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  • Monthly, Founded in 1950
    Supervisor:Chinese Academy of Sciences
    Sponsor:Institute of Zoology,Chinese Academy of Sciences
    The Entomological Society of China
    Domestic postal code: 2-153
    Foreign issuance code: Q61
    ISSN 0454-6296
    CN 11-1832/Q
Table of Content
20 June 2016, Volume 59 Issue 6
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    Transcriptomic analysis of the desert beetle Microdera punctipennis (Coleoptera: Tenebrionidae) in response to short-term cold stress
    Kuerban TUSONG, LU Xue-Ying, LIU Xiao-Ning, MA Ji
    2016, 59(6):  581-591.  doi:10.16380/j.kcxb.2016.06.001
    Abstract ( 1862 )   PDF (2054KB) ( 960 )     
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    【Aim】 Tenebrionid insect Microdera punctipennis is an endemic species in Gurbantünggüt Desert, Xinjiang, China. It has very strong cold hardiness, but the molecular mechanisms by which M. punctipennis responses to low temperature are still unclear. This study aims to enrich the genetic information of M. punctipennis , and to analyze the functional groups of the most up-regulated genes under short-term cold stress and the pathways which they may involve in. 【Methods】 Transcriptomic sequencing reads from M. punctipennis adults exposed to 4℃ as low temperature treatment and 25℃ as the control, respectively, for 3 h were de novo assembled by using Trinity software. Gene annotation was conducted by using Blast2go software. The differentially expressed genes were screened by using DESeq and GFOLD softwares. For the up-regulated genes, GO term enrichment and KEGG metabolic pathway analysis were performed using Blast2go software. 【Results】 In total, 51 712 non-redundant transcripts with the average length of 984 bp were assembled, of which the expression levels of 514 genes were up-regulated and 91 genes down-regulated in response to cold stress. The up-regulated ones were enriched to 35 GO terms and 18 KEGG pathways, showing that stress-related GO terms, biological regulation and immune system are enriched significantly. Pathways such as purine metabolism, thiamine metabolism, and glycolysis/gluconeogenesis are significantly enriched. In the stress responsive GO term, the expression levels of eight abiotic related genes, including heat shock protein (Hsp90) gene, superoxide dismutase (SOD) gene, calnexin gene, etc., were up-regulated significantly. 【Conclusion】 The transcriptomic data of the desert beetle M. punctipennis reveal that the expressions of genes related to the biological processes like metabolic process, stress response, biological regulation and immune system are significantly up-regulated during cold acclimation at 4℃. The metabolic pathways revealed by KEGG enrichment show that low temperature transcriptome of M. punctipennis is much different from those of other species. Our results provide basic data for successive bioinformatics analysis and mining the key genes in cold tolerance for M. punctipennis as well as for elucidation of the related molecular mechanisms.
    Cloning and characterization of an odorant receptor gene AlucOR40 in Apolygus lucorum (Hemiptera: Miridae)
    ZHANG Zhi-Xiang, ZHANG Mei-Ping, WANG Gui-Rong, LIU Yang
    2016, 59(6):  592-601.  doi:10.16380/j.kcxb.2016.06.002
    Abstract ( 1836 )   PDF (5742KB) ( 694 )     
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    【Aim】 This study aims to clone an odorant receptor gene, AlucOR40, from Apolygus lucorum, to characterize its expression profiles in different tissues and to explore its function, so as to provide a theoretical basis for understanding the mechanisms of odor reception in A. lucorum. 【Methods】 The antennal transcriptome sequencing and RT-PCR amplification was used to clone the full-length cDNA of AlucOR40. The relative expression levels of this gene in different tissues of male and female adults of A. lucorum were detected using semi-quantitative RT-PCR. Through the combination of heterologous expression in Xenopus oocytes and two-electrode voltage clamp electrophysiological recording technique, the response of this odorant receptor to 60 odorants including sex pheromone of A. lucorum and plant volatiles was determined. The EAG responses of the 3-d-old adults of A. lucorum to the odorant ligand of AlucOR40 were further measured. 【Results】 AlucOR40 (GenBank accession number: KU886190) was cloned. AlucOR40 encodes 398 amino acids and the deduced protein has 7 transmembrane domains with an intracellular N-terminus and an extracellular C-terminus, which conforms to the typical structure of insect odorant receptors. The results of semi-quantitative RT-PCR showed that AlucOR40 was specifically expressed in the antenna of adults and the expression level was obviously higher in males than females. Two-electrode voltage clamp electrophysiological recording showed that AlucOR40 specifically responded to (E)-2-hexenol among the tested odorants. The EAG experiments showed that the 3-d-old adults had EAG response to (E)-2-hexenol, but the responses of males were significantly higher than those of females. 【Conclusion】 Our results suggest that the AlucOR40 is involved in identification of (E)-2-hexenol and may play an important role in identification of female by male during mating process.
    Molecular cloning, characterization and expression analysis of trehalase genes in the rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)
    TIAN Yu, DU Juan, LI Shang-Wei, LI Jiao, WANG Shuang
    2016, 59(6):  602-612.  doi:10.16380/j.kcxb.2016.06.003
    Abstract ( 1601 )   PDF (5769KB) ( 762 )     
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    【Aim】 Trehalase (Tre) is a key enzyme in trehalose metabolism of insects and plays important roles in the development and regulation of energy. Insects possess two types of trehalases, i.e., soluble trehalase (Tre1) and membranebound trehalase (Tre2). This study aims to clone trehalase gene (CmTre) from the rice leaf folder, Cnaphalocrocis medinalis, to clarify its expression patterns in different tissues and developmental stages, and to analyze the molecular characteristics of the two types of the gene and their products. 【Methods】 Based on the transcriptome data of C. medinalis, the full-length cDNA of CmTre was cloned using the rapid amplification of cDNA ends (RACE)-PCR and analyzed by bioinformatics. CmTre mRNA expression levels in different tissues of adults and developmental stages of C. medinalis were detected by using real-time quantitative PCR (RT-qPCR). 【Results】 We cloned two types of CmTre, i.e., soluble trehalase gene CmTre1 and membrane-bound trehalase gene CmTre2. The full-length cDNA of CmTre1 is 2 364 bp containing a 1 704 bp open reading frame (ORF) that encodes 567 amino acids, while that of CmTre2 is 2 079 bp containing a 1 923 bp ORF that encodes 640 amino acids. Bioinformatics analysis indicated that CmTre includes a signal peptide and CmTre1 has no transmembrane domain, whereas CmTre2 contains a transmembrane domain. Homology and phylogenetic analyses showed that the amino acid sequences of CmTre1 and CmTre2 have the highest identities with those of Tre1 and Tre2 from Omphisa fuscidentalis, which are 74% and 79%, respectively. Homology modeling analysis demonstrated that the tertiary structure of CmTre1 is composed of 19 α-helices and 2 β-sheets while that of CmTre2 is composed of 23 α-helices and without β-sheet. RT-qPCR revealed that CmTre was expressed throughout all developmental stages of C. medinalis, with the highest expression level in adult and a relatively stable expression level in larval stage. CmTre1 was expressed at the lowest level in pupa, while CmTre2 was expressed at the lowest level in the 5th instar larva. CmTre was expressed in all the adult tissues tested (midgut, integument, malpighian tubules, head, ovary, fat body, muscle and testis). CmTre1 was expressed at higher levels in the midgut and integument while CmTre2 was expressed at higher levels in the muscle and midgut. 【Conclusion】 In this study, genes of a soluble and a membrane-bound form of trehalase in C. medinalis were cloned, and their characteristics and expression patterns were analyzed. The findings lay the foundation for further research on the functions of trehalase genes and then using the genes as the targets of pest control.
    Expression and binding characterization of chemosensory protein CSP16 in the silkworm, Bombyx mori
    GUO Xing-Guo, CHEN Ying, XING Qiu-Ting, XING Wan-Jing, HAN Yun-Chao, QIAO Hui-Li
    2016, 59(6):  613-621.  doi:10.16380/j.kcxb.2016.06.004
    Abstract ( 1815 )   PDF (1661KB) ( 513 )     
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    【Aim】 Chemosensory proteins (CSPs) play important roles in reception and recognition of chemical signal, and regulation of insect development. This study aims to explore the function of CSP16 in the silkworm, Bombyx mori. 【Methods】 Real-time quantitative PCR (RT-qPCR) was used to investigate the expression patterns of csp16 gene in different developmental stages and different tissues of the 5th instar larvae of B. mori. Prokaryotic expression system was used to express and purify CSP16. The binding properties of CSP16 were analyzed by fluorescence binding assay. 【Results】 The RT-qPCR results showed that csp16 was regularly expressed in the 1st-5th instar larvae, and highly expressed in molting larvae of each instar. In day-3 5th instar larvae, csp16 was mainly expressed in head, epidermis, testis and ovary. The expression level of csp16 was up-regulated in larvae of different instars and different tissues of the 5th instar larvae after treatment with 20-hydroxyecdysone. Purified CSP16 showed weak affinity to different compounds including alcohols, esters, aldehydes, phenols and benzene. 【Conclusion】 Csp16 is highly expressed in molting larvae of each instar of B. mori, and the expression level is up-regulated by 20-hydroxyecdysone in larvae during feeding, suggesting that CSP16 may be involved in molting process of silkworm larvae.
    Identification and characterization of the CPF family of cuticular protein genes in the genome of Anopheles sinensis (Diptera: Culicidae)
    LIU Bai-Qi, QIAO Liang, XU Bai-Ying, ZHENG Xue-Ling, CHEN Bin
    2016, 59(6):  622-631.  doi:10.16380/j.kcxb.2016.06.005
    Abstract ( 1919 )   PDF (3953KB) ( 812 )     
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    【Aim】 This study aims to identify the CPF family (CPFs) of cuticular protein genes in Anopheles sinensis genome, to analyze their structure and characteristics, to deduce their possible biological functions, and to investigate and compare the CPFs of representative mosquito species so as to provide information frame for the family of genes. 【Methods】 We identified the CPFs in the genomes of An. sinensis, An. gambiae, An. minimus, Aedes aegypti, Culex quinquefasciatus and Drosophila melanogaster using BLASTP, TBLASTN and HMM with An. gambiae CPFs as query, predicted the structure and splicing variation of An. sinensis CPF gene and the signal peptide, transmembrane region, structural domain and 3D structure of An. sinensis CPF proteins using bioinformatics techniques, and constructed phylogenetic relationships using maximum likelihood (ML) method and deduced the origin and evolution of CPFs in these species. 【Results】 There are 4, 4, 4, 3, 3 and 3 CPFs in An. sinensis, An. gambiae, An. minimus, Ae. aegypti, Cx. quinquefasciatus and Dr. melanogaster genomes, respectively. The CPFs in An. sinensis were named as AsCPF1, AsCPF2, AsCPF3 and AsCPF4, respectively. Their full-length cDNA sequences are 736, 2 021, 531, and 1 001 bp, respectively, encoding 219, 345, 148 and 185 amino acids, respectively. AsCPF1, AsCPF2 and AsCPF3 only have one intron, but AsCPF4 contains three introns, which all have phase “0”. There are 3, 2, 1 and 2 selective spicing variants for AsCPF1, AsCPF2, AsCPF3 and AsCPF4, respectively. AsCPF3 has the highest expression quantity, followed by AsCPF4, AsCPF2 and AsCPF1. The theoretical molecular weights of AsCPF1, AsCPF2, AsCPF3 and AsCPF4 are 22.86, 36.47, 15.08 and 18.66 kD, and their isoelectric points are 9.08, 8.97, 9.44 and 9.16, respectively. These AsCPFs contain a 44-amino-acid conserved region and C-terminal region, and all are secretory proteins with signal peptide sequences except for AsCPF2 that is non-secretory protein and lacks a signal peptide sequence. All the four AsCPFs have alpha helix, random coil and extended strand, and only AsCPF4 has a transmembrane region that is located between amino acid 5 to 27. Phylogenetic analysis showed that CPF3 might be the earliest derived CPF gene, CPF1 and CPF2 might originate from a common ancestor and consequently experienced a gene duplication event, and CPF4 might be unique for Anopheles mosquitoes and the latest derived CPF gene. The Ka/Ks ratio of CPFs are all less than 1 in An. sinensis in reference to An. gambiae, suggesting the purification selection of these genes in evolution. 【Conclusion】 The whole-genome identification and characteristics analysis of CPFs in An. sinensis and the comparison of CPFs in representative mosquito species revealed the diversity, structure and amino acid characteristics and the origin and evolution of the CPF family of genes in mosquitoes, which provides a comprehensive information frame for further research and utilization of the CPF gene family.
    Seasonal characteristics of gut bacterial communities associated with carpenter ant Camponotus japonicus (Hymenoptera: Formicidae)(In English
    XU Yang, NAN Xiao-Ning , WEI Cong, HE Hong
    2016, 59(6):  632-640.  doi:10.16380/j.kcxb.2016.06.006
    Abstract ( 2137 )   PDF (1384KB) ( 706 )     
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    【Aim】 Ants have evolved a complicated association with microbes and, in particular, gut bacteria may have a strong impact on dietary evolution and species diversification of ants. Ants of the genus Camponotus are well known for harboring the endosymbiont Blochmannia and some other bacteria, which play an important role in the supplement of host nutrition. However, gut bacterial communities are sensitive to change of the food type, and such information might be useful to investigate the feeding activity of host ants in different seasons. In this study we aim to reveal whether the gut bacterial communities present seasonal characteristics in Camponotus ants. 【Methods】 The gut bacterial communities of foraging workers of C. japonicus collected from two colonies (colony 1 and 2) on four sampling dates (June 12, August 15, and October 10 in 2012, April 15 in 2013) were investigated using the 16S rRNA polymerase chain restriction fragment-length polymorphism (PCR-RFLP) method.【Results】 Seventeen bacterial genera and one unknown bacterium were found, dominated by Blochmannia in all the eight samples (67.1%-98.8%). Bacteria of Pseudomonas and Enterobacter were present in most samples, but other bacteria were distributed only sporadically in one to three samples with very low clone numbers. The gut bacterial communities in the two colonies did not show consistent change trend in the four months, and that they all harbored a low bacterial diversity. In colony 1, the bacterial diversity was relatively higher in April and October, but much lower in June and August, while in colony 2, the bacterial diversity was much higher in August than in April, June and October. The two colonies had similar bacterial communities in June and October, but were distinctly different in August and April.【Conclusion】 The gut bacterial composition and diversity in two colonies of C. japonicus can shift with seasons, but such changes have no consistent trend and no obviously season-specific characteristics have been acquired by this ant.
    Fumigation activities of three essential oils against adult Musca domestica (Diptera: Muscidae)
    ZHANG Jun-Tian, XIONG Guo-Hong, HE Jian-Guo, XU Ying, WEI Li-Xiu, PENG Ying-Hui
    2016, 59(6):  641-646.  doi:10.16380/j.kcxb.2016.06.007
    Abstract ( 1690 )   PDF (944KB) ( 550 )     
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    【Aim】 This research aims to evaluate the fumigation activities of the essential oils (EOs) from Cinnamomum caphora fruits, Cinnamomum pedunculatum leaves and Sabina chinensis var. chinensis leaves, and their mixtures against the adults of the housefly, Musca domestica, so as to provide theoretical basis for developing botanical insecticides. 【Methods】 The fumigation and knock-down toxicities of these EOs extracted by steam distillation and their mixtures against the 5-day-old adults of M. domestica were detected with the erlenmeyer flask method. 【Results】 The median lethal concentration (LC50) values of the three EOs against M. domestica adults were 5.28, 16.86, and 14.54 μg/cm3, respectively, and the median knock-down time (KT50) values in the treatment of LC90 were 12.03, 16.56, and 13.37 min, respectively. The fumigation toxicity of EO from C. pedunculatum leaves against M. domestica adults showed strong synergistic effect when it was mixed with EO from S. chinensis var. chinensis leaves at the LC50 value ratio of 9∶1, with the co-toxicity coefficient (CTC) as high as 367.95. 【Conclusion】 The EO from C. caphora fruits has strong fumigation and knock-down toxicities against M. domestica adults, followed by EO from S. chinensis var. chinensis leaves and then EO from C. pedunculatum leaves. Mixtures of EO from C. pedunculatum leaves and S. chinensis var. chinensis leaves have synergistic effect on their fumigation toxicities against M. domestica adults. The three EOs have the potential to be developed as environmental insecticides.
    Effects of temperature on the development and reproduction of Orius minutus (Hemiptera: Anthocoridae)
    DING Yao, YANG Qun-Fang, LI Qing, JIANG Chun-Xian, WANG Hai-Jian
    2016, 59(6):  647-653.  doi:10.16380/j.kcxb.2016.06.008
    Abstract ( 1820 )   PDF (1008KB) ( 631 )     
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    【Aim】 To illustrate the influences of temperature on the development and reproduction of experimental populations of Orius minutus. 【Methods】 At a temperature gradient (15, 20, 25, 30 and 35℃), the developmental duration, survival rate, fecundity and population parameters of O. minutus on prey Tyrophagus putresceniae were investigated in the laboratory. 【Results】 Within the constant temperatures ranging from 15 to 35℃, the average duration of various developmental stages of O. minutus shortened as the temperature rose gradually. O. minutus needed 52.45 d to complete a life cycle at 15℃, while needed 14.85 d at 35℃. The linear regression analysis showed that the developmental threshold temperature for O. minutus was 8.89℃, and its effective cumulative temperature was 359.20 day-degree. The generation survival rate and the number of eggs laid per female peaked at 25℃, being 17.07% and 41.00, respectively. The population trend index was less than 1 at 15 and 35℃, demonstrating that the mean population growth is negative. On the contrary, the population trend index was greater than 1 at 20-30℃ and reached the maximum (3.92) at 25℃. The net reproductive rate, the intrinsic rate of increase and the finite rate of increase peaked at 25℃, being 3.32, 0.04 and 1.04, respectively. The generation time of the population was the longest (57.76 d) at 15℃, but the shortest (17.50 d) at 35℃. 【Conclusion】 The most suitable temperature range for the development of O. minutus on T. putresceniae is 25-35℃, while the most suitable temperature for its survival, reproduction and the population growth is 25℃. These results provide the basic reference data for artificial rearing of O. minutus using T. putresceniae.
    Age-stage two-sex life tables of Spodoptera litura (Lepidoptera: Noctuidae) at different temperatures
    HAO Qiang, HUANG Qian, LIANG Wei-Bo, GONG Chang-Wei, WANG Xue-Gui
    2016, 59(6):  654-662.  doi:10.16380/j.kcxb.2016.06.009
    Abstract ( 2652 )   PDF (2738KB) ( 1528 )     
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    【Aim】 Spodoptera litura (Fabricius) is a lepidopteran cutworm defoliator and displays different growth and development characteristics under different conditions of hosts and foods. The objective of this study is to determine the effects of temperature on the population growth and reproduction of S. litura. 【Methods】 S. litura was bred and observed under the conditions of three different temperatures (20±1℃, 27±1℃ and 30±1℃), 65%±10% RH and a 14L:10D photoperiod in artificial climate chamber in the laboratory, and age-stage two-sex life tables of S. litura at the three temperatures were constructed. 【Results】 The developmental duration, adult preoviposition period (APOP) and total preoviposition period (TPOP) of S. litura were shortened as the temperature rose in a time-dependent manner. In addition, the average number of eggs laid per female was the highest at 20±1℃ (1 810.36 eggs/female), and lower at 27±1℃ (1 623.51 eggs/female) and the lowest at 30±1℃ (1 084.60 eggs/female). The intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R0) of S. litura were significantly different at different temperatures. The r value and λ value at 20±1℃ was significantly lower than those at 27±1℃and 30±1℃, but the R0 value at 27±1℃ was prominently higher than those at 20±1℃ and 30±1℃. The mean generation time (T) at 20±1℃,27±1℃ and 30±1℃ was 52.51, 30.63 and 26.87 d, respectively, which decreased sequentially and were significantly different between each other (P<0.05). 【Conclusion】 S. litura displays strong reproductive capacity at temperatures of 27±1℃ and 30℃±1℃. The result could provide the theoretical assistance to control S. litura with integrated pest management (IPM) strategy in fields.
    A simplified recording method for insect life table studies: a case study based on Bemisia tabaci (Hemiptera: Aleyrodidae) data
    ZHENG Xiao-Min, QI Xin, CHU Dong
    2016, 59(6):  663-668.  doi:10.16380/j.kcxb.2016.06.010
    Abstract ( 2417 )   PDF (1327KB) ( 987 )     
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    【Aim】 In order to save time and labor in life table studies and still to maintain the accuracy of the analyses involved, we tested the effects of changing the frequency of data sampling for two strains (infected and uninfected by Cardinium) of Bemisia tabaci MED cryptic species (commonly known as biotype Q) on their population parameters. 【Methods】 Based on the raw data recorded daily, the simplified data of the sampling intervals of 2, 4 and 5 d were re-analyzed to test the significant differences between the treatments. 【Results】 No significant differences between the treatments existed in the population parameters including the intrinsic rate of increase (r), finite rate of increase (λ), net reproductive rate (R0), mean generation time (T), preadult duration, fecundity, female longevity, male longevity, and total preoviposition period (TPOP) obtained through daily sampling and the interval-grouped data, except the adult preoviposition period (APOP) and oviposition days. 【Conclusion】 Our results demonstrated that the simplified recording method has no significant effects on the evaluation of the key population parameters and can be used in life table studies with less time and labor. In addition, the simplified recording method may decrease the interference to observed adults due to reduced daily observation. Consequently, the simplified method may be helpful to promote the application of the age-stage, two-sex life table.
    Biological characteristics of Epicephala ancylopa (Lepidoptera: Gracillariidae) on host Glochidion sp. (Phyllanthaceae) and the compositional analysis of its floral scent
    ZHANG Zhen-Guo, TENG Kai-Jian, LI Hou-Hun
    2016, 59(6):  669-681.  doi:10.16380/j.kcxb.2016.06.011
    Abstract ( 2019 )   PDF (18428KB) ( 523 )     
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    【Aim】 To understand the mutualistic stability in the obligate pollination mutualism between Epicephala ancylopa and host Glochidion sp. in biology and chemical ecology. 【Methods】 Based on the field observation and indoor experiment, we investigated the biological characteristics of Glochidion sp. and E. ancylopa, and the benefits that each species obtains from their obligate nursery pollination mutualism. The dynamic headspace technique and gas chromatography-mass spectrometry (GC-MS) were used to analyze the chemical composition of floral volatiles of both female and male flowers from Glochidion sp. The principal component analysis (PCA) was conducted to assess the compositional differences in  floral scent between the sexes of Glochidion sp. 【Results】 E. ancylopa had only one generation per year in Pu’er Sun River National Park, Yunnan Province, and its adults and larvae were active during March-April and August-October, respectively. For Glochidion sp., the rate of ripening was 44.20%, the rate of consumed fruits was 69.94%, the average number of seeds consumed by each larva was 2.55, and the proportion of intact seeds was 83.06% that could keep the stabilization of mutualism in each population. A total of 24 compounds were detected and identified from the floral scent of Glochidion sp., which were dominated by terpenoids and aliphatics compounds. Among them, (Z)-ocimene and β-elemene were found to have the highest amounts in flowers (47.11% and 22.72%, respectively), and inferred as the major scent components attracting E. ancylopa. The PCA revealed that the chemical compositions of male and female floral scents of Glochidion sp. were distinctly different, which was identified as sexual dimorphism in floral scent. 【Conclusion】 One survival strategy of E. ancylopa is to synchronize eclosion with the flowering season of Glochidion sp. by oversummering in the egg stage and overwintering in the pupal stage. Adult of E. ancylopa is active only at night, whereas the floral scent attracting pollinator is only released obviously at night. Sexual dimorphism in floral scent can help Epicephala moths distinguish floral scents between male and female flowers, mediating behaviors of collecting pollen and pollinating. This study provides a new basis for understanding the mechanism to sustain the stabilization in the Glochidion-Epicephala mutualism and the basic data for further electroantennographic detection and bioassay to identify the compounds with the actual physiological activity to Epicephala moths.
    dsRNA delivery methods for RNA interference in insects
    LIU Ji-Sheng, ZHU Wen-Hui, LIAO Wen-Li, BU Xiao-Ling, JIANG Wan-Yi
    2016, 59(6):  682-691.  doi:10.16380/j.kcxb.2016.06.012
    Abstract ( 2257 )   PDF (935KB) ( 1176 )     
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     RNA interference (RNAi) in insects mostly refers to the process that double-stranded RNA (dsRNA) silences the specific target messenger RNA (mRNA) via inhibition of translation or transcription. Recently, RNAi technology has been mainly applied in the research of functional genomics and pest control due to its high specificity. This review focuses on three common ways (microinjection, feeding, and soaking and transfection), which deliver dsRNA in entomological RNAi research. Among these three ways, microinjection can deliver exact amount of dsRNA to target tissues, which is more suitable for gene functional study in the laboratory. Feeding is easy and fast to perform, which is suitable for high-throughput gene screening. Soaking and transfection is also suitable for large-scale gene screening, but more commonly used in cell research.
    Rapid detection of Crithidia bombi (Kinetoplastida: Trypanosomatidae) by using PCR technique
    ZHANG Ti-Yin, LIU Bei, ZHENG Teng, BAI Quan-Yang, TIAN Guo-Ning
    2016, 59(6):  692-698.  doi:10.16380/j.kcxb.2016.06.013
    Abstract ( 1342 )   PDF (5127KB) ( 654 )     
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     【Aim】 As a highly effective pollinator for crops, bumblebee has been widely used in greenhouse and acquired good economical and ecological benefits in foreign countries in recent years. Now many domestic enterprises have imported bumblebees and used them in facility agriculture. Crithidia bombi is one of the most important parasites of bumblebees. Once introduced with importation, it can pose significant risks to domestic bumblebees. So a rapid detection technique for C. bombi should be established urgently. 【Methods】 A PCR detection method including a pair of primers, Cri-F and Cri-R, based on the ITS sequence was established for detection of C. bombi in bumblebee bodies. After preliminarily testing the PCR result, we further optimized the reaction conditions of PCR, including the annealing temperature, primer concentration, cycle number and so on. Meanwhile, the sensitivity, specificity and stability of the PCR method were evaluated as well. 【Results】 The results showed that the PCR method with specific primers from conserved regions of ITS was feasible for detection of C. bombi. The optimized reaction conditions of PCR are the annealing temperature of 59℃, the primer concentration of 0.5 μmol/L, and the amplification cycle number of 35. The sensitivity of the detection method to the total DNA of bumblebee was approximately 13.24×10-5 ng/μL, with a good specificity and stability. When the PCR method was applied in detection of C. bombi, the detection could be finished within 4 h, showing a good applicability. 【Conclusion】 The rapid detection method developed in this study can be applied in detection of C. bombi, which is important for epidemic surveillance of this parasite and the inspection and quarantine of imported bumblebees.
    Contents of Vol. 59 Issue 6
    2016, 59(6):  699. 
    Abstract ( 1079 )   PDF (431KB) ( 336 )     
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