Effects of Diflubenzuron on the Development of Bombyx mori (Lepidoptera: Bombycidae)

ABSTRACT This study was conducted to determine the effects of diflubenzuron (DFB) on the growth and development of the silkworm, Bombyx mori (L.). The DFB treatment extended abnormally the larval duration and affected negatively on larval spinning of the 5th instar. All the larvae treated with high DFB concentration (2.5 × 10^-1 and 2.5x 10^-1μg/μl) lost their spinning capability and finally died, whereas 62% of larvae treated with low DFB concentration (2.5 × 10^-1μg/μl)) spinned cocoon. The larval weights depended sensitively on the DFB treatment period rather than on the DFB concentration. The DFB treatment decreased the larval maturity less than 6% without regard to the concentration and treatment period. All the larva, when treated with DFB before the 5th days of the 5th instar, were not matured.     

Susceptibility of the Bombyx mori cardia cells to Nucleopolyhedrovirus, multiple subgroup, BmMNPV

Bombyx mori entomopathogenic virus infection is a serious problem for silk production in tropical regions. Here, we investigate the susceptibility of the B. mori cardia epithelial cells to B. mori Nucleopolyhedrovirus, BmMNPV. Results show that cardia cells are susceptible to BmMNPV and that the cytopathology is similar to that in other target cells. The infection was detected at 6 day post-inoculation. This infection time, together with the protected cover intima, suggests that the cardia region is a secondary target, infected by budded virus.     

家蚕鞣化激素基因对翅展及繁殖力的调控作用

[目的]昆虫鞣化激素(Bursicon)是由神经系统分泌的一种异源二聚体神经肽,对昆虫表皮鞣化、翅展等功能具有调控作用.本研究旨在探究鞣化激素基因与家蚕Bombyx mori翅发育及对繁殖力相关基因的关系,明确其对翅展和繁殖力的调控作用.[方法]采用RNAi技术,分别注射Bursicon基因的dsRNA(dsBmBur...     

家蚕Bcl-2家族基因 BmBuffy 的克隆及表达分析

【目的】本研究旨在获得家蚕 Bombyx mori Bcl-2家族同源基因,并分析其在不同组织和发育阶段的时空表达模式及功能。【方法】用cDNA末端快速扩增方法（rapid amplification of cDNA ends, RACE）克隆家蚕Bcl-2家族基因BmBuffy,利用SSR分子标记连锁分析确定其染色体定位。同时,用RT-PCR和qPCR技术分析该基因在家蚕幼虫及变态期间不同组织中的表达。【结果】克隆获得全长家蚕Bcl-2家族同源基因 BmBuffy,证明该基因位于第 4号染色体上,开放阅读框长879 bp,编码292 aa,预测其分子量大小为32.4 kDa,等电点为9.94,且第130-231位氨基酸之间存在1个Bcl-2_like Superfamily结构域。系统进化发育树表明,其与黑腹果蝇Drosophila melanogaster的DmBuffy关系最近,氨基酸序列一致性为27%。BmBuffy在幼虫组织中的表达结果显示,其在马氏管中表达量最高,而且在不同组织变态期的关键时间点均有明显变化。【结论】家蚕BmBuffy具有Bcl-2家族典型结构域,BmBuffy定位于第 4号染色体上,BmBuffy在家蚕变态期的组织生理变化中起到一定作用。本文为进一步研究家蚕Bcl-2家族基因的功能奠定基础。     

Fumarate mitigates disruption induced by fenpropathrin in the silkworm Bombyx mori (Lepidoptera): A metabolomics study

The silkworm Bombyx mori L. is a model organism of the order Lepidoptera. Understanding the mechanism of pesticide resistance in silkworms is valuable for Lepidopteran pest control. In this study, comparative metabolomics was used to analyze the metabolites of 2 silkworm strains with different pesticide resistance levels at 6, 12, and 24 h after feeding with fenpropathrin. Twenty-six of 27 metabolites showed significant differences after fenpropathrin treatment and were classified into 6 metabolic pathways: glycerophospholipid metabolism, sulfur metabolism, glycolysis, amino acid metabolism, the urea cycle, and the tricarboxylic acid (TCA) cycle. After analyzing the percentage changes in the metabolic pathways at the 3 time points, sulfur metabolism, glycolysis, and the TCA cycle showed significant responses to fenpropathrin. Confirmatory experiments were performed by feeding silkworms with key metabolites of the 3 pathways. The combination of iron(II) fumarate + folic acid (IF-FA) enhanced fenpropathrin resistance in silkworms 6.38 fold, indicating that the TCA cycle is the core pathway associated with resistance. Furthermore, the disruption of several energy-related metabolic pathways caused by fenpropathrin was shown to be recovered by IF-FA in vitro. Therefore, IF-FA may have a role in boosting silkworm pesticide resistance by modulating the equilibrium between the TCA cycle and its related metabolic pathways.     

C-型咽侧体抑制素(AST-C)和促前胸腺激素释放激素(PTTH)基因在家蚕脑组织中的时空表达分析

【目的】咽侧体抑制素在昆虫体内具有重要的调控功能。本研究比较了家蚕Bombyx mori C-型咽侧体抑制素(allatostatin C,AST-C)与促前胸腺激素释放激素(prothoracicotropic hormone,PTTH)基因在不同发育阶段家蚕脑组织中转录表达的模式及其在脑组织的表达定位,以期为家蚕AST-C的功能研究提供重要的线索。【方法】利用脑组织芯片数据分析比较AST-C和PTTH基因在家蚕脑组织中的发育表达特征,RT-PCR验证其芯片数据,分析AST-C在不同发育阶段家蚕中枢神经系统中的表达模式,并用全组织包埋原位杂交技术对AST-C和PTTH在脑组织的表达进行定位。【结果】AST-C和PTTH在不同发育阶段家蚕脑组织中有相似的转录表达模式,且都在脑组织外侧一对神经分泌细胞中表达。【结论】AST-C可能与PTTH以相同的转录表达模式共同参与家蚕的变态发育调控。     

New Polar Constituents in the Pupae of the Silkworm Bombyx mori L. (II): Developmental Changes of the Constituents

A correlation between the quantitative changes in L-methionine analogs, the ratio of D-serine/L-serine during the pupal stage, and metamorphosis was observed. The glycoside appearing at low blood sugar values during the pupal stage was isolated and characterized as D-glucosyl-L-tyrosine. ¹H-NMR indicated the appearance and increase of this glycoside, and Mirrorcle Ray CV4 equipment was used to take X-ray pictures of the pupal bodies. The results indicate that γ-cyclic di-L-glutamate and L-methionine sulfone might be concerned with ammonia assimilation in the pupae, and that D-glucosyl-L-tyrosine served as a switch for the fatty acid (pupal oil) dissimilation hybrid system.     

Proteomic analysis of nucleopolyhedrovirus infection resistance in the silkworm, Bombyx mori (Lepidoptera: Bombycidae)

Silkworm hemolymph is an important defense tissue to resist bacteria and virus infections. To study the response of silkworm hemolymph in the resistance of Bombyx mori L. nucleopolyhedrovirus (BmNPV), we constructed a near-isogenic silkworm line with BmNPV resistance using highly resistant and highly susceptible parental strains. In this paper, two-dimensional gel electrophoresis (2-DE) and Matrix-Assisted Laser Desorption/Ionization (MALDI)-mass spectrometry were employed to investigate the differences of protein patterns in the hemolymph of the highly resistant, highly susceptible and near-isogenic silkworm strains after BmNPV was administrated to the larvae. A comparison between the proteomes of these three silkworm strains led us to identify two differentially expressed proteins, beta-N-acetylglucosaminidase 2 and aminoacylase. The expression levels of these proteins were higher in the BmNPV resistant strains.     

A digestive beta-glucosidase from the silkworm, Bombyx mori: cDNA cloning, expression and enzymatic characterization

A digestive β-glucosidase cDNA was cloned from the silkworm, Bombyx mori. The B. mori β-glucosidase cDNA contains an open reading frame of 1473 bp encoding 491 amino acid residues. The B. mori β-glucosidase possesses the amino acid residues involved in catalysis and substrate binding conserved in glycosyl hydrolase family 1. Southern blot analysis of genomic DNA suggested the B. mori β-glucosidase to be a single gene. Northern blot analysis of B. mori β-glucosidase gene confirmed larval midgut-specific expression. The B. mori β-glucosidase mRNA expression in larval midgut was detectable only during feeding period, whereas its expression was downregulated during starvation. The B. mori β-glucosidase cDNA was expressed as a 57-kDa polypeptide in baculovirus-infected insect Sf9 cells, and the recombinant β-glucosidase was active on cellobiose and lactose, but not active on salicin, indicating that the B. mori β-glucosidase possesses the characteristics of the Class 2 enzyme. The enzyme activity of the purified recombinant β-glucosidase expressed in baculovirus-infected insect cells was approximately 665 U per μg of recombinant B. mori β-glucosidase. The purified recombinant B. mori β-glucosidase showed the highest activity at 35 °C and pH 6.0, and were stable at 50 °C at least for 10 min. Treatment of recombinant virus-infected Sf9 cells with tunicamycin, a specific inhibitor of N-glycosylation, revealed that the recombinant B. mori β-glucosidase is N-glycosylated, but the carbohydrate moieties are not essential for enzyme activity.     

Effects of cooling and freezing on the motility of Ostrea edulis (L., 1758) spermatozoa after thawing

The aim of this work was to evaluate the effects of temperature, cryoprotectant agents and freezing curves on sperm motility of Ostrea edulis. All phases of cryopreservation were studied (evaluation of semen motility pattern, choice of cryoprotectants and freezing rates) to restore after thawing the motility characteristics distinctive of fresh semen.To assess the temperature effects on sperm motility, semen was activated using four different temperatures (25, 18, 10 and 3 °C). Sperm aliquots were maintained inactive at these temperatures for 1 and 3 h, then activated with FSW at same temperature of conservation. Sperm was activated and incubated to 3 °C with dimethylsulfoxide (Me₂SO), ethylene glycol (EG), 1–2 propylene glycol (PG) (5%, 7%, 10% and 15% final concentrations), glycerol (GlOH; 5%, 10% and 15% final concentrations) and methanol (MetOH; 4% and 10% final concentrations) for 10, 20 and 30 min. A first evaluation of freezing rates was made by testing four freezing curves: −1, −3, −6 and −10 °C/min. Then, an optimization was made by testing four freezing curves: −2.5, −3.0, −3.5 and −4 °C/min.The selected temperature for short term conservation has been 3 °C, because only this temperature has allowed good sperm motility conservation after 3 h of dry-storage; this is a time sufficient to conduct cryopreservation procedures. The sperm showed a particular sensitivity to GlOH and PG to all tested concentrations and to 15% Me₂SO. EG and MetOH to all concentrations and Me₂SO to concentrations lower than 15% have not shown significant toxic effects. The freezing rate −3 °C/min using 15% EG has shown an highest percentage of RVF (rapid, vigorous and forward) spermatozoa (class 3, about 75% of fresh semen) and an highest sperm motility duration.     

中国绮夜蛾属Acontia Ochsenheimer分类学修订（鳞翅目：夜蛾科：绮夜蛾亚科）

首次报道尺蛾科泽尺蛾属Zamarada Moore, 1887和超泽尺蛾Z. excisa Hampson, 1891在我国分布,给出了形态描述和特征图。     

中国优苔螟属一新种（鳞翅目：草螟科：苔螟亚科）

记述采自中国江西武夷山优苔螟属Eudonia 1新种,即截形优苔螟Eudonia abrupta sp. nov.。该种与伊诺优苔螟E. inouei Sasaki,1998 近似,但其雄性外生殖器爪形突的末端与基部等宽,阳茎为抱器瓣长的1.5倍;而伊诺优苔螟的爪形突末端显著窄于基部,阳茎与抱器瓣近等长。文中提供了成虫和外生殖器特征图。模式标本保存在江西农业大学昆虫标本馆。     

Sexual dimorphism in neuronal projections from the antennae of silk moths (Bombyx mori,Antheraea polyphemus) and the gypsy moth (Lymantria dispar)

Summary The antennal lobe of both sexes of the silk moth Bombyx mori contains 55–60 ventrally located antennal glomeruli; in addition, that of the male contains a dorsal macroglomerular complex (MGC). A group of identifiable glomeruli consisting of two lateral large glomeruli (LLG) and four medial small glomeruli (MSG) is present in both sexes, but the LLG are greatly enlarged in the female. A MGC is also present in the male gypsy moth Lymantria dispar and male giant silk moth Antheraea polyphemus. The MGC in all of these species is organized into 3–4 distinct levels of glomeruli. Antennal sensory fibers were stained by cobalt backfills in B. mori, A. polyphemus, and L. dispar. Most fibers stained from cut long hairs (sensilla trichodea) projected to MGC in males and LLG in both sexes of B. mori. The distribution of fibers in the MGC of B. mori was topographically biased in that a majority of fibers from anterior branches projected medially in MGC while most fibers from posterior branches projected laterally or anteriorly. Terminal arborizations of single fibers were each restricted to a single glomerular level of the MGC. Fibers projecting to the posterior antennal center were frequently stained in cut-hair and control preparations, apparently by uptake of cobalt through intact sensilla on flagellar branches.     

中国圆斑小卷蛾属名录及一新种记述（鳞翅目：卷蛾科：新小卷蛾亚科）

记述中国圆斑小卷蛾属Eudemopsis 昆虫10种,其中包括1新种:狭瓣圆斑小卷蛾E. angustivalvae sp. nov.。该新种的体型较小,前翅长仅4.0-4.5 mm,雄性外生殖器中抱器瓣狭长,毛刺稀疏,很容易与属内其他种区分。文中还给出了分种检索表、新种的形态特征图和生殖器图以及该属种类在中国的地理分布图。研究标本保存在南开大学昆虫标本室。     

An L319F mutation in transmembrane region 3 (TM3) selectively reduces sensitivity to okaramine B of the Bombyx mori l-glutamate-gated chloride channel

Okaramines produced by Penicillium simplicissimum AK-40 activate l-glutamate-gated chloride channels (GluCls) and thus paralyze insects. However, the okaramine binding site on insect GluCls is poorly understood. Sequence alignment shows that the equivalent of residue Leucine319 of the okaramine B sensitive Bombyx mori (B. mori) GluCl is a phenylalanine in the okaramine B insensitive B. mori γ-aminobutyric acid-gated chloride channel of the same species. This residue is located in the third transmembrane (TM3) region, a location which in a nematode GluCl is close to the ivermectin binding site. The B. mori GluCl containing the L319F mutation retained its sensitivity to l-glutamate, but responses to ivermectin were reduced and those to okaramine B were completely blocked.     

中国尺蛾科新记录属——泽尺蛾属及其一新记录种记述（鳞翅目：尺蛾科：灰尺蛾亚科）

系统修订了中国绮夜蛾属的种类,将中国记录的11种订正为10种。其中,原记录的白缘绮夜蛾A.nivipicta经鉴定为大理石绮夜蛾A.marmoralis的误订名,迭绮夜蛾A.disrupta被鉴定为斑绮夜蛾A.sexpunctata的同物异名。检视标本均存放于中国科学院动物研究所标本馆。     

Estradiol-17beta: tracing its metabolic significance in female fatbody of fifth instar larvae of silkworm, Bombyx mori L (race: Nistari)

In recent years, various vertebrate peptide and steroid hormones have been identified in invertebrates, estradiol-17beta (E2) being a major one. We have specifically shown NADP-malate dehydrogenase (NADP-MDH) activity in fifth instar larval fatbody of female silkworm, Bombyx mori, as an E2 responsive parameter. Interestingly enough, estradiol-induced increase in the enzyme activity could be counteracted by simultaneous application of specific E2-inhibitor, ICI-182780. Further, a nice correlation was obtained among the E2 titre, specific *E2 binding and expression of NADP-MDH activity in fatbody during different days of normal fifth instar larval development. Though the nature of the binding sites is quite similar to known steroid receptors of vertebrate, the reported absence of estrogen receptor gene in some insects poses a question. A recent finding regarding the presence of an estrogen-related receptor ortholog in fruit fly may provide some answers. The specific effects elicited by estradiol in the female fatbody of this insect support its possibility of having an important metabolic function. This role played by E2, whether hormonal or not, is yet to be identified.     

Parasitism of orangestriped oakworm (Lepidoptera: Saturniidae) eggs in the urban landscape

Orangestriped oakworm, Anisota senatoria (J. E. Smith), has caused widespread defoliation of oak trees in the urban landscape of southeastern Virginia since 1985. Egg masses were collected from 1988 to 1990 to determine the impact of native egg parasites on A. senatoria populations. The most abundant egg parasite was Aprostocetus new sp. and mean egg mass parasitism was 24.6%. The eupelmid Anastatus hirtus (Ashmead), a new host record, parasitized a mean of 11.7% of A. senatoria egg masses. The encyrtid Ooencytrus sp., a new host record, had a mean egg mass parasitism of 0.09%. Inundative releases of Trichogramma minutum (Riley) in 1989 and 1990 did not increase parasitism rates and mean egg mass parasitism was 2.3%. Parasitism of first generation A. senatoria egg masses was higher compared with second generation. The four egg parasites collected in this study parasitized 30% of A. senatoria egg masses and within egg mass parasitism was 7.9%. These relatively low parasitism rates may partially explain the presence of consistently high A. senatoria populations in southeastern Virginia.     

中国雪巢蛾属及一新种记述（鳞翅目：巢蛾科）

记述中国雪巢蛾属Niphonympha Meyrick 3种,其中包括1新种:五指山巢蛾N. wuzhishana sp. nov.。提供了中国雪巢蛾属已知种的检索表以及成虫与外生殖器特征图。     

Derived morphology in a basal moth: The uniquely specialized sternum V glands of Agathiphaga (Lepidoptera: Agathiphagidae)

The sternum V gland is a notable specialization shared by basal Lepidoptera and most Trichoptera; it is generally present in both sexes or in females only. In both orders the secretory cells usually discharge into a sac-like reservoir from which a duct leads to the opening on sternum V. In contrast, the gland in agathiphagid moths is only present in males and it exhibits several unique specialisations: the glandular portion being the apical (morphologically anterior) part of a long tubular cuticular invagination covered with contiguous ‘type-3’ glandular units; together with the following part of the efferent duct it is coiled like a ball of yarn; the duct expands into a fusiform reservoir, and a separate small sac surrounded by a complex muscular coat is present just inside the gland opening. The principal opener muscle of each gland originates on the contralateral side of sternum VI, hence crossing its counterpart in the midline. Characters of the agathiphagid sternum V gland, the function of which remains unclear (though sex pheromone production may appear most likely), add significantly to the suite of autapomorphies exhibited by this otherwise overall remarkably generalized moth taxon.