Effects of applaud on the growth of silkworm (Lepidoptera: Bombycidae)

An experiment was carried out to evaluate the effect of the insecticide Applaud (buprofezin 25% WP) on the silkworm Bombyx mori (L.). This insecticide belongs to the class of insect growth regulators (IGR). The larvae were fed on leaves treated with 3 different concentrations (0.5, 1, 2 g/liter) of Applaud on the 1st d of each instar. Analysis of data with the Tukey-Kramer test at 1% significant level revealed that mortality and larval duration did not differ among the treatments. On the contrary, the larval weight, which was estimated just before mounting (procedure during which the mature larva climbing on a branch or other material to spin the cocoon), differed among the treatments. Also, cocoon weight, shell weight, and cocoon sericin and fibroin content were different among the treatments, except the shell cocoon ratio. Maximum weight was observed in the controls and minimum in the last instar treatments. Our data suggest that supplementation of Applaud through food to larvae does not affect their mortality rate. On the contrary, it affects larval growth and cocoon parameters.     

Xenochironomus canterburyensis (Diptera: Chironomidae) an insectan inquiline commensal of Hyridella menziesi (Mollusca: Lamellibranchia)

Larvae and pupae of the chironomid Xenochironomus canterburyensis (Freeman) are recorded for the first time. They are inquiline commensals of the fresh water mussel Hyridella menziesi (Gray) in Lake Taupo, New Zealand. The chironomid is univoltine and closely dependent on the seasonal growth of the mussel shell for its pupation and adult emergence. The first instar larva is probably a free-swimming, dispersal stage. Second instar larvae appeared in mussels between the outer surface of the mantle and the inner surface of the valve in later summer. They overwintered there as third and fourth instars until spring when they were found at the margin of the valve opposite the inhalent siphon from where they pupated and left the host to emerge at the lake surface. Some mortality of first and second instar larvae occurred as there was an average of 25–5 whole larvae buried in the nacre of each valve of the mussels.     

大分舌蜂不同龄期幼虫及滞育预蛹肠道细菌 群落多样性及其差异

【目的】基于昆虫肠道微生物在宿主健康与调控宿主生长发育中发挥着重要作用,本研究旨在对大分舌蜂Colleles gigas不同龄期幼虫及滞育预蛹肠道细菌群落多样性及其差异进行初步探究。【方法】利用野外采集的大分舌蜂1-5龄幼虫及滞育预蛹,提取肠道内容物细菌DNA进行16S rRNA的V3-V4基因片段PCR扩增,利用IlluminaMiseq二代高通量测序技术进行测序。依据获得的序列数据,利用生物信息学方法分析大分舌蜂幼虫和滞育预蛹肠道细菌的组成、丰度和多样性。【结果】大分舌蜂幼虫肠道细菌菌群共检测到15门,23纲,43目,80科,128属的细菌,其中最主要的门是变形菌门(Proteobacteria)(占93.74%),最主要的目是立克次氏体目(Rickettsiales)(占68.68%),最主要的科是无形体科(Anaplasmataceae)(占68.64%),最主要的属是沃尔巴克氏体属Wolbachia(占68.64%)。Beta多样性分析结果显示,细菌群落变化与幼虫发育有关,显示出低龄组(1-3龄幼虫)、高龄组(4-5龄幼虫)和滞育预蛹组3个水平。Alpha多样性分析显示,滞育预蛹组细菌多样性与高龄组和低龄组的存在显著差异,而高龄组与低龄组间差异不显著。线性判别分析结果显示,滞育预蛹组与低龄组都存在显著优势菌纲,分别是α-变形菌纲(Alphaproteobacteria)和γ 变形菌纲(Gammaproteobacteria),而高龄组不存在显著优势纲。在目水平,肠杆菌目(Enterobacteriales)和假单胞菌目(Pseudomonadales)为低龄组优势目,立克次氏体目(Rickettsiales)为滞育预蛹组的优势目。在科水平,肠杆菌科(Enterobacteriaceae)和莫拉氏菌科(Moraxellaceae)为低龄组的优势科,无形体科(Anaplasmataceae)为滞育预蛹组的优势科。在属水平,低龄组优势属为肠杆菌属 Enterobacter和不动杆菌属Acinetobacter,滞育预蛹组优势属为沃尔巴克氏体属 Wolbachia。功能基因注释结果也显示出这3个组的特点。【结论】大分舌蜂不同龄期幼虫及滞育预蛹肠道细菌群落存在显著差异,从低龄组向高龄组再到滞育预蛹组,细菌多样性逐步降低,这可能与其进食特点以及肠道微生物驱动对肠道环境适应有关。本研究为土壤筑巢独栖野生蜜蜂肠道微生物研究奠定基础,也为此类野生蜜蜂的保护提供新的角度与方向。     

A new stem-borer of the genus Bucculatrix (Lepidoptera: Bucculatricidae) from Japan, with description of the life history

A new species of bucculaticid moth, Bucculatrix hamaboella sp. nov. (Host plant: Hibiscus hamabo , Malvaceae) is described from Wakayama Prefecture, Japan. The feeding habit of the new species is unique in that: (i) the young larva is a leaf miner forming a long red linear mine but in the later instars the larva becomes a stem borer; (ii) later instar larvae undergo double molts within a cocoonet (molting cocoon); and (iii) penultimate and final instars appear on the surface of the leaf as non-feeding stages. The external non-feeding larvae of B. hamaboella undergoing double molts within one cocoonet are considered to be an abbreviated form of the external feeding instars of other bucculatricids typically making first and second cocoonets, undergoing a single molt within each cocoonet. On the basis of morphological characters, this species is related to the species of Sections I and II (Host: Asteraceae) of Braun (1963), rather than to the species of Section VIII (Host: Malvaceae).     

A carotenoid-binding protein (CBP) plays a crucial role in cocoon pigmentation of silkworm (Bombyx mori) larvae

We examined the role of carotenoid-binding protein (CBP) in yellow cocoon pigmentation. First, using yellow or white cocoon races, we investigated the linkage between the yellow pigmentation and CBP expression. CBP was expressed only in the silk gland of the yellow cocoon races, which utilize carotenoids for cocoon pigmentation. Furthermore, CBP expression in the silk glands of day 1-7 fifth instar larvae matched the period of carotenoid uptake into the silk gland. Finally, we gave double-stranded CBP RNA to Bombyx mori (B. mori) larvae to induce RNA interference. The significantly reduced expression of CBP in the silk gland of fifth instar larva was confirmed on day 4 and a decrease in yellow pigmentation was observed in the cocoon. We showed that CBP plays a key role in the yellow cocoon pigmentation caused by carotenoids.     

Infochemicals influencing the host foraging behaviour of Dahlbominus fuscipennis, a pupal parasitoid of the European spruce sawfly (Gilpinia hercyniae)

The chalcidoid wasp Dahlbominus fuscipennis Zett. (Hymenoptera: Eulophidae) parasitises pupae of the European spruce sawfly Gilpinia hercyniae Htg. (Hymenoptera: Diprionidae). In this parasitoid-host system the role of infochemicals for successful parasitisation was studied. Olfactometer experiments and dual choice bioassays in petri dishes showed that D. fuscipennis was not attracted by volatiles from a plant-host complex consisting of spruce, sawfly larvae and pupae. On the other hand, odours from needles of Picea abies (L.), that were collected from the ground, attracted D. fuscipennis to the habitat of the sawfly. Volatiles from host pupae were not attractive, whereas a contact kairomone from sawfly pupae elicited frequent antennal drumming, a behaviour that indicates host recognition. This contact kairomone was extracted with methanol from the outer cocoon layer of the pupa. The kairomone is not produced by the silk glands of the last instar larva spinning the cocoon, since a methanolic extract of silk gland secretion did not elicit any response in D. fuscipennis. Further bioassays revealed that non-volatile components of the integument of the prepupa within the cocoon also elicited antennal drumming. We suggest that chemicals of the prepupal integument diffuse from inside the cocoon to the outside where they inform the parasitoid on the suitability of the host.     

转Bt基因水稻对二化螟绒茧蜂生物学特性的影响

用转Bt基因 (cry1Ab) 水稻KMD1饲喂3龄、4龄和5龄二化螟Chilo suppressalis一定时间后, 作为二化螟绒茧蜂Apanteles chilonis的寄主,研究了转基因水稻经寄主对绒茧蜂生物学特性的影响。结果发现：KMD1处理后,三个龄期幼虫的寄生率都显著下降,其中4龄和5龄达极显著水平;3龄和4龄上的结茧率显著低于对照;蜂蛹历期均短于对照,但仅3龄差异显著;从5龄幼虫所羽化的雄蜂寿命显著短于对照;蜂茧长显著短于对照;而对卵＋幼虫期、茧块茧数、蜂羽化率及性比均无显著影响。     

棉铃虫叉头框蛋白A类似蛋白基因HarmFoxAl的克隆及表达谱分析

【目的】本研究旨在克隆并分析一种棉铃虫Helicoverpa armigera叉头框蛋白A (forkhead box protein A, FoxA)类似蛋白基因HarmFoxAl,探讨2-十三烷酮胁迫下棉铃虫中肠中HarmFoxAl的表达情况,为进一步明确棉铃虫FoxA的功能和参与棉铃虫生长发育的调控通路提供依据。【方法】从棉铃虫幼虫中肠中扩增得到HarmFoxAl的cDNA序列,并对其氨基酸序列和蛋白结构进行分析。将HarmFoxAl的ORF序列连接至pET32a载体并转化大肠杆菌Escherichia coli Transetta菌株,IPTG诱导后检测目的蛋白的表达形式,并利用镍柱亲和层析法纯化融合蛋白。通过qPCR检测棉铃虫不同发育阶段(1-6龄幼虫和预蛹),6龄幼虫不同组织(脂肪体、中肠、体壁和头部)以及10 mg/g 2-十三烷酮处理6龄幼虫不同时间后中肠中HarmFoxAl的表达谱。【结果】HarmFoxAl(GenBank登录号:XM021331806)的开放阅读框为669 bp,编码222个氨基酸,蛋白的相对分子质量和等电点分别为25.03 kD和6.34。氨基酸序列分析表明,HarmFoxAl单体蛋白无信号肽、跨膜区和二硫键,核心区域是由4个α螺旋和3个β折叠组成的球状结构。将重组的Transetta (pET32a-HarmFoxAl)菌株用0.5 mmol/L IPTG在25℃条件下诱导5 h,约45 kD的融合蛋白His-HarmFoxAl能以可溶的形式存在于重组菌中,这与预测的分子量(42.8 kD)相一致。发育阶段特异性表达谱表明,HarmFoxAl在棉铃虫1-3龄幼虫期、6龄幼虫期和预蛹期均有表达,且预蛹期的表达量最高。组织表达谱结果表明,该基因在6龄幼虫的脂肪体、中肠和体壁中表达,且脂肪体内的表达量最高,而在头部中不表达。10 mg/g 2-十三烷酮处理棉铃虫6龄幼虫后中肠中HarmFoxAl的表达量显著降低,但随着时间延长其表达量逐渐升高,处理48 h后表达量显著高于对照。【结论】棉铃虫HarmFoxAl在预蛹期和幼虫脂肪体中表达量最高,2-十三烷酮处理幼虫后HarmFoxAl的表达量急速降低后逐渐升高,推测其在棉铃虫变态发育和解毒代谢过程中发挥重要作用。     

横纹金蛛生物学特性初步研究

横纹金蛛在山东省日照市一年1代,以1龄幼蛛在卵袋内越冬。5月上旬开始出蜇;幼蛛6龄,老熟幼蛛7月中旬羽化。8月上旬产卵,8月下旬孵化。卵期21～45d,幼蛛生活历期311.2d,成蛛寿命52～87d,雌雄比4:1;成蛛日食量0.50609g,日网捕虫30头,每只成蛛期共食虫35.4263g,网捕虫2405头,是一些害虫的重要天敌。该蛛发生与气候、海拔、天敌等关系密切。     

Parasitoid genus‐specific manipulation of orb‐web host spiders (Araneae,Araneidae)

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Susceptibility of Mocis latipes (Lepidoptera: Noctuidae) to Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae)

The susceptibility of larvae, prepupae, and pupae of the grass looper Mocis latipes (Guenée) to the entomopathogenic nematode Heterorhabditis bacteriophora (Poinar) NC strain was evaluated under laboratory conditions. Concentrations of 0, 5, 10, 20, 40, 60, and 120 nematodes per larva, applied in 1 ml of sterile-distilled water, were bioassayed, applying them to groups of 20 individuals of each instar, prepupa or pupa. Mortality was recorded daily for 5 d. All instars and the prepupal stage were the most susceptible to H. bacteriophora. Mortality ranged from 22.5 to 100%. Prepupae had 97.5-100% mortality starting at 10 nematodes per prepupa. Pupal mortality ranged from 27.5 to 41.3% as nematode concentration was increased. H. bacteriophora presented LC50 values that ranged between 5.26 and 37.66 nematodes per larva and LT50 values that ranged between 1.5 and 4.3 d. Results of this study suggest that H. bacteriophora has potential as a biocontrol agent against M. latipes.     

挪威南部地区蚁形郭公虫(Thanasimus formicarius)生活史(英文)

在自然和实验条件下,对蚁形郭公虫（Thanasimusformiyarius（L）生活史及有关生物学进行了研究。基于逐月野外调查结果,报道了蚁形郭公虫群飞期,以及该虫的卵、幼虫、蛹、预蛹和成虫等各发育阶段的历期。研究结果表明,蚁形郭公虫在挪威南部地区的生活史长达2年零4个月。在生活史的第1年里,该幼虫在受害的挪威云杉树基的蛹穴内越冬;成虫于次年羽化并逐步发育成熟。这些成虫将在树干裂缝中越冬,然后于第3年春产卵繁殖。研究揭示,在挪威南部地区,蚁形郭公虫的发生期与多种小蠹虫的危害期吻合。     

外寄生性花绒寄甲的寄生选择及其发育表现

"选择-表现"假说认为,成虫应该选择有利于子代发育的高品质寄主,但在寄主选择中,除了寄主品质外,其他因素也可能影响寄主选择决策。寄主选择研究通常以成虫为对象,而对那些初龄幼虫选择寄主的寄生性昆虫很少关注。以1龄幼虫积极搜寻寄主的寄生性花绒寄甲为模式生物,采用双选试验设计,观察了花绒寄甲初孵幼虫在不同体重青杨天牛幼虫之间、在已被寄生与健康的黄粉虫蛹之间的寄生选择性;然后采用回归设计,观察了花绒寄甲寄生若干不同体重的青杨天牛幼虫后的发育表现。研究结果表明,花绒寄甲1龄幼虫对体型较大的青杨天牛幼虫的选择偏好显著大于对体型较小的寄主幼虫的选择,选择大体型幼虫的比值比是选择小体型幼虫的4.55倍;对已被寄生的寄主黄粉虫蛹的选择偏好显著大于对健康寄主蛹的选择,选择已被寄生寄主的比值比是选择健康寄主的12.57倍。寄生青杨天牛幼虫的花绒寄甲幼虫发育历期平均为11.49 d、蛹历期为26.67 d、幼虫发育至成虫的羽化率50%,这些发育表现与寄生时青杨天牛幼虫的体重没有显著关系。但刚羽化寄甲成虫体重与寄生时寄主的体重存在显著的正直线关系:寄生时的寄主体重每增大0.01 g,羽化出的寄甲成虫体重增大近0.08%;方差分析寄甲成虫体重在不同寄主体重水平之间的差异表明,从体型较大寄主中羽化的寄甲成虫体重显著大于从体型较小寄主中羽化的成虫。研究结果说明,花绒寄甲初孵幼虫在寄主选择决策时,在寄主体型大小与被寄生状态之间可能采取折衷对策,而且对体型大小不同的寄主选择与子代发育适合度表现存在一致性,从而支持"选择-表现"假说。     

Stimuli affecting selection of oviposition sites by female peach twig borer, Anarsia lineatella Zeller (Lepidoptera: Gelechiidae)

In laboratory and field experiments, stimuli were tested that might affect oviposition decisions by female peach twig borer moths, Anarsia lineatella Zeller (Lepidoptera: Gelechiidae). When given a choice between immature green peach fruits, green mature peach fruits and soft-ripe peach fruits, the latter received the fewest eggs. Fuzzy halves of peach fruits received ten times more eggs then shaved hairless halves. Volatiles from both almond and peach shoots induced more oviposition by females than by control stimuli. Similarly, volatiles from immature green peach fruits, mature green or mature hard-ripe peach fruits induced more oviposition than their respective control stimuli. In a choice experiment, volatiles from immature peach fruit stimulated three times more oviposition than those from soft-ripe peach fruit. Discrimination against mature soft-ripe peach fruits as potential oviposition sites may lie in the phenology of A. lineatella and host peach fruits. Larval development to the pupal stage takes 15–27 days. Therefore, any eggs laid on a ripe fruit 14 days before it falls from the tree will not likely develop into adult insects because developing larvae will only reach third or fourth instar before the fruit is decomposed, and only first and second instar larvae can overwinter.     

斜纹夜蛾蜕皮激素合成通路相关CYP450基因的鉴定及表达分析

【目的】本研究旨在探讨蜕皮激素合成通路相关CYP450基因的表达规律,为害虫防治提供潜在的作用靶标。【方法】以家蚕Bombyx mori CYP450基因为查询序列,通过同源比对的方法从斜纹夜蛾Spodoptera litura基因组中鉴定蜕皮激素合成代谢通路中的CYP450基因,并构建其系统进化树;采用qPCR法检测鉴定的CYP450基因在斜纹夜蛾不同发育阶段(6龄幼虫、预蛹和蛹)、这3个发育阶段的不同组织(中肠、表皮和脂肪体)以及4龄幼虫取食不同寄主植物(辣椒Capsicum annuum、黄瓜Cucumis sativus、番薯Ipomoea batatas和花生Arachis hypogaea)叶片后5龄幼虫中肠中的表达量,计算4龄幼虫取食不同寄主植物叶片后发育至蛹的历期;应用PITA, miRanda, microTar和RNAhybrid 4种软件,预测调控鉴定的CYP450基因的微小RNA (miRNA)。【结果】鉴定到斜纹夜蛾蜕皮激素合成通路相关的6个直系同源CYP450基因CYP307A1, CYP306A1, CYP302A1, CYP315A1, CYP314A1和CYP18A1;系统进化树显示,斜纹夜蛾这6个CYP450基因分别归属于CYP2和线粒体CYP两个亚家族。CYP306A1, CYP314A1和CYP18A1分别在6龄幼虫、预蛹和蛹期具有最高的表达量,并且分别在6龄幼虫中肠、预蛹脂肪体和蛹表皮中表达量最高。相比取食人工饲料的对照组,4龄幼虫取食番薯和花生叶片后斜纹夜蛾4龄幼虫发育至蛹的历期显著延长, CYP306A1在5龄幼虫中肠中的表达量显著上调。在CYP307A1,CYP315A1, CYP314A1和CYP18A1上鉴定出了多种miRNA结合位点。【结论】蜕皮激素合成代谢通路中,CYP306A1,CYP314A1和CYP18A1可能分别在斜纹夜蛾幼虫、预蛹和蛹期起着关键的调控作用,同时也参与宿主植物次生代谢产物的解毒代谢,并且受到miRNA的严密调控。研究结果不仅有助于深入理解昆虫变态发育调控的复杂机制,还为将来的害虫防治提供了潜在的作用靶标,有利于斜纹夜蛾等害虫的可持续治理。     

The Immature Stages of Ptilocolepus extensus McLachlan, 1884 (Trichoptera: Hydroptilidae: Ptilocolepinae) with Notes on Biology

The last instar larva and the pupa of Ptilocolepus extensus McLachlan, 1884, an endemic of the Iberian Peninsula, are described for the first time. The final instar larva of Pt. extensus bears close resemblance to those of Ptilocolepus granulatus (Pictet, 1834), but the abdomen has a well-developed dorsal sclerotized plate on segment VIII; this is a morphological feature which has not been observed in any other Ptilocolepinae. Final instar larvae make dorsoventrally flattened, elongate-oval cases of leaf pieces from several moss and liverwort species; such cases are characteristic of the Ptilocolepinae. Some additional data on biology and geographical distribution are given.     

Characterization of Adoxophyes honmai single-nucleocapsid nucleopolyhedrovirus: morphology,structure, and effects on larvae

A nucleopolyhedrovirus (NPV) was isolated from a diseased larva of the smaller tea tortrix, Adoxophyes honmai, collected from a tea field in Tsukuba, Ibaraki, Japan. Electron microscopic observations confirmed that A. honmai NPV (AdhoNPV) was a single-nucleocapsid type virus. The genome size of AdhoNPV was estimated to be 111.6 +/- 0.9kb (mean +/- SE) by restriction endonuclease analysis. AdhoNPV was also infectious to two other Adoxophyes species, the summer fruit tortrix Adoxophyes orana and Adoxophyes dubia. The LD50 values for neonatal, second, third, fourth, and fifth (final) instar larvae of A. honmai were determined as 61, 107, 688, 1,961, and 4,085 occlusion bodies/insect, respectively. Most of the infected larvae died 5-9 days after molting to the final instar, regardless of the timing of inoculation. However, when neonates were exposed to extremely high doses of AdhoNPV (greater than 100 x LD90), larval development was prevented and most of the larvae died in the first instar.     

Impact of the parasitoid Lathrolestes ensator (Hym., Ichneumonidae, Ctenopelmatinae) as antagonist of apple sawfly Hoplocampa testudinea (Hym., Tenthredinidae)

Abstract: Lathrolestes ensator (Brauns) was the only parasitoid species reared from larvae of apple sawfly Hoplocampa testudinea (Klug) collected in Dutch orchards. The life history parameters of apple sawfly and its parasitoid L. ensator were investigated in field and semi‐field experiments. Usually, the adult parasitoids emerge in synchrony with the preferred host stage, the second instar sawfly larvae. Parasitoid females carried 120–175 eggs, but never laid more than half this number. The mortality of sawfly eggs and larvae varied from 75 and 99%. The eggs and mining larvae are most vulnerable, as the older larvae survive for 90% on average. Three factors at least determine how many of the descending larvae become adult sawfly or parasitoid next spring. The failure of descending larvae to construct a cocoon varied from 7 to 31% and was highest in clay soil. Probably a similar mechanical hindrance prohibited more emerging adults, of both sawfly and L. ensator, from attaining the surface in heavier soils next spring. Relatively more sawfly prepupae than parasitoids died in the cocoon stage, from fungus disease or other causes, but more parasitoids than hosts stayed in prolonged diapause. Mainly due to this last factor, the overall result was a decrease of the parasitoid : host ratio during the first season underground. A life table based on survival rates during various life stages yields a net reproductive rate of up to 2.4 daughters per female sawfly. It indicates that 60% of the sawfly larvae need to be parasitized to stop population growth, or correspondingly less when more than 75% of the host eggs and young larvae are killed by predators or other causes.     

The ultrastructure of malpighian tubules and the chemical composition of the cocoon of Aeolothrips intermedius Bagnall (Thysanoptera)

The secretory activity of the two branched malpighian tubules (MTs) of the second‐instar larva in Aeolothrips intermedius is described. MTs of adult thrips have the typical ultrastructure of excretory epithelium with apical microvilli containing long mitochondria and a rich system of basal membrane infoldings. In the second‐instar larva just before pupation, the ultrastructure of MT epithelial cells is dramatically different, and there are numerous huge Golgi systems in the cytoplasm. These cells are involved in an intense secretory activity to produce an electron‐dense product which is released into the MTs lumen. This secretion is extruded from the hindgut and used by the second‐instar larva to build an elaborate protective cocoon for pupation. Electron‐spray‐ionization mass spectrometry analysis of the cocoon revealed the presence of a β‐N‐acetyl‐glucosamine, the main component of chitin, which is also present in the cocoons of Neuroptera and some Coleoptera. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.