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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 September 2010, Volume 53 Issue 9
For Selected: View Abstracts Toggle Thumbnails
  • RESEARCH PAPERS
    Changes in the expression of the CYP3 family genes under the induction of ecdysone in Bombyx mori
    GAO Rui-Na
    2010, 53(9):  943-948. 
    Abstract ( 3294 )   PDF (874KB) ( 1066 )     
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    To study the change in expression of the CYP3 family genes induced by ecdysone, we measured their transcription level by dual spike-in qPCR in the midgut and fat body of Bombyx mori 5th instar larvae fed on mulberry leaves soaked in ecdysone solution (2×10-3μg/μL) with those fed on mulberry leaves not soaked in ecdysone solution as the control. The results showed that compared with the control, the transcription level of CYP302, CYP306 and CYP339 increased by 191.4, 7.4 and 421 times in fat body, respectively, and that of the other genes induced by ecdysone did not change significantly or was not detectable. These results suggest that CYP339 gene may be involved in the metabolism of silkworm ecdysone. This work laid a theoretical foundation for further study on the relationship between P450 genes and the metabolism of endogenous substances.

    Expression profile and function of tyrosine hydroxylase gene BmTh in the silkworm, Bombyx mori
    HAN Yu
    2010, 53(9):  949-956. 
    Abstract ( 3696 )   PDF (7671KB) ( 1101 )     
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    As a key enzyme of catecholamine synthesis, tyrosine hydroxylase is an indispensable enzyme of metabolism in insects, mammals and human. Tyrosine hydroxylase involves in the synthesis of melanin, affects the body color and insect exoskeleton hardening. In order to explore the physiological and biochemical function of tyrosine hydroxylase, in this study, the structure, expression profile and function of tyrosine hydroxylase gene were studied in the silkworm, Bombyx mori, using the combined methods of bioinformatic analysis and experiments. The bioinformatic analysis based on the silkworm genome database and microarray database indicated that the silkworm tyrosine hydroxylase gene BmTh is located on chromosome 1. BmTh includes 8 exons and encodes a protein with 561 amino acid residues. Microarray data in different tissues showed that it has significant expression signals in head and epidermis of day-3 5th instar larvae, which was verified by RT-PCR. In situ hybridization analysis showed that there was remarkable signal on the rim of head and epidermis with paraffin tissue section and RNA probe. Through injecting an inhibitor of tyrosine hydroxylase (3-indole-L-tyrosine, 3-IT) at different concentrations into silkworm larva at the mounting stage, we found that 20 mmol/L 3-IT had no influence on the larval development, 50 mmol/L 3-IT could handicap the larval metamorphosis and pupation, and 100 mmol/L 3-IT could cause the larval death and turn the larval body color into black. These results suggest that tyrosine hydroxylase plays an important role in the processes of metamorphosis and it is an indispensable key enzyme for normal development in the silkworm.

    Fusion expression and purification of pyridoxal kinase of Bombyx mori
    ZHANG Ping-Ping
    2010, 53(9):  957-961. 
    Abstract ( 2970 )   PDF (859KB) ( 988 )     
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    Aim Pyridoxal kinase (PLK, EC 2.7.1.35) is a key enzyme related to vitamin B6 metabolism. In this study, recombinant pyridoxal kinase of Bombyx mori will be expressed, which will lay a foundation for further study on regulation mechanism, catalytic mechanism and structure of PLK in B. mori.Methods After fusion expression vector harboring PLK gene of B. mori was constructed, the recombinant vector was transformed to Escherichia coli Rosetta for induction and expression, and then the characteristics of the purified recombinant protein was analyzed after the expression product was purified using affinity chromatography by Ni2+ column. Results Only one band was observed when the purified PLK was separated on SDS-PAGE. The PLK was enriched by 40fold and its specific activity was 1 800 U/mg. The optimum temperature and pH of this enzyme was about 50, and 5.5-6, respectively, and Zn2+ was the effective activator in enzymatic reaction. Conclusion The recombinant PLK exhibited the same catalytic characteristics as the native protein obtained from tissues of B. mori.

    Molecular cloning, characterization, and expression pattern of chemosensory protein 1 gene (Acer-CSP1) in the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae)
    LI Hong-Liang
    2010, 53(9):  962-968. 
    Abstract ( 3990 )   PDF (4251KB) ( 1031 )     
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    Chemosensory proteins (CSPs) are a class of the most important proteins involved in chemical communications and perception in insects. In this study, one CSP gene from the Chinese honeybee, Apis cerana cerana, was cloned and characterized, which was named Acer-CSP1 (GenBank accession no.: FJ157352). The Acer-CSP1 cDNA was 351 bp in length, encoding 116 amino acid residues with the predicated molecular mass of approximately 13.85 kD and pI of 4.89, of which four conservative cysteines are included. With the general characteristics of the CSP gene family in insects, AcerCSP1 showed high similarity (99.1%) with the CSP1 gene of A. mellifera and moderate similarity (45.3%-68.0%) with the CSP genes of other ten insects analyzed with the deduced amino acid sequences. The expression of the Acer-CSP1 in different sensory organs of A. cerana cerana was compared using both relative (2-ΔΔCt) and absolute quantitative analyses of real-time PCR. The results indicated that the Acer-CSP1 was primarily expressed in the antennae, followed by the head, but least expressed in other organs (wings, legs, thorax, and abdomen). Since the antennae are the major olfactory organs and the head is the major sensory and gustatory organ of the honeybee, the results suggest that the Acer-CSP1 gene is most likely involved in the olfaction, gestation and other chemosensory functions of the insect.
    Effects of cadmium on hemocyte number, encapsulation and morphology in Boettcherisca peregrina (Diptera: Sarcophagidae)
    LI Yan-Min
    2010, 53(9):  969-977. 
    Abstract ( 4095 )   PDF (5298KB) ( 1138 )     
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     In order to explore the effects of heavy metals on insect cellular immunity and its related mechanism, the differences in the number, viability, spreading and encapsulation rates, and morphology of hemocytes between the Cd2+ treated (at the concentration of 150 μg/g) Boettcherisca peregrina and its control were investigated and compared, respectively, using the methods as hemocytes counting, typan blue staining, spreading and encapsulation assays, as well as microstructure and ultrastructure observations. The results indicated that at 24, 48, 72 and 96 h post feeding the neonate larvae of B. peregrina with the artificial diet containing Cd2+, both total hemocyte count and hemocytes viability in the larvae were significantly decreased compared with the control, respectively. In contrast, the in vitro spreading rate and encapsulation capability of hemocytes of the larvae exposed to Cd2+ were significantly lower than those of the control after 72 and 48 h exposure, respectively, and then showed no significant differences with the control. Additionally, the in vivo encapsulation capability of hemocytes of the pupae that developed from the larvae exposed to Cd2+ was also significantly lower than that of the control. The morphological observation results showed that compared with the control, no marked alternation in the microstructure was observed for each type of hemocytes (including prohemocyte, plasmatocyte, granulocyte and oenocytoid), except that some plasmatocytes neither spread nor extended their pseudopods. However, the ultrastructure of each type of hemocytes in the exposed larvae was found to be alternated to various degrees compared to the control, with the main ultrastructural alternation including: damage or break of cell membrane, condensation of chromatin, sharply decreasing of the number of mitochondria, endoplasmic reticulum and other organelles, and the emergence of vacuoles in cytoplasm. The results suggest that heavy metal Cd2+ may have toxic effects on hemocytes of B. peregrina.

    Relationship of WO phage and Wolbachia infection in Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae)
    ZHOU Li-Li
    2010, 53(9):  978-984. 
    Abstract ( 3021 )   PDF (1038KB) ( 920 )     
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     WO phage is the bacterial virus infecting Wolbachia in arthropods. It is supposed that WO phage may participate in controlling genetic changes of the hosts. Laodelphax striatellus planthoppers from four geographic populations of China (Minhang, Shanghai; Puer, Yunnan; Jining, Shandong; Qingtongxia, Ningxia) were bred on rice treated with antibiotics. The infection rates of Wolbachia and WO phage were examined every twenty days, and the infection relationships between Wolbachia and WO phage were analyzed based on the data obtained. The results revealed that the infection rate of Wolbachia declined with time, and so was WO phage. By using Real-time quantitative PCR, we found that the copy numbers of Wolbachia and WO phage varied with the developmental stages of adult female, displaying the highest quantity at 8 d post infection, and WO phage density showed the same pattern as that of Wolbachia. From the results, we inferred that WO phage is the obligate virus infecting Wolbachia, and the WO phage infecting Wolbachia in L. striatellus may be lysogenic.

    Isolation and characterization of the insecticidal constituent acorone from Acorus calamus (Araceae)
    YAO Ying-Juan
    2010, 53(9):  985-992. 
    Abstract ( 2894 )   PDF (1099KB) ( 1447 )     
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    Susceptibilities of the green plant bug, Lygus lucorum Meyer-Dür collected from five cotton-growing regions in Shandong Province, China, to endosulfan, malathion, chlorpyrifos, bifenthrin, methomyl, carbosulfan, imidacloprid and fipronil, were determined by glass-vial bioassay in 2009 in order to choose efficient insecticides for controlling green plant bugs in different regions. Dose-response results indicated that L. lucorum from five cotton-growing regions were susceptible to malathion, chlorpyrifos, carbosulfan, bifenthrin and imidacloprid at LC50 values. Toxicity data indicated that L. lucorum collected in Liaocheng (LC) was more susceptible to methomyl, endosulfan and fipronil than those in other four regions. The results indicated that the LC50 values of Binzhou (BZ) population for methomyl, endosulfan and fipronil were 5.12-, 6.04- and 39.80-fold higher than those of LC population, respectively, and the LC50 values of Qufu (QF) population for methomyl, endosulfan and fipronil were 22.12-, 5.48- and 22.80-fold higher than those of LC population, respectively. The populations collected in BZ and QF was less susceptible to methomyl, endosulfan and fipronil. The toxicity sequence of 8 insecticides to L. lucorum was ranked as follows: fipronilmethomyl, bifenthrin, endosulfanmalathion, chlorpyrifoscarbosulfanimidacloprid. The susceptibilities of L. lucorum adults from Dezhou (DZ) to 8 insecticides were hardly changed during July and October in 2009. The current study may provide a better understanding of insecticide efficacies to L. lucorum and develop efficient pest management strategies in those cotton-growing regions.

    Susceptibilities of Lygus lucorum Meyer-Dür (Hemiptera: Miridae) from five cotton-growing regions in Shandong,China to selected insecticides
    GUO Tian-E
    2010, 53(9):  993-1000. 
    Abstract ( 3949 )   PDF (1046KB) ( 1023 )     
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    Susceptibilities of the green plant bug, Lygus lucorum Meyer-Dür collected from five cotton-growing regions in Shandong Province, China, to endosulfan, malathion, chlorpyrifos, bifenthrin, methomyl, carbosulfan, imidacloprid and fipronil, were determined by glass-vial bioassay in 2009 in order to choose efficient insecticides for controlling green plant bugs in different regions. Dose-response results indicated that L. lucorum from five cotton-growing regions were susceptible to malathion, chlorpyrifos, carbosulfan, bifenthrin and imidacloprid at LC50 values. Toxicity data indicated that L. lucorum collected in Liaocheng (LC) was more susceptible to methomyl, endosulfan and fipronil than those in other four regions. The results indicated that the LC50 values of Binzhou (BZ) population for methomyl, endosulfan and fipronil were 5.12-, 6.04- and 39.80-fold higher than those of LC population, respectively, and the LC50 values of Qufu (QF) population for methomyl, endosulfan and fipronil were 22.12-, 5.48- and 22.80-fold higher than those of LC population, respectively. The populations collected in BZ and QF was less susceptible to methomyl, endosulfan and fipronil. The toxicity sequence of 8 insecticides to L. lucorum was ranked as follows: fipronilmethomyl, bifenthrin, endosulfanmalathion, chlorpyrifoscarbosulfanimidacloprid. The susceptibilities of L. lucorum adults from Dezhou (DZ) to 8 insecticides were hardly changed during July and October in 2009. The current study may provide a better understanding of insecticide efficacies to L. lucorum and develop efficient pest management strategies in those cotton-growing regions.
    Herbivore resistance induced by Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) and its relation to the JA signaling pathway in Chinese cabbage (Brassica campestris L.)
    HU Liu-Cheng
    2010, 53(9):  1001-1008. 
    Abstract ( 3955 )   PDF (1331KB) ( 1053 )     
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    When attacked by herbivores, plants produce defense responses, and the jasmonic acid (JA) signaling pathway plays an important role in this process. So far, however, little was known about the herbivore-induced defense responses in Chinese cabbage (Brassica campestris L.). Therefore, the defense responses of B. campestris attacked by Spodoptera litura (Fabricius) and their relations to the JA signaling pathway were studied through assaying the contents of JA and trypsin protease inhibitors (TrypPIs) in B. campestris plants. The results showed that infestation by S. litura resulted in systemic increases in levels of both JA and TrypPIs in B. campestris plants, of which the level of TrypPIs could also be systemically induced by exogenous application of methyl jasmonate (MeJA). Moreover, body weights of S. litura caterpillars fed on leaves treated by MeJA or infested by the herbivore decreased significantly, only 67.5% and 60.2% of that fed on control leaves, respectively. Treatment with wounding plus the caterpillar oral spit (OS) could cause the increase in JA and TrypPIs level in the treated leaves, but its induction efficacy was similar to that induced by wounding plus water (W) and far lower than that induced by the herbivore infestation. The levels of JA and TrypPIs in OS- or W-treated plants were 68.4% and 62.9%, and 22.4% and 36.9% of those in control plants, respectively. We also found that there was no significant difference in the induction efficacy between the treatments wounding once and wounding multiple times”. The results suggest that the defense responses of B. campestris plants induced by the herbivore are related to the JA signaling pathway, and elicitation of the responses is probably involved in the specific feeding behavior of the herbivore.

     

    Leaf physiological responses in the London plane tree (Platanus×acerifolia) (Platanaceae) to injury by the sycamore lace bug, Corythucha ciliata (Hemiptera: Tingidae)
    JU Rui-Ting
    2010, 53(9):  1009-1014. 
    Abstract ( 3553 )   PDF (867KB) ( 934 )     
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    The sycamore lace bug, Corythucha ciliata, is a newly found invasive insect in China. To examine the effects of piercing-sucking by C. ciliata on its host plants, we measured several physiological and biochemical traits of the London plane tree (Platanus×acerifolia) leaves injured by the bug using photosynthetic apparatus and colorimetric analysis. The results showed that with increasing leaf injury by the bug, the photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr) in Pacerifolia generally decreased. Positive relation existed between leaf injury and intercellular CO2 concentration (Ci) (P< 0.01). After the leaf tissue of Pacerifolia was pierced and sucked by C. ciliata adults, the chlorophyll a, b and carotenoid contents decreased compared with those of the control. There was a negative relation between leaf injury and chloroplast pigment content (P< 0.01). With increasing leaf injury, soluble sugar content decreased, but no consistent pattern of free proline content in leaves was observed. The results suggest that photosynthesis is weakened and some other physiological and biochemical parameters are changed in Pacerifolia leaves injured by C. ciliata.
    Nectarrobbing behavior of Megachile (Xanthosaurus) japonica Alfken (Hymenoptera: Megachilidae) on its nectar plant Thermopsis lupinoides (Leguminosae)
    YANG Jun-Wei
    2010, 53(9):  1015-1021. 
    Abstract ( 3770 )   PDF (4886KB) ( 983 )     
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    【Aim】 To investigate the nectarrobbing behavior of Megachile (Xanthosaurus) japonica Alfken on its nectar plant Thermopsis lupinoides (L.)in Mu Us sandland, Inner Mongolia.【Methods】 During the flowering season of T. lupinoides, we investigated the main pollinators and observed the robbing behavior of M. japonica on T. lupinoides, and recorded the position of holes in the corolla tube chewed by robbers at the base of flowers near the nectar spur. 【Results】 M. japonica is the dominant pollinator of T. lupinoides. As primary robbers, M. japonica bees cut a longitudinal split at the base of flowers by mandibles and obtain nectar through it, while directly obtain nectar without making holes when they act as secondary robbers. They did not show an tendency to seek pre-cut robbing holes, but individuals displayed a preference for one side of the flowers. In 13 plots, the percentage of nectar robbing from blooming flowers was from 95.4% to 100%, and only reached 64.7% in buds. 【Conclusions】 M. japonica is not only the main pollinator but also the primary robber and secondary robber of T. lupinoides in Mu Us sandland.
    Genetic diversity of Conogethes punctiferalis (Guenée) (Lepidoptera: Crambidae) populations from different geographic regions in China
    ZHANG Ying
    2010, 53(9):  1022-1029. 
    Abstract ( 3820 )   PDF (1969KB) ( 942 )     
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    Genetic diversity among eleven geographic populations of Conogethes punctiferalis in China was investigated using InterSimple Sequence Repeat (ISSR) markers in this study. Six of the 34 used primers were chosen for our ISSR polymorphic analysis. The results showed that 209 bands were polymorphic, making up 99.05% of the total 211 amplified bands. The genetic distances between different C. punctiferalis populations were 0.0059-0.0237. The Nei’s index, Shannon information index, and coefficient of genetic (gene) differentiation among populations (Gst) was 0.1750, 0.2966 and 0.053, respectively, and the estimated value of gene flow from Gst was 8.8724. The results suggest that C. punctiferalis populations in China keep a low level of population genetic differentiation due to a considerable gene flow.
    Phylogenetic relationship among the species of Anopheles subgenus Cellia (Diptera: Culicidae) in China: inferred by mitochondrial and ribosomal DNA sequences
    WU Jing
    2010, 53(9):  1030-1038. 
    Abstract ( 3545 )   PDF (1288KB) ( 1109 )     
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    【Aim】 To reconstruct the phylogeny of the known species of Anopheles subgenus Cellia from China based on mtDNA-COⅡ and rDNA-28S-D3 sequences, and to interpret the interspecific genetic relationship among them. 【Methods】 mtDNA-COⅡ and rDNA-28S-D3 genes of 20 species in subgenus Cellia and 2 species in subgenus Anopheles of genus Anopheles were sequenced and analyzed. The trees of neighbor joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) of these species were constructed based on COⅡ, D3 and COⅡ with D3 conjunction dataset, with Anopheles (An.) sinensis and An. (An.) hyrcanus as the outgroup. 【Results】 The length range was 685 bp for mtDNA-COⅡ and 375-410 bp for rDNA-28S-D3, and the level of pairwise genetic distances among Cellia mosquitoes was 0.015-0.117 for COⅡ and 0.003-0.111 for D3 gene. All the phylogeny trees showed that the outgroup was divergent. The Neomyzomyia series was monophyletic, but paraphyletic in COⅡ tree. The relationship between Myzomyia and Neomyzomyia was close. The 50% consensus tree by COⅡ with D3 conjunction dataset showed that the 20 species in subgenus Cellia from China were clustered to 4 major clades, and all members of each series and group were respectively clustered together with above 50% bootstrap value, except Anopheles (Cellia) pseudowillmori not clustered within the maculatus group. 【Conclusion】 This molecular phylogeny clearly showed the interspecies and interseries genetic relationship among Chinese subgenus Cellia mosquitoes, and could be useful for the taxonomy and control measures.
    Effects of flufenoxuron on the expression of chitin synthase gene in Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae) and Oxya chinensis (Thunberg) (Orthoptera: Oedipodidae)
    LIU Xiao-Jian
    2010, 53(9):  1039-1044. 
    Abstract ( 3478 )   PDF (1343KB) ( 1016 )     
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    In order to explore the possible target and mechanism of flufenoxuron, important agricultural pests Locusta migratoria manilensis and Oxya chinensis were used as test insects in this study. A partial fragment of chitin synthase 1 cDNA from O. chinesis (OcCHS1) was amplified using a pair of degenerate primers. With the mid-2nd instar nymphs of O. chinesis and the 1st, 2nd, 3rd instar nymphs of L. migratoria manilensis dipped in flufenoxuron as treatments and acetone treatment as the control, the effects of flufenoxuron on the mRNA expression of chitin synthase genes were analyzed using RT-PCR and real-time quantitative PCR. The partial cDNA fragment of OcCHS1 (GenBank accession number: HM214491) was obtained, which consists of 312 nucleotides that encode 104 amino acid residues. The deduced amino acid sequence of OcCHS1 showed 95% identity with those of LmCHS1. The results of RT-PCR and real-time quantitative PCR analysis showed that the expression of OcCHS1 in the mid-2nd instar nymphs of O. chinesis and LmCHS1 in the 1st, 2nd and 3rd instar nymphs of L. migratoria manilensis exposed to flufenoxuron increased 102%, 34%, 82% and 89%, respectively, compared to the control. Increased expression of CHS1 mRNA may be due to compensation response of the CHS1 gene at the transcriptional level that is caused by the retarded chitin synthesis. It is so inferred that chitin synthase may be one of targets of flufenoxuron.
    Detection and phylogenetic analysis of Wolbachia in the Asian citrus psyllid (Diaphorina citri) (Homoptera: Psylloidea) populations in partial areas in China
    WANG Zhong-Kang
    2010, 53(9):  1045-1054. 
    Abstract ( 3515 )   PDF (1738KB) ( 1074 )     
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     Wolbachia is a common and widespread group of symbiotic bacteria found in arthropods, which is transmitted through the egg cytoplasm and can manipulate reproduction in their hosts in various ways. So, it has important potential significance on the research of evolution of arthropods and may be used in pest bio-control. Taking the Asian citrus psyllid (Diaphorina citri), the vector of the Huanglongbing(HLB) pathogen, as the research object, we detected the natural populations of Diaphorina citri from five areas, i.e., Beihai and Liuzhou in Guangxi, Shenzhen and Yangchun in Guangdong, and Fuzhou in Fujian, based on PCR amplification, cloning and sequencing of 16S rRNA, ftsZ and wsp genes. The obtained sequences were aligned with the published ones in database of NCBI, and the phylogenetic relationships among Wolbachia types found in D. citri and other insect species were analyzed. The results indicated that all the five populations of D. citri were infected by Wolbachia. The three gene sequences of Wolbachia and the phylogenetic tree analysis showed that Wolbachia population in D. citri belongs to supergroup B.  Further phylogenetic analysis based on the wsp gene sequences showed that the Wolbachia of D. citri belongs to Con subgroup. The 16S rRNA, ftsZ and wsp gene sequences were almost identical in the five geographic populations, respectively, and the identity is up to 99%, showing that these three gene sequences of Wolbachia in different geographic populations of D. citri had no significant difference. The results can provide useful information for understanding evolution of citrus psyllids and their integrated control.

    Application of ARIMA and SVM hybrid model in pest forecast
    XIANG Chang-Sheng
    2010, 53(9):  1055-1060. 
    Abstract ( 4235 )   PDF (979KB) ( 2486 )     
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    The data of pest occurrence are complicated and unpredictable time series. The linear or nonlinear features of pest time series can not be captured based on single prediction model. A new hybrid forecasting model based on autoregressive integrating moving average (ARIMA) and support vector machine (SVM) is proposed in this paper. ARIMA model was used to predict the linear component while SVM model was used for the nonlinear residual component of pest time series, and then the hybrid forecasting results were obtained. The prediction performances of the method were tested on Dendrolimus punctatus occurrence area. The results show that the hybrid model, which combines the respective advantages of both linear and nonlinear models, has better accuracy than any single model. Hybrid model is a good and effective method for pest forecasting.