Taxonomy of Iberian Hoplia (Col., Scarabaeoidea,Hopliinae) based on mtDNA analysis

The morphology of some Hoplia species (Scarabaeoidea: Hopliinae) is so variable that parapatric populations have often been considered different species or subspecies. In this study we analyze the nucleotide sequences of a fragment of mitochondrial gene cytochrome c oxidase subunit I (COI) of six species and two subspecies of Palaearctic Hoplia to reexamine the species limits. Based on the analysis of sequences from COI and morphological and ecological observations, we consider Hoplia freyi Baraud to be a junior synonym of Hoplia chlorophana Erichson and H. philanthus ramburi Heyden to be a junior synonym of H. philanthus philanthus (Fuessly). However, complete resolution of relationships among H. philanthus subspecies requires the addition of sequences from genes evolving faster than COI. Phylogenetic relationships among the species studied are discussed.     

Efficacy of the Entomopathogenic Nematodes Heterorhabditis megidis and Heterorhabditis bacteriophora for the control of grubs (Phyllopertha horticola and Aphodius contaminatus) in Golf Course Turf

Liquid culture-produced entomopathogenic nematodes, Heterorhabditis megidis and Heterorhabditis bacteriophora, were applied at 0.5 and 1.5 million dauer juveniles m-2 against Aphodius contaminatus and Phyllopertha horticola on a golf course. The reduction of A. contaminatus was found to be between 40 and 62%. P. horticola reduction reached 70% with H. megidis and 83% with H. bacteriophora. Turf damage caused by birds preying on the grubs was successfully prevented.     

Revealing species-specific trophic links in soil food webs: molecular identification of scarab predators

Soil food webs are particularly important in terrestrial systems, but studying them is difficult. Here we report on the first study to apply a molecular approach to identify species-specific trophic interactions in below-ground food webs. To identify the invertebrate predator guild of the garden chafer Phyllopertha horticola (Coleoptera, Scarabaeidae) whose root-feeding larvae can be highly abundant in grasslands, a specific DNA marker was developed. It allowed detection of P. horticola egg and white grub meals within the gut content of Poecilus versicolor (Coleoptera, Carabidae) larvae for up to 24 h post-feeding. Soil samples from an alpine grassland revealed a diverse below-ground macro-invertebrate community with earthworms, P. horticola larvae, and centipedes as well as beetle larvae as the most abundant detritivores, herbivores, and predators, respectively. Garden chafer DNA was detected in 18.6%, 4.1%, and 4.4% of field-collected Geophilidae (n = 124), beetle larvae (n = 159), and Lithobiidae (n = 49), respectively. We conclude that most of the investigated predators actively preyed on P. horticola, as secondary predation is unlikely to be detected in below-ground systems. Moreover, scavenging most likely contributes only to a small percentage of the revealed trophic links due to the low availability of carrion. Sampling date did not influence prey detection rates, indicating that both P. horticola eggs and larvae were preyed on. Only 2.7% of the below-ground predators tested positive for earthworms, an alternative, highly abundant prey, suggesting that P. horticola represents an important prey source for centipedes and predatory beetle larvae during summer within the soil food web.     

Selection of a highly virulent fungal isolate, Metarhizium anisopliae CLO 53, for controlling Hoplia philanthus

The June beetle, Hoplia philanthus Füessly (Coleoptera: Scarabaeidae), has become a widespread and destructive insect pest of lawns, sport turf, pastures, and horticultural crops in Belgium. The virulence of 34 entomopathogenic fungal isolates from the genera Metarhizium, Beauveria, and Paecilomyces to third-instar H. philanthus was tested in bioassays by dipping larvae in 10(7)conidia/ml suspensions. Two isolates of Metarhizium anisopliae (CLO 53 and CLO 54) caused maximally 90% mortality 10 weeks post-inoculation while other isolates only caused mortalities between 10 and 62%. The virulence of M. anisopliae CLO 53 was further tested by exposing H. philanthus larvae to conidial serial concentrations of 10(4)-10(9)conidia/g sandy soil for up to 11 weeks at 15, 20 or 25 degrees C. Mortality was dependant on the fungal concentration, exposure time, and temperature. Eleven weeks after inoculation, the LC50 values for this isolate ranged from 1.3 to 4.0 x 10(6), 1.0 to 3.2 x 10(5), and 2.5 x 10(4) to 10(5)conidia/g soil at 15, 20, and 25 degrees C, respectively. The LT50 values for this isolate ranged from 3.5 to 21.7, 2.4 to 18.7, and 2.9 to 16.1 weeks at concentrations of 10(9) and 10(4)conidia/g soil at 15, 20, and 25 degrees C, respectively. In glasshouse pot experiment with perennial ryegrass (Lolium perenne L.), the isolate CLO 53 caused mortalities of 50 and 88% of H. philanthus larvae 10 weeks after application of 10(4) and 10(6)conidia/cm(2) soil surface, respectively. The present results suggest that the Belgian isolate CLO 53 has excellent potential for biological control of H. philanthus.     

豌豆彩潜蝇幼期各虫态的形态学研究

【目的】豌豆彩潜蝇Chromatomyia horticola (Goureau)是我国蔬菜上常见的一种重要害虫,目前尚未见该虫幼期卵到蛹各虫态形态特征的系统研究。【方法】本研究采用野外取样和室内饲养的方法收集豌豆彩潜蝇的卵、不同龄期的幼虫以及蛹,对该虫从卵期到羽化成成虫的全过程进行了详细的观察,并通过测量幼虫口钩和头咽骨的长度对幼虫龄期进行了划分。【结果】卵期观察到了豌豆彩潜蝇及其他潜蝇科昆虫中从未记述过的卵的呼吸角。利用口钩和头咽骨长度将幼虫划分为3个龄期,并对不同龄期幼虫的形态特征进行了比较和描述,确定了通过骨化结构对幼虫龄期准确鉴定和通过外部形态快速鉴定的方法。根据豌豆彩潜蝇蛹期发育的形态特征变化将蛹期划分为5个明显的发育阶段。【结论】本研究记述了豌豆彩潜蝇卵到蛹期的形态特征并对幼虫龄期进行了划分,为潜蝇幼虫和蛹的形态学及豌豆彩潜蝇生物学、生理学和防治研究提供基础。     

A review of scarab beetles of the genus <Emphasis Type="Italic">Hoplia</Emphasis> (Coleoptera,Scarabaeidae) from the Russian Far East

A review of the species of the genus Hoplia from the Russian Far East is given. Lectotypes are designated for Anisoplia cincticollis Faldermann and Hoplia djukini Jacobson. Hoplia cincticollis (Faldermann) is recorded from the Russian Far East for the first time. An original key to the species of the genus Hoplia of the Russian Far East fauna is provided.     

Attraction of garden chafer, Phyllopertha horticola, to floral Japanese beetle lure

Field experiments using funnel traps were performed on a golf course to investigate the attractiveness of a floral Japanese beetle lure for adults of the garden chafer, Phyllopertha horticola, a common European turf pest. Traps baited with 10 mg of the floral lure consisting of phenylethyl propionate + eugenol + geraniol (3 : 7 : 3) captured more adult garden chafers of both sexes than non‐baited control traps. In contrast, 1,3‐dimethyl‐2,4‐(1H, 3H)‐quinazolinedione, the sex pheromone of the closely related pale‐brown chafer P. diversa, neither influenced alone nor in combination with the floral lure the behaviour of adult garden chafers. The results suggest that the floral Japanese beetle lure might be used for monitoring or control of P. horticola.     

Phylogenetic relationships of the genus Hoplia Illiger (Scarabaeidae: Hopliinae)

Results of phylogenetic analysis based on 34 morphological characters of 24 species of 11 genera of Hopliinae from Europe, Japan, South Africa, Madagascar, North and Central America, indicates that the genus Hoplia is a monophyletic group with species distributed in Europe, Japan and America. Based in this analysis the Asiatic genus Ectinohoplia is the closest relative of the genus Hoplia, and the South American genus Barybas (Melolonthinae: Macrodactylini) is the sister group of Hopliinae.     

Spatial dynamics of two competing specialist parasitoids in a host metapopulation

1. We investigated spatial dynamics in two specialist larval parasitoids, Cotesia melitaearum and Hyposoter horticola , attacking the Glanville fritillary butterfly, Melitaea cinxia , in Finland.
2. Presence of C. melitaearum in a host larval group significantly reduced the rate of parasitism by H. horticola (18 vs. 33% in groups with and without C. melitaearum ), but there was no significant reverse effect. The parasitism rates at the level of local populations showed a similar trend.
3 Hyposoter horticola females moved frequently among larval groups, whereas C. melitaearum females often spent several days at a single larval group. In agreement with these behavioural observations about movements within populations, H. horticola was found to have a higher colonization rate of host populations than C. melitaearum .
4. At the within-population level, C. melitaearum tended to occupy large larval groups in the centre of the host population, whereas H. horticola parasitized also small and more isolated (peripheral) larval groups, especially in the presence of C. melitaearum . At the metapopulation level, host population size had a significant positive effect on the presence of local populations of both parasitoid species, but isolation had a significant negative effect on the presence of C. melitaearum only.
5. These results suggest that C. melitaearum is a superior competitor, but an inferior disperser to H. horticola , which facilitates the co-existence of the two species both at the level of local populations and at the level of a metapopulation.
6. Data from a 50-patch network showed a decline in the number of host populations from 34 to 13 in 4 years. This decline caused a near-extinction of the superior competitor, but inferior disperser, C. melitaearum , whereas the abundance of the inferior competitor, but superior disperser, H. horticola , remained relatively constant, in agreement with the prediction of a theoretical model.     

Coexistence of competing parasitoids: which is the fugitive and where does it hide?

Most insects harbour a community of parasitoids that coexist in spite of competition for resources. One potential mechanism for coexistence of competitors is a tradeoff between dispersiveness and local competitive ability. Here we present a study of competition between the specialized parasitoids Hyposoter horticola and Cotesia melitaearum sharing the Glanville fritillary butterfly, Melitaea cinxia . Within one host generation, the parasitoid larvae interact inside the host during each of the three C. melitaearum generations. We founds that in the summer when the host is small, the solitary H. horticola is the superior competitor, suppressing the gregarious C. melitaearum as eggs or small larvae. When multiparasitism occurs in the autumn the two parasitoid species engage in physical combat and C. melitaearum is favoured. Finally, a previous study showed that in the third C. melitaearum generation the univoltine H. horticola grows quickly during its final instar, excluding young C. melitaearum simply through limited time and resources. We found that contrary to expectations of the evolution of gregariousness, C. melitaearum , which lives in sibling groups, has biting mandibles in the first instar while the solitary H. horticola has suctorial mouthparts. Previous studies suggest that the two parasitoids co-exist because H. Horticola is dispersive and C. melitaearum is a strong local competitor. However, putting together the results of this experiment and out recent understanding of the adult wasp foraging behaviours and large scale population dynamics, we conclude that H. horticola is both a superior local competitor and more dispersive than C. melitaearum . Cotesia melitaearum has no impact on the population dynamics of H. horticola , persisting as a fugitive using a small fraction the larvae left unparasitized by H. horticola .     

甘肃白银地区潜叶蝇及其寄生蜂的组成和发生调查

【目的】明确严重危害甘肃省白银地区作物的潜叶蝇及天敌寄生蜂的多样性和种群组成。【方法】采用五点取样法田间调查潜叶蝇危害的蔬菜、花卉、杂草并采样,室内对潜叶蝇及其寄生蜂进行形态鉴定和分子鉴定。【结果】在2016-2020年在甘肃白银地区采集的样本中发现6种常见潜叶蝇,包括本地种豌豆彩潜蝇Chromatomyia horticola和葱斑潜蝇Liriomyza chinensis,入侵种南美斑潜蝇L. huidobrensis、三叶草斑潜蝇L. trifolii、番茄斑潜蝇L. bryoniae和美洲斑潜蝇L. sativae。其中,豌豆彩潜蝇为优势种,可危害多科寄主植物;葱斑潜蝇仅在大葱上发现危害;南美斑潜蝇在温室内的蔬菜上发生严重;三叶草斑潜蝇首次在白银市发现;番茄斑潜蝇与美洲斑潜蝇的危害相对较轻。此外,调查的潜叶蝇寄生蜂包括3科12属24种,优势种包括豌豆潜蝇姬小蜂Diglyphus isaea、芙新姬小蜂Neochrysocharis formosa、万氏潜蝇姬小蜂D. wani和西伯利亚离颚茧蜂Dacnusa sibirica。【结论】一方面白银市应加强监测和预警入侵性潜叶蝇的发生,另一方面白银市潜叶蝇的天敌寄生蜂种类丰富,对田间主要危害的潜叶蝇可能发挥着重要的自然防控作用,建议保护利用本地寄生蜂天敌。     

Inhibition of the glutamate uptake in the excitatory neuromuscular synapse of the locust by delta-philanthotoxin; a component of the venom of the solitary wasp Philanthus triangulum F. A high resolution autoradiographic study

Glutamate uptake in insect neuromuscular junctions is inhibited by delta-philanthotoxin. Two other components of the philanthus venom (beta- and gamma-philanthotoxin) do not affect this glutamate uptake. No synergism between the active delta-philanthotoxin and the two inactive components had to be assumed to explain the inhibition by the Philanthus venom.     

蔬菜地潜叶蝇寄生蜂种类研究(Ⅲ)--金小蜂科Pteromalidae和大痣细蜂科Megaspilidae

本文报道蔬菜地潜叶蝇寄生蜂的姬小蜂科 Eulophidae的羽角姬小蜂亚科 Eulophinae和狭面姬小蜂亚科 Elachetinae5属 8种寄生蜂。其中豌豆潜蝇姬小蜂 Diglyphus isaea是蔬菜潜叶蝇寄生蜂优势种。寄主主要是豌豆彩潜蝇、美洲斑潜蝇和葱斑潜蝇等的幼虫和蛹。植物寄主涉及各种蔬菜作物。兰克瑟姬小蜂 Cirrospiluslyncus Walker为中国新记录种 ,另有 5个寄主新记录     

Polymerase chain reaction-restriction fragment length polymorphism assays to distinguish Liriomyza huidobrensis (Diptera: Agromyzidae) from associated species on lettuce cropping systems in Italy

The pea leafminer, Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae), is a serious insect pest infesting open field lettuce plantings in northern Italy. In these cropping systems, it coexists with several other agromyzid species that have negligible economic importance on open field vegetables. The rapid detection of L. huidobrensis is crucial for effective management strategies, but the identification of agromyzids to species can be very difficult at adult as well at immature stages. In this study, a polymerase chain reaction (PCR)-restriction fragment length polymorphism assay is proposed to separate L. huidobrensis from Liriomyza bryoniae (Kaltenbach), Liriomyza trifolii (Burgess), and Chromatomyia horticola (Goureau), which usually occur in the same lettuce plantings. An approximately 1,031-bp region of the mitochondrial genome encompassing the 3' region of cytochrome oxidase I, the whole leucine tRNA, and all of the cytochrome oxidase II was amplified by PCR and digested using the enzymes PvuII and SnaBI separately. Both endonucleases cut the amplicons of L. huidobrensis in two fragments, whereas the original band was not cleaved in the other analyzed species. The presence of Dacnusa spp. DNA does not bias the assay, because the PCR conditions and the primer set here described do not amplify any tract of this endoparasitic wasp genome.     

蔬菜地潜叶蝇寄生蜂种类研究(II)——凹面姬小蜂亚科Entedontinae和啮小蜂亚科Tetrastichinae

本文报道姬小蜂科 Eulophidae的凹面姬小蜂亚科 Entedontinae和啮小蜂亚科 Tetrastichinae的 6属 10种蔬菜地潜叶蝇寄生蜂。底比斯釉姬小蜂 Chrysocharis p entheus Walker、美丽新姬小蜂 N eochrysocharisformosa(Westwood)和潜蝇纹翅姬小蜂 Teleopterus erxias(Walker)是蔬菜地潜叶蝇寄生性小蜂的 3个优势种。曼丹啮小蜂 Tetrastichus mandanis(Walker)和卡拉啮小蜂 Tetrastichuschara Kostjukov为中国新记录种。另有寄生蜂的 2个寄主新记录。     

美洲斑潜蝇SS-PCR检测技术研究

【背景】美洲斑潜蝇是一种严重威胁瓜果蔬菜、烟草、棉花等经济作物和花卉生产的入侵性害虫。由于潜叶蝇类害虫体型较小、生活方式隐蔽、形态相似,本文针对其难以快速准确地进行形态鉴别的问题,以美洲斑潜蝇为研究对象,以菜田常见的4种潜叶蝇类害虫为参照,采用种特异性PCR方法（species-specific PCR,SS-PCR）,研究其快速分子检测鉴定技术。【方法】调用GenBank中一段936bp的美洲斑潜蝇线粒体DNA（mtDNA）细胞色素氧化酶亚基Ⅰ基因（COⅠ）的序列（Gen-Bank登录号为EU219613）,并根据此基因片段的碱基序列设计引物1对,其扩增片段大小为294bp。【结果】种特异性检验结果显示,该引物只对美洲斑潜蝇的COⅠ基因具有扩增能力,对其他种类如南美斑潜蝇、三叶斑潜蝇、葱斑潜蝇、豌豆潜叶蝇等没有扩增能力。该引物不仅对成虫具有良好的扩增效果,对蛹、幼虫以及单粒卵也具有同样的扩增效果,其最低检出阈值为1/3840头成虫。【结论与意义】SS-PCR技术体系可用于美洲斑潜蝇的鉴定识别与检测监测,对阻止其进一步扩散蔓延具有重要意义。     

Susceptibility of Hoplia philanthus (Coleoptera: Scarabaeidae) larvae and pupae to entomopathogenic nematodes (Rhabditida: Steinernematidae, Heterorhabditidae)

Biological control potential of nine entomopathogenic nematodes, Heterorhabditis bacteriophora CLO51 strain (HbCLO51), H. megidis VBM30 strain (HmVBM30), H. indica, Steinernema scarabaei, S. feltiae, S. arenarium, S. carpocapsae Belgian strain (ScBE), S. glaseri Belgian strain (SgBE) and S. glaseri NC strain (SgNC), was tested against second-, and third-instar larvae and pupae of Hoplia philanthus in laboratory and greenhouse experiments. The susceptibility of the developmental stages of H. philanthus differed greatly among tested nematode species/strains. In the laboratory experiments, SgBE, SgNC, HbCLO51 and HmVBM30 were highly virulent to third-instar larvae and pupae while SgBE was only virulent to second-instar larvae. Pupae were highly susceptible to HbCLO51, HmVBM30, SgBE and SgNC (57–100%) followed by H. indica and S. scarabaei (57–76%). In pot experiments, HbCLO51, SgBE and S. scarabaei were highly virulent to the third-instar larvae compared to the second-instar larvae. Our observations, combined with those of previous studies on other nematode and white grub species, show that nematode virulence against white grub developmental stages varies with white grub and nematode species.     

Plasmid R68.45 and S-a transduction by the temperate bacteriophage 59 of Erwinia carotovora 268

Temperature bacteriophage 59 of Erwinia carotovera 268 had transduced extrachromosomal DNA: plasmids of R68.45 and S-a. Before plasmid transduction experiments the suitable donor strains of indicator culture Erwinia horticola 450 harbouring R68.45 and S-a were created. The frequency of plasmid R68.45 transfer from Pseudomonas putida to E. horticola 450-8 by conjugation was equal to 5 x 10(-8) per a donor cell and in the case of S-a--from E. coli C600 for the same recipient cells--was 2 x 10(-6). Bacteriophage 59 has transduced only separate markers of plasmid R68.45, since plasmid S-a is probably transduced by the phage as an intact unit.     

Selection of insect parasitic nematodes for biological control of the garden chafer,Phyllopertha horticola

Larvae ofPhyllopertha horticola L. (Coleoptera: Scarabaeidae) cause increasing problems on sports fields and lawns in NW-Europe. A biological control programme using insect parasitic nematodes is being developed. This paper contains the results of bioassays with various species and isolates of the nematode generaHeterorhabditis andSteinernema. In bioassays in small pots with moist sand, most of the nematode isolates gave 30–60% mortality against each of the three larval stages. The susceptibility of the grubs for nematode infection generally increased with larval development.H. bacteriophora, H. heliothidis, H. megidis, a DutchHeterorhabditis isolate NLH-E87.3 andS. glaseri 326 showed the highest mortality rates, with nearly 100% mortality of third instar grubs. The DutchHeterorhabditis isolate NLH-E87.3 andS. glaseri 326 were selected as candidates for further studies on their potential as biological control agents forP. horticola grubs in the field.     

The identification of vahlkampfiid amoebae by ITS sequencing

We have determined the internal transcribed spacer (ITS) sequences (including the 5.8S ribosomal DNA) of 30 strains of 14 species belonging to eight vahlkampfiid genera. Each previously described species has a specific ITS sequence, except for Tetramitus aberdonicus, Tetramitus thorntoni, and Tetramitus jugosus, which have identical ITS sequences. The latter three may therefore constitute a single species despite their apparent phenotypic differences. The ITS sequence appears to be conserved within a species. The species Willaertia magna appears to be ubiquitous. The 5.8S rDNA sequences of Singhamoeba horticola and Learamoeba waccamwensis indicate that they do not represent different genera, but both belong to the genus Tetramitus. The ITS sequences of 16 undescribed vahlkampfiid isolates were determined. Based on these sequences, seven isolates were identified as belonging to described species, while nine probably represent seven new species. Five of these presumed new species belong to the genus Tetramitus, and one each to the genera Vahlkampfia and Paravahlkampfia.