Storage proteins of the fall webworm,Hyphantria cunea drury

Two storage proteins, storage protein-1 (SP1) and storage protein-2 (SP2), were found in hemolymph and fat body during the development of Hyphantria cunea, the fall webworm. Both storage proteins show similiar quantitative changes during development in males and females; however, SP1 is more abundant. The hemolymph of last instar larvae contains high concentrations of the storage proteins. However, following pupation, the storage proteins accumulate in fat bodies. SP1 peaks in the hemolymph of males and females late in last instar larvae (8-day-old 7th instar larvae). SP1 has a native molecular weight of 460,000 and consists of six identical subunits (Mr = 76,700), while SP2 has a molecular weight of 450,000 and is composed of two different subunits (Mr = 74,100 and 72,400). Both SP1 and SP2 are hexamers and are phosphorylated glycolipoproteins. The pl values of SP1 and SP2 were determined to be 5.70 and 5.50, respectively. Antibodies raised against SP1 react positively with vitellogenin and ovary extract, as well as with proteins in the hemolymph from last instar larvae and proteins in pupal fat bodies. Storage protein synthesis starts in fat bodies of a 4-day-old 7th instar larvae and in female peaks at 6–8 days of the 7th instar.     

Purification and characterization of yolk protein-2 from the fall webworm,Hyphantria cunea drury

Yolk proteins (YP1, YP2, and YP3) of the fall webworm, Hyphantria cunea, are of relatively low molecular weight. Yolk protein-2 (YP2) was purified from gel slices and by KBr density gradient ultracentrifugation followed by ion exchange chromatography. YP2 is composed of one subunit with a molecular weight of 35.5 kDa. YP2 contains neutral lipids (diacylglycerol and triacylglycerol) and phospholipids (phosphatidylcholine and phosphatidylethanolamine). The neutral lipids are largely composed of lauric acid and palmitoleic acid. YP2 contains relatively large amounts of glutamic acid and aspartic acid but small amounts of tyrosine, phenylalanine, and methionine. YP2 is a vitellin (Vn) synthesized by the fat body. Vitellogenin-2 (Vg2), the precursor of YP2, is present in very small amounts in the hemolymph. Lipophorin and storage protein also are found in the ovary of H. cunea, and these proteins do not immunologically cross-react with YP2. YP2 is detected in first instar larvae but completely disappears during the second instar, indicating that YP2 is intensively utilized during postembryonic development. Anti-YP2 antibodies cross-react with ovarial extracts of Bombyx mori but not with those of insects from other orders such as Cletus schmidti (Hemiptera), Lucilia illustris (Diptera), Anechura japonica (Dermaptera), Periplaneta americana (Dictyoptera), and Ducetia japonica (Orthoptera). © 1995 Wiley-Liss, Inc.     

Purification and characterization of a novel sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea

Abstract:  An enzyme that possesses glutathione S -transferase (GST) activity was found in the fall webworm, Hyphantria cunea . The enzyme was purified to homogeneity for the first time by ammonium sulphate fractionation and affinity chromatography. The N-terminal sequence of the purified protein was similar to those of Sigma-class GSTs. The purified GST retained more than 75% of its original GST activity after incubation at pH 5–8. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The enzyme was able to catalyse the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, a universal substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation.     

The complete mitochondrial genome of the fall webworm,Hyphantria cunea (Lepidoptera: Arctiidae)

The complete mitochondrial genome (mitogenome) of the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae) was determined. The genome is a circular molecule 15 481 bp long. It presents a typical gene organization and order for completely sequenced lepidopteran mitogenomes, but differs from the insect ancestral type for the placement of tRNA^Met. The nucleotide composition of the genome is also highly A + T biased, accounting for 80.38%, with a slightly positive AT skewness (0.010), indicating the occurrence of more As than Ts, as found in the Noctuoidea species. All protein-coding genes (PCGs) are initiated by ATN codons, except for COI, which is tentatively designated by the CGA codon as observed in other lepidopterans. Four of 13 PCGs harbor the incomplete termination codon, T or TA. All tRNAs have a typical clover-leaf structure of mitochondrial tRNAs, except for tRNA^Ser(AGN), the DHU arm of which could not form a stable stem-loop structure. The intergenic spacer sequence between tRNA^Ser(AGN) and ND1 also contains the ATACTAA motif, which is conserved across the Lepidoptera order. The H. cunea A+T-rich region of 357 bp is comprised of non-repetitive sequences, but harbors several features common to the Lepidoptera insects, including the motif ATAGA followed by an 18 bp poly-T stretch, a microsatellite-like (AT)₈ element preceded by the ATTTA motif, an 11 bp poly-A present immediately upstream tRNA^Met. The phylogenetic analyses support the view that the H. cunea is closerly related to the Lymantria dispar than Ochrogaster lunifer, and support the hypothesis that Noctuoidea (H. cunea, L. dispar, and O. lunifer) and Geometroidea (Phthonandria atrilineata) are monophyletic. However, in the phylogenetic trees based on mitogenome sequences among the lepidopteran superfamilies, Papillonoidea (Artogeia melete, Acraea issoria, and Coreana raphaelis) joined basally within the monophyly of Lepidoptera, which is different to the traditional classification.     

Comparative analysis of the alkali-liberated components of the Hyphantria cunea and the Diacrisia virginica granulosis viruses

The biochemical and biophysical characteristics of the closely related Diacrisia virginica and Hyphantria cunea granulosis virus isolates were examined. Sucrose gradient sedimentation patterns of alkali-solubilized DGV and HcGV capsules were identical. The top, middle, and bottom fractions from either viral isolate were infectious when injected into susceptible host larvae. Electrophoretic analysis of alkaline-solubilized granulin extracts demonstrated that both viruses contain alkaline proteolytic activity. The major granulin protein (～28,000 daltons) of both isolates comigrated in a SDS-PAGE. Electrophoretic separation of the virus proteins demonstrated some quantitative differences between the two granulosis viruses. The enveloped nucleocapsids and the nucleocapsids of the two viruses were morphologically indistinguishable.     

不同食料植物对美国白蛾生长发育和繁殖的影响

2010年6月至7月,在室内研究了美国白蛾Hyphantria cunea(Drury)第1代幼虫对洋白蜡、欧美107号杨、法桐和白榆的取食量及不同的食料种类对美国白蛾生长发育和繁殖的影响。结果表明,美国白蛾对受试的各树种的取食量以洋白蜡为最多,完成一代平均每头可以取食2.21 g,其次为欧美107号杨1.95 g,再次为法桐和白榆,分别为1.85 g/头和1.68 g/头。3龄后幼虫对不同的寄主呈现出一定的偏好性,特别是5龄和6龄幼虫对不同的寄主的取食量表现出显著差异。不同的食料植物对美国白蛾幼虫和蛹的发育历期、存活率、蛹重、成虫寿命、产卵量等有显著影响。取食法桐对其生长发育和繁殖表现出明显的不利性,主要表现为幼虫期延长、蛹重减轻、单雌产卵量降低。     

重大外来入侵害虫——美国白蛾生物防治技术研究

美国白蛾Hyphantria cunea(Drury)是重大外来入侵害虫,对我国林业和园林绿化造成重大危害,尤其是近年来美国白蛾传入北京,严重威胁着我国“绿色奥运”的顺利举办和首都园林绿化及生态环境安全。根据美国白蛾多发生在居民区、乡镇和城市的特点,作者开展对环境安全的生物防治技术研究。汲取前苏联等国家从美国白蛾的原产地引进天敌进行生物防治没有成功的经验和教训,立足调查和寻找我国本土天敌。经过21年的努力,发现和筛选出白蛾周氏啮小蜂Chouioia cunea Yang(新属新种)这种寄生美国白蛾的特优天敌;研究这种小蜂的行为学、生态学、生物学、解剖学及人工大量繁殖、放蜂防治技术;筛选出人工大量繁殖小蜂的替代寄主,解决小蜂的大量繁殖的瓶颈问题;通过放蜂防治试验,取得良好的防治效果。同时,筛选出防治美国白蛾的高毒力HcNPV病毒毒株,研制出美国白蛾人工饲料,利用人工饲料大量饲养美国白蛾幼虫,然后接种扩增病毒,成功解决HcNPV病毒大量扩增生产和常年生产的技术难题,做到病毒的规模化生产和常年生产,保证大面积生产防治的需要和病毒质量。研究出利用天敌昆虫和病原微生物综合应用的防治模式:在美国白蛾幼虫期喷洒HcNPV病毒,蛹期释放白蛾周氏啮小蜂,达到既控制当代美国白蛾的危害,又有效抑制其下代的种群数量,取得长期的持续控制美国白蛾的防治效果。上海市、大连市、烟台市和青岛市利用该项技术已经完全控制美国白蛾,特别是上海市利用本项生物防治技术防治后,已经连续6年没有再发现美国白蛾。由于该项生物防治技术利用的是自然界原有的控制害虫的生态因子,因而对环境安全,做到既消灭害虫,又保证生态环境和人畜的安全,也保护生物多样性。该项研究成果也为北京采用“绿色”防治技术有效控制美国白蛾提供重要的科技支撑和技术保障。     

Current status of the management of fall webworm,Hyphantria cunea: Towards the integrated pest management development

The fall‐webworm (FWW), Hyphantria cunea, is a highly polyphagous insect pest that is native to North America and distributed in different countries around the world. To manage this insect pest, various control methods have been independently evaluated in the invaded areas. Some of the control methods have been limited to the laboratory and need further study to verify their effectiveness in the field. On the other hand, currently, integrated pest management (IPM) has become a promising ecofriendly insect pest management option to reduce the adverse effect of insecticides on the environment. The development of an IPM for an insect pest must combine different management options in a compatible and applicable manner. In the native areas of the insect pests, there are some recommended management options. However, to date, there is no IPM for the management of the FWW in the newly invaded areas. Therefore, to develop an IPM for this insect pest, compilation of effective management option information is the first step. Thus, believing in the contribution of an IPM to the established management strategies, the chemical, biological, natural enemy, sex pheromone, and molecular studies regarding this insect were reviewed and potential future research areas were delineated in this review study. Therefore, using the currently existing management options, IPM development for this insect pest should be the subject of future research in the newly invaded areas.     

Further studies on the distribution of a nuclear-polyhedrosis virus of the fall webworm,Hyphantria cunea,in soil

The median infectious concentration (IC₅₀) of a nuclear-polyhedrosis virus of the fall webworm, Hyphantria cunea, in soil was 7.14×10⁷ polyhedra per gram of soil. The number of virus-killed larvae equivalent to IC₅₀ decreased with the advance of the larval instar from 22 first-instar to 0.04 seventh-instar larvae per gram of soil. The degree of virus contamination of the surface soil in an area where the virus disease had occurred naturally in host populations was estimated to be as high as IC₅₀ on the basis of soil bioassays. Nearly 1000 virus-killed seventh-instar larvae/m² were necessary to obtain such extensive soil contamination. The possibility of the occurrence of so many virus-killed cadavers in the study area was discussed in relation with the past records of epizootics and host population trends. The distribution of the fallen leaves of large trees favorable to the fall webworm, such as plane and poplar trees, correlated with that of the nuclear-polyhedrosis virus in the soil.     

Protein purification and nucleotide sequence of a lysozyme from the bacteria-induced larvae of the fall webworm,Hyphantria cunea

A protein with lytic activity against Micrococcus luteus was purified from the hemolymph of the fall webworm, Hyphantria cunea, larvae challenged with live E. coli. A bacteriolytic protein of about 14,000 daltons in mass was purified by cation exchange chromatography and reverse-phased HPLC. The optimum pH and optimum temperature range for activity were around pH 6.2 and 50°C, respectively, in a 100 mM phosphate buffer. The aminoterminal amino acid sequence of this protein was determined and the corresponding cDNA was isolated and analyzed. The deduced protein of 142 amino acid residues was composed of a putative leader sequence of 20 residues and the mature enzyme of 122 residues. The cloned lysozyme gene was strongly induced in response to bacterial injection, implying that the enzyme is a part of the immune response of H. cunea. Comparison with other known lysozyme sequences shows that our lysozyme belongs to the chicken lysozyme. Arch. Insect Biochem. Physiol. 35:335–345, 1997.© 1997 Wiley-Liss, Inc.     

Effects of different food plants on the growth, development and reproduction of fall webworm ,Hyphantria cunea, larvae

2010年6月至7月,在室内研究了美国白蛾Hyphantria cunea(Drury)第1代幼虫对洋白蜡、欧美107号杨、法桐和白榆的取食量及不同的食料种类对美国白蛾生长发育和繁殖的影响.结果表明,美国白蛾对受试的各树种的取食量以洋白蜡为最多,完成一代平均每头可以取食2.21g,其次为欧美107号杨1.95g,再次为法桐和白榆,分别为1.85g/头和1.68g/头.3龄后幼虫对不同的寄主呈现出一定的偏好性,特别是5龄和6龄幼虫对不同的寄主的取食量表现出显著差异.不同的食料植物对美国白蛾幼虫和蛹的发育历期、存活率、蛹重、成虫寿命、产卵量等有显著影响.取食法桐对其生长发育和繁殖表现出明显的不利性,主要表现为幼虫期延长、蛹重减轻、单雌产卵量降低.     

Immunological analysis of apolipophorin-III in the haemolymph,ovaries, and testes of the fall webworm,Hyphantria cunea (Drury)

Apolipophorin-III (apoLp-III) was purified from the haemolymph of adult Hyphantria cunea (Drury) by KBr density gradient ultracentrifugation, gel filtration (Sephadex G-100) and ion exchange chromatography (CM-52), and its characteristics, molecular weight, tissue distribution, and sites of synthesis were examined. Molecular weight of apoLp-III was estimated to be 18 kDa. By electrophoretic analysis on 10% gels of male and female haemolymph from diverse developmental stages, apoLp-III was shown to be present in all stages. Western blotting was carried out to show that purified free apoLp-III is identical to apoLp-III associated with adult lipophorin. Immunological analysis also showed that apoLp-III is present in the ovary and the testis and in the case of testis, apoLp-III is heavily accumulated in the cyst. ApoLp-III is synthesized in larval and adult fat body but not in adult testis. Autoradiography following incubation of [¹⁴C]apoLp-III with testis showed that apoLp-III was taken up into testis. © 1996 Wiley-Liss, Inc.     

Immunological analysis of lipophorin in the haemolymph,ovaries, and testes of the fall webworm,Hyphantria cunea (Drury)

Lipophorin (LP) was purified from haemolymph in last instar larvae of Hyphantria cunea (Drury) by KBr density gradient ultracentrifugation and gel filtration. LP is composed of Apo-LP I and Apo-LP II with molecular weights of 230 kDa and 80 kDa, respectively. The level of haemolymph LP in early pupae was somewhat greater than in last instar larvae. In males, this LP concentration is maintained throughout pupal development, whereas the level of haemolymph LP decreases in female pupae beginning at day 7, coincident with the onset of vitellogenesis in the fall webworm. In both male and female adults, haemolymph LP concentrations were dramatically increased in comparison to their pre-adult levels. Actually, LP was found in the ovary by immunodiffusion, tandem-crossed immunoelectrophoresis, and Western blotting. Location of LP in the ovary was also traced by immunogold labelling. Also, LP appeared in small amounts in protein yolk bodies of the ovary at an early stage of vitellogenesis, when nurse cells are bigger than the oocyte, but in greater amounts at those stages when the oocyte is larger than nurse cells—that is, when vitellogenesis is actively taking place. This fact clearly reveals that LP is synthesized by fat body and released into the haemolymph, and then taken up by the growing ovary during vitellogenesis. Also, LP was detected in testes by immunological analysis. Western blotting showed that LP was present in testicular fluid but not in the peritoneal sheath and cysts. To test whether LP is also synthesized in testes, testes and fat body tissues were cultured in vitro, indicating that fat body synthsizes LP but testes do not. The result showed that the haemolymph LP itself is taken up into the testes. © 1994 Wiley-Liss, Inc.     

Mass rearing and augmentative releases of the native parasitoid Chouioia cunea for biological control of the introduced fall webworm Hyphantria cunea in China

The fall webworm, Hyphantria cunea Drury, is an economically important introduced pest in China. A native pupal endoparasitoid, Chouioia cunea Yang (Hymenoptera: Eulophidae, Tetrastichinae) causes considerable mortality of H. cunea pupae in some areas of China. Alternative hosts of C. cunea were tested to find potential substitute hosts for mass rearing the parasitoid. Antheraea pernyi Guerin-Meneville (Lepidoptera: Saturniidae) was chosen as a surrogate host as a single pupa produced an average of 6552 wasps and a maximum of 11,256 per pupa. A mass-rearing technique and successful release program of this parasitoid is described. Parasitism by C. cunea in the areas where it was released averaged 67.74% and was usually over 80%. Other native parasitoids, such as Coccygomimus disparis (Viereck), C. parnasae (Viereck) (Hymenoptera: Ichneumonidae) and Exorita japonica Townes (Diptera: Tachnidae), caused the total average parasitism to exceed 90% on average, reaching a maximum of 96.28% in the release areas. These successful releases indicate that an introduced pest species can also be controlled by the mass-rearing and release of native parasitoids in the country of introduction.     

幼虫密度对美国白蛾生活史参数的影响

美国白蛾Hyphantria cunea原产于北美地区,1979年首次在我国辽宁省丹东地区发现,目前已扩散至江苏省南京市等地区,并有进一步扩散的趋势。由于其寄主范围广,适应能力强,危害严重,因此给我国林业生产造成了重大损失。美国白蛾1~4龄幼虫常聚集危害,4龄后开始破网分散危害。本文旨在探明幼虫密度变化对其生活史参数的影响,为解析美国白蛾种群动态变化规律提供科学依据。于温度25℃±1℃和光周期16 L∶8 D条件下,设置5头、10头、20头、40头和80头幼虫/容器5个密度处理组,待幼虫化蛹和成虫羽化后,分别检测幼虫龄期与成活率、蛹的体型（体长和体宽）与体重及成活率、成虫体型（成虫头幅宽度、前翅长度和后足腿节长度）与寿命及繁殖力等生活史参数。结果表明,幼虫密度增加缩短了幼虫发育历期,同时降低了幼虫成活率。幼虫密度增加对蛹重、蛹长、雌蛹宽度及蛹成活率有明显的负效应,但对雄蛹宽度无显著影响。幼虫密度增加对雌虫头幅宽度、雌虫繁殖力、成虫寿命与卵直径具明显负面影响,对雄虫头幅宽度与雌虫前翅长度无显著影响,对成虫后足腿节长度、雄虫前翅长度则具正面影响。因此,本研究表明幼虫密度对美国白蛾不同阶段的生活史参数具调控作用,其中幼虫密度增加对幼虫成活率、蛹的体型与体重及成活率、雌虫繁殖力与卵直径及成虫寿命的负面影响对其种群适合度可能是不利的;然而,成虫对幼虫密度增加会采取一定的适应策略,高密度条件下羽化的成虫具较长的后足腿节与前翅长度以提高其迁移扩散半径,有助于减轻子代种内竞争压力,进而提高种群适合度。     

Susceptibility of Hyphantria cunea to a granulosis virus isolated from Diacrisia virginica

The dosage-mortality response of Hyphantria cunea larvae to a granulosis virus isolated from Diacrisia virginica was studied. Serial decimal dilutions of the D. virginica granulosis virus were fed to early second-instar H. cunea larvae. the LD₅₀ for this virus (7.9 × 10⁵ capsules/larva) was significantly greater than the LD₅₀ calculated for the H. cunea granulosis virus (7.06 × 10⁴ capsules/larva) against the same instar of H. cunea. Time mortality studies demonstrated that the LT₅₀ values for the D. virginica granulosis were four to six times greater than for comparable dosages of H. cunea granulosis virus. Based on the mortality response of assayed H. cunea, it is possible to distinguish between the D. virginica granulosis virus and the H. cunea granulosis virus.