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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 April 2008, Volume 51 Issue 4
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    Cloning, expression pattern and phylogenetic analysis of cyclophilin A gene in the silkworm, Bombyx mori.
    LI Chun-Feng
    2008, 51(4):  349-356. 
    Abstract ( 3416 )   PDF (1233KB) ( 1429 )     
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    CyP protein family plays an important role in the process of protein folding. In this study, a novel silkworm gene BmCyPA was cloned, which comprises 2 exons and 1 intron. The deduced BmCyPA protein contains 165 amino acids, its molecular mass is 19.4 kD and pI is 8.79. Sequences analysis showed that BmCyPA is highly conserved among different species. The BmCyPA protein has peptidylprolyl cis-trans-isomerase active sites and contains specific amino acids which can bind to the side chain of CsA, which indicated that BmCyPA has potential cis-trans-isomerase activtity and can combine with CsA. RT-PCR of tissue expression pattern and EST analysis revealed that BmCyPA is specifically and highly expressed in silk gland. The potential function of BmCyPA gene was further analyzed by phylogenetic analysis of CyPA genes from different species. The results suggest that BmCyP may be involved in the process of fibroin folding.
    Cloning, sequencing, and expression of cDNA fragments of CYP4G2 from Xylotechus rusticus L. (Coleoptera: Cerambycidae) in Escherichia coli.
    GUAN Hua-Nan
    2008, 51(4):  357-364. 
    Abstract ( 3261 )   PDF (1065KB) ( 1282 )     
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    A pair of primers was designed based on the reported sequences of conservative amino acid regions of CYP4 protein family in insects. The cDNA encoding the cytochrome CYP4G2 protein was isolated from the intestines of Xylotechus rusticus L. by reverse transcription polymerase chain reaction (RT-PCR). The cDNA fragment was constructed to prokaryotic expression vector that named pET-CYP4G2 for overexpression in Escherichia coli JM109.Sequence analysis showed that the full-length of CYP4G2 (GenBank accession no. EF429250) open reading frame (ORF) was 387 bp, encoding 129 amino acid residues;  the predicted MW and pI were 16.9 kD and 5.75, respectively. The deduced amino acid sequence showed high identity to the reported sequences of CYP4 family amino acids from other insects63%86% and shared the typical structural features of cytochrome from other insects. After induction with IPTG, its molecular weight was found to be about 22 kD by checking with SDS polyacrylamide gel electrophoresis. The molecular weight was the same as the prediction of fusion protein. Carbon monoxide (CO) difference spectrum analysis confirmed that the recombinant strain expressed pET-CYP4G2 with biological activity.
    Cloning of HSP70 genes in a desert beetle Mantichorula semenowi Reitter (Coleoptera: Tenebrionidae) and their expression under heat shock
    TANG Ting
    2008, 51(4):  365-371. 
    Abstract ( 3406 )   PDF (1310KB) ( 1418 )     
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    Two different HSP70 gene fragments were cloned and sequenced from a desert beetle Mantichorula semenowi Reitter in Ningxia, which were named MsHSP70 and MsHSC70 respectively. Both of them share high identity with the reported heat shock proteins of other insects. Semi-quantitative RT-PCR analysis showed that 1 h heat shock treatment at 42 caused rapid increase of MsHSP70 mRNA, which reached maximum levels at the end of the heat shock treatment, and decreased gradually after being moved to room temperature. Expression of MsHSC70,however, was inhibited after heat shock treatment: after recovering 2 h and 4 h, the expression levels were only 0.25 and 0.28 times that of the control, respectively. The results suggest that MsHSP70 and MsHSC70 play different roles in cell protection. The present results provide new insights into adaption mechanisms of M. semenowi to extreme environment stress.
    Purification and characterization of vitellin from locust Catantops pinguis (Stål) (Orthoptera: Catantopidae)
    JIN Jian-Ya
    2008, 51(4):  372-378. 
    Abstract ( 3547 )   PDF (327KB) ( 1421 )     
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    The vitellin was purified from eggs of the locust Catantops pinguis (Stål) using distilled water precipitation, gel filtration and gel electrophoresis. Its properties were also analyzed using polyacrylamide gel electrophoresis and different dyeing methods. The results indicated that the vitellin of C. pinguis was a kind of protein containing sugar and fat. The molecular weight of the vitellin was 548 kD, consisting of seven subunits with molecular weight of 147.3, 100.2, 95.9, 59.6, 53.6, 49.0 and 42.2 kD, respectively. From the vitellin, 17 kinds of amino acids and NH3 peaks were detected by HPLC, among which the content of Glu was highest with 13.46% in the vitellin, and the contents of Asp, Leu, Cys and Arg were relatively high, while contents of Pro, His and Lys were low.
    Effects of the mixture of celangulin Ⅳ and Ⅴ from Celastrus angulatus Max. on sodium channels in nerve cells of cotton bollworm (Helicoverpa armigera)
    HU Zhao-Nong
    2008, 51(4):  379-383. 
    Abstract ( 3103 )   PDF (201KB) ( 1249 )     
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    Celangulin and from the root bark of Celastrus angulatus are the major active ingredients of the insecticidal plant which can result in reverse nerve poisoning symptoms, i.e., narcosis and excitation, respectively. The insecticidal efficacy of celangulin can be synergied by celangulin , but celangulin has no synergistic effects on the narcotic efficacy of celangulin . By using patch-clamp technique, the interaction of celangulin and in different ratio (31, 11, 13) mixtures on tetrodotosin (TTX)-sensitive inward sodium channel current (INa) in cultured nerve cell of cotton bollworm (Helicoverpa armigera)was investigated. The results showed that the effects corresponded with the ratio of celangulin and in a mixture. When the ratio of celangulin in a mixture was high, voltage gated sodium channel (VGSC) was blocked and TTX-sensitive INa was decreased. When the ratio of celangulin in a mixture was high, VGSC was activated and TTX-sensitive INa was increased. Different ratio mixtures of celangulin and showed no effect on activated voltage, except the peak voltage shifted to positive potential 10-20 mV. The results suggest that celangulin and share a binding site of VGSC or they bind to separate sodium channel sites which interact altosterically with each other, and they might antagonistically compete each other  to bind the site on VGSC.
    Biochemical mechanisms of resistance to chlorpyrifos and dichlorvos in field populations of Bemisia tabaci (Gennadius) B-biotype
    HE Yu-Xian
    2008, 51(4):  384-389. 
    Abstract ( 3640 )   PDF (191KB) ( 1358 )     
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     Biochemical mechanisms of resistance to chlorpyrifos and dichlorvos were investigated in six field populations of Bemisia tabaci collected from Fujian Province of China with synergist bioassay and biochemical analysis. Compared with the susceptible SUD-S strain, the six field populations showed 54.53–78.43 and 6.23–11.25-fold resistance to chlorpyrifos and dichlorvos, respectively. TPP, PBO and DEF had 3.61–24.94, 1.14–1.76, and 1.04-fold synergistic effect on chlorpyrifos, and 1.67–2.64, 1.33–1.65 and 1.09-fold synergistic effect on dichlorvos, respectively, suggesting that carboxylesterase (CarE)-based detoxification could be an important resistance mechanism to chlorpyrifos in B. tabaci. Km and Vmax of acetylcholinesterase (AChE) of the six field resistant populations were 1.83–4.0 and 0.34–0.62 times those of the SUD-S strain. Activity of AChE was inhibited both in the SUD-S strain and the resistant field populations when their subtrate concentrations were higher than 1.0 mmol/L and 16.0 mmol/L, respectively. AChE of the six field resistant populations was 119.92–161.33 and 10.11–14.24-fold less sensitive to chlorpyrifos and dichlorvos than that of the SUS-S strain, respectively. The results suggest that both insensitive AChE and increased CarE are the major mechanisms conferring chlorpyrifos resistance in B. tabaci, and only insensitive AChE is important for dichlorvos resistance.
    Oviposition repellency and EAG responses of the Oriental fruit fly, Bactrocera dorsalis (Hendel) to mineral spray oils
    OUYANG Ge-Cheng
    2008, 51(4):  390-394. 
    Abstract ( 4045 )   PDF (159KB) ( 1555 )     
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     In order to determine the impact of mineral oils on insect behavior, oviposition behavioural responses of the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) females to deposits of 0.5% (v/v) aqueous emulsions of each of the four HMOs were determined for banana fruit that were dipped in the emulsions. The oils were nC21.5 Total Citrole, nC21 Caltex Lovis, nC23 Ampol D-C-Tron NR and nC24 SK EnSpray 99 Oviposition in fruit dipped in Lovis was significantly lower (85%) than in water-dipped fruit. The other three oils had no impact on oviposition. The results suggested that the influence of mineral oils on the females could be related to the composition of oils. Electroantennogram (EAG) responses of females to volatiles emanating from pieces of filter paper onto which drops of 2% (v/v) emulsions were placed were also determined for each oil. There was no significant correlation between the oviposition repellency of the four HMOs against fruit fly and the EAG responses of female fruit flies to 0.5 hour deposits from which water in the emulsions had not evaporated. However, there was a significant negative correlation between the oviposition repellency and the EAGs responses for 3 hour deposits on filter papers from which all water had evaporated. This suggested that oil volatiles had little effect on repelling female B. dorsalis fruit fly. It was hypothesised that oil deposits may block the release of plant volatiles that female B. dorsalis are attracted to in order to lay their eggs. Ovipositional responses to bananas sprayed or dipped with 0.0% (water), 0.22%, 0.29%, 0.4% and 0.67% (v/v) of Lovis oil were compared in a subsequent experiment. Oviposition in dipped fruit was significantly lower than in the sprayed fruit and the water controls. But there was no difference among the oil-dipped treatments, and in the sprayed treatments, only the 0.67% oil treatment was significantly different from the water sprayed control. These results indicates that for deposits to be effective they must exceed deposits equivalent to or greater than those achieved with the 0.22% dip and 0.67% spray used in the experiments. It was evident that dipping was more efficient than spraying to deposit mineral oils on plant surface, and this further proved that oil film forming on surface of fruits which blocked plant volatiles was related to oviposition repellency against the fruit fly.
    Quantitative risk analysis of invasive alien pests based on the maximum pest limit concept
    LI Bai-Ni
    2008, 51(4):  395-401. 
    Abstract ( 3438 )   PDF (1361KB) ( 1328 )     
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    A quantitative risk assessment is needed for each quarantine insect pest to ensure quarantine security without sacrificing the transparency of international trade. In this paper, a quantitative risk analysis system based on the maximum pest limit concept (MPL) was designed to determine the risk probability of alien insect pests introduced into China. The equations of MPL were constructed to determine the probability of a single pair of bisexual insects surviving in a shipment of commodity which were to be introduced into China. A computer system integrating with SAS V9.0 was developed by PHP program language and MySQL database to assess the risk and released on web through Apache server. The clients just have to submit the form lists including required data after logging into the system, and the computer system will activate the analysis process automatically and show the precise outcome on web. This research is a new attempt in the quantitative risk analysis of alien insect pests, and provides a new analysis tool for the quarantine sector of the government.
    Succession and development of insects on pig carcasses and their significances in estimating PMI in the Pearl River Delta region
    WANG Jiang-Feng
    2008, 51(4):  402-410. 
    Abstract ( 3409 )   PDF (1307KB) ( 1530 )     
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     In order to solve the difficulty of the determination of postmortem interval PMI in criminal cases,  18 pig carcasses were placed in outdoor environment in different seasons to observe and select the critical entomological index in accurately estimating of PMI. The results showed that these indexes, including T1 (larvae begin to appear), T2 (larvae begin to crawl away), T3 (most of the larvae have crawled away), T4 (larvae begin to pupate), T5 (most larvae have pupated), T6 (pupae begin eclosion) and T7 (the end of eclosion), could be used as accurate PMI indicators for their high sensitivity and stability. The carcasses decayed very quickly in spring, summer and autumn, and it took 202±23 h, 202±18 h, 277±20 h for test carcasses to decay from fresh to skeleton in these seasons, respectively. In winter it took a longer time of 1 297±63 h to decay from fesh to skeleton. About 47 insect species were found on carcasses. Most of the necrophagous insects could be found all the year, and there was no marked difference in species composition among the four seasons. The predominant species were very apparent. The fly larvae included mainly Chrysomya megacephala, Achoetandrus rufifacies and Hydrotaea (Ophyra) spinigera. The necrophagous insects could only breed one generation on carcasses before the carcasses become into skeleton in most of the time. Based on these results, critical time points for PMI estimation could be determined.
    The role of foundress relatedness in the offspring sex ratio of fig wasp Diaziella yangi (Hymenoptera: Pteromalidae)
    SONG Bo
    2008, 51(4):  411-416. 
    Abstract ( 3870 )   PDF (221KB) ( 1354 )     
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    Local mate competition (LMC) theory had developed in many aspects, including that the offspring sex ratio should be more female-biased when the foundress oviposited with related females than with unrelated females. We studied the influence of relatedness of Diaziella yangi females oviposited on a fig on sex ratio of offsprings, which was a species of non-pollinator hosting on Ficus curtipes. Treatment 1: The 2 foundresses of D. yangi were from the same fig of maturing tree No. 1 treatment 2:  the 2 foundresses were from the same fig of maturing tree No. 2; treatment 3:  one foundress was from maturing tree No. 1, and the other one was from maturing tree No. 2.The experimental results, however, were not consistent with the predicted values. In the experimental results, the sex ratio of offsprings in two treatments with two foundresses of sister relationship were 0.195±0.028 and 0.189±0.043, respectively; the sex ratio of offsprings produced by two foundresses of non-sister relationship was 0.240±0.030; and there were no significant difference among the three treatments. The results suggest that fig-wasps have no capabilities of kin recognition and adjusting the sex ratio of offsprings accordingly, and this is consistent with many other studies.
    Genetic diversity of geographical populations of four Dendrolimus species (Lepidoptera: Lasiocampidae) in China based on allozyme analysis
    NANGONG Zi-Yan
    2008, 51(4):  417-423. 
    Abstract ( 2849 )   PDF (1922KB) ( 1296 )     
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    Aim Using polyacrylamide gel electrophoresis to find out the genetic diversity among and within nine geographic populations of four Dendrolimus species in China: Dendrolimus punctatus punctatus Walker, D. punctatus spectabilis Butler,D. superans Butler, D. haui Lajonquiere and D. kikuchii Matsumura. MethodsSix allozymes, including lactate dehydrogenase (LDH), malate dehydrogenase (MDH), malic enzyme (ME), alcohol dehydrogenase (ADH), formic acid dehydrogenase (FDH) and glutamate dehydrogenase (GDH), were analyzed. ResultsTen loci and seventeen alleles were recorded with four loci polymorphic, P=40.0%. The average number of alleles per locus A was 1.700, the expected heterozygosity He 0.151, and the mean genetic distance 0.001-0.285. Among six populations of D. punctatus Walker, the coefficient of gene differentiation Fst was 0.265, and the number of migrants per generation Nm 0.692. Genetic relationships among populations and species were assessed by standard genetic distance, and it was found that D. punctatus spectabilis Butler and D. superans Butler had a closer relationship, while D. superans Butler and D. haui Lajonquiere had the farthest relationship. Conclusions According to the heredity parameters, gene flow was low and genetic drift was one of primary factors that led to the genetic differentiation among six geographic populations of D. punctatus Walker. Genetic distances were correlated with geographical distances among populations.
    A taxonomic study on the genus Austrohancockia Gunther (Orthoptera: Tetrigoidea: Cladonotidae), with description of one new species from China
    ZHENG Zhe-Min
    2008, 51(4):  424-429. 
    Abstract ( 3025 )   PDF (219KB) ( 1473 )     
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    The paper deals with 10 species of the genus Austrohancockia  Gunther from China, including one new species from Guangxi, Austrohancockia longidorsalis sp. nov. A key to all species of the genus was provided. Type specimens of new species are kept in the Institute of Zoology, Shaanxi Normal University.
    Baculovirus chitinase: Advances in research and application
    LIU Yan-He
    2008, 51(4):  430-436. 
    Abstract ( 6059 )   PDF (207KB) ( 1934 )     
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    The chitinase gene of baculovirus is the gene that is not essential for the virus replication. It is expressed in the late phase of the virus replication in insect cells, and it is highly conservative. The chitinase contains three functional regions: N-terminal secretion signal peptide, central active region that includes the active site of family 18 chitinases and a functional C-terminal endoplasmic reticulum (ER)-retention sequence. The enzyme possesses high performance of both exo- and endo-chitinase activity, which can hydrolyze inherent chitin in insect body, and promote liquefaction of the host after death. The chitinase may also serve as a molecular chaperone of pro-V-Cath, the precursor of V-Cath, for its proper folding and transportation in the ER. It has an influence on the polyhedra releasing and cell lysis. The chitinase is also relevant to the baculovirus infection. Phylogenetic analyses indicate that baculovirus acquires the chitinase gene from bacterium via horizontal gene transfer, but insect chitinase gene may directly come from baculovirus. For biological control of pests, the baculovirus chitinase can control insect pests as an insecticide, or work together with Bacillus thuringiensis or a baculovirus as its potentiator; the baculovirus chitinase gene can also be introduced into plants to get the transgenic plants that increase resistance against herbivorous pests and fungal pathogens without affecting non-target insects. The baculovirus chitinase ER-retention motif was deleted or modified, or heterologous chitinase was introduced into the baculovirus genome, which made the biological activity of the recombinant virus enhanced. Deletion of the chitinase and v-cathepsin genes from the baculovirus expression vector through genetic engineering has a positive effect on the integral membrane and secreted proteins.
    Molecular cloning and characterization of a cDNA encoding the prophenoloxidase gene in Bombyx mandarina
    ZHANG Yong-Liang, HUANG Zhong, LI Zhi-Qing, ZHANG Chun-Dong, ZHU Yong
    2008, 51(4):  437-442. 
    Abstract ( 3415 )   PDF (363KB) ( 1928 )     
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    Phenoloxidase plays an important role in d efense mechanisms in insect immunity. The complemental deoxyribonuc leic acid (cDNA) of Bombyx mandarina prophenoloxidase (PPO) gen e was cloned by means of reverse transcription polymerase chain reaction (RT-P CR) and rapid amplification of cDNA ends (RACE). The cDNA was 2 134 bp in l ength a nd contained an open reading frame (ORF) of 2 082 bp which encoded 693 amino aci d residues.The deduced amino acid sequence showed a high identity to the report ed sequence of PPO2 from other Lepidopterous insects and shared the typical stru ctural features of PPO from other insects. The results of RT-PCR showed that PP O mRNA was expressed in the integument, haemocyte, head, testis, ovary, fat body and midgut of 5th instar larvae and at different developmental stages of B. m andarina. These results provide molecular basis for further studying the funct ion of ppo gene inB. mandarina.
    Proteome analysis of silkworm (Bombyx mori) midgut proteins related to BmNPV infection resistance or susceptibility
    LIU Xiao-Yong
    2008, 51(4):  443-448. 
    Abstract ( 4065 )   PDF (504KB) ( 1518 )     
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    The midgut epithelium is the first tissue of silkworm (Bombyx mori) which the virus confronts; larvae resist baculovirus infection by selective apoptosis of infected midgut epithelial cells and by sloughing off infected cells from the midgut before they release virions into the hemocoel. To research the mechanism of nucleopolyhedrovirus resistance of silkworm midgut tissue, we constructed a near-isogenic silkworm line with BmNPV resistance through highly resistant and highly susceptible parent strains. In this study, proteome maps and differences of protein patterns of the midgut tissues of the highly resistant, highly susceptible and near-isogenic silkworm strains were investigated by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. We compared the proteomic expression of the midgut in these three silkworm strains, and identified five differently expressed proteins. It is inferred that these proteins may be involved in the resistance or susceptibility of BmNPV.
    Induction of apoptosis by botanical components in Spodoptera litura  cultured cell line
    ZHONG Guo-Hua, SHUI Ke-Juan, HUANG Jin-Fei, JIA Jian-Wen, HU Mei-Ying
    2008, 51(4):  449-453. 
    Abstract ( 5287 )   PDF (405KB) ( 1735 )     
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    In order to determine the apoptotic inductivity of botani cal components toSpodoptera litura cultured cell line SL-1, th e concentration and time dependence of apoptotic-induce ability of nine botanic a l components, including azadirachtin and comptothecin, to SL-1 was investigated by inverted phase contrast microscope. The results showed that a great deal of a poptotic bodies appeared, one of the typical morphological characteristics of ap optosis, 24-48 h after treatments with azadirachtin at a concentration of 0.1-5.0 μg/mL and comptothecin at 0.5-20.0 μmol/L; the same changes, however, di d not appear in SL-1 cells within the whole observation period of 72 h after tre atments with tea saponin, ricinine, sassafras, paeonolum, nicotine, matrine and macleay cordata at concentrations of 0.5-20 μg/mL. Treated by azadirachtin at a concentration of 0.75 μg/mL, the early, mid and late apoptotic stages apoptosis of SL-1 cells appeared during0-36 h, 36-60 h and 60 h af ter treatment, respectively; and treated by comptochetin at a concentration of 5.0 μmol/L, these appeared during 0-24 h, 24-52 h and 54 h after treament, re spectively. It is tentatively concluded that azadirachtin and camptothecin both possess the concentration\|time-dependence induction of apoptosis to SL-1 cel ls.
    Morphology of diapause and nondiapause pupae and their adults of  Sericinus montelus Gray (Lepidoptera: Papilionidae)
    YANG Qiu-Sheng, WANG Xiao-Ping, XU Shu, PAN Yue, LEI Chao-Liang
    2008, 51(4):  454-458. 
    Abstract ( 3546 )   PDF (1293KB) ( 1714 )     
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    Sericinus montelus Gray is an ornamental insect with exploitability, which overw inters as diapause pupae, and adults have polymorphism. In this study, the morph ology of diapause and non-diapause pupae and their adults were inve stigated in this butterfly in terms of color, body and wing size, and the length of spine on the 9th abdomen of pupae. Compared with non-diapause pupae, the co l or of diapuase pupae is darker. The faint yellow of antenna terminal is obviousl y different from the body color. The length of spine on the 9th abdomen of 3-day-old diapause pupae is about four times that of 3-day-old non-diapau se pupae. Therefore, non-diapause and diapause pupae can be identified at this s tage. The wingspan and tail length of adults from diapause pupae are significant smaller than those of adults from non-diapause pupae. There are also obvious differences in pattern of abdomen and wing. These differences are consistent wit h the descriptions of the spring form and summer form in this butterfly. The res ults suggest that the seasonal polymorphism of adult is related to diapause in S. montelus.