Blending synthetic pheromones of cerambycid beetles to develop trap lures that simultaneously attract multiple species

We evaluated attraction of cerambycid beetle species to blends of known cerambycid pheromones to determine whether such blends could be used as effective trap lures for detecting and monitoring multiple species simultaneously. Pheromone-baited traps captured 1,358 cerambycid beetles of which 1,101 (81.1%) belonged to three species in the subfamily Cerambycinae: Neoclytus acuminatus (F.), Neoclytus mucronatus (F.), and Xylotrechus colonus (F.). Beetles of these species were significantly attracted to synthetic blends that contained their pheromone components (isomers of 3-hydroxy-2-hexanone, 2,3-hexanediol, or both), despite the presence of pheromone components of different species, including other isomers of 2,3-hexanediol, (E/Z)-6,10-dimethyl-5,9-undecadien-2-yl acetate, and citral. In some cases, attraction was partially inhibited by the pheromone components of heterospecific species, whereas for N. acuminatus, attraction was completely inhibited when blends contained (2R*,3S*)-hexanediol, the racemic mixture of diastereomers of its pheromone, (2S,3S)-hexanediol. Among the remaining beetles captured were three species in the subfamily Lamiinae: Astyleiopus variegatus (Haldeman), Graphisurus fasciatus (Degeer), and Lepturges angulatus (LeConte). All three lamiine species were previously known to be attracted to (E/Z)-6,10-dimethyl-5,9-undecadien-2-yl acetate and were captured in significant numbers by blends containing that compound. Our results suggest that different types of cerambycid pheromones can be combined to create effective multispecies lures for use in surveillance programs that target exotic cerambycid species.     

天牛成虫信息素及嗅觉感受机制研究进展

评述了天牛成虫信息素及嗅觉感受机制的研究进展。天牛雌、雄成虫均可释放性信息素,迄今已对31种天牛的性信息素进行了研究,其中完成组分鉴定的有13种。天牛性信息素包括长距离、短距离和接触性信息素3种类型。天牛性信息素存在变异现象,同种天牛分布在不同地区,其性信息素组分之间存在差异。触角是天牛感受性信息素的主要器官,也是判别成虫通讯方式的形态指标,性信息素发达的种类其触角常具显著的性二型现象。天牛利用寄主信息素（如萜烯类、醇类和酯类）寻找寄主。性信息素和寄主信息素在林间复合使用可提高诱捕率。天牛信息素还包括异种信息素、忌避信息素和产卵干扰素,能够提高天牛寄主定位效率。天牛触角嗅觉感受的神经细胞（RNs）有3类,气味信息经神经细胞群传输至中枢神经系统,神经信号按标记路线或交叉纤维样式输导。天牛气味结合蛋白（OBP）方面的研究尚未见报道。     

Adults of the cerambycid beetle Megacyllene caryae use both olfactory and visual information to locate mates

Many species of beetles in the family Cerambycidae use volatile pheromones to facilitate the location of mates. Visual cues may also influence the location of mates, as the adults of many species of cerambycids are often brightly patterned and diurnal. Theory predicts that combining signals or cues of different modalities (e.g., chemical, visual) to transmit information will increase the likelihood of an organism responding to this information, compared to when the signal or cue is presented alone. Here, we test the hypothesis that attraction of adults of the cerambycid beetle Megacyllene caryae (Gahan) (Coleoptera: Cerambycidae, Clytini) to their pheromones will be increased when visual cues are present. Consistent with that hypothesis, the number of beetles caught by traps baited with pheromones was increased 3.4‐fold when a dead adult beetle of that species was attached to the trap, relative to those with just pheromone alone. Capture of M. caryae in our study was also influenced by the position of traps within forest stands, with traps at 100 m within stands catching 2.5× as many beetles as traps at the forest edge. These findings suggest that vision and visual cues play an important role in the location of mates by cerambycid beetles and warrant further research. Also, the inclusion of visual cues on traps may enhance the efficacy of trapping cerambycid beetles, such as the detection of species that are non‐native and potentially invasive, or when monitoring species that are native and of conservation concern.     

天牛性信息素、引诱植物和植物性引诱剂的研究与应用

由于目前所拥有的各种天牛防治方法都难以获得理想的防治效果,研发新型天牛控制技术势在必行。其中利用引诱剂是最具潜力的研究领域之一。对天牛的性信息素、引诱植物和植物性引诱剂3个方面的研究及应用作了综述。天牛雌、雄成虫均可释放性信息素进行两性间的通讯联系。天牛性信息素的分泌部位因天牛种类不同而存在着差异。雄、雌成虫触角上均有感受性信息素的感受器,但报道较多的是雄虫感受器。迄今已鉴定出13种天牛性信息素的化学结构。一般而言,天牛成虫的嗜食树种,对天牛具有较强的引诱作用,可作为诱饵树种使用。寄主植物的某些挥发性物质对天牛成虫也有独特的引诱效果。除此之外,对天牛性信息素、引诱植物和植物性引诱剂研究与应用中存在的困难也进行了讨论,并对它们的应用前景做了展望。     

A comparison of trap type and height for capturing cerambycid beetles (Coleoptera)

Wood-boring beetles in the family Cerambycidae (Coleoptera) play important roles in many forest ecosystems. However, increasing numbers of invasive cerambycid species are transported to new countries by global commerce and threaten forest health in the United States and worldwide. Our goal was to identify effective detection tools for a broad array of cerambycid species by testing some known cerambycid attractants and a pheromone in different trap designs placed across a range of habitats. We compared numbers and species richness of cerambycid beetles captured with cross-vane panel traps and 12-unit Lindgren multiple-funnel traps, placed either at ground level (1.5 m high) or canopy level (approximately 3-10 m high), at eight sites classified as either residential, industrial, deciduous forest, or conifer forest. We captured 3,723 beetles representing 72 cerambycid species from 10 June to 15 July 2010. Species richness was highest for the subfamilies Cerambycinae and Lamiinae, which accounted for 33 and 46% of all species captured, respectively. Overall, the cross-vane panel traps captured approximately 1.5 times more beetles than funnel traps. Twenty-one species were captured exclusively in traps at one height, either in the canopy or at ground level. More species were captured in hardwood sites (59 species) where a greater diversity of host material was available than in conifer (34 species), residential (41 species), or industrial (49) sites. Low numbers of beetles (n < 5) were recorded for 28 of the beetle species. The number of species captured per week ranged from 49 species on 21 June to 37 species on 12 July. Cross-vane panel traps installed across a vertical gradient should maximize the number of cerambycid species captured.     

Longhorn beetles and predatory clerid beetles attracted to a blend of longhorn beetle pheromone compounds in a Central European oak forest (Coleoptera: Cerambycidae,Cleridae)

1. A blend of longhorn beetle pheromones was tested as a generic attractant in a Central European oak forest. Overall, 20 cerambycid species totalling 1250 specimens were captured using two trap types.
2. More adults of Phymatodes testaceus and Leiopus nebulosus nebulosus were attracted to pheromone-baited traps compared to solvent controls. Significant numbers of four other species were caught by panel traps but not funnel traps. For the cerambycine Pyrrhidium sanguineum, significantly more beetles were caught in treatment traps than controls. For the cerambycine Anaglyptus mysticus, lepturines Cortodera humeralis and Rhagium sycophanta, the numbers of beetles caught in treatment or control traps were similar, indicating no attraction to the lure blend. Adults of a predatory clerid beetle Clerus mutillarius were caught in significantly larger numbers by both baited trap types in comparison to controls, totalling 1514 specimens.
3. Antennae of both sexes of C. mutillarius showed responses to 3-hydroxyhexan-2-one and 2-methylbutan-1-ol, and males also responded to syn-2,3-hexanediol.
4. Strong attraction of C. mutillarius suggests eavesdropping on the pheromones of cerambycids and that such attractant baits, without traps, might be used to manipulate the local population density of predators in a push-pull biological control program.

     

Ophiostomatoid fungi can emit the bark beetle pheromone verbenone and other semiochemicals in media amended with various pine chemicals and beetle-released compounds

Fungal volatile compounds can mediate fungal-insect interactions. Whether fungi can emit insect pheromones and how volatile chemicals change in response to chemicals the fungi naturally encounter is poorly understood. We analyzed volatiles emitted by Grosmannia clavigera (symbiont of the mountain pine beetle) and Ophiostoma ips (symbiont of the pine engraver beetle) growing in liquid media amended with compounds that the fungi naturally encounter: (−)-α-pinene, (+)-α-pinene, (−)-trans-verbenol, verbenone, or ipsdienol. Nine volatile compounds were identified among the fungal and amendment treatments. Volatiles qualitatively and quantitatively differed between fungal species and among amendment treatments. The bark beetle anti-aggregation pheromone (−)-verbenone was detected from both fungi growing in (−)-trans-verbenol-amended media. G. clavigera and O. ips can emit beetle pheromones and other beetle semiochemicals, suggesting that ophiostomatoid fungi could contribute to the chemical ecology of bark beetles. However, such contributions could be modulated by the presence of other environmental chemicals.     

Beetle responses to artificial gaps in an oceanic island forest: implications for invasive tree management to conserve endemic species diversity

Natural forests are often replaced by invasive alien trees on isolated oceanic islands. Adequate eradication of invasive trees should be conducted with the goal of biodiversity conservation, because islands support many endemic organisms that depend on native forests. An invasive alien tree, Bischofia javanica Blume (Euphorbiaceae), has invaded and replaced natural forests on the oceanic Ogasawara (Bonin) Islands, Japan, in the northwestern Pacific Ocean. To determine how the removal of B. javanica trees affects insect diversity, we examined flying beetles captured using Malaise traps in B. javanica forests on Hahajima. The abundance, species density, and species composition of wood-boring beetles (Coleoptera: Cerambycidae, Elateridae, Mordellidae, and Scolytidae) were compared between closed-canopy sites and gaps created by girdling B. javanica trees in alien forests during two seasons (June–July and October–November 2005). Of the collected beetles, 75.8, 87.5, 90.0, and 0.0% of cerambycid, elaterid, mordellid, and scolytid beetle species, respectively, were endemic to the Ogasawara Islands. More cerambycid, elaterid, and mordellid individuals were captured in June–July than in October–November; the number of scolytid individuals did not differ between seasons. More cerambycid, elaterid, and scolytid individuals were captured in artificial gaps than on the closed-canopy forest floor. Although fewer mordellid individuals were captured in gaps, more endemic mordellids were captured in gaps. More cerambycid and scolytid species were captured in artificial gaps than in closed-canopy areas. The positive responses of beetles to artificial gaps suggest that the removal of B. javanica increases beetle diversity and the abundance of endemic beetles.     

Response of native and exotic longhorn beetles to common pheromone components provides partial support for the pheromone-free space hypothesis

Longhorn beetles are among the most important groups of invasive forest insects worldwide. In parallel, they represent one of the most well-studied insect groups in terms of chemical ecology. Longhorn beetle aggregation-sex pheromones are commonly used as trap lures for specific and generic surveillance programs at points of entry and may play a key role in determining the success or failure of exotic species establishment. An exotic species might be more likely to establish in a novel habitat if it relies on a pheromone channel that is different to that of native species active at the same time of year and day, allowing for unhindered mate location (i.e., pheromone-free space hypothesis). In this study, we first tested the attractiveness of single pheromone components (i.e., racemic 3-hydroxyhexan-2-one, racemic 3-hydroxyoctan-2-one, and syn-2,3-hexanediol), and their binary and tertiary combinations, to native and exotic longhorn beetle species in Canada and Italy. Second, we exploited trap catches to determine their seasonal flight activity. Third, we used pheromone-baited “timer traps” to determine longhorn beetle daily flight activity. The response to single pheromones and their combinations was mostly species specific but the combination of more than one pheromone component allowed catch of multiple species simultaneously in Italy. The response of the exotic species to pheromone components, coupled with results on seasonal and daily flight activity, provided partial support for the pheromone-free space hypothesis. This study aids in the understanding of longhorn beetle chemical ecology and confirms that pheromones can play a key role in longhorn beetle invasions.     

The mode of pheromone evolution: evidence from bark beetles

Sex and aggregation pheromones consist of species-specific blends of chemicals. The way in which different species' blends have evolved has been the subject of some debate. Theoretical predictions suggest that differences between species have arisen not through the accruing of small changes, but through major shifts in chemical composition. Using data on the aggregation pheromones of 34 species of bark beetle from two genera, Dendroctonus and Ips, we investigated how the distributions of the chemical components of their pheromone blends mirror their phylogenetic relationships. We tested whether there were consistent patterns that could be used to help elucidate the mode of pheromone evolution. Although there were obvious differences in pheromone blends between the two genera, the differences between species within each genus followed a less clear phylogenetic pattern. In both genera, closely related species are just as different as more distantly related species. Within Dendroctonus, particularly, most chemical components were distributed randomly across the phylogeny. Indeed, for some chemicals, closely related species may actually be more different than would be expected from a random distribution of chemical components. This argues strongly against the idea of minor shifts in pheromone evolution. Instead, we suggest that, within certain phylogenetic constraints, pheromone evolution in bark beetles is characterized by large saltational shifts, resulting in sibling species being substantially phenotypically (i.e. pheromonally) different from one another, thus agreeing with theoretical predictions.     

Biological invasion into the nested assemblage of tree–beetle associations on the oceanic Ogasawara Islands

Invasion by alien organisms is a common worldwide phenomenon, and many alien species invade native communities. Invasion by alien species is especially likely to occur on oceanic islands. To determine how alien species become integrated into island plant–insect associations, we analyzed the structure of tree–beetle associations using host plant records for larval feeding by wood-feeding beetles (Coleoptera: Cerambycidae) on the oceanic Ogasawara Islands in the northwestern Pacific Ocean. The host plant records comprised 109 associations among 28 tree (including 8 alien) and 26 cerambycid (including 5 alien) species. Of these associations, 41.3% involved at least one alien species. Most native cerambycid species feed on host trees that have recently died. Alien trees were used by as many native cerambycid species (but by significantly more alien cerambycid species) as were native trees. Native cerambycid species used as many alien tree species (but significantly more native tree species) as did alien cerambycids. Thus, we observed many types of interactions among native and alien species. A network analysis revealed a significant nested structure in tree–cerambycid associations regardless of whether alien species were excluded from the analysis. The original nested associations on the Ogasawara Islands may thus have accepted alien species.     

The mode of evolution of aggregation pheromones in Drosophila species

Aggregation pheromones are used by fruit flies of the genus Drosophila to assemble on breeding substrates, where they feed, mate and oviposit communally. These pheromones consist of species-specific blends of chemicals. Here, using a phylogenetic framework, we examine how differences among species in these pheromone blends have evolved. Theoretical predictions, genetic evidence, and previous empirical analysis of bark beetle species, suggest that aggregation pheromones do not evolve gradually, but via major, saltational shifts in chemical composition. Using pheromone data for 28 species of Drosophila we show that, unlike with bark beetles, the distribution of chemical components among species is highly congruent with their phylogeny, with closely related species being more similar in their pheromone blends than are distantly related species. This pattern is also strong within the melanogaster species group, but less so within the virilis species group. Our analysis strongly suggests that the aggregation pheromones of Drosophila exhibit a gradual, not saltational, mode of evolution. We propose that these findings reflect the function of the pheromones in the ecology of Drosophila, which does not hinge on species specificity of aggregation pheromones as signals.     

Differences in endemic insect assemblages among vegetation types on a small island of the oceanic Ogasawara Islands

Natural vegetation is often replaced by invasive alien plants on isolated oceanic islands. To determine how invasive alien plants affect insect diversity, we compared flying insects captured using Malaise traps among different vegetation types on a small island (Nishijima; 0.49 km²) in the oceanic Ogasawara (Bonin) Islands in the north‐western Pacific. The numbers of individuals and species, and the species composition of pollinators (bees), predators (wasps) and wood borers (cerambycid, mordellid and elaterid beetles) were compared among three vegetation types: Casuarina equisetifolia (an invasive alien tree) forest, natural forest and natural grassland (forest edge), during two seasons (June and October–November 2005). In traps, 80.0, 66.7, 87.5, 85.7 and 100.0% of bee, wasp, cerambycid, mordellid and elaterid beetle species, respectively, were endemic to the Ogasawara Islands. Grassland had the highest wasp and bee species richness, whereas natural forest had the highest species richness of wood‐boring beetles. The C. equisetifolia forest had the poorest species richness for most insect groups (except mordellid beetles). More individuals of most insect groups (except bees) were captured in June than in October–November. More individual bees and wasps were captured in grassland than in forests, whereas more individual mordellid and elaterid beetles were captured in forests than in grassland. The number of cerambycid individuals did not differ among vegetation types. Redundancy analysis suggested that most insect species preferred natural forest or grassland to alien forest. Therefore, further invasion of natural grassland and forest by the alien tree C. equisetifolia may negatively affect the endemic insect fauna of Nishijima.     

Suitability of commercially available insect traps and pheromones for monitoring dusky sap beetles (Coleoptera: Nitidulidae) and related insects in Bt sweet corn

Two trap types and pheromone sources for the dusky sap beetle, Carpophilus lugubris Murray (Coleoptera: Nitidulidae), were compared in Bacillus thuringiensis (Bt) and non-Bt sweet corn fields over a 3-yr period. Overall, commercial traps and pheromones were equally effective as experimental traps and pheromones used previously for capturing C. lugubris and other sap beetle species. The commercial trap often caught significantly more Glischrochilus quadrisignatus Say than the experimental trap that had been used in previous studies. Bt corn significantly reduced caterpillar damage to ears compared with the non-Bt isoline and did not adversely affect levels of Orius sp., the most common insect predator. Sap beetle damage was the most common insect damage to Bt sweet corn ears. Sap beetles were detected by traps at population levels below that which are likely to cause economic concern, indicating commercially available traps and pheromone lures for monitoring sap beetles should be suitable for detecting them under commercial growing conditions.     

Midgut and fat body bacteriocytes in neotropical cerambycid beetles (Coleoptera: Cerambycidae)

Xylophagous insects derive nutrients from intractable substrates by producing or ingesting cellulolytic enzymes, or by maintaining associations with symbiotic microbes. Wood-boring cerambycid beetle larvae sometimes house maternally-transmitted endosymbiotic yeasts that are presumed to provide their hosts with nutritional benefits. These are thought to be absent from species in the large subfamily Lamiinae; nevertheless yeasts have been repeatedly isolated from the guts of neotropical lamiines. The objective of this study was to conduct transmission electron microscopy (TEM) studies of cerambycid larval midgut tissues to determine if gut yeasts were intracellular, or simply present in the gut lumen. Nine cerambycid larvae were harvested from two trees in the Brazil nut family (Lecythidaceae) in the rain forest of SE Peru; seven were identified using mtDNA sequence data and processed for TEM. Yeasts cultured from larval frass or exuvia, and identified with rDNA sequence data, were identical or similar to yeasts previously isolated from beetles. In TEM analyses yeast cells were found only in the gut lumens, sometimes associated with fragments of thick-walled xylem cells. Apparent bacteriocytes were found in either midgut or fat body tissue of three larval specimens, including two lamiines. This is the first report of a potential fat body symbiosis in a cerambycid beetle. Future studies of cerambycid symbiosis should distinguish the identities and potential roles of free-living organisms in the gut lumen from those of organisms harbored within gut epithelial or fat body tissue.     

Potential vector switching in the evolution of Bursaphelenchus xylophilus group nematodes (Nematoda: Aphelenchoididae)

To show the importance of vector switching of nematodes in the evolution of the Bursaphelenchus xylophilus group, we tested a hypothesis that “Bursaphelenchus doui (or its ancestor) was transferred by Acalolepta fraudatrix, Acalolepta sejuncta, and/or Monochamus subfasciatus (or their ancestral species) from broad‐leaved trees to conifers, switched vectors from these cerambycid beetles to Monochamus beetles in conifers, and then evolved into the common ancestor of Bursaphelenchus mucronatus and B. xylophilus.” We used a simple nematode‐loading method to beetles and produced 20 binary combinations of five B. xylophilus group species and four cerambycid beetle species in the tribe Lamiini. The affinity of the nematodes for the beetles was examined based on phoretic stage formation of the nematodes. Phoretic stages of B. doui appeared in all beetle species examined, namely Acalolepta luxuriosa, Psacothea hilaris, A. fraudatrix, and Monochamus alternatus, although the affinity of the nematode for M. alternatus was weak. This finding indicates that B. doui could switch vectors to conifer‐using Monochamus beetles after transfer by A. fraudatrix from broad‐leaved trees to conifers. We conclude that vector switching of nematodes could have potentially happened during the evolutionary history of the B. xylophilus group.     

叶甲科昆虫的定殖机制

在复杂的生态环境中,叶甲科昆虫利用寄主植物的挥发物作为嗅觉信号,并协同视觉信号共同作用以提高其远距离搜寻和定位寄主的效率。叶甲科昆虫也释放信息素来调节同种或者异种个体的行为反应;找到寄主植物后,叶子的形态学特性和化学组分的浓度等接触性因素就会影响叶甲科昆虫对寄主植物的最终选择。叶甲对上述这些信号物质的反应也受叶甲本身状态的影响,如生殖、滞育、饥饿、交配以及取食等。叶甲科昆虫对寄主植物的搜寻、辨识和接受、取食以及繁殖的过程受一种或多种因素协同影响。从嗅觉、视觉和触觉等方面对叶甲科昆虫的定殖机制做一综述。     

Toxicity of Chromobacterium subtsugae to southern green stink bug (Heteroptera: Pentatomidae) and corn rootworm (Coleoptera: Chrysomelidae)

Diabrotica spp. (Coleoptera: Chrysomelidae) beetles and southern green stink bugs, Nezara viridula (L.) (Heteroptera: Pentatomidae), are pests on corn, Zea mays L., and soybean, Glycine max (L.) Merr., as well as on cucurbits. Control of these insects has depended on chemicals. An alternative to chemical control is the use of biologicals. Use of bacteria, fungi, viruses, pheromones, and metabolites to control these insects can potentially improve resistance management and reduce pesticide use. Other than Bacillus thuringiensis Berliner, few bacteria have been discovered that are lethal to either of these pests. Chromobacterium subtsugae Martin et al., a newly described bacterium that is known to be toxic to Colorado potato beetle, Leptinotarsa decemlineata (Say), larvae, was found to be toxic to both diabroticite adult beetles and southern green stink bug adults. In laboratory assays, toxins produced by these bacteria kill 80-100% of the adults of two species of diabroticite beetles, Diabrotica undecimpunctata howardi Barber and Diabrotica virgifera virgifera LeConte, and 100% of southern green stink bug adults within 6 d. For green stink bug, live bacteria were not needed for toxicity.     

Small off-shore islands can serve as important refuges for endemic beetle conservation

Although large islands generally support a richer insect fauna than small islands, many large islands, which are more often inhabited, have lost numerous species because of human activities and introduced organisms. To clarify the consequences of endemic insect conservation on small islands near inhabited islands, we compared the species richness, abundance, and composition of two beetle groups (Coleoptera: Cerambycidae and Mordellidae) captured using Malaise traps among three islands (Chichijima, 24.0 km²; Anijima, 7.85 km²; Nishijima, 0.49 km²) in the oceanic Ogasawara (Bonin) Island group in the northwestern Pacific during June–July 2006 and 2007. Chichijima, the largest island, is inhabited, while Anijima and Nishijima are not. The numbers of cerambycid and mordellid species previously recorded were positively correlated with island area. However, the total numbers of cerambycid and mordellid species we captured in Malaise traps were not correlated with island area because we were unable to collect many species previously documented on Chichijima. The numbers of cerambycid and mordellid species per trap did not differ significantly among islands and years, although the deviance was well explained by the island variable. We captured greater numbers of cerambycid and mordellid individuals on Chichijima than on Anijima and Nishijima, and the numbers of cerambycid and mordellid individuals per trap significantly differed among islands and between years. Redundancy analysis (RDA) showed that the species composition of cerambycids and mordellids differed among the three islands. Whereas endangered species were rarely captured on Chichijima, alien or non-endemic species were frequently collected. Cerambycid and mordellid beetles on Chichijima may have been deleteriously affected by recent forest disturbance and introduced organisms. Therefore, conserving insect fauna on uninhabited island “refugia” is important for preserving the insect diversity of the Ogasawara Islands.