Minute pollinators: The role of thrips (Thysanoptera) as pollinators of pointleaf manzanita, Arctostaphylos pungens (Ericaceae).

The feeding habits of thrips on plant tissue, and their ability to transmit viral diseases to their host plants, have usually placed these insects in the general category of pests. However, the characteristics that make them economically important, their high abundance and short- and long-distance movement capability, may also make them effective pollinators. We investigated this lesser-known role of thrips in pointleaf manzanita (Arctostaphylos pungens), a Southwestern US shrub. We measured the abundance of three species of thrips (Orothrips kelloggii, Oligothrips oreios, and Frankliniella occidentalis), examined their pollen-carrying capability, and conducted an exclusion experiment in order to determine whether thrips are able to pollinate this species, and if they do, whether they actually contribute to the reproductive success of the plant. Our data suggest that indeed thrips pollinate and do contribute significantly to reproductive success. Flowers exposed to thrips only produced significantly more fruit than did flowers from which all visitors were excluded. The roles of thrips as antagonists/mutualists are examined in the context of the numerous other floral visitors to the plant.

Florivory and nectar-robbing perforations in flowers of pointleaf manzanita Arctostaphylos pungens (Ericaceae) and their effects on plant reproductive success.

Damage to petals may have varying effects on the reproductive success of the plant. The variation may depend on the kind of damage to the corolla. Whether the damage is limited to the corolla, as is usually the case with nectar-robbing perforations, or extending to the reproductive parts of the flower, as in the case of florivory holes, might determine the extent of the effect on the plant's reproduction. We examined the various perforations in the flowers of Arctostaphylos pungens and correlated their presence with fruiting success. We found that though florivory holes were highly associated with damage to reproductive parts, fruiting success did not differ significantly between flowers with the two kinds of damage. Although nectar-robbing perforations were not associated with reduced number of fruit produced, they were significantly correlated with reduced number of fruit that contained seemingly viable seeds. The implications of our findings are discussed in the context of pollination and antagonism.

Synergy versus potency in the defensive secretions from nymphs of two pentatomomorphan families (Hemiptera: Coreidae and Pentatomidae).

One characteristic of true bugs (Heteroptera) is the presence of dorsal abdominal glands in the immature nymphal stages. These glands usually produce defensive chemicals (allomones) that vary among taxa but are still similar in closely related groups. Knowledge of the chemistry and prevalence of allomones in different taxa may clarify the evolution of these chemical defensive strategies. Within the infraorder Pentatomomorpha, the known secretions of nymphs of Pentatomidae tend to contain the hydrocarbon, n-tridecane, a keto-aldehyde, and an (E)-2-alkenal as the most abundant components. In the Coreidae, the dorsal abdominal gland secretions of nymphs often contain little or no hydrocarbon, and the most abundant keto-aldehyde and (E)-2-alkenal are often of shorter chain-length than those of pentatomids. We hypothesized that the long chain compounds would be less potent than their shorter homologs, and that bugs that carry the former would benefit from a synergistic effect of n-tridecane. To test this hypothesis we used three different behavioral assays with ants. A predator-prey assay tested the deterrence of allomones toward predators; a vapor experiment tested the effectiveness of allomones in the gaseous phase toward predators; and application of allomones onto predators tested the effect of direct contact. The results substantiate the hypothesis of a synergistic effect between n-tridecane and longer chain keto-aldehyde and (E)-2-alkenal in deterring predators. The short chain keto-aldehyde 4-oxo-(E)-2-hexenal was highly effective on its own. Thus, it seems that different groups of the infraorder diverged in their strategies involving defensive chemicals. Implications of this divergence are discussed.

Unusual macrocyclic lactone sex pheromone of Parcoblatta lata, a primary food source of the endangered red-cockaded woodpecker.

Wood cockroaches in the genus Parcoblatta, comprising 12 species endemic to North America, are highly abundant in southeastern pine forests and represent an important prey of the endangered red-cockaded woodpecker, Picoides borealis. The broad wood cockroach, Parcoblatta lata, is among the largest and most abundant of the wood cockroaches, constituting >50% of the biomass of the woodpecker's diet. Because reproduction in red-cockaded woodpeckers is affected dramatically by seasonal and spatial changes in arthropod availability, monitoring P. lata populations could serve as a useful index of habitat suitability for woodpecker conservation and forest management efforts. Female P. lata emit a volatile, long-distance sex pheromone, which, once identified and synthesized, could be deployed for monitoring cockroach populations. We describe here the identification, synthesis, and confirmation of the chemical structure of this pheromone as (4Z,11Z)-oxacyclotrideca-4,11-dien-2-one [= (3Z,10Z)-dodecadienolide; herein referred to as "parcoblattalactone"]. This macrocyclic lactone is a previously unidentified natural product and a previously unknown pheromonal structure for cockroaches, highlighting the great chemical diversity that characterizes olfactory communication in cockroaches: Each long-range sex pheromone identified to date from different genera belongs to a different chemical class. Parcoblattalactone was biologically active in electrophysiological assays and attracted not only P. lata but also several other Parcoblatta species in pine forests, underscoring its utility in monitoring several endemic wood cockroach species in red-cockaded woodpecker habitats.

Venom alkaloid and cuticular hydrocarbon profiles are associated with social organization, queen fertility status, and queen genotype in the fire ant Solenopsis invicta.

Queens in social insect colonies advertise their presence in the colony to: a) attract workers' attention and care; b) gain acceptance by workers as replacement or supplemental reproductives; c) prevent reproductive development in nestmates. We analyzed the chemical content of whole body surface extracts of adult queens of different developmental and reproductive stages, and of adult workers from monogyne (single colony queen) and polygyne (multiple colony queens) forms of the fire ant Solenopsis invicta. We found that the composition of the most abundant components, venom alkaloids, differed between queens and workers, as well as between reproductive and non-reproductive queens. Additionally, workers of the two forms could be distinguished by alkaloid composition. Finally, sexually mature, non-reproductive queens from polygyne colonies differed in their proportions of cis-piperidine alkaloids, depending on their Gp-9 genotype, although the difference disappeared once they became functional reproductives. Among the unsaturated cuticular hydrocarbons characteristic of queens, there were differences in amounts of alkenes/alkadienes between non-reproductive polygyne queens of different Gp-9 genotypes, between non-reproductive and reproductive queens, and between polygyne and monogyne reproductive queens, with the amounts increasing at a relatively higher rate through reproductive ontogeny in queens bearing the Gp-9 b allele. Given that the genotype-specific piperidine differences reflect differences in rates of reproductive maturation between queens, we speculate that these abundant and unique compounds have been co-opted to serve in fertility signaling, while the cuticular hydrocarbons now play a complementary role in regulation of social organization by signaling queen Gp-9 genotype.

Ants (Formicidae): models for social complexity.

The family Formicidae (ants) is composed of more than 12,000 described species that vary greatly in size, morphology, behavior, life history, ecology, and social organization. Ants occur in most terrestrial habitats and are the dominant animals in many of them. They have been used as models to address fundamental questions in ecology, evolution, behavior, and development. The literature on ants is extensive, and the natural history of many species is known in detail. Phylogenetic relationships for the family, as well as within many subfamilies, are known, enabling comparative studies. Their ease of sampling and ecological variation makes them attractive for studying populations and questions relating to communities. Their sociality and variation in social organization have contributed greatly to an understanding of complex systems, division of labor, and chemical communication. Ants occur in colonies composed of tens to millions of individuals that vary greatly in morphology, physiology, and behavior; this variation has been used to address proximate and ultimate mechanisms generating phenotypic plasticity. Relatedness asymmetries within colonies have been fundamental to the formulation and empirical testing of kin and group selection theories. Genomic resources have been developed for some species, and a whole-genome sequence for several species is likely to follow in the near future; comparative genomics in ants should provide new insights into the evolution of complexity and sociogenomics. Future studies using ants should help establish a more comprehensive understanding of social life, from molecules to colonies.

GC-MS for characterization and identification of ant semiochemicals.

Living in a predominantly dark environment, ants rely mostly on chemical signals for communication. Trail and alarm pheromones are the most widely studied and best characterized of all ant semiochemicals, but other such compounds can influence a variety of other behaviors, including reproductive activities, sexual development, nest mate and caste recognition, and defense. A typical worker body contains more than 10 different semiochemical-producing glands, and the surface of the cuticle is covered with lipids that serve as recognition signals. The methods of choice for collection and identification of ant semiochemicals should be determined based on results of behavioral analyses. These can indicate the source (e.g., glandular, cuticular) and the nature (volatile vs. nonvolatile) of the chemical. This protocol presents a number of different methods for collecting lipid semiochemicals. These can be followed by gas chromatography (GC) coupled with mass spectrometry (MS) to better characterize, and possibly identify, the semiochemical in question.

Cuticular hydrocarbons as maternal provisions in embryos and nymphs of the cockroach Blattella germanica.

Cuticular hydrocarbons of arthropods serve multiple functions, including as barriers to water loss and as pheromones and pheromone precursors. In the oviparous German cockroach, Blattella germanica, long-chain hydrocarbons are produced by oenocytes within the abdominal integument and are transported by a blood lipoprotein, lipophorin, both to the cuticular surface and into vitellogenic oocytes. Using radiotracer approaches, we tracked the location and metabolic fate of 14C- and 3H-labeled hydrocarbons through vitellogenic females and their embryos and nymphs. A considerable amount ( approximately 50%) of radiolabeled maternal hydrocarbons was transferred to oocytes and persisted through a 20-day embryogenesis and the first two nymphal stadia. The maternal hydrocarbons were not degraded or lost during this protracted period, except for significant losses of cuticular hydrocarbons starting with the first-to-second instar molt. Thus, although embryos and nymphs can produce their own hydrocarbons, maternal hydrocarbons provide a significant fraction of the cuticular and hemolymph hydrocarbons of both stages. These results show, for the first time in any insect, that a mother provides a significant complement of her offspring's cuticular hydrocarbons. Further research will be needed to determine whether provisioning hydrocarbons to eggs is a general strategy among insects and other arthropods or if this strategy is limited to taxa where eggs and early instars are susceptible to desiccation.

New contact sex pheromone components of the German cockroach, Blattella germanica, predicted from the proposed biosynthetic pathway.

Upon contacting the cuticle of a sexually mature female, a male German cockroach exhibits a characteristic courtship behavior: he turns away from the female and raises his wings, thereby exposing tergal glands. The glandular secretion stimulates the female to mount the male and feed, thus positioning her appropriately for copulation. A multi-component contact sex pheromone produced by females is responsible for eliciting courtship behavior. The most abundant pheromone components are 3,11-dimethylnonacosan-2-one and 3,11-dimethylheptacosan-2-one, oxidation products of the abundant hydrocarbon analogs 3,11-dimethylnonacosane and 3,11-dimethylheptacosane, respectively. The C(29)-dimethyl ketone is thought to be further metabolized to two less abundant pheromone components, 29-hydroxy-3,11-dimethylnonacosan-2-one and 29-oxo-3,11-dimethylnonacosan-2-one. Based on this proposed biosynthetic pathway of pheromone production, we hypothesized that 3,11-dimethylheptacosan-2-one also would be oxidized to give two candidate pheromone components, 27-hydroxy-3,11-dimethylheptacosan-2-one, and 27-oxo-3,11-dimethylheptacosan-2-one. By using bioassay-guided fractionation and chemical analyses of cuticular extracts of virgin females and synthesis of the (3S,11S)-isomer of each of the two predicted pheromone components, we showed that the epicuticle of the German cockroach does indeed contain these two compounds. The contact sex pheromone of the female German cockroach, thus may consist of at least six biosynthetically related components.

Identification of cuticular lipids eliciting interspecific courtship in the German cockroach, Blattella germanica.

The cuticular surface of sexually mature females of the German cockroach contains a sex pheromone that, upon contact with the male's antennae, elicits a characteristic species-specific courtship behavior. This female-specific pheromone is a blend of several long-chain methyl ketones, alcohols and aldehydes, all derived from prominent cuticular hydrocarbons found in all life stages of this cockroach. We found that contact with the antennae of 5 out of 20 assayed cockroach species elicited courtship behavior in German cockroach males. The heterospecific courtship-eliciting compounds were isolated by behaviorally guided fractionation of the active crude extracts and compared to the native sex pheromone components. We identified two active compounds from the cuticular extract of the Oriental cockroach, Blatta orientalis -- 11-methylheptacosan-2-one and 27-oxo-11-methylheptacosan-2-one; the former compound was confirmed by synthesis and proved to independently stimulate courtship in German cockroach males. These compounds share common features with, but are distinct from, any of the known contact sex pheromone components. This suggests that sex pheromone reception in the male German cockroach is unusually promiscuous, accepting a wide range of compounds that share certain features with its native pheromone, thus resulting in a broad spectrum of behavioral response to other species. We propose that several characteristics of their mating system -- chiefly, absence of closely related species in the anthropogenic environment, resulting in relaxation of selection on sexual communication, and a highly male-biased operational sex ratio -- have driven males to respond with extremely low thresholds to a wide spectrum of related compounds.

Behavioral activity of stereoisomers and a new component of the contact sex pheromone of female German cockroach, Blattella germanica.

(3S,11S)-3,11-Dimethylnonacosan-2-one is a major component of the courtship stimulating, contact sex pheromone of the female German cockroach. Although the four synthetic stereoisomers of this compound have been tested in behavioral assays, their relative activity remains unresolved. Using isolated male antennae dosed with synthetic test compounds to assay male behavior, we found that at high doses all four stereoisomers elicited responses from 100% of the males. However, at physiologically relevant doses similar to those found on the female antenna, the (3S,11S)-isomer was the least effective of the four stereoisomers at eliciting courtship responses in males. This is the first example of a natural stereoisomer having less bioactivity than related stereoisomers that do not occur naturally. Another component of the sex pheromone blend, 3,11-dimethylheptacosan-2-one, was previously purified from the female's epicuticule and behaviorally assayed, but its activity was not confirmed through synthesis. We now confirm that synthetic (3S,11S)-3,11-dimethylheptacosan-2-one elicits behavioral responses, but less so than its C29 homolog.