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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 January 2009, Volume 52 Issue 1
For Selected: View Abstracts Toggle Thumbnails
  • RESEARCH PAPERS
    Molecular cloning, sequence analysis and tissue expression of serpin-6 gene in Bombyx mori
    2009, 52(1):  1-9. 
    Abstract ( 3164 )   PDF (818KB) ( 1314 )     
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    The ORF and the 5UTR sequences of serpin were cloned from Bombyx mori using RT-PCR and rapid amplification of cDNA ends (RACE) strategies. We named this gene as serpin-6 of B. mori, which was deposited in GenBank with the accession no. EU159447. In order to study the mechanism of immunity of serpin-6 from B. mori, we performed the tissue expression and detected the immunity response. The results of semi-quantitative RT-PCR showed that the mRNA of this gene was highly expressed in head and gonads, and lowly expressed in the midgut, fat body, silk glands and haemolymph. When day-3 fifth instar larvae were injected with LPS, the RT-PCR results showed that this gene was highly induced and it was highly expressed in the fat body and haemolymph, and the highest expression was at 6 h after injection. It is inferred that this serpin gene may play an important role in immunity of B. mori.
    Molecular cloning and sequencing of cDNAs of two peritrophic membrane proteins from Holotrichia oblita (Coleoptera: Melolonthidae)
    2009, 52(1):  10-16. 
    Abstract ( 3152 )   PDF (1743KB) ( 1166 )     
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    Peritrophic membrane (PM) proteins of a coleopteran insect Holotrichia oblita were studied by constructing a H. oblita larval midgut cDNA expression library with the cDNA Synthesis Kit of Stratagene Company and screening with a PM protein polyclonal antiserum from Helicoverpa armigera according to the theory of modern immunology. The titer of the primary cDNA expression library was 1.9×106 pfu/mL and the recombination rate was 99.97%. Two positive cDNA clones named Ho-Peritrophin1 and Ho-Peritrophin2, with the sizes of 2 385 and 1 633 bp, were screened and sequenced from the library, respectively. The longest open reading frame of Ho-Peritrophin1 and Ho-Peritrophin2 coded for 729 and 477 amino acids respectively, both of which showed the highest similarity to CBP2 of Trichoplusia ni, though their similarities to the latter were only 21.9% and 19.1%, respectively. Ho-Peritrophin1 and Ho-Peritrophin2 contained nine and six chitin binding domains (CBDs) respectively with a little O-linked glycosylation sites and no mucinlike domain was found in the two protein sequences. The cleavage sites of trypsin and chymotrypsin are mainly located inside CBDs of the Ho-Peritrophin1 and Ho-Peritrophin2, but the two proteins are protected by the intradomain disulfide bonds, so they can resist the enzymes and exert physiological function in the midgut. Compared with the normal CBD, the CBD of carboxylic terminal of the two proteins contains only four cysteines, which form two pairs of disulfide bonds between the 1st and 3rd cysteines and the 4th and 5th cysteines, lacking the one between the 2nd and 6th cysteines. It is inferred that there are one signal peptide and unknown CBDs in the N end of Ho-Peritrophin1 and Ho-Peritrophin2 respectively and these require further study.
    Cloning, sequence analysis and developmental expression of a cDNA encoding nicotinic acetylcholine receptor α subunit from Plutella xylostella (L.) (Lepidoptera: Plutellidae)
    2009, 52(1):  17-26. 
    Abstract ( 2456 )   PDF (1720KB) ( 1110 )     
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    Nicotinic acetylcholine receptors (nAChR) mediate the fast action of excitatory neurotransmitter at cholinergic synapses in the central nervous system of insects and are also the target site for neonicotinoids and spinosad. By using RT-PCR and RACE technique, the full-length cDNA of a novel nAChR α subunit (Pxα8) was cloned from Plutella xylostella (GenBank accession no. EU914853). Sequence analysis showed that the full-length cDNA is 1 744 bp in length and contained an open reading frame (ORF) of 1 602 bp which encodes 534 amino acid residues. The deduced amino acid sequence of Pxα8 shared typical features of nAChR α subunits and showed 77%-96% amino acid similarity with insect nAChR α8 subunit orthologues. Interestingly, it also showed 76% amino acid similarity with the nAChR β2 subunit of Drosophila melanogaster. There are many single nucleotide polymorphism (SNP) sites in the ORF of Pxα8, some of which resulted in amino acid substitution. There are more SNPs in female 4th-instar larvae than in male 4th-instar larvae, and the SNP sites are different between the female and male 4th-instar larvae. Semi-quantitative RT-PCR analysis indicated that the mRNA expression level of Pxα8 was higher at adult stage than at either pupal or 4th-instar larval stage. These findings provide important basis for further study on diversity of nAChR subunits and mechanism of target resistance to spinosad in P. xylostella.
    Purification and partial biological functions of β-1,3-glucan recognition protein from Tenebrio molitor (Coleoptera: Tenebrionidae) larvae
    2009, 52(1):  27-32. 
    Abstract ( 2979 )   PDF (851KB) ( 850 )     
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    One kind of β-1,3-glucan recognition protein (Tm-βGRP) was purified from Tenebrio molitor larvae by such traditional means as neutral salt precipitation and gel chromatography, etc., and its biological functions in pro-phenoloxidase activation system were primarily studied. The results showed that the molecular weight of Tm-βGRP was approximately 70 kDa, and the protein mainly existed in plasma. Purified Tm-βGRP could only specifically recognize β-1,3-glucan, but not peptidoglycan. During the process of pro-phenoloxidase activation induced by β-1,3-glucan, with the proceeding of pro-phenoloxidase activation, the amount of endogenous Tm-βGRP decreased significantly. The polyclonal antibody against Tm-βGRP could inhibit phenoloxidase activity in the hemolymph of T. molitor induced by β-1, 3-glucan in a dose-dependent manner. This research may help us deeply elucidate the biological function of β-1, 3-glucan in pro-phenoloxidase activation system in the hemolymph of T. molitor.
    Isolation and identification of lipid rafts from the larval midgut of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)
    2009, 52(1):  33-38. 
    Abstract ( 2867 )   PDF (1172KB) ( 1032 )     
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    Lipid raft is a detergent-resistant microdomain that enriches in cholesterol, sphingolipids and glycosylphosphatidylinositol-anchored proteins. It is considered as the platform that pore-forming toxins aggregate and form oligomer on the surface of cell membrane. In order to investigate the relationship between lipid raft and the formation of membrane pore by Bt β-endotoxins, the preparation and identification of lipid raft from the larval midgut of Helicoverpa armigera (Hübner) was performed. According to the insolubility of lipid raft in detergent at 4, brush border membrane vesicle BBMV of the larval midgut of H. armigera was treated with Triton X-100 to dissolve non-raft membrane fraction, and Triton insoluble fraction (lipid raft) was then isolated by OptiPrep gradient centrifugation. GM1 gangliosidoses, the specific component in lipid raft, was used as the hallmark of lipid raft. The distribution and expression of GM1 were probed by horse radish peroxidase (CTB-HRP) coupled to cholera β-subunit, and revealed by chemiluminiscent HRP substrates. The results indicate that the isolation method established in this study is simple and easy to operate, and also time saving compared with traditional sucrose density centrifugation.
    Ultrastructure of olfactory sensilla on the antenna of Proagopertha lucidula (Coleoptera: Scarabaeidae)
    2009, 52(1):  39-45. 
    Abstract ( 3231 )   PDF (4384KB) ( 955 )     
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    The ultrastructure of olfactory sensilla on the antenna of Proagopertha lucidula (Faldermann) adult was studied using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results showed that the olfactory sensilla concentrated on antennal lamellae and all these sensilla were situated in cuticle or in the sunken cavity formed by antennal cuticle depressions. Five types of olfactory sensilla in lamellae were found. There are mainly sensilla placodea (S. pl) in lamellae of P. lucidula. In addition to S. pl, there are sensilla basiconca (S. ba), sensilla cavitica (S. ca), sensilla coeloconica (S. co) and sensilla trichodea (S. tr). Based on the differences of plate and cone shape, S. pl are further classified into four subtypes, S. ba into two subtypes, and S. co into two subtypes. There are a plenty of pores and pore tubules in the cuticle of olfactory sensillum. The number of neurons in olfactory sensilla is inconsistent, including one olfactory receptor neuron (ORN), or two ORNs. The total number of sensilla in the male is 1.8 times that in the female. The number of S. bal in male is two times that in female, the number of S. co in female is four times that in male, while the number of S. pl in male is two times that in female. There is no significant difference in the number of S. co between male and female.
    Histopathological characteristics of a densovirus-like from silkworm, Bombyx mori.
    2009, 52(1):  46-51. 
    Abstract ( 2785 )   PDF (5956KB) ( 1882 )     
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    A small icosahedral nonenveloped DNA virus with the main biological and biophysical properties of densoviruses (DNVs) was isolated from silkworm Bombyx mori with abnormal larval mortality. Histopathological studies suggested that the virus first parasited on columnar cell of midgut, and then the nucleus of columnar cell became extremely inflated, or even broken. In situ hybridization results also demonstrated the virus was first reproduced in nucleus of columnar cells among midgut epithelial cells. It can also infect and reproduce in the goblet cells and most of the larval tissues at the last stage of infected silkworm.
    Establishment of baseline susceptibility data to various insecticides for aphids Rhopalosiphum padi(Linnaeus)and Sitobion avenae(Fabricius)(Homoptera: Aphididae) by the method of residual film in glass tube
    2009, 52(1):  52-58. 
    Abstract ( 3272 )   PDF (982KB) ( 1211 )     
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    Aim The objective of this research was to establish susceptible toxicity baselines of two wheat aphids Rhopalosiphum padi and Sitobion avenae to 22 insecticides. Methods The method of the residual film in glass tube with 3 h exposure to insecticides was employed for determining toxicity of insecticides to both wheat aphid species, which were originally collected from different provinces of China during 2005-2007 and reared on wheat seedlings in the laboratory for more than 30 generations. Results In R. padi, the LC50 values of susceptible toxicity baselines were 0.02 and 0.007 μg/cm2 for neonicotinoid insecticides, imidacloprid and acetamiprid respectively; 0.124 μg/cm2 for pymetrozin; 0.0026-0.70 μg/cm2 for carbamate insecticides, carbosulfan, thiodicarb, methomyl, pirimicarb and carbaryl; 0.005-0.065 μg/cm2 for organophosphate insecticides, triazophos, profenofos, omethoate, dimethoate, malathion, phoxim, dichlorvos and chlorpyrifos; 0.033-0.240 μg/cm2 for pyrethroid insecticides, lamba-cyhalothrin, beta-cypermethrin, deltamethrin, bifenthrin, fenvalerate and cypermethrin.In S. avenae, the LC50 values of susceptible toxicity baselines were 0.15 μg/cm2 and 0.12 μg/cm2 for neonicotinoid insecticides, imidacloprid and acetamiprid respectively; 0.41 μg/cm2 for pymetrozin; 0.005-0.76 μg/cm2 for carbamate insecticides carbosulfan, thiodicarb, methomyl, pirimicarb and carbaryl; 0.018-0.36 μg/cm2 for organophosphate insecticides, triazophos, profenofos, omethoate, dimethoate, malathion, phoxim, dichlorvos and chlorpyrifos and 0.20-2.94 μg/cm2 for pyrethroid insecticides, lambacyhalothrin, beta-cypermethrin, deltamethrin, bifenthrin, fenvalerate and cypermethrin. Conclusion Susceptible toxicity baselines of both R. padi and S. avenae to 22 insecticides established in this study could be used as a reference for resistance monitoring or other related researches. The insecticide susceptibility is higher in R. padi than in S. avenae for most of insecticides.
    Effects of selection by several insecticides on resistance levels and kdr allele frequency of housefly, Musca domestica (Diptera: Muscidae)
    2009, 52(1):  59-64. 
    Abstract ( 3462 )   PDF (966KB) ( 1018 )     
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    The high-level resistant strain (Cyh-R) of Musca domestica to cyhalothrin was selected by using deltamethrin and pirimiphos-methyl to study the effects of several insecticides on Cyh-R housefly strain and explore the methods of housefly resistance management. The knockdown resistance (kdr) allele frequencies of these housefly strains were detected with PCR amplification of specific allele PASAtechnology in order to study the relationship of kdr allele frequency and resistance level. The results showed that the toxicity of cyhalothrin to Cyh-R strain after selection with pirimiphos-methyl tended to decline. The LD50 value dropped from 2.8434 μg/housefly in F0 to 0.4404 μg/housefly in F8. However, the toxicity of cyhalothrin to Cyh-R strain after selected with deltamethrin tended to rise. The LD50 value increased from 2.8434 μg/housefly in F0 to 24.3249 μg/housefly in F16. These results suggest that the high-level resistance of housefly strain to cyhalothrin could decline with the application of organophosphate pesticide or without insecticides, while increase with the application of pyrethroids. The results of PASA showed that kdr allele frequency of Cyh-R strain was 88.8%. The kdr allele frequency decreased to 69.7% without insecticide selection and 78.8% after pirimiphos-methyl selection, respectively, while increased greatly after deltamethrin selection, amounting to 98.9%. The results further indicate that there is a positive correlation between kdr allele frequency and LD50 value of cyhalothrin.
    Analysis of spatio-temporal dynamics of Laodelphax striatellus (Fallén) (Homoptera: Delphacidae) and spiders in paddy fields based on geostatistics
    2009, 52(1):  65-73. 
    Abstract ( 2954 )   PDF (1993KB) ( 1094 )     
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    Field studies were conducted to compare the spatio-temporal dynamics of small brown planthopper (SBPH) Laodelphax striatellus (Fallén) and their important natual enemy spiders by using the geo-statistical methods. By fitting the models of semivariogram of them and analyzing the patterns, it was found that the nugget range of macropterous SBPH and larval SBPH were approaching to that of spiders, as well as the range of structural intensity, but the nuggets of SBPH were not approaching to that of spiders, neither the structural intensity. The spatial pattern of larval SBPH and spiders were aggregation, but that of macropterous SBPH was random sometimes. We drawn the predicted surface maps with the Ordinary Kriging method and illustrated in the time order to provide a visible presentation of the spatio-temporal distributions. By analyzing those maps, we found that the spatial distribution of spiders in the paddy field was similar to that of macropterous SBPH and larval SBPH, and was more similar to larval SBPH, and showed an obvious pursuing phenomenon between them, while the pursuing had a close relationship with the temporal dynamics of SBPH number. But the pursuing phenomenon was complex and influenced by factors such as the weather, the environmental conditions in paddy fields, other enemies of SBPH and other foods of spiders.
    Molecular phylogenetic analysis of Elateridae (Insecta: Coleoptera) based on 28S rDNA gene fragments
    2009, 52(1):  74-83. 
    Abstract ( 3316 )   PDF (1581KB) ( 1087 )     
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    Aims To study the molecular phylogenetic relationships of click beetles (Elateridae) based on partial sequences of nuclear 28S ribosomal DNA and comparison with traditional taxonomy results. All these could lay the foundation for the confirmation and further amendment of classification of Elateridae. Methods The fragments of 28S rDNA gene were sequenced from 10 samples for 9 species (including two geographical populations), of Elateridae, and the sequences for other 32 species of Elateridae were downloaded from GenBank. Molecular phylogenetic trees were reconstructed using software DNA Star Lasergene v 7.1.0 and MEGA4.0 (NJ, MP and ME methods). Results Sequence analysis showed that there were 477 conserved sites, occupying 56.1% of all sites, and 291 parsim-informative sites, occupying 34.2% of all sites. The average content of G+C was 63.9%, obviously higher than A+T, showing that the base compositions were biased in favor of G+C. Transition value is slightly higher than transversion. The genetic distances within subfamilies ranged from 0.000 to 0.130, which were obviously smaller than that among subfamilies. The molecular phylogenetic trees indicated that the 10 subfamilies analyzed were divided into four clades: cladeincluded Elaterinae and Melanotinae, cladeincluded Pyrophorinae, Agrypninae and Conoderinae, clade included Cardiophorinae and Negastriinae, while clade included Denticollinae, Oxynopterinae and Pityobiinae. All ingrou Ptaxa formed two lineages: lineage 1 (clade) and lineage 2 (clades++). Senodonia quadricollis always represented a single lineage, which was separated obviously from other click beetles. Conclusions This study confirms that traditional taxonomy based on the morphological characteristics of adults and larvae is feasible. At first, the family Elateridae has been proved to be a monophyletic group. The second, the 10 subfamilies of Elateridae can be distinctly divided into four clades. The third, the subfamily Cardiophorinae has been proved to be a monophyletic group, but the other subfamilies were found to be paraphyletic groups, and especially the assignment of Senodonia quadricollis needs further study. The analysis of 28S rDNA gene sequence is proved to be a good way to study the phylogenetic relationships of click beetles.
    Autophagy and apoptosis in insect cells during metamorphosis
    2009, 52(1):  84-94. 
    Abstract ( 4210 )   PDF (5240KB) ( 1832 )     
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    During insect metamorphosis, degeneration or dissolution of larval tissues was involved in programmed cell death PCD including autophagy and apoptosis induced by ecdysteroids. Autophagy, which usually responds to hungry to avoid cell death, appears as a low-level Cvt vesicle and autophagosome  to swallow and dissolve partial cytosolprotein aggregates and cellular organelles. PCD pathway is initiated by ecdysteroids and regulated by genetic cascade system during the period of insect metamorphosis. The initiation of PCD pathway by ecdysteroids turns the low-level normal autophagy to high-level one and triggers apoptosis at the same timeand leads to irreversible death. Subsequently larval tissues degenerate or dissolve completely. The most important discoveries of PCD in fruit fly Drosophila metamorphosis include: 1In the molecular pathways of PI3KI-Tor and PI3KIII, high-level autophagy initiated by Atg1 autophagy-related protein 1 can induce apoptosis; 2 Transcription factors induced by ecdysteroids, including βFTZ-F1, E93, BR-C and E74A, can not only activate the apoptosis pathway, but also induce autophagy pathway.
    Research progress in insect alkaline phosphatases
    2009, 52(1):  95-105. 
    Abstract ( 4271 )   PDF (1857KB) ( 1353 )     
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    Insect alkaline phosphatases are distributed in such insect organs and tissues as head, salivary gland (saliva), intestine, Malpighian tube, cuticule, haemolymph, fat body, reproductive system and appendage. They may be involved in the process of insect development, nerve conduction, hormone synthesis, substance metabolism and diapause, and caste formation in social insects, etc. Alkaline phosphatases can make insect more resistant to insecticides, especially to Bt toxins. They can also be the target enzymes for some insecticides. Some biological substances can affect the enzyme activity, so can some viruses and fungi. Studies on the insect alkaline phosphatases may help the understanding of biochemical mechanism and metabolic process in insects, and also provide new ideas for pest management and the rearing of resource insects. Research progress in insect alkaline phosphatases is reviewed, and the biochemical properties of the enzymes and their relationship with physiological function are also described.
    Localization of the specific allergens in the body of Blomia tropicalis (Acari: Echymiopodidae) by immunohistochemistry
    2009, 52(1):  106-109. 
    Abstract ( 3553 )   PDF (1848KB) ( 842 )     
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    【Aims】 To localize the specific allergens in the body of Blomia tropicalis by immunohistochemistry. 【Methods】 Paraffin sections of B. tropicalis mites stained by hatmatoxylin-osin (HE) were observed by optical microscope. Serum specific IgE antibodies from patients who were sensitive to the house dust mite served as probe, and the specific allergens in the body of B. tropicalis were localized. 【Results and Conclusion】 Immunohistochemistry staining revealed that the specific allergens were located in the gut wall, gut contents and the reproductive system, but strongly bound to the midgut wall and contents.
    Fumigation toxicity of eucalyptus oil and its active compound against dried fish pest, Dermestes maculatus Degeer(Coleoptera: Dermestidae)
    2009, 52(1):  110-115. 
    Abstract ( 4247 )   PDF (816KB) ( 1335 )     
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    To assess the insecticidal activity of eucalyptus oil and α-pinene against dried fish pest Dermestes maculates. Mortalities of its larvae and adults under different doses or treatment time were investigated by bioassay method. The results showed that the tested oil and its active ingredient (α-pinene) affected significantly (P<0.001) on the mortality of both the larvae and adults of D. maculatus. Responses varied with compound, dose and exposure time. Over 90% mortality at 72 h after treatment was achieved with the eucalyptus oil against larvae at a dose of 32 μL/cm3. Eucalyptus oil at 32 μL/cm3, were highly toxic to adults of D. maculatus 72 h after treatment. α-Pinene also showed highly toxic effects on both larvae and adults of the beetles at the same exposure time and doses. It is also interesting to mention that the adults of D. maculatus were found to be more tolerant than larvae. In probit test eucalyptus oil was much more effective than α-pinene. These naturally occurring materials could be useful for managing populations of D. maculatus.
    A preliminary study on the ultrastructure and chemical components of egg surface of the Chinese honeybee, Apis cerana cerana
    2009, 52(1):  116-120. 
    Abstract ( 3061 )   PDF (1942KB) ( 1086 )     
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    To understand the worker policing mechanism of honeybees, we compared the surface ultrastructure of queen-laid diploid and haploid eggs of Apis cerana cerana to that of worker-laid eggs using SEM and conducted a detailed comparative analysis on chemical components of newly laid (0 h) and 5 h eggs of various types by GC-MS. The results show that the egg surface is covered with an outer layer typified by hexagonal structures filled with small nodules and there are no differences between different types of eggs. The chemical components coating queen eggs, diploid and haploid, are more abundant than those of workers. C27∶2 and C27∶1 may be the chemical pheromones on the egg surface.