metabolism, phototransduction, temperature adaptation, and ovarian development were identified. Two co-selected genes, ISL-1 and FOXO, were found in the populations of Southern Tibet Plateau and Northern Yunnan Plateau to be mainly involved in insulin secretion in response to cellular stress, suggesting that they play important roles in the adaption of A. cerana to the habitat of the southeastern Qinghai-Tibetan Plateau. 【Conclusion】 The genetic diversity of A. cerana in the Qinghai-Tibet Plateau is highly abundant, and the colonies of Southern Tibet Plateau and Northern Yunnan Plateau on the southeastern edge and Western Sichuan Plateau and Northern Sichuan Plateau on the eastern edge can be obviously distinguished. The four plateau colonies are geographically isolated after the diffusion of non-plateau populations in adjacent areas, resulting in population differentiation. The 
potential genes of plateau environmental adaptability of A. cerana were identified by preliminary screening. This study lays a basis for further exploring the molecular evolutionary mechanism of the adaptation of A. cerana to plateau habitat.
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Effects of exogenous juvenile hormone on the ovarian development and transcription levels  of the reprodution-related genes in Harmonia axyridis (Coleoptera: Coccinellidae)  
HAN Hui, FENG Zhao-Yang, ZHANG Shuo, HE Yun-Zhuan
Acta Entomologica Sinica    2022, 65 (9): 1090-1097.   DOI: 10.16380/j.kcxb.2022.09.002
Abstract195)      PDF(pc) (7691KB)(128)       Save
【Aim】 This study aims to clarify the effects of exogenous juvenile hormone (JH)  on the ovarian development and transcriptional levels of the key genes in the reproduction -related signaling pathways of Harmonia axyridis. 【Methods】 The 2-day-old female  adults of H. axyridis fed with artificial diet were exposed to different doses (80, 120 and  160 ng/individual) of JHⅢ by topical application method for 1, 3, 5, 7 and 9 d (the female  adults fed with the turnip aphid, Lipaphis erysimi, were used as the blank control group,  and those treated with acetone solution of the same volume by topical application method as  the vehicle control group), the ovaries were dissected, and the ovarian length and the  length and width of their first egg chamber were photographed and measured. qPCR was used  to analyze the expression levels of the key genes including JH receptor  methoprene-tolerant (Met) gene, krüppel homolog 1 (Kr-h1) gene, vitellogenin genes (Vg and Vg2) and vitellogenin receptor (VgR) gene in the reproduction-related signal pathways  of the female adults of H. axyridis exposed to the optimum dose (120 ng/individual) of JHⅢ  for 1, 5, and 9 d. 【Results】 Compared with the vehicle control group, treatments with 80  and 120 ng/individual of JHⅢ by topical application method could promote the ovarian 
development of H. axyridis adults. After the 2-day-old female adults were exposed to 120  ng/individual of JHⅢ for 7 d, a large amount of yolk deposition appeared in the ovary,  which was consistent with the ovarian developmental state in the blank control group fed 
with the turnip aphid, and the ovarian length and the length and width of the first egg  chamber were significantly higher than those in the vehicle control group. However, the  treatment with 160 ng/individual of JHⅢ inhibited the ovarian development of H. axyridis  adults. The qPCR results showed that the expression levels of HaMet, HaKr-h1, HaVg1, HaVg and HaVgR in female adults at 5 and 9 d after treatment with 120 ng/individual of JHⅢ  increased as compared to the those in the control group (acetone treatment group): those of  HaMet, HaVg1 and HaVgR in female adults at 5 d after treatment were 2.78, 13.14 and 8.28  times, respectively, and those in female adults at 9 d after treatment were 2.36, 1.85,  and 2.01 times, respectively, as high as those in the control group, showing extremely  significant difference. 【Conclusion】 JHⅢ at the dose of 120 ng/individual can promote  the ovarian development of H. axyridis adults and upregulate the expression levels of  HaMet, HaKr-h1, HaVg and HaVgR in female adults at 5 and 9 d after treatment. It is  speculated that these key genes play important roles in the regulation of JH on the ovarian  development and yolk formation of H. axyridis.
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Cloning of nicotinic acetylcholine receptor α6 and α7 subunit genes in Spodoptera  frugiperda (Lepidoptera: Noctuidae) and their response to spinetoram stress
GAO Zu-Peng, JING Da-Peng, HUANG Xiao-DAN, WANG Zhen-Ying
Acta Entomologica Sinica    2022, 65 (9): 1098-1105.   DOI: 10.16380/j.kcxb.2022.09.003
Abstract195)      PDF(pc) (6443KB)(124)       Save
【Aim】 Nicotinic acetylcholine receptors (nAchRs), as the most crucial  neurotransmitter receptors in the central nervous system of insects, are the important  targets of spinetoram. This study aims to clone the nAchR genes of Spodoptera frugiperda  and investigate their response to spinetoram stress. 【Methods】 Genes of nAchR α6 and α7  subunits of S. frugiperda were cloned via RTPCR and RACE based on the previous  transcriptome database of S. frugiperda. After the 3rd instar larvae of S. frugiperda were  exposed to spinetoram (0.400 mg/L) for 48 h, the changes of expression levels of nAchR α6  and α7 subunit genes were assayed by RT-qPCR. 【Results】 The full-length ORF of nAchR  α6 subunit gene (GenBank accession no.: MT951400) is 1 506 bp in length, encoding 502  amino acids with transmembrane region and signal peptide. The full-length ORF of nAchR α7  subunit gene (GenBank accession no.: MW557608) is 1 524 bp in length, encoding 508 amino  acids with transmembrane region and signal peptide. nAchR α6 and α7 subunits of S.  fruiperda have the typical characteristic of nAchR α family according to amino acid  sequence multiple alignment analysis. After the 3rd instar larvae of S. frugiperda were  exposed to 0.400 mg/L spinetoram for 48 h, the expression level of their nAchR α6 subunit  gene increased significantly, while that of nAchR α7 subunit gene did not change  significantly. 【Conclusion】 Spinetoram may have an impact on the nAchR α6 subunit of S.  fruiperda. This study lays a preliminary foundation for future in-depth research on the  underlying target resistance mechanisms of S. frugiperda in response to spinetoram.
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Effects of CO2 narcosis on the ovarian development of virgin queens of Apis cerana cerana  (Hymenoptera: Apidae)
ZHOU Jian-Yang, SONG Hua-Li, DENG Ping, TAO Kun-Ling, TANG Xiang-You, YAN Le-Le, ZHANG Xiao-Yan, SHI Peng, XU Jin-Shan
Acta Entomologica Sinica    2022, 65 (5): 577-586.   DOI: 10.16380/j.kcxb.2022.05.005
Abstract193)      PDF(pc) (19278KB)(134)    PDF(mobile) (19278KB)(16)    Save
【Aim】 To clarify the dynamic changes of reproductive system of queens of the  Chinese honeybee (Apis cerana cerana) after emergence, and to explore the effects of CO2  narcosis on the ovarian development of their virgin queens. 【Methods】 Through artificial  breeding technology and storage of the queens, we obtained different day-old virgin queens  of A. cerana cerana, observed the morphological characteristics of their reproductive  system after emergence, and determined the morphological indexes of their reproductive  system including the body weight, ovarian weight, diameter of spermatheca, and number of  ovarioles. After the virgin queens were narcotized with different concentrations (50%, 75%  and 90%) of CO2 in two consecutive days (at the 5- and 6-d-old, respectively) for 8 min,  and different day-old virgin queens were respectively narcotized with 75% CO2 for 1-3  times (each time for 8 min), the morphological characteristics of the ovaries of the 10-d -old queen were observed by paraffin section and HE staining, and their ovarian weight and  the length of the largest oocyte of ovaries were detected. Meanwhile, the relative  expression level of vitellogenin gene (Vg) in the head of the 10-d-old queens was  measured by real-time fluorescence quantitative PCR (RT-qPCR). 【Results】 The number of  ovarioles and the diameter of spermatheca of the queens of A. cerana cerana did not  fluctuate obviously with the age increasing. However, the ovarian weight of queens  increased obviously during the 6-12-d-old and then tended to be stable. As compared to  the control (not subjected to CO2 narcosis), narcosis with 50%, 75% and 90% CO2 resulted in  the increase of the ovarian weight and the length of the largest oocyte of queens of A.  cerana cerana and the upregulation of the relative expression level of Vg gene in the  queen head. Moreover, narcosis with 75% CO2 twice significantly promoted the ovarian  development of A. cerana cerana queens. 【Conclusion】 The ovaries of A. cerana cerana  queens present obvious developmental changes after emergence, and CO2 narcosis can  accelerate their ovarian development.
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Gonadal development and gene expression related to the reproductive plasticity of workers  of the termite Reticulitermes labralis(Isoptera: Rhinotermitidae)
YE Chen-Xu, SONG Zhuan-Zhuan, ZHANG Wen-Xiu, WU Tao-Yu, LIU He, XING Lian-Xi, SU Xiao-Hong
Acta Entomologica Sinica    2022, 65 (6): 657-667.   DOI: 10.16380/j.kcxb.2022.06.001
Abstract193)      PDF(pc) (9365KB)(112)       Save
【Aim】 In order to investigate the characteristics of ovarian development and  the starting point for resumption of oocyte development during the differentiation of  female workers into neotenic reproductives (NRs) of Reticulitermes labralis. 【Methods】  The dynamic changes of ovary and oocyte during the development of female workers (from the  3rd instar to the 6th instar) and the differentiation of female workers into NRs of R.  labralis were observed. Based on the transcriptome data of the development of primary  reproductives and NRs from nymphs of R. labralis, genes involved in oocyte growth were  identified and their expression levels during the differentiation of workers into NRs were  detected using qRT-PCR method. 【Results】 The size of ovaries in workers of R. labralis  increased gradually from early instar to late instar, the length and width of the ovaries  of pre-neotenic reproductives (pre-NRs) were approximately 2- and 3-fold as long as  those of workers, respectively, but did not increase significantly after the pre-NRs  developed into NRs. After the workers developed into pre-NRs, there was no significant  change in the size (long diameter) of oocytes and the thickness of the layer of follicle  cells. However, after the pre-NRs developed into NRs, the size of oocytes and the  thickness of the layer of follicle cells increased significantly. The expression levels of  six genes involved in oocyte growth in NRs were extremely significantly increased. The  expression levels of the six genes cyclin-dependent kinase 1, cell division cycle protein  20, G2/mitotic-specific cyclin-B3, G2/mitotic-specific cyclin-A, aurora kinase A and  serine/threonine-protein kinase polo in NRs were approximately 34-, 62-, 91-, 36-, 57-  and 106-fold as high as those in pre-NRs. After the workers developed into pre-NRs, only  the expression levels of G2/mitotic-specific cyclin-B3, G2/mitotic-specific cyclin-A  and aurora kinase A increased significantly, being approximately 3-, 3- and 2-fold in  pre-NRs as high as in workers. 【Conclusion】 The ovarian development of R. labralis  workers is arrested at late instar stage, and the oocyte development and meiotic resumption  begin after the workers develop into NRs.
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Analysis of differential proteins in Reticulitermes perilucifugus (Isoptera:  Rhinotermitidae) workers fed with different diets
SU Li-Juan, ZHAO Peng-Fei, DU Yun-Liang, GUAN Yu-Liang, GUO Rui-Yao, YIN Zheng-Xing, SONG An-Dong
Acta Entomologica Sinica    2022, 65 (8): 986-998.   DOI: 10.16380/j.kcxb.2022.08.007
Abstract193)      PDF(pc) (5614KB)(173)       Save
【Aim】 This study aims to provide the basis at the protein level for exploring the mechanism of nutrient digestion and absorption in termites by comparing the differences in protein composition and expression in the fore-midgut and hindgut contents of  Reticulitermes perilucifugus workers fed with diets with different lignocellulose contents. 【Methods】 R. perilucifugus workers were fed with three diets (pine, straw and filter paper) with different lignocellulose contents. The proteins in contents of different parts of their gut were analyzed by two dimensional (2D) electrophoresis, and the differiential proteins were sequenced and bioinformatically analyzed by MALDI-TOF/MS. 【Results】 The results of 2D electrophoresis showed that the number of visible protein spots in the fore-midgut of R. perilucifugus workers fed with the same diet was significantly higher than that in the hindgut, while those in the fore-midgut and hindgut of workers fed with pine were the highest, those in the fore-midgut and hindgut of workers fed with filter paper followed, and those in the fore-midgut and hindgut of workers fed with straw were the least. The sequencing results of 115 protein points showed that the mainly differential proteins between the fore-midgut and hindgut were proteins with catalytic activity including enzymes related to amino acid metabolism, pyruvate metabolism, carbon metabolism, nitrogen metabolism, TCA cycle, glycolysis, gluconeogenesis and cellulose degradation and proteins involved in cellular components and signal transduction. 【Conclusion】 The differential proteins in the gut of R. perilucifugus workers fed with different diets are 
mainly proteins with catalytic activity, and proteins involved in cell composnents and signal transduction, indicating that different diets affect the composition of intestinal proteins of R. perilucifugus. These results provide data for revealing the degradation mechanism of lignocellulose in termites.
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Structures of the midgut and Malpighian tubules in the cave cricket, Tachycines asynamorus  (Orthoptera: Rhaphidophoridae)
ZHANG Li-Dan, GONG Xu-Ping, PENG Xin-Fei, SHI Fu-Ming, CHANG Yan-Lin
Acta Entomologica Sinica    2022, 65 (6): 695-707.   DOI: 10.16380/j.kcxb.2022.06.005
Abstract191)      PDF(pc) (35850KB)(66)       Save
【Aim】 Taking the cave cricket, Tachycines asynamorus, as an example, this study  aims to explore the structural adaptation relationship of digestive system and excretory  system of cave crickets to living environment. 【Methods】 The structures of midgut and  Malpighian tubules of T. asynamorus were explored by anatomical methods, paraffin section,  frozen section and ultrathin section. 【Results】 Three gastric caeca of midgut of T.  asynamorus extend forward to enclose a proventriculus. The midgut epithelia consist of  regenerative cells, columnar cells, and endocrine cells, with typical niches of  regenerative cells. Closed endocrine cells are usually located in the periphery of the  niches and a large number of secretory granules are gathered in the basal region of  endocrine cells. There are two kinds of large secretory granules in columnar cells, linear  clump-like ones and spherical ones with high electron density. In the lumen of the midgut,  there is a distinct peritrophic matrix. The base of the midgut is composed by the basal  lamina and muscle layer. Malpighian tubules are joined to the digestive tract at the  junction of the midgut and the hindgut. The transverse section of the Malpighian tubules  contains 3-5 cells. There are a vast number of long microvilli in the apical side of the  cells, facing the lumen. Numerous concentrically layered spherites with dense electrons  were observed in the cells. In the basal regions of the epithelial cells, there are  numerous basement membranes which form membrane labyrinth by infolding. 【Conclusion】In  the midgut of T. asynamorus, the linear clump-like secretory granules in columnar cells  are wrapped by microfilaments. The endocrine cells from the niche stem cell produce  endocrine granules, and then excrete the endocrine granules into the hemocoel. The midgut  basal lamina including glycoconjugates and microfilaments develops well and provides  support for intestinal peristalsis by supporting midgut epithelial cells. A large number of  particles and concentrically layered spherites exist in the Malpighian tubule cells of T.  asynamorus, implying their functions of storage and excretion.
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Cloning, identification and functional analysis of the peroxidase gene NlPOD1 in the brown  planthopper, Nilaparvata lugens(Hemiptera: Delphacidae)
LI Mu-Yu, WANG Yan-Dan, WANG Zheng-Liang, YU Xiao-Ping
Acta Entomologica Sinica    2022, 65 (8): 958-966.   DOI: 10.16380/j.kcxb.2022.08.004
Abstract190)      PDF(pc) (2070KB)(140)       Save
 【Aim】 To clarify the molecular characteristics, expression patterns and  biological function of the peroxidase gene NlPOD1 from the brown planthopper, Nilaparvata  lugens. 【Methods】 Based on the transcriptome data of N. lugens, the full-length cDNA  sequence of NlPOD1 was cloned by PCR, and its nucleotide and protein sequences were  subsequently characterized using bioinformatics tools. The expression patterns of NlPOD across different developmental stages (egg, 1 st-5th instar nymphs and newly emerged female  and male adults), in different tissues (head, fat body, hemolymph and gut) of the 5th  instar nymphs, and in the 5th instar nymphs at different time post injection of different  microbes (Escherichia coli, Staphylococcus aureus and Metarhizium anisopliae) at the  concentration of 1×107/mL were determined by qRT-PCR. The target gene NlPOD1 in the 5th  instar nymphs of N. lugens was further silenced by RNAi, and the survival rates and the  median lethal time (LT50) values of the nymphs after target gene silencing and infection  with M. anisopliae (1×108 conidia/mL) were determined by bioassay. 【Results】 The  full-length cDNA sequence of NlPOD1 (GenBank accession no.: MZ682107) was successfully  cloned from N. lugens. Its open reading frame (ORF) is 2 049 bp in length, encoding 682  amino acids with a typical animal heme peroxidase domain (An_ peroxidase domain) and a  predicted signal peptide consisting of 29 amino acid residues at the N-terminus. The  phylogenetic analysis showed that NlPOD1 is closely related to the PODs of other hemipteran  insects, and has the highest homology with the POD of Halyomorpha halys. Developmental  expression profiling showed that NlPOD1 was expressed in various developmental stages of N.  lugens, with the lowest and highest expression levels in the eggs and the 5th instar  nymphs, respectively. Tissue expression profiling revealed that NlPOD1 was expressed in  different tissues of the 5th instar nymphs of N. lugens, with significantly higher  expression levels in the gut and haemolymph than in the head and fat body. The expression  patterns under microbial induction showed that the expression level of NlPOD1 in the 5th  instar nymphs of N. lugens was significantly up-regulated within 48 h post injection with  E. coli as compared to the control group injected with PBS. However, the expression level  of NlPOD1 increased firstly and then stabilized when the 5th instar nymphs of N. lugens  were challenged by S. aureus and M. anisopliae. The RNAi results showed that the expression  levels of NlPOD1 could be significantly inhibited by microinjection of dsNlPOD1. Inhibition  of NlPOD1 expression by RNAi caused no changes in the survival rate of the 5th instar  nymphs of N. lugens.  However, the LT50 of the combined treatment with dsNlPOD1 injection  and M. anisopliae infection to the 5th instar nymphs of N. lugens was 4.5 d, which was  significantly shorter than that of the control group (5.4 d) of combined treatment with  dsGFP injection and M. anisopliae infection, indicating that silencing of NlPOD1 could  significantly decrease the resistance of N. lugens to the infection with M. anisopliae.  【Conclusion】 NlPOD1 plays important roles in the pathogen defense of N. lugens and can be  used as a potential target in the development of N. lugens biocontrol technology mediated  by RNAi and entomopathogenic fungi.
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Biological characteristics of overwintering nymphs of the horned-gall aphid,  Schlechtendalia chinensis (Hemiptera: Aphididae)
XU Xin, WEI Hong-Yuan, FENG Guo-Rui, SHAO Shu-Xia, YANG Zi-Xiang
Acta Entomologica Sinica    2022, 65 (6): 730-736.   DOI: 10.16380/j.kcxb.2022.06.008
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【Aim】 The horned-gall aphid, Schlechtendalia chinensis, is the major  productive species of Chinese gallnuts. Understanding the biological characteristics and  population dynamics of its overwintering nymphs under the condition of soilless moss in the  field will provide a basis for further reducing the mortality of overwintering nymphs and  increasing the yield of Chinese gallnuts. 【Methods】 The overwintering nymphs of S.  chinensis were reared in the field on the host moss Plagiomnium maximoviczii which was  planted on the non-woven fabrics. Mosses with S.chinensis at different developmental  stages were collected regularly and brought back to the laboratory. The behavioral habits,  morphological characteristics, population dynamics and instar distributions of S. chinensis  nymphs during overwintering were investigated using a digital microscopic system. The  thickness and coverage rate of P. maximoviczii duringtheoverwinteringperiodwere measured as  well. 【Results】 Autumn migrants of S.  chinensis reproduce ovoviviparously. The nymphs of S. chinensis excrete wax at the base of  phyllidia of P. maximoviczii and wrap themselves individually to form wax balls where they  feed and overwinter. Usually there is a single nymph in a wax ball. The body color of  nymphs from the 1st instar to the 4th instar deepens gradually from light yellow to dark  brown. The body length and width of nymphs increased with the nymphal instar, from 552.92± 16.95 and 294.70±11.52 μm of the 1st instar nymphs to 1 205.25±10.75 and 593.15±7 .66 μm of the 4th instar nymphs, respectively. During the overwintering period, the  population density of nymphs dropped from 131 000 aphids/m2 in mid-October to 10 500  aphids/m2 in next March. The total mortality rate was as high as 91.98%. The developmental  progress of overwintering nymphs was irregular and closely related to the changes of local  temperature. The thickness and coverage rate of the moss layer increased gradually during  overwintering of S. chinensis nymphs. 【Conclusion】 The body length and body width may be  used as the main indicators for the identification of different instars of overwintering  nymphs of S. chinensis. The total mortality rate of overwintering nymphs of S. chinensis in  the field is very high, and the mortality rates during the early overwintering stage and  the middle host-transferring stage are higher than those during the other stages.
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Green synthesis of silver nanoparticles using plant extracts and their toxicity to  Coptotermes formosanus  (Blattodea: Rhinotermitidae)
XIE Xiao-Jun, YANG Gui-Ying, YU Bao-Ting, HU Yin, MO Jian-Chu
Acta Entomologica Sinica    2022, 65 (4): 480-489.   DOI: 10.16380/j.kcxb.2022.04.008
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【Aim】 This study aims to make green synthesis of silver nanoparticles (AgNPs) using aqueous extracts of multiple medicinal plants, and to detect and analyze their toxicity and action mechanism against the Formosan subterranean termite, Coptotermes formosanus, so as to explore the potential of AgNPs in termite control and expand its future application in agriculture. 【Methods】 The aqueous extracts of four medicinal plants including root of rhubarb ( Rheum palmatum), plants of decumbent bugle herb ( Ajuga nipponensis), root of lightyellow sophorn ( Sophora flavescens) and plants of houttuynia ( Houttuynia cordata) were used as the raw materials to synthesize AgNPs. The formation of AgNPs was characterized by UV-visiblespectroscopy (UV-vis), scanning electron microscopy (SEM), transmission electronmicroscopy (TEM), X-ray energy dispersive spectrum(EDS) and nanoparticle size analyzer to assess the size, shape and aggregation degree of AgNPs. Then, the toxicity of AgNPs to C. formosanus workers was measured in the laboratory condition and the action mechanism of AgNPs on termites was explored by determining the soluble protein content, acetylcholinesterase (AchE) activity and filter paper activity (FPA) in C. formosanus workers at 7 d after treatment with 800 mg/L AgNPs.【Results】 The green synthesized AgNPs with the aqueous extracts of four medicinal plants were spherical particles, with an average particle diameter of 69-180 nm. The LC 50 values of AgNPs against C. formosanus workers in 7 d were 150, 340, 342 and 309 mg/L, respectively. The soluble protein contents, AchE activities and FPA in C. formosanus workers at 7 d after treatment with 800 mg/L AgNPs were significantly decreased compared with those in the control. 【Conclusion】 AgNPs synthesized by four plant extracts show high toxicity to C. formosanus workers, and the survival of termites can be affected by AgNPs by reducing the soluble protein content, AchE activity and FPA, suggesting that AgNPs have great potential in control of C. formosanus.
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Effects of Tomato chlorosis virus on the host preference and feeding behaviors of Bemisia tabaci MED (Hemiptera: Aleyrodidae) adults on different host plants
FAN Xiao-Fan, LIU Yong, TAN Xin-Qiu, ZHANG Zhan-Hong, ZHANG Zhuo, SHI Xiao-Bin, ZHANG De-Yong
Acta Entomologica Sinica    2022, 65 (8): 1018-1025.   DOI: 10.16380/j.kcxb.2022.08.010
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【Aim】 Tomato chlorosis virus (ToCV) is transmitted by whiteflies and breaks out  heavily. At present, ToCV has spread to most regions of the world, and can harm a variety  of crops and cause serious damage to agricultural and forestry economic production. Virus  infection can influence the host preference and feeding behaviors of vector insects, thus  influencing virus transmission. The aim of this study is to investigate the effects of ToCV  on the host preference and feeding behaviors of Bemisia tabaci MED adults on different host  plants. 【Methods】 The host selectivity of ToCV-infected and non-infected B. tabaci MED  female adults was determined by Y-tube olfactometer, and the feeding behaviors between  ToCV-infected and ToCV-non-infected female adults of B. tabaci MED on four healthy host  plants including tomato, pepper, cotton and cowpea were compared with electrical  penetration graph (EPG). 【Results】 The preference of ToCV-non-infected female adults of  B. tabaci MED to tomato and pepper was the highest, that to cotton followed and that to  cowpea was the least. The preference of female adults of B. tabaci MED infected by ToCV to  tomato, pepper and cotton was higher than that to cowpea. The numbers of probes of the ToCV -infected female adults of B. tabaci MED on the four plants were significantly increased,  their first time to probe phloem was significantly delayed, and their total feeding time  and phloem feeding time significantly decreased as compared with those of the  ToCV-non-infected female adults. 【Conclusion】 ToCV significantly changes the host  preference and feeding behaviors of B. tabaci, thus increases the probability of its own  transmission among host plants.
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A survey of the composition and occurrence of agromyzid leafminers and their parasitoids in Baiyin, Gansu, northwestern China
HE Jing, DU Su-Jie, CHENG Xin-Fei, WANG Qi-Jing, GUO Jian-Yang, WANG Zuo-Wei, WANG Fu-Lian, LIU Wan-Xue
Acta Entomologica Sinica    2022, 65 (4): 533-540.   DOI: 10.16380/j.kcxb.2022.04.013
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【Aim】 The purpose of this study is to reveal the diversity and population composition of the agromyzid leafminers attacking crops and their parasitoids in Baiyin, Gansu province, northwestern China. 【Methods】 The vegetables, flowers, and weeds damaged by the agromyzid leafminers were surveyed and sampled in fields by five-point sampling method, and the agromyzid leafminers and their parasitoids were identified by morphological and molecular methods. 【Results】 In total six species of agromyzid leafminers were found in the collected samples in Baiyin, Gansu province during 2016-2020, including native species Chromatomyia horticola and Liriomyza chinensis, and invasive species L. huidobrensis, L. trifolii, L. bryoniae, and L. sativae. Among them, C. horticola was the dominant species which damaged multi-family host plants. L. chinensis was only found on Allium fistulosum . L. huidobrensis occurred seriously on vegetables in the greenhouse. Furthermore, it was the first time to find L. trifolii in Baiyin. Besides, L. bryoniae and L. sativae had less damage than other agromyzid leafminers. In addition, the surveyed parasitoids included 24 species, 12 genera and 3 families, and the dominant species were Diglyphus isaea, Neochrysocharis formosa, D. wani, and Dacnusa sibirica. 【Conclusion】 On the one hand, monitoring and early warning of the occurrence of the invasive agromyzid leafminers should be strengthened in Baiyin. On the other hand, the parasitoids of the agromyzid leafminers are abundant in Baiyin and may play an important role in natural control of the main damage by the agromyzid leafminers in the field, so it is recommended to protect and apply of local parasitoids.
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Functional analysis of survivin gene in the mitosis of BmN4 cells of Bombyx mori
ZHANG Bing, WANG Yi, LI Na, LI Dan-Dan, KAN Yun-Chao
Acta Entomologica Sinica    2022, 65 (5): 541-547.   DOI: 10.16380/j.kcxb.2022.05.001
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【Aim】 To clarify the function of survivin gene in the mitosis of BmN4 cells of Bombyx mori. 【Methods】 The expression levels of BmSurvivin in different tissues (silk gland, midgut, Malpighian tubules, testis, ovary, fat body, epidermis and cuticle) of the day-3 5th instar larvae of B. mori were detected by qRT-PCR. The fusion vector pIZT/V5-His-BmSurvivin-GFP (BG) was constructed and transfected into BmN4 cells. The localization of BmSurvivin and Ser10 phosphorylated histone H3 (H3Ser10ph) in different mitotic stages of BmN4 cells was detected by immunofluorescence. 【Results】 The expression level of BmSurvivin was the highest in the Malpighian tubules of the day-3 5th instar larvae of B. mori, followed by that in the silk gland and midgut. pIZT/V5-His-BmSurvivin-GFP vector was successfully constructed. The immunofluorescence results showed that obvious GFP signal could be seen in the interphase nucleus of BmN4 cells, covering the nuclear and cytoplasmic regions indicating the location of BmSurvivin. At the mitotic phase, BmSurvivin was co-localized with chromatin indicated by GFP signal. When the BmN4 cells formed obvious double orientation, GFP signal was also visible in the spindle region. At the anaphase stage, with the separation of sister chromatids, BmSurvivin was located in the chromatin and cytokinesis region indicated by GFP signal. At the telophase stage, with the formation of two new daughter cells, BmSurvivin was located only in the cytokinesis region indicated by GFP signal. H3Ser10ph was located at the site of chromatin condensation at the prometaphase stage of BmN4 cells, and the signal intensity was the strongest at the metaphase, coinciding with the whole chromosome. The signal disappeared at the anaphase stage. 【Conclusion】 BmSurvivin is associated with the dynamic changes of chromatin and spindle during mitotic cycle.
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Circadian rhythms of the sex pheromone biosynthesis and release and the calling and mating  behaviors of adult Sesamia inferens(Lepidoptera: Noctuidae)
GUO Qian-Shuang, ZHUO Fu-Yan, ZHU Jing-Quan, CHEN Qing-Hua, ZHU Feng, HUANG De-Chao, DU Yong-Jun
Acta Entomologica Sinica    2022, 65 (7): 877-885.   DOI: 10.16380/j.kcxb.2022.07.009
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【Aim】 To explore the circadian rhythms of the synthesis and release of sex  heromones Z11-16∶Ac and Z11-16∶OH and the calling and mating behaviors of adult  Sesamia inferens, and their relationships with sex pheromone trapping in the field.  【Methods】 The sex pheromone titers of Z11-16∶Ac and Z11-16∶OH of female moths of S.  inferens were analyzed by solvent extraction and solid phase microextraction (SPME), and  the circadian rhythms of sex pheromone biosynthesis and release and the calling and mating  behaviours of moths were investigated by behavioral observation and real-time counting of  moth catches by sex pheromone trapping in the field in multiple localities. 【Results】 The  detectable time of sex pheromone Z11-16∶Ac and Z11-16∶OH in the pheromone glands of  female moths of S. inferens started at 1 h before scotophase, and increased rapidly at 4 h  into scotophase, reaching the first peak at 8 h into scotophase and the 2nd peak at 1 h  into photophase. The sex pheromone compounds were significantly detected at 5 h into  photophase. The detectable time for sex pheromone compounds emitted outside of the glands  started at 6 h into scotophase and peaked at 10 h into scotophase, and the sex pheromone  Z11-16∶Ac titer still remained 96.9±20.9 ng/female even at 1 h into photophase. The  ratios of Z11-16∶Ac and Z11-16∶OH obtained by solvent extraction were not significantly  different in scotophase and photophase, with averages of 2.8±1.9 and 2.5±0.9,  respectively, while the difference in the ratios of Z11-16∶Ac and Z11-16∶OH obtained by  SPME was statistically significant in scotophase and photophase, with averages of 8.5±1.2  and 5.7±0.6, respectively. The full extrusion of ovipositor occurred at 6-8 h after  scotophase, and lasted 80.8±4.4 min on average. The mating of S. inferens occurred at 4-10  h into scotophase, and lasted 83.4±5.0 min on average. The real-time hourly and daily  counting of moth catches in the field by automatic counting of pheromone trapping in the  four provinces of Guangdong, Sichuan, Zhejiang and Jiangsu showed that the number of moths  trapped was relatively concentrated in time in the overwintering generation, but was  relatively scattered in the following generations. The circadian rhythms of male moth  trapping in the field were affected by such factors as geographical environment, season and  generation. 【Conclusion】 This study reveals that the circadian rhythms of mating and sex  pheromone release of S. inferens are inconsistent and the mating time is in the earlier  period after scotophase. The range of effective release time of sex pheromone of female  moths is shorter than that of response time of male moths to sex pheromone. The extrusion  of the ovipositor is associated with the elevated release rate and the disperse range of  the sex pheromones released by female moths.
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Effects of RNAi-mediated gene silencing of  GdHsp 60 and GdHsp70 on the cold hardiness of  Galeruca daurica (Coleoptera: Chrysomelidae) larvae
ZHANG Hong-Ling, REN Hao, LI Kai-Xuan, TIAN Yu, ZHANG Heng, LI Yan-Yan, LI Ling, PANG Bao-Ping, TAN Yao
Acta Entomologica Sinica    2022, 65 (7): 807-817.   DOI: 10.16380/j.kcxb.2022.07.003
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【Aim】 This study aims to compare the efficiency of different RNAi methods for  silencing the heat shock protein genes (GdHsp60 and GdHsp70) of Galeruca daurica, so as to  select a method for efficiently reducing the expression level of target gene expression for  the functional study to clarify the function of these two genes in the cold hardiness of G.  daurica larvae. 【Methods】 GdHsp60 and GdHsp70 in the 1st and 2nd instar larvae of G.  daurica were silenced by RNAi via feeding and microinjection, respectively, and their  silencing efficiencies were detected by qPCR. After RNAi of GdHsp60 and GdHsp70 by  microinjection for 24 h, the super-cooling point and freezing point of the 2nd instar  larvae of G. daurica were determined with thermocouple method, and the median lethal  temperature (Ltemp50) for the 2nd instar larvae exposed to different low temperatures (-6 --14℃) for 2 h and the median lethal time (Ltime50) for exposed to -5℃ for different p eriods were determined by bioassay. 【Results】 GdHsp60 and GdHsp70 could be silenced by  RNAi via both feeding and microinjection, but microinjection-based RNAi had higher  silencing efficiency. After microinjection of dsGdHsp60 and dsGdHsp70 into the 2nd instar  larvae of G. daurica for 24 h, the expression levels of GdHsp60 and GdHsp70 decreased to  the lowest point, being decreased by 84.15% and 92.38%, respectively, as compared to that  in the control (microinjected with dsGFP).  In the 2nd instar larvae of G. daurica, the  super-cooling point, freezing point, and Ltemp50 and Ltime50 values after microinjection  of dsGdHsp60 for 24 h were -10.56±0.42℃, -7.66±0.56℃, -8.33℃ and 49.25 h,  respectively, while those after microinjection of dsGdHsp70 for 24 h were -9.08±0.23℃,  -6.09±0.28℃, -8.20℃, and 52.21 h, respectively, and those in the control were 14.71±0 .11℃, 13.94±0.09℃, -10.63℃ and 87.13 h, respectively. Compared with the control  (microinjected with dsGFP), the super-cooling point, freezing point and Ltemp50 in the 2nd  instar larvae of G. daurica microinjected with dsGdHsp60 and dsGdHsp70 for 24 h  significantly increased, while the Ltime50 value significantly shortened. 【Conclusion】  Microinjection method can be used to efficiently silence the Hsp-related genes in G.  daurica. Silencing GdHsp60 and GdHsp70 can significantly decrease the cold hardiness of G.  daurica larvae.
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Analysis of the midgut transcriptome and ABC gene expression in Spodoptera frugiperda  (Lepidoptera: Noctuidae) after feeding Cry1Ab and Cry1Fa proteins 
HUANG Xiao-Dan, JING Da-Peng, ZHANG Tian-Tao, WANG Zhen-Ying
Acta Entomologica Sinica    2022, 65 (9): 1127-1135.   DOI: 10.16380/j.kcxb.2022.09.006
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【Aim】 To reveal the relationship between the feeding of Bt proteins by  Spodoptera frugiperda larvae and the changes in the expression levels of related ABC (ATP- binding cassette transporter) genes in the midgut. 【Methods】 Artificial diets containing  Cry1Ab (LC70=240.2 μg/g) or Cry1Fa (LC70=270.0 μg/g) activated crystal protein were  used to feed the 4th instar larvae of S. frugiperda for 48 h, respectively. Transcriptome  sequencing of midgut and bioinformatics analysis were used to screen differentially  expressed genes in the midgut after treatment. RT-qPCR was used to verify the expression  levels of the differentially expressed ABC genes. 【Results】 A total of 1 305 and 1 202  differentially expressed genes were detected in the midgut transcriptome of the 4th instar  larvae of S. frugiperda fed with the artificial diet containing 240.2 μg/g Cry1Ab and  270.0 μg/g Cry1Fa, respectively, compared with those fed with the normal artificial diet  (the control group). There were 994 and 912 differentially expressed genes between the  Cry1Ab and Cry1Fa treatment groups and the control group, respectively, and were annotated  by GO function into three categories biological process, molecular function and cell  component. Among the nine differentially expressed ABC family genes screened, there were  four differentially expressed ABC genes (three up-regulated and one down-regulated)  between the Cry1Ab treatment group and the control group and five differentially expressed  ABC genes (two up-regulated and three down-regulated) between the Cry1Fa treatment group 
and the control group. The expression levels of two ABC genes (LOC118267200 and  LOC118267201) in the Cry1Ab and Cry1Fa treatment groups were significantly up-regulated as  compared to those in the control grpup. RT-qPCR validation results showed that the  expression levels of three and two ABC genes in the Cry1Ab treatment group were extremely  significantly up-regulated and down-regulated, respectively, and those of five ABC genes  and one ABC gene in the Cry1Fa treatment group were up-regulated and down-regulated,  respectively, as compared to those in the control grpup. 【Conclusion】 The intake of  Cry1Ab and Cry1Fa proteins could affect the expression levels of certain ABC family genes  in the midgut of S. frugiperda larvae, and the expression level changes of these genes are  related to pest resistance. After comparison, we found that the expression levels of ABCC  family and ABCG8 genes changed significantly. This study provides a theoretical basis for  further clarifying the role of ABC transporter proteins in the insecticidal mechanism of Bt  proteins in S. frugiperda and the rational use of Bt proteins for controlling S. frugiperda  and delaying resistance.
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