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  • Monthly, Founded in 1950
    Supervisor:Chinese Academy of Sciences
    Sponsor:Institute of Zoology,Chinese Academy of Sciences
    The Entomological Society of China
    Domestic postal code: 2-153
    Foreign issuance code: Q61
    ISSN 0454-6296
    CN 11-1832/Q
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Research progress in pigmentation and the formation mechanisms of black stripes and spots on the body in insects
GAO Yun, LIANG Yan-Ting, LIU Yi-Qin, XU Yu-Song, WANG Hua-Bing
Acta Entomologica Sinica    2020, 63 (10): 1268-1275.   DOI: 10.16380/j.kcxb.2020.10.012
Abstract645)      PDF(pc) (3193KB)(188)    PDF(mobile) (3193KB)(38)    Save
 Insects are constantly attacked by predators and exhibit adaptive traits for defense against predators during their growth and development. One of the defense strategies is an adaptive stripe pattern. The various coloration patterns of insects represent warning signals or mimic host plants to escape detection by predators and hasten avoidance learning. Furthermore, insect pigmentation patterns play important roles in such processes as mate preference, geographical adaptation, thermoregulation, and ultraviolet resistance. Thus, multifarious pigmentation patterns are highly researched. Lepidopteran insects include multiple species that are widely distributed. The adaptive stripe pattern is often observed in lepidopteran insects; thus, its biological roles are more evident than those in other insects. Recent research suggests that pigmentary color and schemochrome are the main determinants of insect color patterns, and ommochromes, melanin, and peridines significantly influence visual color patterns. In addition, hosts, environmental factors, and hormones are also thought to dramatically affect the diversity of color patterns in insects. In recent years, many genes that are crucial to pigmentation have been identified through positional cloning strategies, classical linkage maps, RNA interference, and high-throughput sequencing technologies combined with genome editing techniques. Recent studies suggest that TH, DDC, yellow, laccase2, ebony, AA-NAT, tan, and GTPCHI all play key roles in pigment synthesis. Furthermore, the pleiotropic genes spz3, apt-like, and wnt1 as well as the 20E-inducible transcription factors E75A and spalt target the melanin synthesis pathway, affecting the synthesis and deposition of melanin. In this article, we reviewed the progress in the research of the formation and influencing factors of the diversity of insect body color and markings, the type and material basis of insect coloration, and the regulation of black spots and markings on insects, hoping to provide a theoretical basis for the utilization of pigmentation-related genes and new insights for pest control.
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Ultrastructure of sensilla on the antennae, proboscis and tarsi of adult Achelura yunnanensis (Lepidoptera: Zygaenidae)
LI Gen-Ceng, ZHAO Yu-Jie, LI Jia-Li, LU Guo-Yan, LIU Nai-Yong
Acta Entomologica Sinica    2020, 63 (11): 1385-1398.   DOI: 10.16380/j.kcxb.2020.11.011
Abstract626)      PDF(pc) (1540KB)(104)    PDF(mobile) (1540KB)(32)    Save
【Aim】 This study aims to ascertain the types and differences of sensilla on the main chemosensory organs, including the antennae, proboscis and tarsi, of female and male adults of Achelura yunnanensis, so as to lay a basis for further studies associated with chemosensory reception of this moth. 【Methods】 The morphological characteristics of the antennae, proboscis and tarsi of female and male adults of A. yunnanensis and the types and ultrastructure of sensilla on them were observed under light microscopy and scanning electron microscopy. 【Results】 Seventeen subtypes of seven types of sensilla were identified on female and male antennae of A. yunnanensis adults, including five sensilla basiconica (SB), three sensilla trichodea (ST), three sensilla chaetica (SCH), two Bhm bristles (BB), two sensilla coeloconica (SCO), one sensilla auricillica (SAU) and one sensilla squamiformia (SSQ). Among them, BBI is present only on male antennae, and ST-II, ST-III and SCO-I show sexual dimorphism in length. Five subtypes of two types of sensilla were identified on female and male proboscises, including four SB and one SCH. Of these sensilla, SB-III is distributed only on female proboscis. In addition, 13 subtypes of five types of sensilla were found on female and male tarsi, including four ST, four SB, two SSQ, two BB and one SCH. Notably, ST-I is present only on female tarsi, while SB-II and SB-III are present only on male tarsi. The lengths of SB-IV and BB-I on female and male tarsi show sexual dimorphism. 【Conclusion】 There are 17, 5 and 13 subtypes of sensilla on the antennae, proboscis and tarsi of female and male adults of A. yunnanensisrespectively. Some of these sensilla exhibit sexual dimorphism in the type, length or number.
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Molecular cloning, prokaryotic expression and expression profiling of the laccase gene MaLac1 in Monochamus alternatus (Coleoptera: Cerambycidae)
CHEN Hao, LUO Qiao-Yu, MA Qiu-Yu, CHU Xu, YUAN Chao, LIU Ying, YU Wei, WU Song-Qing, WANG Rong, LIANG Guang-Hong, ZHANG Fei-Ping, HU Xia
Acta Entomologica Sinica    2020, 63 (4): 381-389.   DOI: 10.16380/j.kcxb.2020.04.001
Abstract596)      PDF(pc) (2548KB)(246)    PDF(mobile) (2548KB)(66)    Save
【Aim】 This study aims to clone and identify the laccase gene MaLac1 from the Japanese pine sawyer, Monochamus alternatus, and to analyze its expression levels in different developmental stages of the beetle, so as to provide some clues for further study of the function of MaLac1. 【Methods】 Based on the transcriptome sequencing data of M. alternatus gut, the full-length cDNA of MaLac1 was cloned from M. alternatus using the RACE method and analyzed by bioinformatic software. MaLac1 sequence was then inserted into the prokaryotic expression vector pET-32a(+) to construct a recombinant plasmid pET-MaLac1, which was then transformed into Escherichia coli Rosetta (DE3) to express MaLac1. The expression patterns of MaLac1 in the gut of M. alternatus at different developmental stages (early instar larva, mature larva, pupa, female adult, and male adult) were assayed by qPCR. 【Results】 The full-length cDNA of MaLac1 from M. alternatus (GenBank accession no.: KY073340) were obtained. Its open reading frame is 2 067 bp in length, encoding a protein of 688 amino acids, with an estimated molecular mass of 78.34 kD and an isoeletric point of 5.30. MaLac1 was predicted to contain an N-terminal signal peptide with 15 amino acids by SignalP 4.1 Server. Sequence alignment analysis showed that MaLac1 shows the typical characteristics of insect laccase genes, with 93% amino acid sequence identity with the laccase gene of Tribolium castaneum. SDS-PAGE results showed that IPTG induced a specific protein band of about 78 kD, which was consistent with the estimated size. The qPCR results showed that MaLac1 was expressed in the gut of M. alternatus at various developmental stages, with the highest expression level in the female adult gut, followed by that in the male adult gut, and the lowest expression level in the larval gut. 【Conclusion】 The expression level of MaLac1 in adults of M. alternatus is significantly higher than that in larvae, and this may be relevant to the feeding differences between adults and larvae. Further study is needed to reveal the exact function of MaLac1 in M. alternatus
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Research progress in odorant binding proteins of insects
DU Ya-Li, XU Kai, ZHAO Hui-Ting, LIU Yu-Ling, NIU Qing-Sheng, JIANG Yu-Suo
Acta Entomologica Sinica    2020, 63 (3): 365-380.   DOI: 10.16380/j.kcxb.2020.03.013
Abstract566)      PDF(pc) (4377KB)(305)    PDF(mobile) (4377KB)(76)    Save
Abstract: Insects depend critically on their sophisticated and sensitive chemoreception system to recognize and distinguish a variety of semiochemical signals from the environment. Olfaction is a sensory modality responsible for the olfactory signal transduction, which can evoke insect survival and reproduction behaviors, such as foraging, oviposition, capulation, and avoiding predators. As essential proteins in the normally functioning olfactory system, odorant binding proteins (OBPs) initially interact with external lipophilic chemicals in the process of olfactory sensing, and transfer them to chemical receptor neurons to activate olfactory receptors (ORs) distributed on the surface of dendritic membrane. In recent years, with the rapid development of high-throughput sequencing and molecular biology technologies, more and more OBPs have been identified in different species and their biological functions have been explored. Insect OBPs are a class of soluble small molecular proteins, and generally contain a stable and compact hydrophobic binding pocket consisting of 6 αhelices, where the conformational change varies with insect species and ligand structures. The expression of OBPs is not limited to olfactory organs, but also in non-olfactory tissues such as mouthparts, legs, midgut and glands. Insect OBPs can perform different physiological roles in olfaction recognition, gustatory perception, nutrient transport, pheromone synthesis and release, and histological development and differentiation. It is reasonable to suspect that the common property linking their very different functions is the ability of OBPs to bind and solubilize small hydrophobic compounds including pheromone components, common odor molecules, and nonvolatile substances. The stability and versatility of OBPs suggest their extensive application potentials in many fields including pest control, biosensors, analytical chemistry and ecology. In this article we reviewed the relevant studies of insect OBPs over the last 20 years to provide theoretical references for the in-depth study on the functions of OBPs.
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Determination of larval instars and the observation of head morphology and sensilla of the final instar larva of Callosobruchus chinensis (Coleoptera: Bruchidae)
CUI Xiao-Lin, WANG Hong-Min, ZHANG Jing, ZHANG Xian-Hong, ZHENG Hai-Xia
Acta Entomologica Sinica    2020, 63 (4): 512-522.   DOI: 10.16380/j.kcxb.2020.04.014
Abstract562)      PDF(pc) (7066KB)(136)    PDF(mobile) (7066KB)(53)    Save
【Aim】 This study aims to clarify the number of larval instars and to understand the types, morphology and distribution of sensilla on the final instar larval head of the adzuki bean beetle, Callosobruchus chinensis. 【Methods】 The body length, head capsule 
width and mandible width of C. chinensis larvae were measured. Based on the frequency histograms, regression relationship and Dyar’s rule of the obtained data, the optimal morphological index for determining the larval instars of C. chinensis was determined. The obtained result was verified by Crosby growth rule and linear regression method. The morphology and sensilla on the head of the final instar larva of C. chinensis were observed under scanning electron microscope. 【Results】 The frequency histograms of the body length, head capsule width and mandible width of C. chinensis larvae showed four obvious peaks, and it is so inferred that C. chinensis larvae have four instars. The four instars had the body length range of 1.581-2.556, 2.406-3.381, 3.381-4.281 and 4.206-4.881 mm, the head capsule width range of 0.444-0.689, 0.654-0.934, 0.934-1.179 and 1.144-1.389 mm, and the mandible width range of 0.080-0.256, 0.234-0.344, 0.322-0.542 and 0.542-0.652 mm, respectively. The body length, head capsule width and mandible width are in accordance with Dyar’s rule and Crosby growth rule, and show obvious linear relationship. Therefore, the body length, head capsule width and mandible width can be used as important indexes for the division of larval instars of C. chinensis. The Crosby ratio of head capsule width is smaller than those of body length and mandible width, and the correlation coefficient between the logarithmic value of head capsule width and body length and larval instar is higher than that between mandible width and larval instar, so the head capsule width can be used as the optimal index for the division of larval instars. Seven types of sensilla including sensillum basiconicum, sensillum trichodeum, sensillum lageniform, sensillum chaeticum, sensillum placodeum, sensillum styloconicum and sensillum ampullaceum were observed on the head and are mainly located on the antennae, maxillary palp, labrum and mandible. 【Conclusion】 The morphological indexes of the division of larval instars and the observation of head morphology and of the C. chinensis provide a theoretical basis for the study of behavior and integrated control.
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Plant pathogen-insect vector interactions: research progress and prospects
YAN Feng-Ming
Acta Entomologica Sinica    2020, 63 (2): 123-130.   DOI: 10.16380/j.kcxb.2020.02.001
Abstract560)      PDF(pc) (1449KB)(592)    PDF(mobile) (1449KB)(108)    Save
 Most plant viruses and some plant pathogenic bacteria are transmitted by insect vectors. Understandings on plant pathogen-insect vector interactions can facilitate the recognition of key issues in effective management of plant pathogens and their vectors. Therefore, pathogen-vector interaction has become one of the core topics in the studies of pathogen transmission mechanisms. This article reviewed the recent research progress in plant pathogen-insect vector interactions, introduced the main contents of papers in this issue, and based on them proposed the following three directions for future research from ecological and evolutionary perspectives: (1) the roles of the interactions between plant pathogens and insect vectors in reshaping ecological systems; (2) the inter-relationships between different modes of plant virus transmission by insect vectors, and issues related to co-evolution among viruses, vectors and plants; and (3) the mechanisms of interactions between plant pathogens and insect vectors in nature. Studies on plant pathogen-insect vector interactions are not only fundamentals in ecology and evolutionary biology, but closely related to effective management of plant pathogens and their insect vectors as well.
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Identification of the IKK related genes and their antiviral roles in response to Rice stripe virus infection in Laodelphax striatellus (Hemiptera: Delphacidae)
LU Yan-Hua, LU Gang, QI Yu-Hua, YE Zhuang-Xin, LI Jun-Min, CHEN Jian-Ping
Acta Entomologica Sinica    2020, 63 (2): 131-141.   DOI: 10.16380/j.kcxb.2020.02.002
Abstract541)      PDF(pc) (2733KB)(132)    PDF(mobile) (2733KB)(38)    Save
【Aim】 This study aims to identify the IκB kinase (IKK) related genes in the small brown planthopper, Laodelphax striatellus, and to investigate its antiviral role in this insect, so as to further understand the innate immune mechanisms in insect vectors responsive to plant viruses. 【Methods】 The IKK related genes in the L. striatellus genome were identified and characterized using bioinformatics approach. The healthy and the rice stripe virus (RSV) infected planthopper were used as the materials. The expression levels of IKK related genes in various developmental stages (egg, 1-5 instar nymphs, male and female adults) and adult tissues (gut, salivary gland, hemolymph, fat body, ovary and testis) of the RSV-free planthoppers were detected by RT-PCR. The relative expression levels of IKK related genes in the above different developmental stages and adult tissues between the RSV-free and RSV-infected planthoppers were assayed by qRT-PCR. In addition, the transcription levels of coat protein (CP) gene indicating the accumulation levels of RSV were determined after RNAi by injection of the double-stranded RNA of IKK related genes in the 3rd instar nymph of RSV-infected planthoppers. 【Results】 Two IKK related genes, IKKα (GenBank accession number: MK903504) and TANK-binding kinase 1 gene TBK1 (GenBank accession number: MN124506), were identified in the genome of L. striatellus. The open reading frame (ORF) of IKKα is 2 379 bp in length, encoding 792 amino acids, while the ORF of TBK1 is 1 551 bp in length, encoding 516 amino acids. The proteins coded by both genes have a conserved serine/threonine kinase domain and a ubiquitin folding domain. The RT-PCR results confirmed that IKKα and TBK1 were ubiquitously expressed in various developmental stages and different adult tissues of RSV-free L. striatellus. Significant expression differences of IKKα and TBK1 in various developmental stages and different adult tissues between the RSV-free and RSV-infected planthoppers were observed. The accumulation level of RSV was significantly increased after RNAi of IKKα and TBK1 in the 3rd instar nymphs of the RSV-infected planthoppers, respectively. 【Conclusion】 The results indicate that IKKα and TBK1, the important genes of NF-κB signaling pathway, are widely expressed in L. striatellus and may play important roles in L. striatellus in response to RSV invasion. These results will contribute to the further investigation on antiviral immune pathway in insect vectors against plant virus.
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Nanoinsecticides and their application in agricultural insect pest management
ZHANG Da-Xia, PAN Shou-He, BAI Hai-Xiu, DU Jiang, LIU Feng, HOU You-Ming
Acta Entomologica Sinica    2020, 63 (10): 1276-1286.   DOI: 10.16380/j.kcxb.2020.10.013
Abstract537)      PDF(pc) (2035KB)(588)    PDF(mobile) (2035KB)(55)    Save
 Nanotechnology has received great attention in the field of agriculture, and it is expected to improve the utilization rate and application effect of pesticides and fertilizers. In recent years, a number of experiments have been carried out to verify the potential for nanoinsecticides controlling agricultural insect pests and laid the foundation for the green agriculture, modern agriculture and intelligent agriculture. In this article, we reviewed the research status of the advantages of nanoinsecticides and the mechanisms of their synergistic effects. The advantages of nanoinsecticides lie in that: nanocarriers may damage the body wall, causing water loss or disturbing the normal physiological function of pests; the active ingredients can be delivered to the target position by the functionalized nanocarriers for improving utilization rate; the adhesion of insecticides on plant surface and their absorption by plants can be improved due to the functional groups and nanoscale effects of nanocarriers; and nucleic acid pesticides can be transported into plants by nanocarriers to regulate the expression of target genes in plants or pests. Although nanoinsecticides show many advantages, there are still some problems to be studied: (1) the uptake of nanoinsecticides by plants depends on the size and type of carrier, so suitable scales and carriers should be selected according to the application scenarios to improve the utilization rate of pesticides while reducing pesticide residues; (2) it is of great significance to comprehensively evaluate the environmental risks of nanoinsecticides by studying their degradation, transfer, and enrichment behavior in the environment as well as the influence caused by the difference in carriers; (3) at present, the preparation process of most of nanoinsecticides is too complex and sophisticated to be suitable for industrialization; and (4) the standard of nano-preparation and the evaluation criteria of environmental risks should be established to provide the basis for pesticide registration. In addition, the potential of nanosensors in agricultural pest monitoring is also worth attention.
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Wing morph differentiation of plant-pathogen borne vector insects  
YU Jin-Ting, CHEN Xiao-Fang, REN Ying-Dang, CUI Feng
Acta Entomologica Sinica    2020, 63 (2): 229-234.   DOI: 10.16380/j.kcxb.2020.02.013
Abstract530)      PDF(pc) (1085KB)(124)    PDF(mobile) (1085KB)(24)    Save
Wing polymorphism is one of insect polyphenism phenomena, including short-winged or apterous morph without flight ability and long-winged or alate morph for long distance migration. Wing polymorphism is often observed in vector insects that transmit plant pathogens, and therefore affects the spatial-temporal distribution and outbreak of plant diseases. In this article we reviewed the research progress of wing dimorphism in aphids and planthoppers from the genetic laws, induction factors, molecular mechanisms, and other physiological modification accompanying wing morph differentiation. Wing dimorphism is mainly induced by abiotic factors, such as temperature, humidity and photoperiod, and biotic factors, such as population density, host nutrition and viruses. The underlying molecular mechanisms are involved in the insulin/insulin-like growth factor signaling (IIS) pathway, c-Jun NH2-terminal kinase (JNK) signaling pathway, Wingless and olfactory receptor SaveOrco. Wing morph differentiation of insects is accompanied by changes in physiological status. The short-winged insects have stronger reproduction ability, while the long-winged insects contain richer flight muscle components. So far, the research of wing dimorphism is not comprehensive, and many problems need to be solved, such as identifying the target genes in the insulin/insulin-like growth factor signaling pathway, the regulation mechanisms of JNK on wing dimorphism and the molecular mechanisms of wing dimorphism mediated by viruses in vector insects. This review may shed light on the control of vector-borne pathogens and the research of wing polymorphism of other insects.
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Diversity analysis and function prediction of gut microbiota in three springtail species
CHEN Wei, CHEN Xia, LI Juan, MA Xin-Ran, CUI Wei, QU Feng-Tian, XIE Gui-Lin, ZHAO Hong-Qing
Acta Entomologica Sinica    2020, 63 (4): 450-461.   DOI: 10.16380/j.kcxb.2020.04.008
Abstract517)      PDF(pc) (8308KB)(81)    PDF(mobile) (8308KB)(41)    Save
【Aim】 Springtails make a significant contribution to the soil ecosystem. This study aims to investigate the gut microbial structure and diversity and their potential functions in three springtail species, Sinella (Coecobrya) oligoseta, Proisotoma minuta and Tomocerus missus. 【Methods】 Microbes in the adult intestinal contents of the three springtail species were analyzed and compared by 16S rDNA amplicon sequencing, and the function prediction of gut microbial genes was conducted using Tax4Fun method. 【Results】Among the three springtail species, T. missus showed the highest adult gut microbial diversity, while S. (C.) oligoseta showed the lowest adult gut microbial diversity. At the phylum level, Proteobacteria, Firmicutes and Bacteroidetes were the most predominant microbes in the adult gut microbiota of the three springtail species, followed by Actinobacteria. At the genus level, the abundance of Pseudomonas in the gut of S. (C.) oligoseta (16.21%) was significantly higher than those in the guts of P. minuta (0.87%) and T. missus (1.37%). The abundance of Vibrio in the gut of P. minuta (25.81%) was significantly higher than that in the gut of S. (C.) oligoseta (3.35%) and T. missus (0.004%). Additionally, based on KEGG pathway annotations, we predicted that the genes in adult gut microbiota of the three springtail species may be involved in the metabolism of carbohydrates and amino acids, infectious diseases and drug resistance. 【Conclusion】 In the intestines of the three springtail species, S. (C.) oligoseta, P. minuta and T. missusthe core microbiota are the same at the phylum level, but obviously different at the genus level. The influencing factors should include the genetic traits and the abundance and diversity of microbes in the habitat of each species. Actinomycetes in springtail guts are conducive to the discovery of new strains and the application of their metabolites. Drug-resistant and pathogenic bacteria likely exist in the intestines of the three springtail species, implying new directions of functional studies on gut microbiota in springtails.
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Research progress in ionotropic receptors and their functions in insects
GUO Jin-Meng, DONG Shuang-Lin
Acta Entomologica Sinica    2020, 63 (11): 1399-1410.   DOI: 10.16380/j.kcxb.2020.11.012
Abstract514)      PDF(pc) (1624KB)(145)    PDF(mobile) (1624KB)(25)    Save
 As the largest group in the animal kingdom, insects have evolved complex sensory systems in response to various environmental stimuli, in which the chemosensation (olfaction and gustation) is the most important. The odorant receptors (ORs) and gustatory receptors (GRs) play a key role in olfaction and gustation, respectively. However, in recent years, it has been found that ionotropic receptors (IRs) are also involved in chemosensation as well as non-chemosensation such as sensation of temperature and humidity. The IR family is evolved from ionotropic glutamate receptors (iGluRs) which contain three typical transmembrane domains. Similar to ORs, each IR needs to be co-expressed with at least one of the four identified co-receptors (IR8a, IR25a, IR76b and IR93a) for its normal function. Two or more IRs can be co-expressed in a same olfactory neuron, and different IR combinations lead to their different response profiles in neurons. One insect species usually contains several to over one hundred IRs, with some IRs showing antennaespecific expression patterns, but most IRs remaining unknown in the function and the mechanism of action. In this article, the research advances in the structure and evolution of IRs, and the identification, expression and function of IR genes in insects were reviewed, hoping to provide some references for the further studies on the function and mechanisms of action of IRs, and the potential use of insect IRs as targets in pest control.
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Comparative analysis of the transcriptomes of male adult and the 5th instar larva of Spodoptera frugiperda (Lepidoptera: Noctuidae)
LIAO Yong-Lin, LI Yan-Fang, YANG Bin, ZHAO Hua, GAO Yan, CHEN Wei-Ping, CAI Shi-Ke, WANG Ji-Hua
Acta Entomologica Sinica    2020, 63 (5): 523-532.   DOI: 10.16380/j.kcxb.2020.05.001
Abstract512)      PDF(pc) (12663KB)(248)    PDF(mobile) (12663KB)(97)    Save
【Aim】 The fall armyworm, Spodoptera frugiperda, is a newly invaded important pest in China. This study aims to comparatively analyze the transcriptomes of two developmental stages, male adult and the 5th instar larva, of S. frugiperda. 【Methods】 High-throughput sequencing technology was used to sequence and assembly the transcriptomes of male adult and the 5th instar larva of S. frugiperda, and the functional annotation and comparative analysis of transcriptome data were performed. 【Results】 After a de novo assembly, a total of 209 002 transcripts were obtained, with an average length of 687.55 bp and an N50 of 982 bp. A total of 46 198 (57.43%) unigenes were annotated in at least one database, among which 1 713 (2.13%) unigenes were annotated in all databases. In the GO database, 205 269 unigene annotations were obtained, mainly including 68 functional classifications. A total of 3 408 unigenes were annotated in KEGG database, involving 277 metabolic pathways. A total of 424 olfaction-related genes were identified and showed differential expression in male adults and the 5th instar larvae of S. frugiperda. By comparative transcriptome analysis, 9 162 up-regulated differentially expressed genes (DEGs) and 6 399 down-regulated DEGs were identified in male adult. Functional enrichment analysis revealed that the up-regulated DEGs were significantly enriched in the pathways involved in pheromone and signal transduction, while the down-regulated DEGs were significantly enriched in the pathways related to detoxification. 【Conclusion】 These transcriptome data provide some valuable information for exploring the functional genes related to the growth and development and olfaction and candidate molecular targets in S. frugiperda.
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Honeydew excretion mechanisms in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae): progress and prospects
SHU Ben-Shui, WU Zhong-Zhen, CUI Yang, LIN Jia-Hao, LIN Jin-Tian
Acta Entomologica Sinica    2020, 63 (2): 235-244.   DOI: 10.16380/j.kcxb.2020.02.014
Abstract503)      PDF(pc) (1960KB)(101)    PDF(mobile) (1960KB)(29)    Save
The Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae), is a general pest causing economic losses in orchards, mainly feeds on the highly osmotic sap of citrus phloem and transmits pathogen Candidatus Liberibacter asiaticus to induce a devastating disease of citrus named huanglongbing (HLB). The osmotic pressure of phloem sap is a challenge for hemipteran insects, which is regulated by a set of osmoregulation mechanisms evolved in insects. The excessive sugars are converted into long chain oligosaccharides and excreted outside of insects as honeydew. In this article, the behavior of honeydew excretion, composition of honeydew and factors affecting honeydew excretion of D. citri are summarized, and the functions of osmoregulatory genes involved are overviewed. The studies show that the behavior of honeydew excretion and the color, texture and composition of honeydew were different among male and female adults and nymphs of D. citri. The host plants, insecticides, pathogenic microorganisms and compounds secreted by natural enemies were demonstrated as the main factors affecting the excretion behavior of D. citri. In addition, the pivotal osmoregulatory genes encoding α-glucoside hydrolases, aquaporins and sugar transporters were considered as the potential genes involved in the regulation of honeydew excretion. This review may provide a reference for further studying honeydew excretion behavior of D. citri and exploring the new targets for developing new insecticides for the control of this insect.
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Analysis of alkaloid components in the soil from the nest of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae)
LI Xiao-Ling, CHEN Li, FANG Shou-Guo
Acta Entomologica Sinica    2020, 63 (4): 494-501.   DOI: 10.16380/j.kcxb.2020.04.012
Abstract498)      PDF(pc) (1588KB)(87)    PDF(mobile) (1588KB)(7)    Save
 【Aim】 The red imported fire ant, Solenopsis invicta, inhabits the soil environment rich in bacteria and fungi. Although they are vulnerable to infection of bacterial and fungal diseases, fire ants with defensive and antibacterial venom alkaloids can survive in such soil environment. This study aims to establish an optimal extraction method of alkaloids from the soil of the red imported fire ant nest and then to quantify the major components of venom alkaloids. 【Methods】 In the recovery experiment, S. invicta venom was added to soil obtained 3 m away from S. invicta nests. Venom alkaloids were then extracted from soil using a filtration method. GC-FID was used for quantitative analysis of venom alkaloids, screening of the optimal extraction solvent, and determination of the optimal volume of triethylamine. The optimized method was used for quantitative analysis of venom alkaloids from the soil of ant nest. 【Results】 Among the five extraction solvents including n-hexane, dichloromethane, ethyl acetate, acetone and methanol, the recovery rate of alkaloids by n-hexane extraction was relatively higher. The best extraction efficiency was achieved by adding 1 mL or more triethylamine. Among the venom alkaloids in the soil of S. invicta nest, the content of trans-C15∶1 was the highest, followed by that 
of trans-C13∶1. The content of total alkaloids in S. invicta nest soil was about 22 μg/g. 【Conclusion】 Triethylamine is very effective for extracting venom alkaloids from the soil of ant nests. The concentration of alkaloids in the soil of S. invicta nest is relatively high, which may have an important impact on the microbial community in the nest environment.
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Observation of antennal sensilla of flower-visiting insect Bibio rufiventris (Diptera: Bibionidae) with scanning electron microscope
YUAN Ke, ZHU Hui, QU Ye-Kuan, REN Bing-Zhong, YOU Yang
Acta Entomologica Sinica    2020, 63 (4): 439-449.   DOI: 10.16380/j.kcxb.2020.04.007
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【Aim】 This study aims to clarify the types and distribution of antennal sensilla of flower-visiting insect Bibio rufiventris. 【Methods】 The type, quantity and morphology of antennal sensilla of B. rufiventris were observed with scanning electron microscope (SEM), and their differences between male and female adults were compared. 【Results】 The female and male adult antennae of B. rufiventris include three parts, scape (Sc), pedicel (P) and flagellum (F), and the flagellum consists of eight flagellomeres. The average lengths of antennae of female and male adults are 862.556±78.662 and 880.361±83.253 μm, respectively. There is significant difference in the length of the 8th flagellomere, while the lengths of other flagellomeres are almost equal. Four types of sensilla including sensilla chaetica (Sc), sensilla basiconca (Sb), sensilla trichodea (St) and Böhm bristles (Bb) were observed on the female and male antennae. The antennal sensilla of female adults have six subtypes, i.e., sensilla chaetica (Sc), sensilla trichodea subtype 2 (St.2), sensilla basiconca subtypes 1, 2 and 4 (Sb.1, Sb.2 and Sb.4) and Böhm bristles (Bb), while those of male adults have five subtypes, i.e., sensilla chaetica (Sc), sensilla trichodea subtype 1 (St.1), sensilla basiconca subtype 2  (Sb.2) and sensilla basiconca subtype 3 (Sb.3) and Böhm bristles (Bb). 【Conclusion】 The types, amounts and characteristics of antennal sensilla of B. rufiventris adults show some 
differences between females and males. This study provides a morphological basis for studying the physiological functions of these sensilla and the molecular mechanisms of behavioral activities of B. rufiventris
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Morphology, histology and ultrastructure of the alimentary canal of the adult mute cicada, Karenia caelatata (Hemiptera: Cicadidae) (In English)
ZHONG Hai-Ying, ZHANG Ya-Lin, WEI Cong
Acta Entomologica Sinica    2020, 63 (4): 421-432.   DOI: 10.16380/j.kcxb.2020.04.005
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【Aim】In order to better understand the morphology and function of alimentary canal of insects in Cicadidae, the alimentary canal of the mute cicada, Karenia caelatata, was investigated at the morphological, histological and ultrastructural levels. 【Methods】 The gross morphology of alimentary canal, ultrastructure of associated organs including oesophagus, filter chamber (including anterior and posterior extremities of the midgut, internal proximal extremity of the ileum, and internal proximal extremities of the Malpighian tubules), conical segment, a midgut loop, and hindgut (ileum and rectum), and histology of filter chamber of male adult of K. caelatata were observed using light and transmission electron microscopy. 【Results】 The results showed that the alimentary canal of K. caelatata consists of the oesophagus, filter chamber, external midgut section and hindgut. The elongated oesophagus is lined with a cuticle containing an epicuticle and an endocuticle. The filter chamber is enveloped by a thin filter chamber sheath, which contains the anterior and posterior extremities of the midgut, internal proximal extremity of the ileum and internal proximal extremities of the Malpighian tubules. Cells of the anterior and posterior extremities of the midgut possess well-developed basal infoldings and densely-packed apical microvilli. Numerous mitochondria and electron-dense secretory granules were observed in the posterior extremity of the midgut. The external midgut section contains a conical segment and a midgut loop. The conical segment is dilated, and consists of two types of cells: the first is featured by the well-developed basal infoldings, and the second lacks basal infoldings. The midgut loop is divided into three distinct regions, i.e., the anterior segment containing numerous secretory granules, mitochondria, rough endoplasmic reticulum and lysosomes in the cytoplasm, the mid-segment containing secretory granules in the cytoplasm, and the posterior segment with cells possessing numerous electron-lucent secretory granules and smooth endoplasmic reticulum. Ferritin-like granules scatter in the cells of the anterior segment and mid-segment of the midgut loop. Filamentous materials coat the microvilli of the cells of the conical and anterior segments of the midgut loop. The hindgut is lined by a layer of cuticle. Microorganisms reside in the oesophagus, mid-segment of the midgut loop and rectum cells. 【Conclusion】 Findings of the morphological, histological  and ultrastructural characteristics of the gut alimentary canal of K. caelatata  in this study improve our understanding of the functional differentiation of alimentary canal in this species. The discovery of microorganisms in the alimentary canal is formative to future study of coevolution between the Cicadoidea and related bacteria.
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Comparison of feeding behaviors and pathogen acquisition rates of adults and the 5th instar nymphs of Diaphorina citri (Hemiptera: Liviidae) on huanglongbing-infected citrus plants
WU Tian-Yu, ZHANG Xu-Ying, George A. C. BEATTIE, CEN Yi-Jing
Acta Entomologica Sinica    2020, 63 (2): 166-173.   DOI: 10.16380/j.kcxb.2020.02.006
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【Aim】 The Asiatic citrus psyllid (ACP), Diaphorina citri, is the major vector of ‘Candidatus Liberibacter asiaticus’ (‘CLas’) (α-Proteobacteria), the pathogen that causes the Asiatic form of huanglongbing (HLB), the most devastating disease of citrus. The aims of this study are to determine whether the feeding behaviors of adults and the 5th instar nymphs of D. citri differ, and if so, whether these differences influence pathogen acquisition efficiency, and whether HLB-infected plants influence feeding by the 5th instar nymphs. 【Methods】 The feeding behavior of adults and the 5th instar nymphs of D. citri on immature shoots of HLB-infected Citrus reticulata cv. Sunki was recorded for 10 h with DC-EPG giga-4. The acquisition efficiency of the pathogen by individual adults and nymphs from HLB-infected shoots was determined by qPCR. The feeding behaviors of the 5th instar nymphs on pathogen-free and HLB-infected shoots were also recorded and compared by EPG. 【Results】 The feeding behaviors of adults and the 5th instar nymphs of D. citri on C. reticulata cv. Sunki shoots infected with HLB differed significantly. The 5th instar nymphs started to ingest xylem and phloem sap sooner than adults. Durations of the phloem salivation and ingestion phases of nymphs were significantly longer than those of adults. When feeding on HLB-infected shoots, 37.5% of the 5th instar nymphs acquired ‘CLas’ in contrast 20% of adults. The frequencies of probing and salivation and phloem sap ingestion by the 5th instar nymphs feeding on HLB-infected shoots were significantly lower than those on healthy shoots. However, HLB-infected plants did not influence the duration of phloem salivation and sap ingestion by the 5th instar nymphs. Feeding also commenced sooner in phloem of HLB-infected shoots than in phloem of healthy shoots. 【Conclusion】 The 5th instar nymphs of D. citri feed for longer intervals, ingest more phloem sap, and acquire higher titers of ‘CLas’ than adults. This may be due to that nymphs require more nutrients for their growth and development than adults. They are therefore less inclined to withdraw their stylets from, and re-insert their stylets into ‘CLas’-infected leaf tissues. ‘CLas’-induced changes in host-plant phytochemistry promote feeding by the 5th instar nymphs of D. citri.
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Transcriptomic analysis reveals the molecular mechanism underlying Nosema ceranae infection of Apis mellifera ligustica
GENG Si-Hai, ZHOU Ding-Ding, FAN Xiao-Xue, JIANG Hai-Bin, ZHU Zhi-Wei, WANG Jie, FAN Yuan-Chan, WANG Xin-Rui, XIONG Cui-Ling, ZHENG Yan-Zhen, FU Zhong-Min, CHEN Da-Fu, GUO Rui
Acta Entomologica Sinica    2020, 63 (3): 294-308.   DOI: 10.16380/j.kcxb.2020.03.006
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【Aim】 This study aims to reveal the molecular mechanism underlying Nosema ceranae infection of Apis mellifera ligustica at the transcriptome level by investigation of differentially expressed genes (DEGs), virulence factors and infection-associated factors. 【Methods】 Following the criteria of P≤0.05 and |log2(Fold change)|≥1, DEGs within NcCK vs NcT1, NcCK vs NcT2 and NcT1 vs NcT2 comparison groups were screened out via the comparative analysis based on our previously obtained high-quality transcriptome datasets from purified spores of N. ceranae (NcCK) and N. ceranae in the midgut of A. m. ligustica workers at 7 d and 10 d post infection (NcT1 and NcT2, respectively). Venn analysis, GO classification and KEGG pathway enrichment analysis of DEGs were conducted using the related bioinformatic software, virulence factors and infection-associated factors of N. ceranae were summarized and analyzed based on Nr database annotations, KEGG database annotations and related literature documentations. The transcriptomic data and expression trends of DEGs were verified by RT-qPCR. 【Results】 In total, 1 397, 1 497 and 52 DEGs were identified in NcCK vs NcT1, NcCK vs NcT2 and NcT1 vs NcT2, respectively. Venn analysis showed that 10 up-regulated genes and one down-regulated gene were shared by various comparison groups. GO functional classification results showed the largest functional terms of DEGs in NcCK vs NcT1 and NcCK vs NcT2 were metabolic process, cellular process, single-organism process, cell, cell part, organelle, catalytic activity and binding, while DEGs in NcT1 vs NcT2 were mostly enriched in metabolic process, cellular process, single-organism process, catalytic activity and binding. In addition, KEGG pathway enrichment analysis indicated that DEGs within NcCK vs NcT1 and NcCK vs NcT2 were enriched in 80 and 79 pathways, respectively. The number of up-regulated genes enriched in glycolysis/gluconeogenesis and MAPK signaling pathway was more than that of down-regulated genes. Investigation of virulence factors displayed that spore wall protein 9 encoding gene and spore wall protein 12 encoding gene were down-regulated in NcCK vs NcT1 and NcCK vs NcT2, while spore wall protein 8 encoding gene was down-regulated only in NcCK vs NcT1. In additional, the expression levels of spore wall protein precursor encoding gene, spore wall and anchoring disk complex protein encoding gene, chitinase encoding gene, polar tube protein encoding gene, and ricin B lectin encoding gene were all up-regulated in NcCK vs NcT1 and NcCK vs NcT2. Moreover, the infection-associated factor analysis demonstrated that in NcCK vs NcT1 and NcCK vs NcT2 three key enzyme genes engaged in glycolytic pathway were up-regulated, while in NcCK vs NcT1 and NcCK vs NcT2 three ATP/ADP transferase-associated genes were up-regulated but one down-regulated. Two ABC transporterassociated genes were up-regulated in NcCK vs NcT1 and NcCK vs NcT2, while four down-regulated. Finally, the RT-qPCR results verified the authenticity of the transcriptomic data and expression trends of DEGs. 【Conclusion】 In this study the transcriptomic dynamics of N. ceranae infecting the A. m. ligustica worker was deciphered through the comparative analyses. Our findings revealed that genes encoding virulence factors including spore wall protein, spore wall and anchoring disk complex protein, chitinase, polar tube protein and ricin B lectin, and genes encoding infection-associated factors such as hexokinase, pyruvate kinase, 6-phosphofructokinase, ATP/ADP transferase and ABC transporters, are likely to play key roles in pathogen proliferation, providing a basis for clarifying the infection mechanism of N. ceranae.
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Projection of major pheromone tuned olfactory receptor neurons of Helicoverpa armigera (Lepidoptera: Noctuidae) revealed by using single sensillum recording and staining
MA Bai-Wei, LIU Xiao-Lan, CHANG Ya-Jun, XIE Gui-Ying, CHEN Wen-Bo, LIU Yang, ZHAO Xin-Cheng, WANG Gui-Rong
Acta Entomologica Sinica    2020, 63 (4): 413-420.   DOI: 10.16380/j.kcxb.2020.04.004
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【Aim】 To characterize the function and morphology of major pheromone tuned olfactory receptor neurons and their central projection to the brain in the cotton bollworm, Helicoverpa armigera. 【Methods】 Single sensillum recording was used to record the response of major pheromone tuned olfactory receptor neurons to sex pheromones, and staining with a fluorescent dye used to label the neurons. Immunohistochemistry was used to label the antennal lobe glomeruli. Laser scanning confocal microscope was used to acquire the images of antennal lobes with stained neurons, and the image software ZEN and Amira 4.1.1 was used to analyze the data. 【Results】 Obvious electrophysiological responses of trichoidea sensilla on the antennae of male adults of H. armigera to the major pheromone component Z11-16∶Ald were recorded, and the olfactory receptor neurons housed in the sensilla were successfully labeled by staining. Two olfactory receptor neurons were observed in Z11-16∶Ald-responsive sensllium. The axons of these two olfactory receptor neurons project via the antennal nerve to the cumulus and an ordinary glomerulus in the antennal lobe, respectively. 【Conclusion】 The combination of single sensillum recording and staining with the fluorescent dye can be applied in the identification of the function and morphology of olfactory receptor neurons housed in the antennal sensilla of insects and their central projection pathway to the antennal lobe glomeruli. Compared with the cobalt-lysine, the staining with fluorescent dye is more practical and facilitates the image acquisition and three-dimensional reconstruction. Such method provides technical support for the investigation on the projection pathway of other olfactory receptor neurons of H. armigera to determine the relationship between the peripheral odor receptor perception and the central nervous system.
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Cloning of juvenile hormone esterase and juvenile hormone epoxide hydrolase genes and their expression dynamics during diapause and metamorphosis in Sitodiplosis mosellana (Diptera: Cecidomyiidae)
WANG Jing-Jing, LIANG Ting-Ting, CHENG Wei-Ning, ZHU Ke-Yan
Acta Entomologica Sinica    2020, 63 (4): 401-412.   DOI: 10.16380/j.kcxb.2020.04.003
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【Aim】 Juvenile hormone (JH) plays important roles in diapause induction, maintenance and post-diapause quiescence of the wheat midge, Sitodiplosis mosellana, an insect species undergoing obligatory larval diapause. Juvenile hormone esterase (JHE) and juvenile hormone epoxide hydrolase (JHEH) are important JH degradation enzymes that play key roles in regulating insect JH titers. This study aims to explore the potential roles of JHE and JHEH in the diapause and metamorphosis of S. mosellana. 【Methods】 The full-length cDNAs of JHE and JHEH were cloned from pre-diapause larvae of S. mosellana by RT-PCR and RACE technologies. The sequence characteristics of nucleotide and amino acid sequences of the cloned JHE and JHEH were analyzed by bioinformatics software. The 
expression levels of JHE and JHEH in the 3rd instar larvae of S. mosellana at different diapause stages (pre-diapause, diapause, post-diapause quiescent and post-diapause development) and different developmental stages from the 1st instar larva to adult  (the 1st-2nd instar larva, pre-pupa, early pupa, middle pupa, late pupa, male adult and female adult) were determined by qPCR. 【Results】 The full-length cDNA sequences of JHE and JHEH  were obtained from S. mosellana and named SmJHE and SmJHEH (GenBank accession number: MG876768 and MG876769, respectively), with the length of 3 102 and 1 980 bp, respectively. SmJHE and SmJHEH contain 1 740 and 1 371 bp ORFs, respectively, which encode the proteins of 579 and 456 amino acids with the predicted molecular weights of 65.67 and 51.65 kD, respectively. SmJHE contains five specific conserved motifs of JHE protein family, and all of the conserved domains of JHEHs including the residues Asp228, Asp404 and His431 in the catalytic triad, and two Tyr residues (Tyr299 and Tyr374) and the HGWP motif corresponding to the oxyanion hole. Multiple sequence alignment and phylogenetic analysis showed that SmJHE and SmJHEH have higher amino acid sequence identity and the closest relationship to the corresponding homologues from Nematocera insects of Diptera. The expression levels of SmJHE and SmJHEH showed no significant change at the pre-diapause stage (the 1st, 2nd and early 3rd instar larval stages), remained stable after entering diapause, but were the lowest at the post-diapause quiescence phase (from December to the next January). The expression of SmJHE increased gradually after development resumption, reached the peak in the pre-pupal stage, and was significantly lower in female adults than in male adults. The expression of SmJHEH, however, reached the highest in female adults and the lowest in pre-pupae. 【Conclusion】 SmJHE and SmJHEH are involved in diapause of S. mosellana. Their reduced expression during diapause might be related to accumulation of JH titers in post-diapause quiescence. Increased expression of SmJHE during the development process of S. mosellana might play a role in metamorphosis from larva to pupa, and its reduced expression might be related to reproductive development.
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Analysis of the antennal transcriptome and olfaction-related genes of Agrilus zanthoxylumi (Coleoptera: Buprestidae)
GONG Xue-Fang, XIE Shou-An, YANG Ping, GUO Li, CHEN Di, CHE Xian-Rong, WANG Yan-Lai, LÜ Shu-Jie
Acta Entomologica Sinica    2020, 63 (10): 1159-1170.   DOI: 10.16380/j.kcxb.2020.10.001
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【Aim】 To establish the antennal transcriptome database of Agrilus zanthoxylumi adult and to explore target genes related to olfaction, so as to provide the theoretical basis for the future study of the chemosensory mechanism of its antenna and its biological control. 【Methods】 High-throughput sequencing platform Illumina NovaSeq 6000 was used for transcriptome sequencing of the antennae of female and male adults of A. zanthoxylumiTrinity software was used for sequence splicing and assembly of the high-quality reads obtained. BLAST software was used to compare the antennal transcriptome data with public databases NR, NT, Swiss-Prot, GO, KEGG, BLASTX, eggNOG, Pfam, TmHMM, SignalP, KO, Map, BLASTP and RNAMMER. Phylogenetic analysis was carried out using MEGA software based on the nucleotide sequences of the preliminarily screened candidate odorant binding proteins (OBPs) and chemosensory proteins (CSPs) of A.zanthoxylumi, and homologous proteins from other coleopteran insects. The RPKM (reads per kilobase per million mapped reads) value was used to evaluate the expression levels of olfaction-related genes. 【Results】 By sequencing of the antennal transcriptome of female and male adults of A. zanthoxylumi, a total of 36 209 unigenes and 90 982 transcripts were obtained with the N50 of 2 103 and 2 523 bp, respectively, indicating the high completeness of assembly. Most unigenes (41.62%) were annotated to the NR database, with the highest proportion (19%) of genes similar to those of Tribolium castaneum. In the GO database, 11 614 unigenes were annotated and divided into 57 branches of three functional categories, cellular component, molecular function and biological process, among which binding (70.57%) and catalytic activity (45.51%) related unigenes in the category of molecular function were the most. The KEGG metabolic pathway analysis indicated that 7 427 unigenes are involved in five categories of metabolic pathways, with the most unigenes (815) involved in signal transduction. Seven candidate OBP genes and 5 CSP genes with full-length open reading frame were screened, and their encoded proteins have the typical characteristics of chemosensory protein family. The phylogenetic analysis showed that the OBPs and CSPs of A. zanthoxylumi have the highest sequence identity with the OBPs and CSPs of A. mali, respectively. The RPKM values showed that AzanOBP1 and AzanOBP2 were not expressed in the antennae of female adults, but slightly expressed in the antennae of male adults, and AzanOBP3 was expressed with high abundance in the antennae of male adults. 【Conclusion】 For the first time, the antennal transcriptome data of A. zanthoxylumi were obtained, and the olfaction-related OBP, CSP, OR, IR and SNMP genes of A. zanthoxylumi were screened. It is speculated that OBPs expressed with high abundance in the antennae play a key role in identifying pheromones released by female adults of the same species or the volatiles released by host plants. The results can lay a molecular basis for further studying the functions of chemosensory genes and olfactory sensitivity mechanism of A. zanthoxylumi.
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Genetic analysis of the non-diapause red egg mutant Re-nd in the silkworm, Bombyx mori
ZHANG Yu-Jing, ZHANG Hai-Yan, SHEN Guan-Wang, WU Jin-Xin, JIA Lin-Bang, MAO Xue-Qin, JIAO Meng-Yao, LIN Ying
Acta Entomologica Sinica    2020, 63 (11): 1287-1294.   DOI: 10.16380/j.kcxb.2020.11.001
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【Aim】 Re-nd, a non-diapause red egg mutant of the silkworm, Bombyx mori, is a new egg color mutant. It was obtained by mutagenesis of the C108 strain induced by the chemical mutagen N-methyl-N-nitrosourea (MNU), belonging to serous membrane mutation. This study aims to analyze the mechanism of mutant phenotype by traditional genetic methods, so as to lay a foundation for further cloning, functional research and application of the mutant genes of Re-nd. 【Methods】 The hybridizing, sib mating and backcrossing between non-diapause mutant Re-nd and the wild-type diapause bivoltine Dazao strain and non-diapause polyvoltine N4 strain of B. mori, respectively, and the genetic analysis were carried out. Meanwhile, the hybridizing, sib mating and backcrossing between Re-nd and diapause egg color mutants with the known mutants such as white egg mutant w-2, peach red eye white egg mutant pe and red egg mutant re of B. mori, respectively, and the genetic analysis were also carried out. 【Results】 The results showed that the non-diapause eggs of F1 hybrids of Re-nd with Dazao and N4 were both in light red. The non-diapause eggs of F2 generation of sib mating were in red, light red and light yellow with the ratio of the number of red eggs (including light red eggs) to the number of light yellow eggs basically conforming to 3∶1. The nondiapause eggs of backcross B1 generation were in red and light yellow, with the ratio of the number of red eggs to the number of the light yellow eggs being basically consistent with 1∶1. The diapause eggs of F1 hybrids of Re-nd with w-2 and pe were in light brown. The diapause eggs of F2 hybrid of Re-nd with w-2 were in normal dark brown, light brown, yellow and light yellow with slightly red, while the non-diapause eggs of F2 hybrid were in red, light yellow with slightly red, orange-yellow and light yellow. The F2 hybrid eggs of Re-nd with pe were all diapause eggs in normal dark brown, light brown, light yellow with slightly red and red. The hybrid diapause eggs of Re-nd with re in F1were in light brown, and those of in F2 were all diapause eggs in normal dark brown, light brown and red. The diapause eggs in re parents were in red, and no new egg color separation was found in the hybrid eggs of Re-nd with re in F2. 【Conclusion】 Through the hybridization and genetic analysis, it was confirmed that Re-nd mutant is an independent dominant inheritance, and the mutant gene of Re-nd should be located in the downstream of the mutant genes of w-2 and pe and may be involved in the process of further metabolism of 3-hydroxy-caninuric acid to ommatins at the later stage of ocular pigment biosynthesis, which is different from the wild-type egg pigment metabolism branch.
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Advances in landscape genomics and its application in integrated pest management
CHEN Yan-Ting, YOU Shi-Jun, KE Fu-Shi, LIU Tian-Sheng, LI Jian-Yu, YOU Min-Sheng
Acta Entomologica Sinica    2020, 63 (11): 1411-1430.   DOI: 10.16380/j.kcxb.2020.11.013
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The recent development of novel sequencing technologies and bioinformatics has greatly facilitated the fundamental and applied studies on insect genomes with the steady increase of accomplishments. Studies that integrate variables of agricultural landscape with the genomic structure, genetically adaptive variation, and gene functions aid in better understanding of the molecular mechanisms associated with the adaptability and infestation of a target pest in a specific landscape framework, providing new thoughts and directions for pest management. Landscape genomics is an interdisciplinary subject combining landscape ecology with population genomics and aims to examine the genetic basis of adaptation of a given species to changing environments. Landscape genomics mainly focus on the investigation and analysis of adaptive evolution in species as well as gene flow and genetic drift among populations that are under environmentassociated selection. The research methodology of landscape genomics can be generalized as the design of spatial sampling strategy, utilization of molecular markers, and data collection and analysis. In this article we reviewed the research status of landscape genomics including the major findings and achievements in recent years. Further, we elaborated the application and prospects of landscape genomics in the aspects of insecticide resistance management, conservation biological control, and integrated pest management in the context of global climate change, with the ultimate goal of reducing the use of insecticides and promoting the safe production of crops.
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Gustatory perception and feeding preference of Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae to four stimuli
HOU Wen-Hua, SUN Long-Long, MA Ying, SUN Hui-Wan, ZHANG Jia-Jia, BAI Run-E, ZHAO Xin-Cheng, TANG Qing-Bo
Acta Entomologica Sinica    2020, 63 (5): 545-557.   DOI: 10.16380/j.kcxb.2020.05.003
Abstract437)      PDF(pc) (9139KB)(129)    PDF(mobile) (9139KB)(29)    Save
【Aim】 The aim of this study is to screen effective phagostimulants and deterrents, and to explore the gustatory perception mechanism of Spodoptera frugiperda larvae, so as to provide theoretical and practical references for the ecological control of the pest. 【Methods】 The single sensillum recording technique was used to investigate the electrophysiological characteristics of the medial sensillum styloconicum and the lateral sensillum styloconicum located on the maxillary galea of mouthparts of the day-2 5th instar larvae to four stimuli of sucrose, sinigrin, tannic acid, and quinine hydrochloride at different concentrations. The two-choice feeding assay was used to investigate the feeding choice of S. frugiperda larvae to these stimuli. 【Results】 Both the medial sensillum styloconicum and the lateral sensillum styloconicum of S. frugiperda larvae consist of gustatory receptor neurons sensitive to sucrose, sinigrin and tannic, with the responding activities of neurons changing based on stimuli and concentrations. However, the quinine hydrochloride only elicited electrophysiological responses of the medial sensillum styloconicum but not the lateral sensillum styloconicum. The responses of gustatory receptor neurons in both the medial and lateral sensilla styloconica to sucrose and sinigrin showed typical dose-dependent patterns, while the response of quinine hydrochloride to the medial sensillum styloconicum showed a reverse dose-dependent pattern. The results of behavioral choice showed that sucrose significantly induced the feeding choice of S. frugiperda larvae, while sinigrin, tannic and quinine hydrochloride inhibited their feeding choice in a dose-dependent pattern. 【Conclusion】 Gustatory receptor neurons housed in both the medial and lateral sensilla styloconica of S. frugiperda larvae respond to phagostimulants and deterrents. The two types of sensilla demonstrate different responding patterns in response spectrum and responding activities. Sucrose can be regarded as one candidate compound of phagostimulants, while quinine hydrochloride, sinigrin and tannic acid can be regarded as the candidate compounds of deterrents. The gustatory responding profiles of larvae may contribute to determining their feeding preference for these compounds. This study provides gustatory information for the ecological control of S. frugiperda.
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Evaluation of laboratory toxicities of nC22 agricultural mineral oil and its mixture with imidacloprid against the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae)
GAO Jing, YANG Qiong-Yu, FENG Yao-Heng, MAO Run-Qian
Acta Entomologica Sinica    2020, 63 (2): 191-198.   DOI: 10.16380/j.kcxb.2020.02.009
Abstract429)      PDF(pc) (1277KB)(48)    PDF(mobile) (1277KB)(10)    Save
【Aim】 The objective of this study is to evaluate the toxicities of nC22 mineral oil against the Asian citrus psyllid, Diaphorina citri, when used alone or mixed with imidacloprid in the laboratory, and to screen the mixing ratios that show synergistic effect on imidacloprid, so as to provide scientific support for the application of mineral oil against D. citri. 【Methods】 In the laboratory, the lethal effect of nC22 mineral oil and the positive controls nC23 and nC28 mineral oils against eggs, and the early instar nymphs, late instar nymphs, and adults of D. citri were determined by immersion method and spraying method, respectively, and the toxicities were calculated based on the LC50 values (7 d after treatment for eggs, and 1 d after treatment for nymphs and adults). The nC22 mineral oil was mixed with imidacloprid at various ratios, and their lethal effects on the early instar nymphs of D. citri were determined. The synergistic effect of nC22 mineral oil on imidacloprid was evaluated by interactive determination, co-toxicity factor (CTF), and co-toxicity coefficient (CTC) methods. 【Results】 When used alone, the LC50 value of nC22 mineral oil against D. citri eggs was significantly lower than those of nC23 and nC28 mineral oils. The LC50values of nC22 mineral oil against both the early and late instar nymphs were similar to those of nC23 mineral oil, and lower than those of nC28 mineral oil. The LC50 value of nC22 mineral oil against adults was similar to that of nC28 mineral oil, and lower than that of nC23 mineral oil. The mixtures of nC22 mineral oil and imidacloprid at the ratios of 3∶7, 4∶6 and 7∶3 showed remarkable synergistic effect on imidacloprid: the actual mortalities of the early instar nymphs of D. citri were above the equivalent line in the interactive determination method, the CTF value were 34.807, 22.655, and 40.798, and the CTC values were 187.430, 183.876 and 222.936, respectively. The mixture of nC22 mineral oil and imidacloprid at the ratio of 7∶3 showed the strongest synergistic effect on imidacloprid. 【Conclusion】 nC22 mineral oil shows higher toxicity against D. citri than the imported nC23 and traditional nC28 mineral oils. When mixed with imidacloprid at proper ratios, nC22 mineral oil shows strong synergistic effect, and theoretically it can significantly reduce the use of chemical insecticide, which could be further tested in fields.
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Genetic differences among geographical populations and rapid early detection of a nymphal parasitoid of Lycorma delicatula (Hemiptera: Fulgoridae)
XIN Bei, Atif MANZOOR, CAO Liang-Ming, WANG Xiao-Yi
Acta Entomologica Sinica    2020, 63 (2): 218-228.   DOI: 10.16380/j.kcxb.2020.02.012
Abstract428)      PDF(pc) (7356KB)(132)    PDF(mobile) (7356KB)(41)    Save
【Aim】 This study aims to determine the genetic differences among parasitoids of Lycorma delicatula nymphs from different geographical populations, and to rapidly identify the parasitism of parasitoids on L. delicatula nymphs at the early parasitization stage so as to evaluate the control effects of parasitoids on L. delicatula populations. 【Methods】 The DNA barcoding method was used to sequence COI and 28S rDNA genes of parasitoids of L. delicatula nymphs from different geographical populations. The genetic distances between the parasitoids from different geographical populations were calculated using K2P model, and a phylogenetic tree was constructed using neighbor-joining (NJ) method. The species-specific PCR (SS-PCR) primers were designed to determine whether L. delicatula was parasitized by Dryinus sinicus by amplification of DNA from L. delicatula nymphs using SS-PCR method. Visual assessment method and SS-PCR amplification were used to determine the parasitism rates of the parasitoids on L. delicatula nymphs from different sampling localities. 【Results】 The parasitoids of L. delicatula nymphs were identified as D. sinicus. A total of 16 haplotypes and four haplotypes were detected in COI and 28S rDNA sequences of D. sinicus from different geographical populations, respectively. The genetic distance among D. sinicus from different geographical populations was 0.00691-0.01310. The phylogenetic tree constructed by NJ method showed that D. sinicus samples from different geographical populations were clustered in one branch. SS-PCR primers based on COI sequence could produce good amplification results for both adult and larva of D. sinicus, with the detection threshold of 0.000005 ng/μL DNA. The parasitism rates of D. sinicus on L. delicatula nymphs from different sampling localities were 22.54%-60.00% detected by SS-PCR, significantly higher than those by visual assessment method (5.63%-36.98%). 【Conclusion】 The genetic differences of D. sinicus from different geographical populations are quite low. SS-PCR primers can be used for rapid detection of the early parasitism of L. delicatula by D. sinicus.
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Cloning, prokaryotic expression and tissue expression profiling of odorant binding protein gene AzanOBP3 from Agrilus zanthoxylumi (Coleoptera: Buprestidae)
GONG Xue-Fang, YANG Ping, WANG Yan-Lai, GUO Li, CHEN Di, XIE Shou-An, LÜ Shu-Jie
Acta Entomologica Sinica    2020, 63 (4): 390-400.   DOI: 10.16380/j.kcxb.2020.04.002
Abstract427)      PDF(pc) (2563KB)(121)    PDF(mobile) (2563KB)(47)    Save
【Aim】 The objective of this study is to clone the coding sequence of odorant binding protein (OBP) gene Azan OBP3 in Agrilus zanthoxylumi and to analyze its structure properties and expression profile, so as to further understand the process of identifying 
odorant substances in A. zanthoxylumi adult antennae and to provide a new fundamental evidence for the pest management and control. 【Methods】 The cDNA sequence of AzanOBP3 was amplified from A. zanthoxylumi with RT-PCR, and the nucleotide and deduced amino acid sequences of the gene were analyzed using different bioinformatics software. The recombinant expression vector pET-28a(+)/AzanOBP3 was constructed and transformed into Escherichia coli BL21(DE3) competent cells for expression by IPIG iduction. The recombinant protein was identified by SDS-PAGE and Western blotting. The expression profiles of AzanOBP3 in different female and male adult tissues (head, thorax, abdomen, leg and wing) of A. zanthoxylumi were detected by qPCR. 【Results】 We cloned the full-length cDNA sequence of AzanOBP3 (GenBank accession no.: MT318832) from A. zanthoxylumi. Its ORF is 414 bp in length, encoding 137 amino acids, with a predicted molecular weight of 16.038 kD and the isoelectric point of 4.79, and a signal peptide sequence of 29 amino acids at the N terminus. The encoded protein has six conserved cysteines belonging to the typical insect OBPs. Homologous sequence alignment analyses showed that AzanOBP3 has the highest amino acid sequence identity with AmalOBP2 from Agrilus mali and AplaGOBP from Agrilus planipennis (74.45% and 76.92%, respectively). The recombinant expression vector pET-28a(+)/AzanOBP3 was successfully constructed and the target protein was stably expressed in host bacteria in the form of fusion protein after culture at 37℃ 180 r/min in a shaker incubator and induction with 1 mmol/L IPTG for 4 h. The qPCR results revealed that AzanOBPwas expressed in various tissues of male and female adults of A. zanthoxylumi, with the highest expression level in the male adult leg. 【Conclusion】 In this study, the nucleotide and amino acid sequences of AzanOBP3 and the physiochemical properties of its encoded protein were clarified. The tissue expression profile suggests that AzanOBP3 may not be limited to the recognition of olfactory odorant, also have physiological functions in non-olfactory tissues, and especially may play important roles during insect feeding and host locating. Its functions need further study. This study provides a basis for exploring the molecular structure of odorant binding proteins and their functional mechanisms in the chemical sensing system of A. zanthoxylumi.
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Feeding and oviposition preference and fitness of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), on rice and maize
QIU Liang-Miao, LIU Qi-Quan, YANG Xiu-Juan, HUANG Xiao-Yan, GUAN Rui-Feng, LIU Bi-Pao, HE Yu-Xian, ZHAN Zhi-Xiong
Acta Entomologica Sinica    2020, 63 (5): 604-612.   DOI: 10.16380/j.kcxb.2020.05.010
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【Aim】 This study aims to clarify the feeding and oviposition preference and fitness of the fall armyworm, Spodoptera frugiperda, on host plants rice and maize and then to analyze its damage risk to rice. 【Methods】 The feeding and oviposition preference of S. frugiperda were tested by free choice method. The development, survival rate and fecundity of S. frugiperda fed on maize and rice seedlings were investigated, and the fitness of S. frugiperda on rice was evaluated by the population growth trend index (I). 【Results】 There were significant differences in the feeding and oviposition preference of S. frugiperda to rice and maize. The choice rate of larvae to feed on maize increased with time during 2-48 h post infestation, while that on rice decreased with time. At 48 h post infestation, the choice rates of the 2nd and 4th instar larvae to feed on maize were 93.06% and 59.72%, respectively, which were 66.95 and 21.48 times as high as those to rice. In terms of oviposition preference, the number of eggs laid on maize was 8.64 times as high as that on rice after a free oviposition period of 6 d. The individuals of S. frugiperda fed on rice seedlings had longer developmental duration, lower egg hatching rate, larval growth rate, pupation rate, adult emergence rate and proportion of females, shorter female longevity and less eggs laid per female as compared to those fed on maize seedlings. The population growth trend index (I) of S. frugiperda fed on maize seedlings was 165.93, and the relative fitness of S. frugiperda on rice seedlings was 0.21. 【Conclusion】 S. frugiperda prefers to feed and lay eggs on maize rather than on rice, and shows a higher fitness on maize. Rice is a non.preferable host to S. frugiperda that has invaded China, but it can feed on rice seedlings to grow and develop normally and finish its life cycle. The results suggest that S. frugiperda may become a potential threat to rice production when it has a high population density and its preferable host plants are absent.
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Effects of intestinal bacteria Citrobacter freundi and Klebsiella oxytoca on the development and substance metabolism of Drosophila suzukii (Diptera: Drosophilidae) 
GAO Huan-Huan, QIN Dong-Yun, DAI Xiao-Yan, LIU Jie, YU Yi
Acta Entomologica Sinica    2020, 63 (4): 462-469.   DOI: 10.16380/j.kcxb.2020.04.009
Abstract425)      PDF(pc) (1918KB)(63)    PDF(mobile) (1918KB)(19)    Save
 【Aim】 To understand the effect of the intestinal symbiotic bacteria Citrobacter freundi and Klebsiella oxytoca on the development and substance metabolism of the spotted wing drosophila, Drosophila suzukii. 【Methods】 The egg hatching rates, body weight of the 3rd instar larvae and pupation rates of the normal D. suzukii strain, sterile D. suzukii strain, C. freundi infected D. suzukii strain and K. oxytoca infected D. suzukii strain were determined. The contents of metabolites including protein, amino acids, glycogen and 
free fatty acids and the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in the 3rd instar larvae of these D. suzukii strains were detected. 【Results】 The egg hatching rate, body weight of the 3rd instar larvae, pupation rate and 
protein content in the 3rd instar larvae of the normal D. suzukii strain were all higher than those of other strains, while those of the sterile D. suzukii strain were the lowest. The 3rd instar larvae of D. suzukii strains infected by C. freundi or K. oxytoca had lower 
contents of amino acids and glycogen than the sterile and normal D. suzukii strains. The contents of free fatty acids in the 3rd instar larvae of D. suzukii infected by C. freundi were also lower than those in the other D. suzukii strains. In the 3rd instar larvae of D. 
suzukii strains infected by C. freundi or K. oxytoca, the POD activity was significantly higher than those in the sterile and normal strains, while the CAT activity was significantly lower than those in the sterile strain. 【Conclusion】 The development of D. 
suzukii delays in the absence of intestinal symbiotic bacteria. However, the development of D. suzukii can be enhanced in some extent after adding C. freundi or K. oxytoca in the food, and this is closely related to the changes of metabolites in D. suzukii after adding 
intestinal bacteria.
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RNAi of the calcium-binding protein gene and its biological effects in Bemisia tabaci MED (Hemiptera: Aleyrodidae)
GUO Lei, PAN Zheng-Yuan, LIU Jia-Yin, HAN Ming-Xuan, CHU Dong
Acta Entomologica Sinica    2020, 63 (2): 142-148.   DOI: 10.16380/j.kcxb.2020.02.003
Abstract425)      PDF(pc) (1188KB)(115)    PDF(mobile) (1188KB)(41)    Save
【Aim】 Our previous study revealed that two calcium-binding proteins (BtCaBP1 and BtCaBP2) are involved in the stress response of Bemisia tabaci MED to cyantraniliprole. This study aims to systematically reveal the biological effects of calcium-binding proteins on B. tabaci MED by RNAi of their genes. 【Methods】 After knocking down the calcium-binding protein genes BtCaBP1 and BtCaBP2 in B. tabaci MED adults by feeding dsRNA, the expression levels of the two genes were determined by qPCR. The biological parameters including the longevity (female and male) and fecundity of adults, and the egg hatching rate and pre-adult duration of offspring of B. tabaci MED between the control group (feeding on dsEGFP) and the treatment group (feeding on dsBtCaBP1 and dsBtCaBP2, respectively) were observed and compared after RNAi for 3 d. 【Results】 After feeding on dsBtCaBP1 and dsBtCaBP2, respectively, for 3 d, the expression levels of the target genes BtCaBP1 and BtCaBP2 of B. tabaci MED adults decreased significantly as compared with those in the control group. Compared with the control group, the dsBtCaBP2 treated-group had significantly increased the longevity (female: 15.46±1.24 d vs 13.25±0.58 d; male: 13.84±0.38 d vs 12.67±0.65 d), significantly decreased the number of eggs laid per female (39.53±3.04 vs 76.06±4.76), significantly decreased the egg hatching rate of offspring (81.58%±4.42% vs 87.22%±3.21%), and significantly shortened the pre-adult duration of offspring (24.42±1.09 d vs 27.52±1.73 d). However, feeding on dsBtCaBP1 had no significant effect on the above biological parameters of B. tabaci MED. 【Conclusion】 The RNAi of BtCaBP1 and BtCaBP2 has different biological effects on B. tabaci MED, and this finding will be helpful to reveal the function of CaBPs.
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Cloning and functional characterization of eight odorant receptor genes in Apolygus lucorum (Hemiptera: Miridae)
LI Bin, ZHANG Sai, WANG Chen-Rui, WANG Gui-Rong, LIU Yang
Acta Entomologica Sinica    2020, 63 (9): 1048-1058.   DOI: 10.16380/j.kcxb.2020.09.002
Abstract425)      PDF(pc) (2347KB)(165)    PDF(mobile) (2347KB)(76)    Save
【Aim】 This study aims to clone odorant receptor (OR) genes in Apolygus lucorum and to assay their expression levels in different adult tissues and their electrophysiological responses to host plant volatiles, so as to provide the theoretical basis for understanding the mechanism of host plant recognition in A. lucorum. 【Methods】 Based on the sequencing and analysis of the antennal transcriptome of A. lucorum, the full-length sequences of eight OR genes with intact ORF were cloned by PCR. The expression levels of these genes in different adult tissues (antenna, head without antenna, thorax, abdomen, leg and wing) were detected by using qPCR. The electrophysiological responses of these ORs to 56 odor compounds were tested in Xenopus oocytes as a heterologous expression system using two-electrode voltage clamp recording. 【Results】 The full-length cDNA sequences of 8 OR genes AlucOR9, AlucOR16, AlucOR38, AlucOR53, AlucOR55, AlucOR56, AlucOR57 and AlucOR58 (GenBank accession no.: MN905538-MN905545)of A. lucorum were cloned. These ORs contain seven transmembrane domains, with the intracellular N-terminus and extracellular C-terminus, consistent with the typical characteristics of insect ORs. The qPCR results showed that all the eight OR genes were highly expressed in adult antennae and lowly expressed in all the other tissues of A. lucorum. Moreover, the expression of the other seven OR genes except AlucOR38 in the antennae showed significant differences between male and female. The results of two-electrode voltage clamp recording showed that only AlucOR57 responded to 15 odor compounds [benzaldehyde, (-)-caryophyllene oxide, heptanal, (E)-2-hexenal, benzyl acetate, (1R)-(-)-myrtenal, 4-ethylbenzaldehyde, nonyl acetate, 3,7-dimethyl-3-octanol, tridecane, (E)-3-hexenol, butyl acrylate, butyl propionate, octyl acetate and amyl acetate], while the other seven ORs did not respond to all the odor compounds tested. 【Conclusion】 AlucOR57 responds to partial odor compounds tested, suggesting that it might be involved in the recognition of host plant volatiles in A. lucorum. The other seven ORs fail to respond to all the tested odor compounds, and their
functions need further study.
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Ovary structure and development of Spodoptera frugiperda (Lepidoptera: Noctuidae) from fields in different regions of China
FENG Bo, GUO Qian-Shuang, MAO Bi-Peng, ZHONG Ling, SONG Jian-Hui, XIAO Ming-Hui, DU Yong-Jun
Acta Entomologica Sinica    2020, 63 (5): 639-648.   DOI: 10.16380/j.kcxb.2020.05.013
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【Aim】 This study aims to explore and understand the ovary developmental status of Spodoptera frugiperda and its variation in fields so as to provide accurate monitoring basis for the occurrence and migration dynamics of S. frugiperda populations. 【Methods】 The morphological differences of female ovary of S. frugiperda at different ages (day-old) and from fields of different regions in China were compared by systematic dissection of female reproductive system. The relationship between ovarian maturation and mating of female moths was analyzed. 【Results】 The reproductive system of female moth of S. frugiperda is composed of ovaries, oviducts, copulatory pouch, afferent duct, spermatheca, accessory glands and ovipositors. According to the morphological characteristics, eggs of field female moths can be divided into three stages: pre-vitellogenesis, vitellogenesis and maturation. The eggs in the ovaries of 1-day old female moths barely reach the middle stage of vitellogenesis with the ovarian stalk empty. The ovaries start to differentiate at the 3-day old female moth stage. Some ovaries in the 3-day old female moths are still similar to those in the 1-day old moths, but some ovaries have mature eggs. The differentiation of ovary lasts till the 11-day old female moth stage. Some female moths from fields of Jiangcheng County and Mile County of Yunnan Province, Tianyang County of Guangxi Province, and Rui′an County and Zhenhai County of Zhejiang Province had immature ovaries until death, which accounted for 61.5%, 51.7%, 41.7%, 42.1% and 35.5% of the total ovaries, respectively. A proportion of 39.6% of the first generation female moths reared indoors still had immature ovaries at the time of death, which was similar to that of the field female moths. The ovaries of mated female moths could be mature and immature, but the proportion of immature ovaries was only 18.0%±5.0%. 【Conclusion】 The results suggest that field female moths of S. frugiperda have a mixture of migratory and non-migrating individuals in the same generation, and their proportions vary with the location and season. The results of this study explain to some extent the uncertainty of occurrence and damage of S. frugiperda populations and provide a basis for monitoring the migratory status and green control of S. frugiperda in the field.
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Sequencing and analysis of the complete mitochondrial genome of Carpoglyphus lactis (Acari: Carpoglyphidae)
ZHAO Ya-Nan, LI Chao-Pin
Acta Entomologica Sinica    2020, 63 (3): 354-364.   DOI: 10.16380/j.kcxb.2020.03.012
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【Aim】 To sequence and analyze the complete mitochondrial genome of Carpoglyphus lactis, and to explore the phylogenetic position of C. lactis in Acariformes based on the mitochondrial genome sequence, so as to provide a scientific basis for the classification of Acariformes and the study of mitochondrial genome of Carpoglyphidae. 【Methods】 Genomic DNA of C. lactis was initially extracted from adult mites of C. lactis bred in the laboratory using conventional phenol-chloroform and commercial DNA extraction kit. Partial sequences of mitochondrial genes including cox1, cob, rrnS and nad4-nad5 were amplified by PCR using universal primers for arthropods or mites. Species-specific primers were designed for long-PCR amplification and primer-walking sequencing, and the complete mitochondrial genome of C. lactis was sequenced. Bioinformatic software SeqMan, SEQUIN 9.0 and tRNAscan were used to analyze the genetic structure of complete mitochondrial genome. Finally, the phylogenetic relationship was constructed by using maximum likelihood method (ML) based on the nucleotide sequences of concatenated proteincoding genes (PCGs) of 17 species of Acariformes. 【Results】 The complete mitochondrial genome of C. lactis (GenBank accession no.: MN073839) is 14 060 bp in length. It is a classical closed circular double-stranded DNA molecule, and comprises 37 coding genes, including 13 proteincoding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes, and one large non-coding region (LNR). The results of phylogenetic analysis showed that C. lactis belongs to Acaroidea and clustered into one branch with Aleuroglyphus ovatus. Acaroidae and Histiostomatoidae clustered into a group, forming a sister group to Psoroptidia. 【Conclusion】 We successfully sequenced and analyzed the complete mitochondrial genome of C. lactis. The relationship between C. lactis and A. ovatus is close.
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Research advances in symbiotic microorganisms in insects and their functions
WANG Wei-Xia, ZHU Ting-Heng, LAI Feng-Xiang
Acta Entomologica Sinica    2021, 64 (1): 121-140.   DOI: 10.16380/j.kcxb.2021.01.013
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Symbiotic microorganisms can account for 1%-10% of insect biomass, including acteria, fungi, archaea and viruses. Insects and symbiotic microorganisms co-evolve to form holobionts. Symbiotic microorganisms play an important role in the biological characteristics, diversity formation, ecological adaptability and stress resistance of insects. Crop pest insects seriously affect agricultural production. In this article, the research advances in the diversity, research methods, and functional mechanisms of insect symbiotic microorganisms, the interaction between symbiotic microorganisms and their application in pest control since 2000 were reviewed and prospected. With the continuous development and application of advanced research methods such as molecular microbial ecology and metagenomic sequencing, breakthroughs have been made in the research of symbiotic microorganisms of agricultural pest insects. It was found that symbiotic microorganisms mainly affect host insects in the following ways: (1) Synthesis of nutrients or production of digestive enzymes to promote host growth and development and to expand host ecological niche; (2) Production of protective metabolites to directly protect the host against stress or indirectly protect the host by regulating the defense response of host plants; and (3) Production of active substances to regulate host propagation, mating, aggregation and movement. The abundance and community composition of insect symbiotic microorganisms maintain dynamic changes in a certain spatial-temporal range and have an important impact on host phenotype, which is the result of the benefit trade-off among host, environment, and interactive microorganisms. We suggest that future research should focus on clarifying the molecular mechanisms underlying the formation and maintenance of symbionts, ascertaining the complex interactions among symbiotic microorganisms, host insects, plants, natural enemies and the environment in more spatial-temporal dimensions, and designing green and efficient pest control strategies through targeted regulation of pest insect symbionts.
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Toxicity of spinetoram and its effects on the detoxifying enzyme and acetyl cholinesterase activities in Spodoptera frugiperda(Lepidoptera: Noctuidae) larvae
GAO Zu-Peng, GUO Jing-Fei, HE Kang-Lai, WANG Zhen-Ying
Acta Entomologica Sinica    2020, 63 (5): 558-564.   DOI: 10.16380/j.kcxb.2020.05.004
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【Aim】 The study aims to evaluate the toxicity and mechanism of action of spinetoram against the fall armyworm, Spodoptera frugiperda. 【Methods】 The LC50 and LC90 values of spinetoram against the 2nd, 3rd and 4th instar larvae of S. frugiperda were determined by diet-overlay bioassays with chlorantraniliprole as the control. The activities of mixed-functional oxidase (MFO), glutathione-S-transferase (GST), carboxylesterase (CarE) and acetyl cholinesterase (AchE) in the 3rd instar larvae of S. frugiperda treated by different concentrations of spinetoram (0.127, 0.183, 0.250, 0.400 and 0.572 mg/L) at 48 h post treatment were determined by enzyme-linked immunosorbent assay (ELISA). 【Results】 Compared with chlorantraniliprole, spinetoram had higher toxicity to S. frugiperda larvae. The LC50 values of spinetoram against the 2nd, 3rd and 4th instar larvae at 48 h post treatment were 0.21, 0.34 and 0.59 mg/L, and the LC90 values were 0.59, 0.75 and 2.01 mg/L, respectively. When the 3rd instar larvae were treated with spinetoram, the MFO and AchE activities increased significantly as the treatment concentration increased, and reached the highest (52.23 and 23.98 U/mg pro, respectively) at the concentration of 0.572 mg/L. When the 3rd instar larvae were exposed to the low concentrations of spinetoram (0.127 and 0.183 mg/L), the CarE activity showed no significant change as compared with the vehicle control (0.1% Tween-80), but significantly increased as the concentration increased to 0.400 and 0.572 mg/L. The GST activity increased significantly as the concentration of spinetoram increased, but showed no significant difference between the treatment concentrations of 0.400 and 0.572 mg/L. 【Conclusion】 Spinetoram is highly effective against S. frugiperda, especially to its 4th instar larvae, as compared with chlorantraniliprole. The activities of CarE, MFO and AchE in S. frugiperda larvae increase after exposure to different concentrations of spinetoram.
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Screening and evaluation of semiochemical mixtures attracting Holotrichia oblita (Coleoptera: Melolonthidae)
LI Xiao-Feng, LI Jin-Qiao, CAO Ya-Zhong, YIN Jiao, ZHANG Shuai, QIN Jian-Hui, WANG Si-Yu, LU Jun-Jiao, LI Ke-Bin
Acta Entomologica Sinica    2020, 63 (4): 482-493.   DOI: 10.16380/j.kcxb.2020.04.011
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【Aim】 To find effective attractant formulations for trapping Holotrichia oblita for ecological prevention and control. 【Methods】 Five plant volatiles, two aggregation pheromones and one sex pheromone were selected. According to the optimal concentration of each chemical, these compounds were mixed in pairs. A total of 17 binary attractant formulations were obtained, including plant volatiles plus aggregation pheromones, plant volatiles plus sex pheromones and aggregation pheromones plus sex pheromones. Based on the electro-antennogram (EAG) and taxis, the formulations with high attractiveness to H. oblita adults were gradually screened. Then, the trapping efficacies of these formulations and their single components on H. oblita adults were compared, and the attractiveness of these formulations was tested by trapping test in the indoor cage. 【Results】 Three binary attractant formulations cis-3-hexenyl acetate+(Z)-9-octadecene acetate, 1-hexanol+(Z)-9-octadecene acetate, and dibutyl phthalate+methyl glycate with strong attractiveness to H. oblita adults were screened from 17 attractant formulations by EAG and taxis tests, causing the EAG responses of 5 981%, 5 796% and 5 722%, respectively, in females, and 3 983%, 4 712% and 3 809%, respectively, in males. The formulation dibutyl phthalate+methyl glycate caused the highest selective response rate (71.21%) in H. oblita adults, followed by the formulations 1-hexanol+(Z)-9-octadecene acetate and cis-3-hexenyl acetate+(Z)-9-octadecene acetate (70.84% and 70.72%, respectively), causing significantly higher selective response rate than other formulations. The results of comparison of attractiveness between three formulations and their single components showed that the formulation cis-3-hexenyl acetate+(Z)-9-octadecene acetate showed significant synergistic effect on cis-3-hexenyl acetate but showed no significant synergistic effect on cis-9-octadecyl acetate. Both the formulations 1-hexanol+(Z)-9-octadecene acetate and dibutyl phthalate+methyl glycate showed significant synergistic effect on their respective single components. The results of indoor caged trapping showed that the numbers of adults trapped by the formulations cis-3-hexenyl acetate+(Z)-9-octadecene acetate, 1-hexanol+(Z)-9-octadecene acetate and dibutyl phthalate+methyl glycate were significantly higher than that of the control (paraffine oil). The formulation cis-3-hexenyl acetate+(Z)-9-octadecene acetate caused the highest selective response rate (74.06%) in H. oblita adults, while the formulations 1-hexanol+(Z)-9-octadecene acetate and dibutyl phthalate+methyl glycate caused the selective response rates of 65.89% and 64.25%, respectively. The selective response rates in H. oblita adults caused by the three formulations, however, showed no significant difference. 【Conclusion】 The binary attractant formulations cis-3-hexenyl acetate+(Z)-9-octadecene acetate, 1-hexanol+(Z)-9-octadecene acetate, and dibutyl phthalate+methyl glycate show strong attractiveness to H. oblita adults and can be used as effective attractants to ecologically control this insect.
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Impact of transgenic herbicide-resistant soybean on the diversity of arthropods and weeds in soybean fields
CHEN Yan-Jun, LIU Lai-Pan, GUAN Xiao, LIU Biao
Acta Entomologica Sinica    2020, 63 (11): 1366-1376.   DOI: 10.16380/j.kcxb.2020.11.009
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【Aim】 With the increasing of planting areas of transgenic crops, more and more attention has been paid to the ecological security issues. The study of the impact of the transgenic crops on biodiversity is not only an important component for the environmental release of transgenic crops, but also an important tool to objectively evaluate their risks. This study aims to analyze the impacts of herbicideresistant soybean on the diversity of arthropods and weeds in soybean fields. 【Methods】 From June to September 2019, we investigated the number of arthropods (the number of insects per 100 plants) and their diversity indexes (Margalef index, Shannon-Wiener index, Simpson index, dominance concentration index and Pielou index), the species and quantity of pests and natural enemies, and the density, species, quantity and diversity indexes of weeds in soybean fields planted with soybean cultivars of four treatments, i.e., the herbicide-resistant transgenic g10-epsps soybean ZUTS-33 developed in China, the receptor soybean Huachun 3 (HC-3) and the main soybean cultivar Zhonghuang 13 (ZH-13) sprayed with water, and the transgenic soybean ZUTS-33 sprayed with the glyphosate herbicide Roundup (3 000 mL/hm2), respectively, in Hefei, Anhui Province, eastern China. 【Results】 In the early and middle growth stages of soybean in 2019, the numbers of insects per 100 plants in the four treatments exhibited similar changing trends except that the number of insects per 100 plants in the treatment of ZUTS-33 sprayed with water on August 13 was significantly lower than those in the other three treatments. The number of arthropods and their various diversity indexes showed no significant difference among the four treatments at the early and middle growth stages of soybean, and the numbers of six pest groups including whiteflies, aphids, thrips, leaf beetles, leafhoppers and Helicoverpa armigera and the numbers of six natural enemy groups including spiders, ladybirds, lacewing, flower bugs, plant bugs and Anomis flava showed similar dynamics among the four treatments during the investigation period. There were 10 weed species belonging to 8 families in the investigated soybean fields. The density and number of weeds in the treatment of ZUTS-33 sprayed with Roundup were significantly lower than those in the other three treatments, and their Margalef index, Shannon-Wiener index, Simpson index, and Pielou index in the treatment of ZUTS-33 sprayed with Roundup were significantly different from those in the other three treatments at the late growth stage of soybean. 【Conclusion】 The planting of the herbicide-resistant transgenic g10-epsps soybean ZUTS-33 has no significant effect on the diversity of arthropods and weeds in soybean fields.
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Cloning and expression profiling of vitellogenin gene and its response to UV-A stress in Ostrinia furnacalis (Lepidoptera: Crambidae)
LIU Fang, MENG Jian-Yu, YANG Chang-Li, ZHANG Chang-Yu
Acta Entomologica Sinica    2020, 63 (3): 255-265.   DOI: 10.16380/j.kcxb.2020.03.002
Abstract401)      PDF(pc) (3201KB)(111)    PDF(mobile) (3201KB)(24)    Save
【Aim】 This study aims to explore the effects of UV-A stress on the vitellogenin (Vg) gene expression and fecundity of the Asian corn borer Ostrinia furnacalis by cloning Vg and analyzing its expression pattern. 【Methods】 The full-length cDNA of Vg gene was cloned from O. furnacalis with RT-PCR and RACE techniques and analyzed by bioinformatic methods. The relative expression levels of Vg gene in different developmental stages (egg, 1st-5th instar larva, pupa and adult), different tissues (head, leg, cuticle, ovary, midgut and fat body) of female adults, and female adults of O. furnacalis exposed to UV-A for different time (0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4 h) were detected by RT-qPCR. The reproduction and development of the F1 generation of O. furnacalis adults after exposure to UV-A for 0, 1, 2, 3 and 4 h/d were measured. 【Results】 The complete sequence of Vg gene from O. furnacalis was cloned and named as OfVg (GenBank accession no.: MK782978). Its full-length cDNA is 5 760 bp and contains a 5 332 bp open reading frame (ORF), encoding 1 776 amino acids with the predicted molecular weight of 202.10 kD and isoelectric point of 9.06. OfVg contains three functional domains (Vg-N, DUF 1943D and VWD) but has no transmembrane region. The phylogenetic analysis showed that OfVg was most closely related to Vg proteins of other lepidopteran insects. Developmental expression profiling showed that the expression level of OfVg in female adult was significantly higher than those in other developmental stages, reaching the peak in 24 h-old female adult. Tissue expression profiling revealed that OfVg was specifically expressed in the fat body of female adults. UV-A stress induced the expression of OfVg in female adults, with the expression level decreasing firstly and then increasing with the increase of exposure time, reaching the maximum at 3.5 h post exposure to UV-A, and then decreasing abruptly. When the adults were exposed to UV-A for 1, 2 and 3 h/d, their fecundity increased, and the developmental duration of larvae of F1 generation was prolonged significantly. 【Conclusion】 The expression level of OfVg varies in different developmental stages, different tissues of female adults and female adults of O. furnacalis exposed to UV-A for different time. This study lays a foundation for further studying the effect of UV-A stress on the development and reproduction of O. furnacalis.
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Cloning of heat shock protein gene SfHsp90 and its expression under high and low temperature and UV-A stresses in Spodoptera frugiperda (Lepidoptera: Noctuidae)
ZHOU Lü, MENG Jian-Yu, YANG Chang-Li, LI Jin, HU Chao-Xing, ZHANG Chang-Yu
Acta Entomologica Sinica    2020, 63 (5): 533-544.   DOI: 10.16380/j.kcxb.2020.05.002
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【Aim】 This study aims to explore the molecular mechanism of Spodoptera frugiperda in response to high and low temperature and UV-A stresses. 【Methods】 The heat shock protein Hsp90 gene of S. frugiperda was cloned by RT-PCR technique, and its sequence characteristics were analyzed using bioinformatics methods. The relative expression levels of Hsp90 in different developmental stages (egg, 1st-6th instar larva, pupa and adult), different adult tissues (head with antennae and compound eyes removed, thorax, abdomen, antenna, compound eye, leg, wing, midgut, testis and ovary) and adults exposed to high temperature of 36℃, low temperature of 4℃ and UV-A radiation for different time (0, 30, 60, 90, 120 and 150 min) were detected by RT-qPCR. 【Results】 The Hsp90 gene was cloned from S. frugiperda and named SfHsp90 (GenBank accession no.: MN832694). Its open reading frame (ORF) is 2 154 bp in length, encoding 717 amino acids, with the relative molecular weight of 82.52 kD and the isoelectric point (pI) of 5.01. The C-terminal sequence contains the conserved motif EEVD, suggesting that the protein is a cytoplasmic heat shock protein. Phylogenetic analysis showed that insect Hsp90 proteins are highly conserved. Developmental expression profile showed that the expression level of SfHsp90 was the highest in the 1st instar larva, and the tissue expression profile revealed that the expression levels of SfHsp90 were significantly higher in the antenna, compound eyes and head with antennae and compound eyes removed of male and female adults than in other tissues. High and low temperature stresses induced the expression of SfHsp90 obviously. The expression level of SfHsp90 in adults exposed to high temperature of 36℃ was significantly higher than that of the control group. After the adults were exposed to high temperature of 36℃, the expression level of SfHsp90 increased at first and then decreased with the increase of exposure time, and reached the maximum at 60 min after exposure in male adults, while its expression level in female adults increased gradually with the prolonging of exposure time. After the adults were exposed to low temperature of 4℃, the expression level of SfHsp90 increased firstly and then decreased in male adults with the prolonging of exposure time, and reached the maximum at 30 min after exposure, while its expression level in female adults increased gradually with the prolonging of exposure time. The expression of SfHsp90 in female and male adults was induced by UV-A stress. After the adults were exposed to UV-A radiation, the expression level of SfHsp90 increased firstly and then decreased with the increase of exposure time, and reached the highest at 90 min in male adults and at 60 min in female adults, respectively. 【Conclusion】 The differential expression of SfHsp90 in S. frugiperda adults under high and low temperature and UV-A stresses suggests that this gene plays an important role in the molecular mechanism in S. frugiperda in response to environmental stresses.
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Comparison of the feeding and growth characteristics of larvae of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), on corn and wheat
LÜ​ Liang, LI Yu-Qing, CHEN Cong-Liang, CHANG Xiang-Qian, ZHANG Shu, XU Dong, WAN Peng
Acta Entomologica Sinica    2020, 63 (5): 597-603.   DOI: 10.16380/j.kcxb.2020.05.009
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【Aim】 To assess the risk of damage and outbreak caused by the fall armyworm, Spodoptera frugiperda, shifted to wheat. 【Methods】 The feeding and growth characteristics and population life table parameters of S. frugiperda on corn and wheat under 23℃ in the laboratory were determined and compared by indoor breeding and survey statistics. 【Results】 S. frugiperda could complete its life cycle on wheat, and the food intake and body weight of larvae after the 3rd instar fed on wheat were significantly higher than those fed on corn at the same time after treatment, but the food utilization efficiency, larval survival, larval duration, and egg hatching rate fed on wheat were significantly lower than those fed on corn. There were no significant differences in such indexes as the average pupal weight, pre-oviposition period, and number of eggs laid per female of S. frugiperda fed on corn and wheat. In addition, the comparison of life table parameters showed that there were no significant differences in the mean generation time (T), intrinsic rate of increase (rm), and finite rate of increase (λ) of S. frugiperda fed on corn and wheat, but the net reproductive rate (R0) of S. frugiperda fed on corn was 303.55±2.04, which was significantly higher than that fed on wheat. 【Conclusion】 S. frugiperda fed on wheat grows and develops fast and can complete the life cycle. However, the food utilization efficiency and population reproductive ability of S. frugiperda fed on wheat are lower than those on corn, suggesting that S. frugiperda is more suitable for feeding on corn. There is a risk of shifting to damage wheat by S. frugiperda. But considering the source of the pest, natural temperature and other conditions, S. frugiperda is less likely to have an outbreak during damaging wheat. The results of this study provide basic data for clarifying the risk of S. frugiperda damage and outbreak on wheat and the scientific control of S. frugiperda.
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Identification of circadian clock genes Gmper and Gmtim and their contributions to the emergence rhythm in Grapholita molesta (Lepidoptera: Tortricidae)
FANG Hai-Bo, ZHANG Jing, LIU Xiao-Xia, ZHANG Qing-Wen, LI Zhen
Acta Entomologica Sinica    2020, 63 (6): 655-666.   DOI: 10.16380/j.kcxb.2020.06.001
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【Aim】 The purpose of this study is to investigate the molecular characteristics and expression patterns of two circadian clock genes Gmper and Gmtim in the oriental fruit moth, Grapholita molesta, and to explore their regulatory roles in the emergence rhythm, so as to provide potential new targets for controlling G. molesta. 【Methods】 Based on the transcriptome data of G. molesta, the full-length cDNAs of two circadian clock genes Gmper and Gmtim were cloned by PCR. The expression levels of these two genes in the head, thorax, abdomen and leg of adults and their daily expression patterns in the pupal head were determined by RT-qPCR. Then the regulatory roles of Gmper and Gmtim in the emergence rhythm of G. molesta were explored by RNAi with siRNAs. 【Results】 The full-length cDNAs of Gmper (GenBank accession no.: MN862636) and Gmtim (GenBank accession no.: MN862637) were cloned from G. molesta. The open reading frame of Gmper is 2 862 bp in length, encoding 953 amino acids with two PAS domains and a PAC domain. The open reading frame of Gmtim is 3 048 bp in length, encoding 1 015 amino acids. The expression levels of Gmper and Gmtim in heads were higher than those in other tissues of both male and female adults. In pupal head, the expression levels of the two genes were significantly higher in the scotophase than in the photophase. The emergence rhythm changed after RNAi of both Gmper and Gmtim, and G. molesta in the RNAi group showed a more dispersed emergence pattern than that in the control group, and the amount of emerged adults during the emergence peak period decreased after RNAi. 【Conclusion】 Both Gmper and Gmtim in G. molesta show differential expression patterns among different tissues and between day and night, and they may play important regulatory roles in the emergence rhythm of G. molesta. The results provide clues to the development of monitoring and control technologies for G. molesta based on behavioral rhythm regulation.
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Transcriptomic analysis of insecticide target and detoxification related genes in Adoxophyes orana (Lepidoptera: Tortricidae)
SUN Li-Na, ZHANG Huai-Jiang, LIU Xiao-He, QIU Gui-Sheng
Acta Entomologica Sinica    2020, 63 (4): 470-481.   DOI: 10.16380/j.kcxb.2020.04.010
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 【Aim】 To establish the transcriptome database of Adoxophyes orana, and to explore insecticide target and detoxification related genes. 【Methods】 The transcriptome of A. orana was sequenced by Illumina HiSeqTM 2000 sequencing platform to explore insecticide target genes. The expression levels of six insecticide target genes from A. ornan at different developmental stages including egg, larva, pupa and adult were identified and verified using qPCR. The metabolic pathway and evolutionary relationship of 
detoxification related genes in the transcriptome of A. orana were analyzed. 【Results】 A total of 48 610 unigenes (GenBank accession no.: GGMW00000000) were identified by assembling valid sequences. One hundred and fifty-five insecticides target genes were identified. The qPCR results confirmed that there was difference in the expression levels of six insecticide target genes including one ecdysone receptor (ECR) gene, two acetylcholinesterase (AChE) genes, one chloride channel (CLC) gene, one chitinase (CS) gene 
and one ryanodine receptor (RyR) gene at different developmental stages of A. ornanAdditionally, 69 carboxylesterase (CarE) unigenes, 66 glutathion S-transferase (GST) unigenes and 205 cytochrome P450 unigenes were identified. A total of 20 CarE, 32 GST and 30 P450 unigenes were classified into the metabolism categories related to toxic substance metabolism. The results of cluster analyses based on the amino acid sequences of unigenes with complete ORF showed that 9 of 12 CarEs were classified to G class (Lepidoptera neotenin), 10 AoGSTs into subfamily Delta and another 10 AoGSTs into subfamily Epsilon, and all of 18 P450s into CYP3 group. 【Conclusion】 This study contributes to the discovery of insecticide target genes and the research of pesticide resistance in A. orana.
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Cloning and functional characterization of the peptidoglycan recognition protein gene BdPGRP-SB1 in Bactrocera dorsalis (Diptera: Tephritidae)
ZHANG Ying-Xin, CHEN Dong, ZHANG Su-Yun, WEI Dong, WANG Jin-Jun
Acta Entomologica Sinica    2020, 63 (9): 1070-1080.   DOI: 10.16380/j.kcxb.2020.09.004
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【Aim】 To explore the role of a peptidoglycan recognition protein (PGRP) gene, BdPGRP-SB1, in the immunity of the oriental fruit fly, Bactrocera dorsalis. 【Methods】 The full-length cDNA sequence of BdPGRP-SB1 of B. dorsalis was cloned by PCR. The nucleotide and amino acid sequence characteristics of this gene were analyzed using bioinformatics software. The relative expression levels of BdPGRP-SB1 in different developmental stages (egg, larva, pupa and adult) and tissues (midgut, Malpighian tubules, hindgut, fat body, ovary and testis) of the 5-day-old adult of B. dorsalis were analyzed by RT-qPCR. The expression levels of BdPGRP-SB1 in the 5-day-old female adults of B. dorsalis injected with the peptidoglycan PGN-EB from Escherichia coli 0111:B4 and PGN-SA from Staphylococcus aureus, respectively, were detected by RT-qPCR. After the expression of BdPGRP-SB1 was suppressed by RNAi, the mortality of female adults of B. dorsalis post injection of E. coli and S. aureus and the expression levels of three antimicrobial peptide (AMP) genes including attacin-A, defensin and diptercin in female adults of B. dorsalis post infection ofi  E. colwere assayed. 【Results】 The full-length cDNA sequence of BdPGRP-SB1 (GenBank accession no.: MN892482) was successfully cloned, and its ORF is 558 bp in length, encoding a protein of 185 amino acid residues with a predicted molecular weight of 21.45 kD and a theoretical pI of 8.57. Sequence analysis indicated that BdPGRP-SB1 is a secreted protein with a signal peptide and a conserved PGRP domain but without transmembrane domain, and has the Zn2+-dependent amidase activity and the recognition sites of DAP-type peptidoglycan. Phylogenetic analysis indicated that BdPGRP-SB1 is the most closely related to PGRP-SB1 of B. latifrons, sharing 96% amino acid sequence identity. The developmental expression profile revealed that BdPGRP-SB1 was highly expressed in the 3-day-old larva and adult of B. dorsalis, and the tissue expression profile showed that it was expressed in various tissues of the 5-day-old adults, with the highest expression level in fat body. Both PGN-EB and PGN-SA induced the expression of BdPGRP-SB1 in female adults of B. dorsalis. After the suppression of BdPGRP-SB1 expression by RNAi, E. coli infection resulted in significantly higher mortality and significant up-regulation of attacin-A, defensin and diptercin in female adults of B. dorsalis. 【Conclusion】 The results suggest that BdPGRP-SB1 is involved in the recognition of gram-negative bacteria and may participate in Imd pathway to regulate immune response in B. dorsalis.
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Cloning and expression profiling of the serine protease inhibitor gene Nlserpin4 in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)
WANG Zheng-Liang, ZHU Hang-Feng, PAN Hai-Bo, YU Xiao-Ping
Acta Entomologica Sinica    2020, 63 (7): 779-787.   DOI: 10.16380/j.kcxb.2020.07.001
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【Aim】 To clone and characterize the serine protease inhibitor gene Nlserpin4 from the brown planthopper (BPH), Nilaparvata lugens, and to determine its spatio-temporal expression profiles and its expression pattern induced by entomopathogenic fungi. 【Methods】 Based on the transcriptome and whole genome data of BPH, the full-length cDNA of Nlserpin4 from BPH was cloned by PCR, and its nucleotide and protein sequences were subsequently characterized using bioinformatics tools. The expression patterns of Nlserpin4 across different developmental stages (egg, 1st-5th instar nymphs and newly emerged female and male adults), in different tissues (fat body, gut, hemolymph and carcass) of the 5th instar nymphs, and in the 5th instar nymphs at different time post injection of the entomopathogenic fungus Metarhizium anisopliae were determined by qRT-PCR. 【Results】 The full-length cDNA of Nlserpin4 (GenBank accession no.: MN822802) was successfully cloned from BPH. The open reading frame (ORF) of Nlserpin4 is 1 227 bp in length, encoding 408 amino acids with the predicted molecular weight of 45.91 kD and isoelectric point (pI) of 6.23. The amino acid sequence analysis revealed that Nlserpin4 protein has no putative N-glycosylation site, but contains a predicted signal peptide consisting of 23 amino acid residues at the N-terminal region and a typical RCL region of the serpin family at the C-terminal region with an active cleavage site which can be recognized by target protease. The phylogenetic analysis showed that Nlserpin4 is closely related to the serpins of other hemipteran insects, with the highest homology with Sipha flava serpin4. The qRT-PCR results showed that the expression of Nlsperpin4 had obvious temporospatial characteristics. The expression level of Nlsperpin4 in adults was significantly higher than those in other developmental stages, and the highest expression level was observed in male adult. Nlsperpin4 was expressed in the fat body, gut, haemolymph and carcass, with the highest expression level in the carcass of the 5th instar nymphs. The expression of Nlserpin4 in BPH was significantly down-regulated after infection with M. anisopliae within 48 h post injection, but the expression level of Nlserpin4 was gradually increased with the increase of infection time. 【Conclusion】 The Nlserpin4 of BPH is differentially expressed in different developmental stages, different tissues and different time after infection with the entomopathogenic fungus M. anisopliae. The results of this study provide a theoretical basis for further studying the functions of Nlserpin4 in the growth, development and immune regulation of BPH.
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Effects of three insecticides at the sublethal dose on the expression of cytochrome P450 genes in Spodoptera frugiperda (Lepidoptera: Noctuidae)
ZHANG Bai-Zhong, SU Xu, LU Liu-Yang, ZHEN Cong-Ai, ZHU Bin, LI Ya-She, DONG Wen-Yang, WANG Geng, XU Yan-Bo, KONG Fan-Bin, LIU Run-Qiang, CHEN Xi-Ling, GAO Xi-Wu
Acta Entomologica Sinica    2020, 63 (5): 565-573.   DOI: 10.16380/j.kcxb.2020.05.005
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【Aim】 To determine the effects of insecticides at the sublethal dose on the expression of cytochrome P450 genes in the fall armyworm, Spodoptera frugiperda. 【Methods】 The toxicity of three insecticides (chlorantraniliprole, emamectin benzoate and Bacillus thuringiensis) against the 2nd instar larvae of S. frugiperda was assayed by leaf-dipping method, and the transcriptional levels of 16 P450 genes in the 2nd instar larvae of S. frugiperda treated with the sublethal dose (LC10) of the three insecticides were measured by real-time quantitative PCR (RT-qPCR) at 48 h post treatment. 【Results】 The LC10 values of chlorantraniliprole, emamectin benzoate and B. thuringiensis against the 2nd instar larvae of S. frugiperda were 0.931, 0.283, and 1 089.688 mg/L, respectively. In the 2nd instar larvae exposed to the LC10 of chlorantraniliprole, 13 P450 genes including CYP4G75, CYP6AB12, CYP6B50, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP9A59, CYP9A58, CYP6AE44, and CYP6AE43 were up-regulated, and the transcription level of CYP6AE44 was 34.60-fold as high as that of the control. In the 2nd instar larvae exposed to the LC10 of emamectin benzoate, 11 P450 genes including CYP4G75, CYP6AB12, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP9A58, CYP6AE44, and CYP6AE43 were up-regulated, and the expression level of CYP321B1 was 28.70-fold as high as that of the control. In the 2nd instar larvae exposed to the LC10 of B. thuringiensis, 11 P450 genes including CYP4G75, CYP6AB12, CYP6AN4, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP6AE44, and CYP6AE43 were up-regulated, and the expression level of CYP6AE44 was 40.80-fold as high as that of the control. 【Conclusion】 Multiple P450 genes are up-regulated in the 2nd instar larvae of S. frugiperda in response to the sublethal dose of the three insecticides, and the expressions of CYP4G75, CYP6AB12, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP321B5, CYP6AE44, and CYP6AE43 are induced by all the three insecticides.
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Effects of Varroa destructor on the respiratory metabolism of Apis mellifera ligustica workers
TAN Jing, WU Jiang-Li, TU Yang-Yang, Tessema AYNALEM, YU Hui-Min, LI Nan-Nan, LI Xiao-Ying, XU Shu-Fa
Acta Entomologica Sinica    2020, 63 (11): 1325-1332.   DOI: 10.16380/j.kcxb.2020.11.005
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【Aim】 The Italian honey bee, Apis mellifera ligustica, is one of the most economically important pollinators in the world, and plays important roles in maintaining the ecological balance, improving the output of agricultural products and ensuring food safety. However, the bee suffers from huge annual loss worldwide due to infestation of Varroa destructor. The purpose of this study is to investigate the effects of V. destructor on the respiratory metabolism of A. mellifera ligustica workers, so as to lay a foundation for further studying the interaction between the mite and the bee. 【Methods】 The oxygen consumption, CO2 emission and respiration quotient of V. destructor-infected and healthy workers of A. mellifera ligustica at different developmental stages (4-day-old larva, 5-day-old pupa, newly emerged adult and 6-day-old adult) were measured using SSI multi-channel insect respiration instrument, and the characteristics of their changes were analyzed. Meanwhile, the expression levels of respiratory metabolism-related genes (Coq7, COXⅠ, CytB, CytC and IF-2mt) in V. destructor-infected and healthy adult worker bees were analyzed using real-time fluorescence quantitative PCR (RT-qPCR). 【Results】 The oxygen consumption and CO2 emission of A. mellifera ligustica worker bees first increased and then decreased after infection by V. destructor. After infection by V. destructor, the oxygen consumption decreased from 0.0244 mL/min in the 5-day-old pupal stage of A. mellifera ligustica to 0.0093 mL/min in the 6-day-old adult stage, and the CO2 emission decreased from 0.0174 mL/min in the 5-day-old pupal stage to 0.0040 mL/min in the 6-day-old adult stage, which was significantly lower than that of the healthy worker bees of the same developmental stage. The respiratory quotient of healthy worker bees of A. mellifera ligustica increased gradually with larval age, and reached the peak (0.9169) at the 6-day-old adult stage, while the respiratory quotient of the V. destructor-infected worker bees decreased, and was the lowest (0.4424) at the 6-day-old adult stage, which was significantly lower than that of the healthy worker bees of the same developmental stage. The RT-qPCR results showed that the relative expression levels of CytB and CytC genes in the V. destructor-infected adult worker bees of A. mellifera ligustica were 1.7676 and 1.9929-fold as high as those of the healthy adult worker bees, respectively. 【Conclusion】 V. destructor infection can directly affect the respiratory metabolism of A. mellifera ligustica workers, leading to the disorder of their normal metabolic pathway, and seriously damaging the normal growth and development of bee colonies.
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Identification and expression analysis of miR-2769 target gene in Bombyx mori
SUN Yan-Yan, WANG Jing-Ya, WANG Lu, LI Xiao-Zhe, KAN Yun-Chao, QIAO Hui-Li
Acta Entomologica Sinica    2020, 63 (3): 245-254.   DOI: 10.16380/j.kcxb.2020.03.001
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【Aim】 MiRNAs play very important roles in insect metamorphosis. The study of miRNAs and their target genes in Bombyx mori will help to elucidate the molecular mechanism of miRNAs involved in the metamorphosis of this insect. 【Methods】 The expression of miR-2769 in the fat body of B. mori was detected by qRT-PCR after injecting molting hormone, 20-hydroxyecdysone (20E), into the hemolymph of the day-2 5th instar larvae. The target genes of miR-2769 of B. mori were predicted by bioinformatics methods. The interaction between miR-2769 and its predicted target gene BmE75B was analyzed by dual luciferase reporter assay system. The expression levels of miR-2769 and different splice isoforms of its target gene BmE75 at different developmental stages (larva, pupa and adult) and larval tissues (head, epidermis, silk gland, fat body, testis, ovary, Malpighian tubules, midgut and hemolymph) of B. mori were detected by qRT-PCR. 【Results】 The results showed that miR-2769 could significantly inhibit the expression of luciferase reporter gene by interacting with the 3′UTR binding site of BmE75B of B. mori. The qRT-PCR results showed that miR-2769 and BmE75A/BmE75B presented the opposite expression trends in the fat body of B. mori post induction by 20E. The spatio-temporal expression analysis showed that miR-2769 and different splice isoforms of the target gene BmE75 had specific expression characteristics in different developmental stages and larval tissues of B. mori. The expression levels of miR-2769 and BmE75A were low in B. mori at different developmental stages; however, the expression level of BmE75B was relatively high in the pupal stage and that of BmE75C was extremely high in the late 5th instar larvae and early pupae. Furthermore, the expression of miR-2769 was negatively correlated with that of different splice isoforms of BmE75. miR-2769 promoted the expression of BmE75A in the hemolymph and BmE75B in the fat body of the 5th instar larvae of B. mori, while it inhibited the expression of different splice isoforms of BmE75 in other tissues. 【Conclusion】 MiR-2769 can negatively regulate the expression of different splice isoforms of BmE75 by interacting with the 3′UTR region of E75B in B. mori.
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Low temperature adaptability of Sitobion avenae (Hemiptera: Aphididae) and its spring source in wheat fields in Yangling, Shaanxi, northwestern China
XU Xiang-Li, HE Le-Le, YU Xiao-Ting, WU Jun-Xiang
Acta Entomologica Sinica    2020, 63 (3): 309-316.   DOI: 10.16380/j.kcxb.2020.03.007
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【Aim】 This study aims to clarify the survival rate and response to rapid cold acclimation in the grain aphid, Sitobion avenae, under extremely low temperature so as to provide a basis for the study of cold tolerance and accurate prediction of this insect. 【Methods】 The supercooling point and freezing point of the laboratory population of S. avenae at various developmental stages were determined, the survival rates of the 1st instar nymphs and the apterous adults exposed to extremely low temperature from -7℃ to -11℃ for 3 h and those exposed to lethal temperature for 3 h after cold acclimation at 0℃ for 1-5 h, respectively, and then transferred to 15℃ for 72 h, were assessed, and the population dynamics of S. avenae was investigated during the whole wheat growth period in Yangling, Shaanxi, northwestern China. 【Results】 The supercooling points of the 1st and 2nd instar nymphs of S. avenae showed a small fluctuation from -27.4℃ to -19.2℃ and from -27.3℃ to -18.3℃, respectively, while those of the 3rd and 4th instar nymphs and adults showed a large fluctuation from -27.4℃ to -10.7℃, from -26.7℃ to -12.5℃, and from -26.7℃ to -11.2℃, respectively. The supercooling point and freezing point of S. avenae increased significantly with the increasing instars. The supercooling point of adults was significantly higher than those of the 1st and 2nd instar nymphs. There were no significant differences in the supercooling points of the 3rd and 4th instar nymphs and adults between the alate and apterous morphs at the same developmental stage. The analysis of survival rate at low temperature showed that the lethal temperatures (80% mortality) for the 1st instar nymphs and apterous adults remained around -10.5℃ and -8.1℃, respectively. The rapid cold acclimation at 0℃ improved significantly the survival rate of the 1st instar nymphs and apterous adults at extremely low temperature, with the highest survival rate after cold acclimation for 2 h. The results of field investigation from 2018 to 2019 revealed that S. avenae could overwinter as parthenogenetic nymphs and adults in Yangling, Shaanxi. 【Conclusion】 S. avenae shows strong low temperature adaptability. Its parthenogenetic aphids can overwinter successfully in Yangling, Shaanxi. These results suggest that aphids in the wheat field in early spring may source partly from the survived locally overwintering individuals of this aphid.
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Phylogeny of hydradephagan water beetles (Coleoptera: Adephaga) inferred with mitochondrial genome sequences
TIAN Tian, YUAN Huan, CHEN Bin
Acta Entomologica Sinica    2020, 63 (8): 1016-1027.   DOI: 10.16380/j.kcxb.2020.08.012
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【Aim】 To clarify the general features of mitochondrial genomes of hydradephagan water beetles, and to analyze the phylogenetic relationship of hydradephagan water beetles based on the mitochondrial genome sequences. 【Methods】 Based on the Illumina HiSeq X Ten sequencing technology, we sequenced and annotated the complete mitochondrial genomes of Dineutus mellyi and Eretes sticticus, and predicted the secondary structures of tRNA genes. Combined with the published mitochondrial genomes of 17 species of hydradephagan water beetles, comparative genomics analyses including the AT content, codon usage bias, and selection pressure were performed on the mitochondrial protein-coding genes (PCGs) of the total 19 species. Based on the amino acid dataset and nucleotide dataset of 13 PCGs, the phylogenetic relationships of hydradephagan water beetles were reconstructed by using maximum likelihood (ML) and Bayesian (BI) methods, respectively, and the systematic position of Noteridae and Meruidae was further evaluated by FcLM analysis. 【Results】 The mitochondrial genomes of D. mellyi and E. sticticus are 16 123 bp (GenBank accession no.: MN781126) and 16 196 bp (GenBank accession no.: MN781132) in length, respectively. Each mitochondrial genome contains 13 PCGs, 22 tRNA genes, 2 rRNA genes and a D-loop region (control region). Within the 19 species of hydradephagan water beetles, the nucleotide composition biases show a higher A+T content, and the codon use bias analysis also found that there is a serious bias towards using AT-rich codons. The evolutionary rates of the 13 protein-coding genes showed a purifying selection. The phylogenetic relationship of hydradephagan water beetles based on the amino acid sequences of the 13 PCGs was: (Gyrinidae+(Haliplidae+((Aspidytidae+(Amphizoidae+Dytiscidae))+(Hygrobiidae+(Meruidae+Noteridae))))). 【Conclusion】 Our results indicate that Gyrinidae is the base group of hydradephagan water beetles, followed by Haliplidae and Dytiscoidea, Noteridae and Meruidae form a sister-group branch within Dytiscoidea, and a closer relationship exists between Amphizoidae and Dytiscidae.
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Analysis of gene co-expression patterns in different tissues of Aedes aegypti (Diptera: Culicidae)
LIU Jie, LI Bo, CHEN Xiao-Jie, CHEN Bin
Acta Entomologica Sinica    2020, 63 (10): 1171-1182.   DOI: 10.16380/j.kcxb.2020.10.002
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【Aim】 To explore the co-expression patterns of genes in different tissues of Aedes aegypti by using weighted gene co-expression network analysis (WGCNA). 【Methods】 Paired sequencing data of representative nine adult tissues (antenna and brain of female and male, proboscis, maxillary palp and ovary of female, and foreleg, midleg, hindleg and abdominal tip of male) were selected from the transcriptome data of different tissues of A. aegypti in the NCBI SRA database. After removing missing values and calculating variance, 5 000 genes with the highest variance were chosen, and the WGCNA package in R software was used to establish the gene co-expression networks in different adult tissues of A. aegypti and divide modules. Then the clusterProfiler package was used to conduct GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis of the genes in the tissue-specific modules, and CytoHubba plug-in in Cytoscape software was adopted to screen the hub genes in co-expression modules. 【Results】 A total of 11 gene co-expression modules in different adult tissues of A. aegypti were identified, and one specific expression module was identified in every tissue of the antenna, proboscis, ovary and maxillary palp of female adult, and the brain and abdominal tip of male adult. There was no specific expression module in tissues of the foreleg, midleg and hindleg of male adults. Genes in six tissue-specific expression modules were annotated to biological functions of the corresponding tissues. Among them, genes in the female antenna specific green module have functions such as odor binding and olfactory receptor activity. Genes in the female proboscis specific purple module have functions such as serine-type endopeptidase activity and serine hydrolase activity. Genes in the male brain specific blue module play a major role in regulation of biological process, signal transduction, and nervous system processes. The hub genes with high connectivity in selected tissue-specific modules, including AAEL010426, AAEL002896, AAEL002600, AAEL000961, AAEL007784 and AAEL006429, were identified by using CytoHubba. 【Conclusion】 In this study, based on the transcriptome data of different tissues of A. aegypti we discovered many important co-expressed gene modules by using the WGCNA method. The results provide new ideas and a method basis for the analysis of mosquito gene co-expression patterns and have reference value for exploring the unique gene resource information and bioinformatical studies of functional genes in different tissues of mosquitoes.
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