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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 December 2014, Volume 57 Issue 12
For Selected: View Abstracts Toggle Thumbnails
  • RESEARCH PAPERS
    Prokaryotic expression of TAT-Cloan-DH-EGFP fusion protein and its transduction in larvae of the gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae) by oral administration
    ZHOU Zhou, LI Yong-Li, YUAN Chun-Yan, QU Liang-Jian
    2014, 57(12):  1361-1367. 
    Abstract ( 1830 )   PDF (3528KB) ( 797 )     
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    【Aim】 In order to explore the biological effects of diapause hormone by insect feeding, a fusion protein is designed to accelerate its transmembrane transport and overcome the degradation in insect digestive tract. 【Methods】 The diapause hormone from Clostera anastomosis L. (Cloan-DH) was over-expressed as a fusion protein with a transactivator of transcription (TAT) protein of HIV-1 and an enhanced green fluorescent protein (EGFP). Escherichia coli BL21 cells expressing TAT-Cloan-DH-EGFP and EGFP were added to artificial diets respectively to rear the larvae of the gypsy moth, Lymantria dispar. Paraffin-embedded sections of the abdominal segments containing the midgut were collected from larvae fed with the artificial diets for 0, 8, 24, 48, 72 and 90 h, respectively. Then cellular transduction was evaluated by observing thin sections of gastric tissues to detect intracellular EGFP. 【Results】 After induction with 0.2 mmol/L IPTG at 37℃, the TAT-Cloan-DH-EGFP fusion protein was over-expressed and reached up to 18.1% of total cellular proteins mainly in inclusion bodies. Strong green-fluorescent signals of EGFP were detected in the inner layer of the midgut, muscle and cuticle sections of larvae fed with TAT-Cloan-DH-EGFP. It was found that the fluorescence intensity of EGFP increased with the quantity of the fusion protein uptake by the larvae. At 72 h after feeding the artificial diets containing TAT-Cloan-DH-EGFP, the fluorescence intensity of EGFP was up to the highest. 【Conclusion】 The results indicate that TAT-Cloan-DH-EGFP can transfer across intestine and enter other tissues by oral applications, and the quantity of transferred protein increases in proportion to the intake by the larvae.
    Expression levels of three miRNAs in the brain of different day-old workers of Apis mellifera ligustica (Hymenoptera: Apidae)
    SHI Teng-Fei, LIU Fang, YU Lin-Sheng, WANG Tian-Shu, QI Lei
    2014, 57(12):  1368-1374. 
    Abstract ( 1990 )   PDF (1075KB) ( 905 )     
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    【Aim】 The Italian honey bee, Apis mellifera ligustica, is an eusocial insect and the workers show age-dependent behavioral changes, and the mechanisms of the behavioral changes have been the hot research topic. In this study, we investigated the expression of 3 candidate miRNAs, i.e., ame-let-7, ame-miR-13b and ame-miR-279, in A. mellifera ligustica workers at different age, to confirm the differentially expressed miRNAs that may be involved in age-dependent behavioural changes in workers of the honeybee. 【Methods】 We tested the expression levels of the three miRNAs in the brain of A. mellifera ligustica workers at different age (4, 8, 12, 17, 22, 26 and 30 day-old) by RT-qPCR respectively, and then analyzed the differential expression among them. 【Results】 The expression level of ame-let-7 was reduced with the increasing age of the workers and tended to be stable after 17 day-old. The expression level of ame-miR13b increased with age of workers and kept stable in 26 and 30 day-old workers. The expression level of ame-miR-279 was near to the normal distribution, peaked in the 12 day-old workers and kept stable in over 17 day-old workers. However, the expression of all the 3 kinds of miRNAs showed a high degree of temporal specificity. 【Conclusion】 Our findings show that the levels of miRNAs expression are related with the age-dependent behavioral changes in honey bees, and the conclusions provide important information for exploring the action sites of miRNAs in the A. mellifera ligustica workers.
    Evaluation of endogenous reference genes in Bactrocera minax (Diptera: Tephritidae)
    WANG Jia, ZHAO Jing, LIU Ying-Hong
    2014, 57(12):  1375-1380. 
    Abstract ( 1915 )   PDF (1144KB) ( 935 )     
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    【Aim】 The Chinese citrus fly, Bactrocera minax, is a devastating pest of citrus. Our study aims to screen out the stably expressed endogenous reference genes in B. minax under specified conditions to assure the reliability of quantitative real-time PCR (qRT-PCR) analysis of target genes. 【Methods】 Ten candidate reference genes were selected for qRT-PCR, and five softwares were adopted to determine the stability of candidate reference genes in different developmental stages (low-instar larva, 3rd instar larva, 1 day-old pupa, 80 day-old pupa, 160 day-old pupa, male adult, and female adult) and body parts of male and female adults (head, thorax, abdomen, and whole body) by analyzing the Ct value. 【Results】 In different developmental stages and body parts of adults of B. minax, the Ct values of candidate reference genes were between 15 and 30. The disparity of the Ct value indicated that the expression levels of genes were different. Based on the comprehensive analysis of stability ranking of candidate reference genes with the five softwares and the optimal reference gene number calculated by geNorm, UBQ, GAPDH and GST were recommended as the reference genes for different developmental stages, and TUB, GAPDH and GST for different body parts of adults. 【Conclusion】 To obtain the precise expression patterns of target genes under specified conditions, the combination of reference genes is suggested to use. Our study is conducive to investigating the target gene expressions in B. minax under specified conditions.
    Effects of methoxyfenozide on the activities and gene expression levels of glutathione S-transferases (GST) in the methoxyfenozide-resistant and homologous control populations of Helicoverpa armigera (Lepidoptera: Noctuidae)
    XU Xi-Bao, ZHANG Jing, RUI Chang-Hui
    2014, 57(12):  1381-1388. 
    Abstract ( 2258 )   PDF (2127KB) ( 657 )     
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    【Aim】 In order to develop more effective methods for molecular detection of insecticide resistance, the molecular mechanisms of resistance to methoxyfenozide related to glutathione S-transferases (GSTs) in Helicoverpa armigera (Hübner) were studied. 【Methods】 The early 3rd instar larvae of the methoxyfenozide-resistant (R-methoxyfenozide) and the homologous control (S-methoxyfenozide) populations of H. armigera were treated with LC40 of methoxyfenozide, and the specific activities of GST and the expression of five GST genes were measured before and after treatment. The coding sequence of one delta GST gene (GSTd1) was compared in the two populations. 【Results】 The coding sequences of GSTd1 in the R-methoxyfenozide and S-methoxyfenozide populations of H. armigera were identical, suggesting that no structural change occurrs in the isozyme. The GST activity in R-methoxyfenozide population of H. armigera was higher than that in the S-methoxyfenozide population before treatment. After treatment, the GST activity decreased in both populations, but fast recovered in the R-methoxyfenozide population. Except GSTe2, GST genes had a higher expression in the R-methoxyfenozide population than in the S-methoxyfenozide population before treatment. Effects generated by methoxyfenozide treatment on the expression of GST genes were quite different in the two populations. After treatment, the expression of GST genes, except GSTs1, initially decreased in the S-methoxyfenozide population, and then the expression of GSTd2 and GSTe2 recovered gradually. The expression of GSTs1 showed no obvious change within 36 h and increased at 48 h after treatment. But in the R-methoxyfenozide population, the expression of the five GST genes after treatment decreased initially, and then fast recovered. Except GSTe2, the initial and final expression of GST genes in the R-methoxyfenozide population were remarkably higher than that in the S-methoxyfenozide population. 【Conclusion】 The results suggest that resistance to methoxyfenozide in H. armigera is related to enhanced GST activities which are rooted from the over-expression of GST genes.
    Effects of fraxinellone on the peritrophic memebrane of Mythimna separata (Lepidoptera: Noctuidae)
    Lü Min, CHEN Su-Ying, HAN Li-Bo, HU Jun-Yun, XU Hui, WU Wen-Jun
    2014, 57(12):  1389-1394. 
    Abstract ( 1977 )   PDF (1884KB) ( 659 )     
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    【Aim】 Isolated from the rootbark of Diatamnus dasycarpus Turcz., fraxinellone exhibits stomach toxicity against pests. This study aims to clarify the insecticidal mechanism of fraxinellone by investigating the effects of fraxinellone on the peritrophic membrane of the 6th instar larvae of Mythimna separata.【Methods】 Through biochemical methods and SEM observation, the content of carbohydrates, the contents and components of proteins and the surface structure of the peritrophic membrane of the 6th instar larvae of M. separata after in vivo and in vitro treatments with fraxinellone were studied. 【Results】 In vivo treatment of fraxinellone (20 mg/mL) resulted in a drop of the protein content in the 6th instar larvae of M. separata. However, after M. separata larvae were subjected to in vivo (20 mg/mL fraxinellone) and in vitro (8 mg/mL fraxinellone) treatments with fraxinellone, the contents of carbohydrates of both treatments increased, which were 1.75 and 2.17 folds as high as those of the control, respectively. SDS-PAGE result showed that partial proteins were hydrolyzed in the 6th instar larvae of M. separata subjected to in vivo and in vitro treatment with fraxinellone. SEM observation showed that the surface structure of peritrophic membrane changed after the 6th instar larvae of M. separata were treated with fraxinellone in vivo and in vitro. 【Conclusion】 These results showed that fraxinellone affected the contents of carbohydrates and the contents and components of proteins, and altered the surface structure of the peritrophic membrane of M. separata larvae. This study provides a basis for further research on the insecticidal mechanisms of fraxinellone against pests.
    Effects of UV-B radiation intensity and duration on the growth, development and fecundity of Sitobion avenae (Hemiptera: Aphididae)
    DU Yi-Min, WANG Ping, YANG Jie, ZHOU Dong, LI Guang-Yu, ZHAO Hui-Yan, HU Zu-Qing, HU Xiang-Shun
    2014, 57(12):  1395-1401. 
    Abstract ( 2223 )   PDF (1628KB) ( 988 )     
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    【Aim】 This study aims to explore the effects of different radiation intensity and radiation duration of UV-B on the growth, development and fecundity of Sitobion avenae (Fabricius), and the interactions between radiation intensity and radiation duration. 【Methods】 Following different UV-B treatments (0.20 mW/cm2 and 0.75 mW/cm2 UV-B intensity, 3 h, 9 h and 15 h radiation duration, respectively) on S. avenae, the age-specific life table of S. avenae was established, the body weight was measured, and the mean relative growth rate, life table population parameters, fecundity parameters, age-specific survival rate and age-specific fecundity were calculated. 【Results】 The results revealed that in the same radiation duration, the intrinsic rate of increase (rm), the net rate of increase (R0) and fecundity (F) of S. avenae exposed to the high intensity UV-B (0.75 mW/cm2) were significantly lower than those exposed to the low intensity UVB (0.20 mW/cm2) and the control (UV-B absent), with the mortality increasing faster in a short period. Under the same intensity UV-B, the rm, R and F values of S. avenae also decreased significantly with the radiation durations, and the period of the fastest decline of the survival rate shifted to an earlier date. The UV-B intensity and the radiation duration had significant interactions. However, under low intensity UV-B for 3 h, the rm, R0 and F values of S. avenae were higher than those in the control. The MRGR of S. avenae exposed to high intensity UV-B for 15 h decreased significantly compared with the control, but were not significantly different among other treatments. The number of alataes increased when S. avenae was exposed to high intensity UV-B (0.75 mW/cm2) for long radiation duration (15 h) as compared with that in the control. 【Conclusion】 The growth and development of S. avenae are affected by the UV-B radiation and change with the intensity and duration of UV-B radiation, and these two factors (radiation intensity and duration) also have significant interactions.
    Control effects of aqueous extracts of tobacco on grape phylloxera (Daktulosphaira vitifoliae) infection and damage
    ZHANG Lei, SUN Yong-Jiang, SUN Lu-Long, DU Yuan-Peng, ZHAI Heng*
    2014, 57(12):  1402-1407. 
    Abstract ( 1523 )   PDF (1881KB) ( 784 )     
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    【Aim】 Grape phylloxera (Daktulosphaira vitifoliae) has been found in some vineyards in China and caused serious damage. Our previous research found that aqueous extracts of tobacco could control the damage of D. vitifoliae on excised roots. The aim of this research was to evaluate the control efficacy of aqueous extracts of tobacco towards grape phylloxera infection and grape growth. 【Methods】 Two year-old Kyoho grapes were chosen as the test materials in the greenhouse, and the root activity, overground growth amount, number of nodosities and photosynthesis rate of the grape were determined. 【Results】 The results showed that aqueous extracts of tobacco had certain alleviating effects on phylloxera infection. Both the aqueous extracts of tobacco at the concentration of 20 mg/mL and 50 mg/mL had an inhibition to phylloxera infection. At 21 d after the aqueous extracts of tobacco were applied, the number of nodosities of the grape in the treatments with 20 mg/mL and 50 mg/mL aqueous extracts of tobacco decreased by 42.9% and 52.8% as compared with that of the phylloxera infected grapes (the infection control). Compared to the infection control, plant growth amount in the treatments with 20 mg/mL and 50 mg/mL aqueous extracts of tobacco increased by 57.1% and 7.7%, respectively. Grape phylloxera infection decreased the microorganism amount in the soil; however, the aqueous extracts of tobacco decreased the amount of fungi, which could decrease the secondary infection possibility of microorganisms. 【Conclusion】 Aqueous extracts of tobacco have a control effect on grape phylloxera and can ease the damages caused by phylloxera infection in potted tests.
    Age-stage two-sex life tables of the experimental population of Problepsis superans (Lepidoptera: Geometridae) on three Ligustrum species
    HU Liang-Xiong, HE Zheng-Sheng, ZHANG Xiao-Gu
    2014, 57(12):  1408-1417. 
    Abstract ( 2069 )   PDF (2834KB) ( 845 )     
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    【Aim】 Problepsis superans is a pest, which feeds the leaves of the garden plant Ligustrum quihoui Carr. No research has been done on other host plants and their effects on life history of P. superans. The study can help understand the influence of L. quihoui and two potential host plants, Ligustrum × vicaryi Hort. and Ligustrum lucidum Ait., on the development and reproduction of P. superans. 【Methods】 Age-stage two-sex life tables of the experimental population of P. superans on three ligustrum species, i.e., L. × vicary, L. quihoui, and L. lucidum were constructed under the conditions of 25±1℃, 75%±5% R. H. and 16L∶8D photoperiod. 【Results】 The results showed that there were significant differences in larval longevity and fecundity of P. superans on the three ligustrum species, however, no significant differences were found in egg and pupal duration, adult preoviposition period (APOP), and male survival time. The total larval duration of P. superans on L. × vicaryi (21.33 d) was significantly shorter than that on L. quihoui (23.46 d) and L. lucidum (27.28 d); however, the fecundity on L. × vicaryi (529.1 eggs/female) was significantly higher than that on L. quihoui (442.5 eggs/female) and L. lucidum (339.7 eggs/female). The intrinsic rate of increase (r), the finite rate of increase (λ) and the net reproductive rate (R0) of P. superans on L. × vicaryi and L. quihoui were significantly higher than that on L. lucidum; however, there was no significant difference between on L. × vicaryi and on L. quihou. The mean generation time (T) of P. superans on the three species differed significantly and ranked as L. × vicaryi < L. quihoui < L. lucidum. However, there were no significant differences in the gross reproduction rates (GRR) of P. superans on the three species. 【Conclusion】 L. vicaryi and L. quihoui provide better quality food for P. superans than L. lucidum, and L. vicaryi and L. lucidum may serve as hosts or temporary hosts of P. superans.
    Relationship of overwintering larvae and adult occurrence of the oriental fruit moth, Grapholita molesta (Lepidoptera: Tortricidae) in orchards
    LI Li-Li, GUO Ting-Ting, ZHAO Nan, MEN Xing-Yuan, ZHUANG Qian-Ying, ZHOU Xian-Hong, XU Yong-Yu, YU Yi
    2014, 57(12):  1418-1423. 
    Abstract ( 1794 )   PDF (827KB) ( 934 )     
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    【Aim】 The overwintering site of the oriental fruit moth, Grapholita molesta (Busck) and the relationship between the number of overwintering larvae and adult occurrence were investigated in different orchards to provide the scientific basis for forecasting and controlling this insect pest. 【Methods】 Six typical orchards located in Jinan, Tai’an, Feicheng, Guangrao and Laiwu of Shandong Province were selected for systematic investigation and analysis. The method of scraping bark was used to investigate the survival rate of overwintering larvae of G. molesta and the overwintering locations on tree branches, and closed gauze was used to investigate the numbers of overwintering larvae in the soil. Meanwhile, the occurrence of overwintering adults was monitored by sex pheromone traps and the sugar and vinegar liquid, and the relationship between the numbers of overwintering larvae and adult occurrence was analyzed.【Results】 The survival rate of overwintering larvae of G. molesta was as high as 62.99%, and the numbers of overwintering larvae were significantly different among different orchards (P<0.001). More overwintering larvae were found in main trunk (28.48%) and bough (44.24%) located in lower (45.10%) and middle (46.28%) parts of fruit tree, while less overwintering larvae were found on the upper part of fruit tree (8.62%) and the lateral branches (27.28%). There were no significant differences in the number of overwintering larvae among four directions of tree branches (east: 27.57%; west: 26.13%; south: 23.76%; north: 22.54%). No significant correlation was found between the numbers of the overwintering larvae and overwintering generation adults caught in the same orchard. 【Conclusion】 G. molesta overwinters as mature larvae in main trunk and bough located in lower and middle parts of fruit tree or in the soil. In the same orchard, the population of overwintering larvae of G. molesta is not suitable to be the reference basis for the forecast and control of overwintering adults, because the surrounding environmental conditions also affect adult occurrence of this moth.
    Population genetic differentiation analysis of the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae) based on mtDNA COI gene
    WANG Meng, XU Lang, ZHANG Run-Jie, ZHANG Gui-Ming, YU Dao-Jian
    2014, 57(12):  1424-1438. 
    Abstract ( 2231 )   PDF (2359KB) ( 965 )     
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    【Aim】 This study aims to infer the invasion routes of the oriental fruit fly, Bactrocera dorsalis Hendel in China and the invasion source of its recent introduction region. 【Methods】 mtDNA COI (1 496 bp, 97.3% of full length) of 192 individuals of 31 populations from China, Thailand, Japan, Laos, Bangladesh and USA was sequenced. The genetic diversity of each population and genetic differentiation between populations were analyzed by DnaSP 5.0, MEGA 6.0 and Arlequin 3.5, as well as haloptype analysis. 【Results】 High nucleotide diversity (0.00663) and haplotype diversity (0.98069) were detected overall from the 31 populations of B. dorsalis. Genetic differentiation between populations analyzed using F-statistics method revealed that lower genetic differentiation were detected between populations of China, different levels of genetic differentiation were detected between populations from China and populations from Thailand, Japan, Laos, Bangladesh and USA, and the highest genetic differentiation was detected between populations from China and populations from USA and Japan. Mantel test showed that genetic differentiation among 31 populations from China, Thailand, Japan, Laos, Bangladesh and USA bore a relation to spatial distance (R=0.670, P<0.0001), but spatial distance was not responsible for genetic differentiation among populations from China (R=0.038, P=0.534). There was no shared haloptype among populations from China, Thailand, Japan, Laos, Bangladesh and USA. The phylogenetic tree showed that original populations from China clustered into two groups, i.e., the southwest group and the southeast group. Neutrality test and mismatch distribution analysis revealed that B. dorsalis populations had experienced a large-scale expansion. 【Conclusion】 It is inferred that there are two main invasion routes to inland China, one is from Southeast China and the other from Southwest China, and Guangdong, Fujian, Guangxi and Guizhou are the influential invasion sources. Based on the results of genetic diversity, genetic differentiation and haplotype network, it is inferred that the main sources of B. dorsalis introduced to Anhui (Hefei) are from Fujian (Changle), Guangdong (Zhuhai) and Shanghai.
    REVIEW ARTICLES
    Characteristics and functions of DNA methylation in insects
    LIANG Shi-Ke, ZHANG Mei, LIANG Zi-Qiang, LI Guang-Hong, WANG Fang-Hai
    2014, 57(12):  1439-1446. 
    Abstract ( 2501 )   PDF (711KB) ( 899 )     
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    DNA methylation, one important form of epigenetic modifications, plays an important role in gene regulation in animals. With more attentions paid to DNA methylation in insects in recent years, it was found that besides certain similarities between insects and mammals, there are also some unique features and functions in DNA methylation of insects. Here, we reviewed the advances in researches on DNA methylation in insects and focused on its main characteristics and functions so as to provide some references for further exploring DNA methylation in insects. The type and nature of DNA methyl-transferase vary widely across insect species, and the DNA methylation mainly occurs in gene areas in insects and the level is low. The functions of DNA methylation in insects mainly involve the regulation of embryonic development, participation in genomic imprinting, regulation of caste and wing differentiation, sex determination, and involvement in pesticide resistance.
    CONTENTS
    Contents of Vol. 57 Issue 12
    2014, 57(12):  1447. 
    Abstract ( 1092 )   PDF (707KB) ( 738 )     
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    General content of Volume 57(1-12)
    2014, 57(12):  1450. 
    Abstract ( 1044 )   PDF (780KB) ( 720 )     
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