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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 March 2012, Volume 55 Issue 3
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    Cloning, expression and identification of GRD and LCCH3 subunits of GABA receptor in Drosophila melanogaster
    ZHANG Xiao-Min, MA Cai-Xia, REN Tian-Rui, WANG Feng
    2012, 55(3):  259-266. 
    Abstract ( 3816 )   PDF (5671KB) ( 1332 )     
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    γ-Aminobutyric acid receptor (GABAR) is one of the most important insecticide targets. In this study, the fragments of genes of GRD and LCCH3 subunits in Drosophila melanogaster were amplified by RT-PCR and cloned into pET-32a expression vector. Homologous analysis showed that they have more than 99% identity with the published amino acid sequences of the homologous genes in GenBank, without frameshift mutation. The recombinant plasmids were transformed into Escherichia coli and expressed under IPTG induction. LCCH3 gene was expressed successfully, but GRD gene failed. The expressed LCCH3 subunit was purified by the processes of inclusion body washing, denaturation, Ni2+ affinity chromatography and renaturation. The secondary structure of LCCH3 subunit was determined by circular dichroism (CD) spectroscopy, the result showed that this subunit was rich in β-strand. The results provide important information for the study of relationship between the structure and function of GABA receptor.
    Identification of the gene MdSOD3 and its role in resisting heavy metal stress in housefly (Musca domestica)
    TANG Ting, GAO Yi-Fu, LIU Yan-Juan, GE Xu-Dong, LI Xiang, LIU Feng-Song
    2012, 55(3):  267-275. 
    Abstract ( 3529 )   PDF (9170KB) ( 1584 )     
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    As a member of the antioxidant enzymes, superoxide dismutase (SOD) plays a key role in insect antioxidant protection system. In this study, the full-length cDNA of a SOD gene MdSOD3 (GenBank accession no. JQ408979) from the housefly, Musca domestica, was cloned and characterized by RT-PCR, which is 817 bp in length, the open reading frame is 534 bp encoding 177 amino acid residues. The homologous alignment and phylogenetic tree indicate that the MdSOD3 from housefly belongs to Cu/ZnSOD family. The temporal expression profiling revealed that MdSOD3 was present in the fat body, gut, hemocytes and cuticle. The expression of MdSOD3 was enhanced by Cd stress. The highest expression level was found in treatment at 5 mmol/L Cd2+ within a 24 h period. Quantitative PCR (qPCR) showed that mRNA level of MdSOD3 gene was reduced compared to the control by RNAi. High mortality was observed in the larvae treated with dsRNA of MdSOD3 for 60 h at 5 mmol/L Cd stress for 24 h. The RNAi of MdSOD3 gene resulted in the larval abdominal apex to become black. The results suggest that MdSOD3 may play an important role in adapting to heavy metal stress in housefly.
    Molecular cloning, prokaryotic expression and expression at different developmental stages of cathepsin B gene in mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)
    LUO Mei, DONG Zhang-Yong, BIN Shu-Ying, LIAO Hong-Zhi, LIN Jin-Tian
    2012, 55(3):  276-283. 
    Abstract ( 3305 )   PDF (5119KB) ( 1371 )     
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    Cathepsin B plays an important role in insect metabolism. In this study, cathepsin B gene was cloned from mealybug Phenacoccus solenopsis and named as PsCb (GenBank accession no. JQ727999). This cDNA contains an open reading frame (ORF) which is 927 bp in length encoding 308 amino acids. Homology analysis shows that the N-terminal sequence of PsCB has larger variation, while the C-terminal sequence is conserved. Phylogenetic tree shows that PsCB is separated with other branches. Prokaryotic protein expression test showed the expressed product has the MW of 35 kDa, nearly equal to the predicted. Real-time PCR analysis revealed that PsCb mRNA was expressed in various developmental stages of P. solenopsis. The expression level was relatively lower at the egg stage, reached a peak at the 2nd instar nymph and then declined. Our study provides a theoretical basis for further study of the gene function, the development of cathepsin inhibitors as ovicides and inhibitors of embryonic development of P. solenopsis.
    Cloning and expression profiling of the DNA methyltransferase Dnmt3 gene in the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae)
    LIU Ting-Ting, LIU Jun-Feng, WANG Wen-Xiang, WANG Huan, WANG Zi-Long, ZENG Zhi-Jiang, YAN Wei-Yu
    2012, 55(3):  284-290. 
    Abstract ( 4645 )   PDF (5805KB) ( 2380 )     
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    To explore the pattern of methylation in the Chinese honeybee, Apis cerana cerana, the DNA methyltransferase 3 (Dnmt3) gene in A. cerana cerana was cloned by using RT-PCR, and the quantitative analysis of the expression level of Dnmt3 mRNA in different developmental stages of worker (4 d-old pupa, 1, 7 and 30 d-old workers, and laying worker) and queen (4 d-old pupa, 1 d-old queen and laying queen) were conducted using real-time qPCR. The full-length cDNA of Dnmt3 gene (GenBank accession no. JQ740768) is 2 277 bp, encoding 758 amino acids, and the predicted MW and pI are 88.24 kD and 7.85, respectively. Based on the comparison of the domain and the phylogenetic tree of the amino acids of Dnmt3 in A. cerana cerana and other species, the sequence obtained has up to 99% identity with that of A. mellifera. The Dnmt3 transcript was clearly detected in different developmental stages of worker and queen, and it was expressed significantly higher in 30 d-old worker than in 1 d- and 7 d-old worker (P<0.05), while no difference existed between 1 d- and 7 d-old worker (P>0.05). The Dnmt3 transcript was expressed higher in queen pupae than in worker pupae (P<0.05), and was higher in 1 d-old queen than in 1 d-old worker (P<0.05). The expression level of Dnmt3 gene between laying worker and laying queen had no significant difference. The results suggested that Dnmt3 may be involved in the division of labor in workers and ovary development in honeybees.
    Extraction and identification of cuticular semiochemical components of Cyrtotrachelus buqueti Guerin-Meneville (Coleoptera: Curculionidae)
    MANG Ding-Ze, LUO Qing-Huai, SHU Min, WEI Wei
    2012, 55(3):  291-302. 
    Abstract ( 4065 )   PDF (3546KB) ( 1445 )     
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    The aim of this study is to identify the semiochemicals of the female weevil, Cyrtotrachelus buqueti Guerin-Meneville, an important bamboo forest weevil pest in Southern China. The behavioral responses of male weevils to the crude cuticular extracts from virgin female weevils in a flight tunnel was analyzed, and the results showed that male weevils had positive response to the cuticular extracts as well as to virgin female weevils. The sensitive reaction of male weevil’s antenna to the crude cuticular extracts from virgin female weevils was detected by GC-EAD, and the components of the cuticular extracts that consistently elicited antennal responses were identified by GC-MS. The results indicated that male weevil’s antenna had sensitive reaction to synthetic standard chemicals of five components in the female extracts, i.e., phenol, ethyl hexanoate, 2-nonanone, nonanal, and pentadecanoic acid methyl ester. Flight tunnel and GC-EAD studies showed that the five synthetic chemicals were also bioactive to male weevils, suggesting that phenol, ethyl hexanoate, 2-nonanone, nonanal, and pentadecanoic acid methyl ester are semiochemicals of C. buqueti.
    EAG and behavioral responses of workers of the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae) to its trail pheromones
    ZHU Hui-Yan, WAN Shu-Qing, CHEN Li
    2012, 55(3):  303-308. 
    Abstract ( 3201 )   PDF (7085KB) ( 1136 )     
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    【Aim】 This study attempted to test electroantennographic (EAG) and behavioral responses of the workers of the red imported fire ant (RIFA), Solenopsis invicta Buren, to its trail pheromones.【Methods】 Dufour’s gland of RIFA workers was dissected, and then extracted with hexane followed by GC-MS analysis. EAG and recruitment behavioral responses of the RIFA workers to Dufour’s gland extract, extract of workers, and synthesized farnesenes were tested. 【Results】 In comparison to farnesenes synthesized from (±)-nerolidol, the major component of Dufour’s gland extract was not Z, E-α-farnesene as previously identified. Dufour’s gland extract, extract of workers and different doses of synthesized farnesenes could elicit significant EAG responses in RIFA workers and there was no significant difference among Dufour’s gland extract, extract of workers and 100 μg farnesenes. Dufour’s gland extract and extract of workers elicited a significant recruitment behavior, but no recruitment response was elicited by synthesized farnesenes at various doses including 10, 1, 0.1 and 0.01 μg. 【Conclusion】 Z, E-α-farnesene is not the major component of tail pheromones of S. invicta. Dufour’s gland extract and extract of workers could elicit significant EAG and behavioral responses in RIFA workers.
    Organization and postembryonic development of glial cells in the visual system of the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae)
    LI Zhao-Ying
    2012, 55(3):  309-315. 
    Abstract ( 3158 )   PDF (14583KB) ( 1144 )     
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    Glial cells exist throughout the visual system, and play essential roles in various aspects of neural development and function. The type and development of glial cells of larval and pupal visual system in the Chinese honeybee, Apis cerana cerana, were comparatively studied by using tissue anatomy and immunohistochemical method (5-bromo-2-deoxyuridine, BrdU incorporation). The results show that in the visual system of A. cerana cerana, glial cells are classified into three categories based on the position of their cell bodies and cell morphology, i.e., surface glia, cortex glia, and neuropil glia. The glia have at least two different sources, including the optic stalk and the glial precursor centre (GPC) in the developing optic lobe. The increase in the number of glial cells is most likely due to the migration of glial cells and glial proliferation during postembryonic development. These results lay an essential foundation for studying glia development and function in insect brain.
    Virulence of Metarhizium flavoviride 82 to different developmental stages of the brown planthopper, Nilaparvata lugens (Hemiptera:Delphacidae)
    LI Mao-Ye, LIN Hua-Feng, LI Shi-Guang, JIN Li
    2012, 55(3):  316-323. 
    Abstract ( 3695 )   PDF (3535KB) ( 1179 )     
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    At present there still lacks ideal commercial microbial insecticides to control the brown planthopper (BPH), Nilaparvata lugens (Stal), an important pest of rice in Asia. Entomopathogenic fungi can penetrate the cuticle of insect hosts and thus have the advantage of controlling sucking pests. In this study, laboratory bioassays were carried out on 12 isolates of three entomopathogenic fungal species, Metarhizium anisopliae, Metarhizium flavoviride and Beauveria bassiana, which were originally obtained from various hosts and geographic origins, for their virulence against BPH adults. The results showed that at the concentration of 1 100 conidia/mm2, the strain Mf82 had the highest virulence to adults with the corrected mortality reaching 83.5% within 10 d after treatment and the LT50 value 4.65 d. The pathogenicity of Mf82 to adults, nymphs at different instars and eggs of BPH were further assessed at a series of conidial concentrations. This strain was most virulent to adults, moderately virulent to old nymphs, and least virulent to young nymphs. It was also found that Mf82 could infect both oviposition marks and eggs in rice seedlings, with the infection rates of 66.7% and 51.2%, respectively, at 10 d after treatment. The younger the eggs, the higher the infection rate, with the 0.5 d-old eggs to be the most easily vulnerable. The results suggest that Mf82 is a promising biocontrol agent for BPH.
    Effects of phylloxera infestation on the root ultrastructure of grape cultivars with different resistance
    DU Yuan-Peng, JIANG En-Shun, ZHAI Heng
    2012, 55(3):  324-329. 
    Abstract ( 3226 )   PDF (10712KB) ( 1168 )     
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    【Aim】 To investigate the effects of phylloxera infestation on the root ultrastructure of grape cultivars with different resistance. 【Methods】 Excised tertiary roots of rootstock 140Ru (Vitis rupestris×V. berlandieri) and Vitis vinifera cv. Cabernet Sauvignon and potted 140Ru were inoculated with phylloxera eggs. After two weeks, tissues at the probing sites were collected to make ultrathin section and the transmission electronic microscope was used to observe the changes of ultrastructure, and potted 140Ru nodosities in different stages were collected to examine the phenol content. 【Results】 The outermost periderm cell wall in 140Ru (1 031.25 nm) was thicker than that in Cabernet Sauvignon (543.75 nm), and the number of periderm cell layers in 140Ru (6-7 layers) was more than that in Cabernet Sauvignon (3-4 layers ). The total phenol content of 140Ru tertiary roots was 35% higher than that of Cabernet Sauvignon. After phylloxera infestation, the cell wall of 140Ru periderm cells from the third layer became thicker, and much more polyphenol materials were accumulated in the periderm cells. Total phenol content of 140Ru nodosities kept increasing after phylloxera infestation, which was 2.4 times as high as the control at 20 d after inoculation. After phylloxera infestation, the cytoplasm concentration of Cabernet Sauvignon became higher, and the numbers of starch granule, mitochondria and endoplasmic reticulum also increased.【Conclusion】Periderm structure of 140Ru impedes the penetration of phylloxera stylet compared to that of Cabernet Sauvignon, and the phylloxera infestation causes the changes unfavorable to phylloxera feeding.
    Influence of separate rearing on adult longevity and fecundity of the emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae)
    LU Ji-Fang, WANG Xiao-Yi, YANG Zhong-Qi, LIU En-Shan, BAO Jin-Mei, WANG Jun-Ping, LIU Yun-Cheng, ZHANG Xian-Wen
    2012, 55(3):  330-335. 
    Abstract ( 3506 )   PDF (5410KB) ( 1356 )     
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    The emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is an important international quarantine insect pest. To understand the influence of different rearing conditions and intraspecific individual communications on biological parameters of the EAB adults, the adult longevity, food consumption, oviposition and oviposition duration were determined by group rearing, couple rearing, single female rearing without separation, single female rearing under visual separation and single female rearing under olfactory separation in laboratory. The results showed that separate rearing reduced the longevity and fecundity of EAB adults. The adult longevity and fecundity differed significantly (P<0.05) among different treatments, but there was no significant difference in oviposition (P>0.05). No separation for single female rearing gave the longest average longevity (32.40 d), while group rearing caused the shortest average longevity (20.77 d). The average longevity in the treatment of single female rearing was longer than those in couple rearing and group rearing. For single female rearing, adults with visual separation consumed the maximum ash leaf (156.16 mm2 per day), while those with olfactory separation consumed the least ash leaf (107.35 mm2 per day). No separation for single female rearing caused the highest oviposition and the longest oviposition duration, which was 98.33 d and 21.33 d, respectively. It is concluded that block of intraspecific individual communications significantly influences the feeding, development and fecundity of EAB adults, and olfactory plays the most important roles in intraspecific individual communications and vision follows. In addition, disturbance or competition among individuals may also have some effects on the longevity and fecundity of EAB adults. These findings provide theoretical supports for the EAB management using separation measures such as planting mixed tree species.
    Establishment and application of DNA barcoding technology for identification of the immatures and adult debris of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)
    LIU Shen-Si, ZHANG Gui-Fen, WU Qiang, ZHANG Ai-Bing, WANG Jin-Jun, WAN Fang-Hao
    2012, 55(3):  336-343. 
    Abstract ( 3281 )   PDF (2890KB) ( 1221 )     
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    Many species in the family Tephritidae are quarantine pests worldwide. Usually, identification of the tephritid fruit flies is mainly based on external morphological characteristics of adults. In this research, the immatures (including egg, larva and pupa) and adult debris (including leg, wing, head, thorax and abdomen) of Bactrocera dorsalis (Hendel) were used to develop a rapid identification technique for tephritid fruit flies based on DNA barcoding technology. The other four tephritid fruit fly species, i.e., B. correcta, B. cucurbitae, B. tau and B. minax, were used to verify the feasibility of the fruit fly identification technique developed. The results showed that the nucleotide sequence identity of the partial COⅠ gene between the immatures or adult debris of B. dorsalis and the target gene from GenBank database is 99.51%-99.84%, while between other four tephritid fruit fly species and the target gene from GenBank database it is 100%, 100%, 99.81%-99.83% and 100%, respectively. Neighbor-joining tree was established based on the analysis of COⅠ gene sequences. The target species and the corresponding species in the database cluster in the same branches. All bootstrap values of the original divergence within one species are 100%. The intra- and inter-species genetic distances were calculated with MEGA version 5.0 software using the Kimera 2-Parameter model. The intraspecies genetic distances are 0.0000-0.0041, with an average of 0.0019. The interspecies genetic distances are 0.0597-0.2363, with an average of 0.1693. There is no overlap between intra- and inter-species genetic distances. The results indicated that the developed DNA barcoding identification techniques based on the partial COⅠ gene can provide a rapid and accurate method for identification of immatures or adult debris of tephritid fruit fly species.
    Application of DNA barcoding technology for species identification of common thrips (Insecta: Thysanoptera) in China
    QIAO Wei-Na, WAN Fang-Hao, ZHANG Ai-Bing, MIN Liang, ZHANG Gui-Fen
    2012, 55(3):  344-356. 
    Abstract ( 4646 )   PDF (6413KB) ( 1544 )     
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    Thrips (Insecta: Thysanoptera) are of a species-rich order with small size, which makes them hard to be identified quickly and accurately by traditional morphological identification methods. In this study, the mitochondrial cytochrome c oxidase subunit Ⅰ(COⅠ) genes of 25 thrips species common in China were amplified using DNA barcoding universal primers. The obtained partial COⅠ fragments (about 650 bp) were sequenced. The phylogenetic tree was established by a neighbor-joining (NJ) method. The intra- and inter-species genetic distances were calculated with MEGA version 4.0 software using the Kimura 2-Parameter model. The results indicated that the identification results based on the phylogenetic tree obtained was consistent with those based on morphological characteristics for the 25 thrips species. In these species, the lengths of intra-species branches were much shorter than those of the interspecies branches. Each branch corresponded to one species and all bootstrap values of the original divergence among different haplotypes within one species were 100%. The genetic distance between species for these 25 species was 102.1 times as high as the corresponding values within species (0.2757 vs. 0.0027). There was no overlap between inter- and intra- species genetic distances. The results suggest that the DNA barcoding based on partial COⅠ gene can provide rapid and accurate identification of these thrips species.
    Assay and comparison of chemical constituents of crude methanol extracts from different developmental stages of darkling beetle Zophpbas morio (Coleoptera: Tenebrionidae)
    CHEN Wen-Ting, LIU Hai-Li, YAO Li-Gong, YE Zhu-Liang, GUO Yue-Wei
    2012, 55(3):  357-360. 
    Abstract ( 3209 )   PDF (5015KB) ( 1161 )     
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    To study the chemical changes of darkling beetle Zophobas morio during metamorphosis, the crude methanol extracts of Z. morio in three developmental stages (larva, pupa and adult) were assayed by modern chromatographic techniques, 1H NMR and GC-MS and compared. The results showed that pupae contained characteristic chemical constituents. The methanol extract of the pupae was systematically studied and ten fatty acids were identified. 10-oxohexadecanoic acid and 10-oxooctadecanoic acid were found from pupae of Z. morio, which were separated from insects for the first time. (8E, 11E)-8, 11-octadecenoic acid, (9Z, 12Z)-9, 12-octadecenoic acid, (9Z, 12Z, 15Z)-9, 12, 15-octadecenoic acid, 8-(3-octyl-2-oxiranyl)octanoic acid, and 8-(2-octylcyclopropyl)octanoic acid were also found from Z. morio for the first time. These results offer the new theoretical basis for the utilization of darkling beetle at different developmental stages.
    Effects of larval density on food utilization efficiency and digestive enzyme activity of the beet webworm, Loxostege sticticalis (Lepidoptera: Pyralidae)
    KONG Hai-Long, LUO Li-Zhi, JIANG Xing-Fu, ZHANG Lei, YANG Zhi-Lan, HU Yi
    2012, 55(3):  361-366. 
    Abstract ( 3419 )   PDF (967KB) ( 1103 )     
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    To understand the larval feeding pattern of the outbreak population, Loxostege sticticalis, the variation of larval food utilization parameters and digestive enzyme activity of L. sticticalis among three larval densities (1, 10 and 30 larvae/jar of 650 mL) were studied in the laboratory (22±1℃, 70% RH). The results showed that the highest and lowest midgut relative mass was found in the larvae reared at the density of 10 larvae per jar and 1 larva per jar, respectively. Larval relative consumption rate and excretion dry weight increased, but body weight and the efficiency of conversation of ingested food (ECI) declined as larval density rose. The relative consumption rate and dry weight of excretion of larvae reared at the density of 30 larvae per jar was significantly higher than that of larvae reared at the density of 1 larva per jar and 10 larvae per jar, while the ECI of larvae reared at the density of 30 larvae per jar was significantly lower than that of larvae reared at the density of 1 larva per jar. The approximate digestibility (AD) was significantly declined with increasing larval density. Furthermore, larval density had a significant influence on the activity of protease enzyme. The activity of total protease and leucine aminopeptidase enzyme was significantly higher in the larvae reared at the density of 10 than that at the densities of 1 and 30 larvae per jar, but the activity of amylase enzyme was not affected by larval density. The change trend of relative mass of larval midgut among three densities was consistent with the activity of total protease and leucine aminopeptidase enzyme. The variation of digestive enzyme activity may be relevant to the midgut relative mass. Thus, larval population density is one of the major factors affecting feeding behavior feature of L. sticticalis. These results provide evidence for clarifying the different damage feature between the outbreak and common populations.
    Spatial distribution pattern of Eupolyphaga sinensis Walker (Blattaria: Polyphagidae) and the effects of soil properties on its survival
    WANG Yan-Yang, LIANG Guang-Wen
    2012, 55(3):  367-370. 
    Abstract ( 3245 )   PDF (883KB) ( 1087 )     
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    To study the characteristics of the spatial distribution of Eupolyphaga sinensis Walker in the natural environment and the effects of soil properties on its survival, Morisita’s index of dispersion was used to examine the type of distribution. The results showed that the nymphae of E. sinensis were clustered in the natural environment and the adults were randomly distributed outdoors, from ground surface to the depth of 40 cm. Testing under soil conditions with four different initial water contents ranging from 16% to 29% showed that the egg hatch rate in soils with the initial water contents of 21% and 24% was significantly higher than that in soils with other two water contents, and the survival rate of nymphae in soil with the initial water content of 29% was significantly lower than those in soils with other three water contents. Investigation using soils of five different sources as the habitat showed that the compost and habitat soil from the foot of a mountain was more suitable for survival of E. sinensis. The results provide scientific data for collecting, rearing and further research of E. sinensis.