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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 2012, Volume 55 Issue 9
For Selected: View Abstracts Toggle Thumbnails
    Identification and expression profiling of regulatory molecules involved in immune homeostasis in the silkworm, Bombyx mori
    WANG Fei, LI Ya-Ming, HUA Xiao-Ting, XIA Qing-You
    2012, 55(9):  999-1007. 
    Abstract ( 3161 )   PDF (14725KB) ( 1435 )     
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    Maintenance of insect immune homeostasis requires prompt activation and down-modulation of the key transcriptional factors: Dorsal/Dif in Toll signaling pathway or Relish in IMD signaling pathway. Several regulatory molecules which modulate the stability or activity of the transcriptional factors in immune response have been identified in Drosophila and some other insects. Mutation or silencing of these molecules leads to over activation of immune system. So far there is no report about the regulatory molecules limiting immune signaling in the silkworm, Bombyx mori. In this study, several molecules which are predicted to be involved in immune homeostasis, including Wnt family members, Ubc9, FAF and POSH, were identified in the silkworm genome by comparative genomic analysis. The expression patterns of these molecules in multiple tissues after microbial infection were recorded, and the results showed that their expression levels generally decreased after microbial infection. Although an increase of more than 1.5-folds in expression level was observed in certain tissues, a rapid decrease followed and the high level was not maintained. Interestingly, the correspondence between the expression patterns of these molecules and the particular signaling pathway that microbes induced varied in different tissues. This is the first report of the regulatory molecules involved in silkworm immune homeostasis, which provides reference for further investigation on the molecular mechanism of immuno-negative modulation in the silkworm.
    Temporal and spatial expression profiles and the microbial sensitivity of antimicrobial peptide genes Cecropin and Moricin from Spodoptera litura (Lepidopetera: Noctuidae)
    XU Xiao-Xia, WANG Shuang, ZHANG Wen-Qing, SUN Qiang, WANG Chao, JIN Feng-Liang
    2012, 55(9):  1008-1013. 
    Abstract ( 3710 )   PDF (5507KB) ( 1260 )     
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    Antimicrobial peptides (AMPs) are small molecular weight proteins with broad spectrum antimicrobial activity against bacteria, viruses, and fungi. In the present study, the real-time PCR and semi-quantitative RT-PCR were used to check the temporal and spatial expression patterns and the microbial sensitivity of genes of two very important antimicrobial peptides, moricin and cecropin, from Spodoptera litura to microbes. The real-time PCR result indicated that Moricin and Cecropin were expressed during the whole developmental stages, and the expression level increased with the development, with the highest expression level in the 6th instar larvae. The semi-quantitative RT-PCR result showed that both Cecropin and Moricin were expressed in the seven tissues including trachea, testis, Malpighian tubes, integument, midgut, fat body and hemocyte, and Cecropin had the highest expression level in fat body while Moricin had a relatively higher expression level in hemocyte. The temporal expression result detected by semi-quantitative RT-PCR showed that the expression of Cecropin and Moricin increased significantly within 48 h after induction with Escherichia coli, and reached the peak at 8 h after induction. Real-time PCR result showed that the transcript level of Moricin and Cecropin increased significantly after induction with different microbes (E. coli, Staphyloccocus aureus and Beauveria bassiana), respectively, and most sensitive to the induction of E. coli. The results of this study lay the foundation for further function research of Cecropin and Moricin from S. litura.
    Cloning and expression profiling of the esterase gene AlucEST1 in the green plant bug, Apolygus lucorum (Hemiptera: Miridae)
    SHEN Chen, GU Shao-Hua, WU Hong-Zhen, HAN Rong, ZHANG Yong-Jun, GUO Yu-Yuan
    2012, 55(9):  1014-1021. 
    Abstract ( 2751 )   PDF (4684KB) ( 1137 )     
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    Odorant degrading enzymes (ODEs) can degrade the excess odor molecules so as to prevent olfactory receptors from the sustained stimulation. Esterase is a kind of odorant degrading enzyme, and plays an important role in the process of insect olfactory behavior. In this study, a novel putative ODE gene AlucEST1 (GenBank accession number: JQ715614) from the green plant bug, Apolygus lucorum was cloned by using rapid-amplification of cDNA ends (RACE) technique. Sequence analysis showed that AlucEST1 is 1 810 bp in length with the open reading frame of 1 713 bp, and encodes 571 amino acids. The N-terminal hydrophobic region contains a 17 amino acid signal peptide. Semi-quantitative RT-PCR and Real-time qPCR were conducted to explore the AlucEST1 expression in different tissues of A. lucorum. The results showed that AlucEST1 was expressed in various tissues including antennae, head, thorax, abdomen, legs and wings. AlucEST1 had the highest expression level in thorax and abdomen, and was also abundantly expressed in the antennae. It is so inferred that the gene plays an important role in the degradation of odor molecules and is also involved in a variety of metabolic regulation in A. lucorum.
    Cloning and expression profiling of aminopeptidase N encoding gene in the larval midgut of Sesamia inferens (Lepidoptera: Noctuidae)
    WANG Xing-Yun, MA Wen-Jing, HAN Lan-Zhi, HOU Mao-Lin
    2012, 55(9):  1022-1030. 
    Abstract ( 3058 )   PDF (7360KB) ( 1271 )     
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    Aminopeptidase N (APN) is located in the brush border membrane vesicles (BBMV) of insect midgut. It is the major receptor of insecticidal crystal proteins from Bacillus thuringiensis (Bt) and is closely related to the resistance of insects to Bt toxin. To clarify the relationship of APN and the resistance of Sesamia inferens to Bt toxin, the full-length APN-encoding cDNA sequence was cloned from BBMV of S. inferens midgut by degenerate PCR and RACE techniques and named as SiAPN3 (GenBank accession no.: HQ636624). The relative expression levels of SiAPN3 in different larval gut tissues and instars of S. inferens were determined by Quantitative Real-time PCR (qPCR) assays. The results showed that SiAPN3 is 3 411 bp in full-length and consists of an opening reading frame of 3 018 bp that encodes a protein of 1 006 amino acid residues. The predicted molecular weight and isoelectric point are 114 kDa and 4.95, respectively. The putative protein sequence includes one N-linked and twelve O-linked glycosylation sites, a zinc metal binding site motif of HEXXHX18E that is typical of the active sites of zinc-dependent metalloproteases, and a highly conserved GAMEN motif that forms part of the active site. In addition, it contains a cleavable N-terminal signal peptide with 18 amino acids, as well as a glycosylphosphatidylinositol (GPI) anchor signal peptide with 22 amino acids at the C-terminus, which are also the typical characteristics of lepidopteran APN proteins. The expression profiles in different portions of gut of the 4th instar larva revealed that the expression level of SiAPN3 was significantly higher in larval midgut and hindgut compared with that in foregut (P<0.05). The expression profiles in different instar larvae showed that SiAPN3 was expressed in all instars of S. inferens larvae, with the highest level in the 3rd instar larvae and the lowest level in the 1st instar larvae. The expression levels of SiAPN3 at the 3rd and 4th instars were obviously higher than those at other instars, despite lack of statistically significant differences; no significant difference was found in the expression levels among the 1st, 2nd and 5th instars (P>0.05). The results provide experimental evidence for revealing the function of APN gene and the molecular mechanism of resistance to Bt toxin in S. inferens.
    Chilling enhances the NAD levels and cytosolic malate dehydrogenase activity in diapause eggs of the silkworm, Bombyx mori
    WANG Qi-Long, WAN Hua-Xing, YAO Jin-Mei, SI Ma-Yang-Hu, ZHAO Lin-Chuan
    2012, 55(9):  1031-1036. 
    Abstract ( 2813 )   PDF (2845KB) ( 1086 )     
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    To investigate whether chilling at 5℃ changes the metabolism of NAD in diapause eggs of the silkworm, Bombyx mori, the levels of NADH and NAD+ along with the activities of lactate dehydrogenase (LDH) and cytosolic malate dehydrogenase (cMDH) in diapause eggs incubated at 25℃ and 5℃, respectively, were determined using HPLC and spectrophotometric methods. NAD (NADH + NAD+) levels and the cMDH activity in diapause eggs incubated at 5℃ increased by 106% and 53%, respectively, which were significantly higher than those in diapause eggs incubated at 25℃ (P<0.01). However, no significant difference in the NADH/NAD+ ratio and the LDH activity was observed between the incubation treatments at 25℃ and 5℃ (P>0.05). It is so inferred that the ability for NAD+ synthesis and regeneration in Bombyx diapause eggs is enhanced by chilling.
    Differential allocation of resources to flight muscles and ovaries in different morphs of the wing-dimorphic cricket Velarifictorus ornatus (Orthoptera: Gryllidae)
    ZHAO Lu-Quan, ZHU Dao-Hong, ZENG Yang
    2012, 55(9):  1037-1045. 
    Abstract ( 2831 )   PDF (4985KB) ( 1063 )     
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    The wing-dimorphic cricket Velarifictorus ornatus exists physiological trade-offs between flight muscles and ovarian development. In this study, we quantitatively analyzed the contents of protein, glycogen and total lipids in flight muscles and ovaries between the long-winged and short-winged females by coomassie brilliant blue G-250, anthrone method and sulphophosphovanillin method, respectively. At 10 d after adult emergence, there was no difference in body weight (P>0.05) but the short-winged females loaded more eggs than long-winged females (P<0.05). De-alation could stimulate the long-winged females to produce more eggs than the intact long-winged females (P<0.05). There was no variation in contents of protein, glycogen and total lipids in light muscles of short-winged females after adult emergence but the protein content in flight muscles of long-winged females reached the maximum (564.4±87.5 μg/♀) at day 3, while the contents of glycogen and total lipids also reached the maximum (85.2±21.7 μg/♀, 5 284.7±1 267.4 μg/♀, respectively) at day 5, and then the contents of protein, glycogen and total lipids decreased from day 5. The contents of protein, glycogen and total lipids in flight muscles of long-winged females were much more than those of short-winged females at 1, 3, 5 and 10 d after emergence (P<0.05), while the contents of protein, glycogen and total lipids in ovaries of short-winged females were more than those of long-winged females after adult emergence (P<0.05). Age had obvious effect on the allocation of protein, glycogen and total lipids between the flight muscles and ovaries (P<0.05). De-alation stimulated the increase in the contents of protein, glycogen and total lipids in ovaries of long-winged females, while de-alation elicited the decrease in the contents of protein, glycogen and total lipids in flight muscles of long-winged females. At 10 d after de-alation, the contents of total lipids in flight muscles decreased to 2 394.9±1 461.8 μg/♀, which was only half of the maximum and similar to that of the short-winged females. Application of juvenile hormone Ⅲ (JH Ⅲ) induced the increase in the contents of protein, glycogen and total lipids (P<0.05) in the ovaries of long-winged females but had no effect on their contents in flight muscles (P>0.05). Application of precocene Ⅰ had no effect on the contents of protein, glycogen and total lipids in ovaries of short-winged females (P> 0.05). These results indicate that long-winged females use resources preferentially for the development of flight muscles, but short-winged females use them for the development of the ovaries first, and application of juvenile hormone can change resource allocation between flight muscles and ovaries in long-winged females.
    Characterization of the antifungal peptide MAF-1 from the hemolymph of Musca domestica larvae and its antifungal activity
    TU Tao-Tian, JI Heng-Qing, GUO Guo, FU Ping, WU Jian-Wei
    2012, 55(9):  1046-1053. 
    Abstract ( 3166 )   PDF (9346KB) ( 1259 )     
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    To study the antifungal mechanism of Musca domestica antifungal peptide, MAF-1 was successfully prepared by solid phase extraction (SPE) and RPHPLC from the hemolymph of M. domestica larvae. MAF-1 exists mainly as α-helices and β-sheets in aqueous solution, as revealed by analysis of circular dichroism spectra. The results detected by scanning electron microscopy showed that the membranes of some Candida albicans fungi were depressed, shrinked and ruptured at 1 h after treatment with MAF-1. The results of single-cell gel electrophoresis showed that the “comet”-like cells were observed in the experimental group; however, typical round fungi were found in the normal control group. SDS-PAGE analysis showed that some protein bands were significantly weaker, the number of bands decreased and some bands even disappeared when C. albicans was treated with MAF-1. The results suggest that MAF-1 may have a unique antifungal mechanism.
    Quantitative calculation of the influence of the molecular association between terpenoid repellents and CO2 on their repellency against mosquitoes
    LIAO Sheng-Liang, SONG Jie, WANG Zong-De, CHEN Jin-Zhu, CHEN Shang-Jian, FAN Guo-Rong, JIANG Zhi-Kuan, HAN Zhao-Jiu
    2012, 55(9):  1054-1061. 
    Abstract ( 2690 )   PDF (7998KB) ( 1160 )     
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    This study attempted to demonstrate the molecular association between terpenoid repellents and CO2, and investigated its influence on their repellency against mosquitoes. Association energy was calculated using computational chemistry software: three-dimensional structures of CO2, 22 terpenoid repellents and their complexes that are associated with CO2 were built and optimized using Gaussian View and Gaussian 03W, respectively, and the association energy were obtained in Ampac 8.16. Dependence of repellency on the molecular association was studied by quantitative structure-activity relationship method. Significant activity-affecting parameters were screened from structural descriptors of 22 terpenoid repellents and their complexes, as well as descriptors of characteristic fragments of complexes, which were all calculated by Codessa 2.7.10. The quantitative structure-activity relationship (QSAR) model was obtained to analyze the relationship between the structural descriptors and the logarithm value of the corrected repelling time against Aedes albopictus. Association energies between terpenoid repellents and CO2 were thus calculated, and the results showed that the molecular association between them was strong enough to form the complexes. A statistical QSAR model with four parameters and with R2 of 0.9643 was built, and the most significant activity-affecting parameters were COM-WNSA-3 Weighted PNSA (PNSA3*TMSA/1 000) [Zefirov’s PC], f-TerCO2-Min e-n attraction for a C-O bond, M-Max 1-electron reactivity index for an O atom, M-Min (>0.1) bond order of an H atom, respectively. The results of computational chemistry calculation show that the molecular association between terpenoid repellents and CO2 is present, and the association can affect the repellency greatly.
    Effects of methanol extracts of Mikania micrantha on the growth and development of the rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Dynastidae)
    ZHONG Bao-Zhu, 吕Chao-Jun , WANG Dong-Ming, LI Hong, QIN Wei-Quan
    2012, 55(9):  1062-1068. 
    Abstract ( 3032 )   PDF (2110KB) ( 1017 )     
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    【Aim】 To explore the potential of Mikania micrantha on ecological control of the rhinoceros beetle, Oryctes rhinoceros. 【Methods】 The effects of methanol extracts of M. micrantha on O. rhinoceros was studied using dipping method and feeding method. 【Results】 M. micrantha extracts showed good activity on feeding amount, hatchability, pupation, emergency and larval development of O. rhinoceros. M. micrantha extracts had antifeedant activity against the 3rd larvae of O. rhinoceros, with a positive correlation between antifeedant rate and the concentration of extracts. The egg hatchability decreased significantly and the hatching time was delayed after O. rhinoceros was treated with M. micrantha extracts, with the hatchability of 66.66% and the hatching time being prolonged 3 d compared with the control. The mortality was 40.43% when the 1st instar larvae hatched from eggs treated with 10 mg/mL extracts. The body weight gain of the larvae was also restrained by the M. micrantha extracts, being 3.83 g and 4.53 g at 90 d after the M. micrantha extracts were added to the diets at the concentrations of 10 mg/mL and 5 mg/mL, respectively, while in the blank, the body weight gain was 6.87 g. M. micrantha extracts also inhibited the pupation of O. rhinoceros. The pupation time was delayed, and the number of deformed pupae increased after the larvae were treated with the M. micrantha extracts. M. micrantha extracts also decreased the emergence of O. rhinoceros and prolonged the emergence time. The deformity of the treated adults was increased, manifested as deformed wings, smaller body size, disorder in walking, etc. 【Conclusion】 M. micrantha extracts show good prospects to be exploited as botanical insect growth regulators and can be applied in O. rhinoceros control.
    Effects of female body size on oviposition strategy in the gregarious parasitoid Pteromalus puparum (Hymenopera: Pteromalidae)
    XIA Shi-Yang, MENG Ling, LI Bao-Ping
    2012, 55(9):  1069-1074. 
    Abstract ( 2920 )   PDF (822KB) ( 1069 )     
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    Abstract: Host body size, one of host quality attributes, is often a major focus of the study of foraging behavior in parasitoid behavioral ecology. But little emphasis was placed on parasitoid body size. To investigate the effects of female body size on oviposition strategy in the gregarious parasitoid, Pteromalus puparum, experiments were conducted in laboratory under the stringent control of host (Pieris rapae pupae) body size to observe the oviposition behavior and examine the changes in the clutch size, sex ratio, and body size of the progeny parasitoids. The results indicated that the residence time of females on host pupae decreased with increase of their body size, but increased with host body size. Larger females gave birth to more offsprings with more eggs reserved after a bout of oviposition. Sex ratio and body size of offspring parasitoids were not affected by female body size, but the body size of female offsprings increased with host body size. The results confirm that female body size may affect part of the oviposition strategy in P. puparum, and should therefore be incorporated into models of oviposition strategies for gregarious parasitoids as an important variable.
    DNA barcoding based on the mitochondrial COⅠgene sequences for Ips species (Coleoptera: Scolytidae)
    CHANG Hong, HAO De-Jun, XIAO Rong-Tang, LIU Yong, QIAN Lu, AN Yu-Lin, YANG Xiao-Jun
    2012, 55(9):  1075-1081. 
    Abstract ( 4517 )   PDF (1094KB) ( 1297 )     
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    Species of the genus Ips (Coleoptera: Scolytidae) are often intercepted and captured in quarantine at port. To explore the feasibility of identifying Ips species rapidly and accurately by DNA barcode of special sequence in mitochondrial cytochrome c coxidase subunit Ⅰ (CO Ⅰ), the 462 bp sequence of mitochondrial CO Ⅰ gene in Ips spp. was measured and analyzed. The sequence analysis results revealed that there are 259 mutation sites, 203 conserved sites, 181 parsimony-informative sites and 78 singletons in this gene. Among all sites, the average proportion of base A, G, C and T are 35.8%, 17.2%, 16.5% and 30.5%, respectively. A+T base pair accounts for 66.3% of the mitochondrial CO Ⅰ gene, which is significantly higher than that of G+C base pair and shows an obvious preference to A+T in the sequence. The numbers of A and T are equal, conforming to the basic feature of base composition in insect mitochondria. The result of transition and transversion showed that the sequence did not reach saturation and an evolution analysis can be achieved accurately. Model Kimura 2-parameter was used to analyze the genetic distance, and the results showed that the genetic distance among the same species varied from 0.002 to 0.007, while that among different species ranged from 0.056 to 0.431 with the average value of 0.199, confirming that the genetic distance can be used to discriminate different species. Neighbor-joining phylogenetic tree was built based on the CO Ⅰ gene sequences, and the result indicated that the same species was gathered in the same branch, with the bootstrap value of 100% and the related species could also gather in the same branch, with a high confidence (≥97%). The results suggest that DNA barcoding based on mitochondrial CO Ⅰ gene sequences is applicable in the classification and identification of Ips species.
    Species identification of Noctuidae moths (Insecta: Lepidoptera) from Baihuashan, Beijing, China with DNA barcoding
    YANG Cong-Hui, HAN Hui-Lin, CHI Mei-Yan, JIN Qian, WU Chun-Sheng, ZHU Chao-Dong, ZHANG Ai-Bing
    2012, 55(9):  1082-1092. 
    Abstract ( 5995 )   PDF (4849KB) ( 1763 )     
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    To explore the feasibility of DNA barcoding in the identification of Noctuidae moths, the COI genes of 75 samples belonging to 43 species from Noctuidae in Baihuashan, Beijing, were amplified using universal barcoding primers. The intraspecies and interspecies genetic distances were calculated using the Kimura-2-parameter model, the phylogenetic trees were reconstructed by neighbor-joining (NJ) and maximum parsimony (MP) method, and the identification of MOTU was performed using sequence divergence threshold technique. The results showed that all of Noctuidae species were successfully distinguished by the phylogenetic tree. The average intraspecific genetic distance (0.02%) was much less than the average interspecific genetic distance (11.28%). Seventy-five Noctuidae samples were divided into 42 MOTUs based on 1% sequence divergence threshold and the accuracy rate was 95%. Most of MOTUs (41/42) were classified into their corresponding morphospecies with only one exception (MOTU04). The study indicates that the species from Noctuidae family can be well distinguished with the commonly used COI barcodes, which are potentially well utilized in moth species identification.
    Pheromone receptors and their function in moths
    ZHENG Kai-Di, DU Yong-Jun
    2012, 55(9):  1093-1102. 
    Abstract ( 4431 )   PDF (6944KB) ( 2026 )     
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    The pheromone receptors in moths were first identified from Heliothis virescens and Bombyx mori. Dozens of pheromone receptor genes were cloned from 19 moth species and clustered into a subgroup in the phylogeny of olfactory receptors. Moth pheromone receptors can be detected in the pupal stage and many receptors are exclusively or mainly expressed in the olfactory neurons of male antennae. Some moth receptors are also expressed in the female antennae, other neurons of male antennae or other tissues. The majority of moth pheromone receptors respond to more than one pheromone compound. Some moth pheromone receptors can be excited by non-pheromone compounds. The ligands of several identified moth receptors are still unknown. The olfactory receptors only expressed in antenna of female moth to detect pheromones secreted by male moths have been identified. In the process of pheromone recognization, pheromone binding proteins not only transport the moth pheromone molecules to the dendrites of olfactory neurons by selectively binding to pheromone, but also improve the combination between pheromones and receptors. OrCo-like is very important in the process of moth pheromone recognization and always co-expressed with pheromone receptor in the same neuron. However, the termination of neuron response to moth pheromone is not controlled by pheromone receptors, but by ordorant-degrading enzymes. Many questions about the function of pheromone receptors need to be answered and the process of insect pheromone recognization is complexed than we known.
    Progress in the insect symbiont Rickettsia
    PAN Hui-Peng, ZHANG You-Jun
    2012, 55(9):  1103-1108. 
    Abstract ( 3290 )   PDF (1009KB) ( 1182 )     
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    Rickettsia species are intracellular symbionts of eukaryotes that are well known for infecting and causing serious diseases in humans and other vertebrates. All known vertebrate associated Rickettsia bacteria are vectored by arthropods as part of their life-cycle, and many other Rickettsia species are found exclusively in arthropods without any secondary host. For convenience, we refer to the former as vertebrateRickettsia  and the latter as arthropod Rickettsia. Previous research efforts mainly focused on medically important vertebrate Rickettsia. The biological studies of arthropod Rickettsia were limited. In recent years, more efforts have been made on the insect symbiont Rickettsia and considerable achievements have been obtained. The insect symbiont Rickettsia   is distributed among insects, and two different localization patterns exist in the same insect species. It can transmit vertically through eggs, and horizontally through parasitic wasps and host plants. It can influence the host reproduction by male-killing and parthenogenesis. It has beneficial or detrimental effects on its hosts and can increase the tolerance of its hosts to high temperature and parasitic wasps, and is correlated with its hosts’ susceptibility to insecticides. The insect symbiont   Rickettsia has a reduced genome and might undergo considerable further reduction.
    Species, control status and outbreak causes of main pest insects in mangrove ecosystems in China
    LI Zhi-Gang, DAI Jian-Qing, YE Jing-Wen, XU Hua-Lin, HAN Shi-Chou
    2012, 55(9):  1109-1118. 
    Abstract ( 5116 )   PDF (1809KB) ( 1489 )     
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    Mangrove wetlands are one of the key ecosystems in southeast coast of China. A lot of pest outbreaks have occurred in mangroves in recent years, and thus sustainable uses of mangrove wetlands face serious challenges. Species and control status of main pest insects in mangrove ecosystems in China are summarized in this article. The main pest insects include Acrobasis sp., Lasiognatha cellifera, Latoia lepida, Chalioides kondonis, Chalia larminati, Acanthopsyche subferalbata, etc. Biological control, including biological insecticide, insect growth regulators, and natural enemies are the major measures at present. The outbreak causes of main pest insects are discussed preliminarily in terms of ecosystem health and diversity of insect community. Furthermore, in order to achieve the goal of sustainable pest control, the strategies of habitat regulation in mangrove ecosystem are proposed, which would provide important theoretical references and scientific guidance for improving the level of pest management of mangrove wetlands as well as promoting the sustainable development of mangrove.