Social closure,aggressive behavior,and cuticular hydrocarbon profiles in the polydomous antCataglyphis iberica (hymenoptera,Formicidae)

Nestmate recognition was studied in the polydomous antCataglyphis iberica (Formicinae) in the laboratory. The study examined six colonies collected from two different populations 600 km apart in the Iberian peninsula (Barcelona and Murcia). Introduction of an alien worker into an allocolonial arena always ended in death to the intruder, demonstrating that in this species societies are extremely closed. Dyadic encounters composed of individuals from different colonies in a neutral arena confirmed the existence of high aggression between allocolonial individuals. We also investigated variability in the composition of the major cuticular hydrocarbons between the colonies used in the behaviorial tests. There were marked quantitative differences between the profiles of ants from the two populations, suggesting that the populations are completely segregated. Cuticular profiles within a population tended to be more similar, but were nevertheless colony specific. The degree of colony closure inC. iberica seemed to be independent of geographic distance since aggression between the colonies was always at its maximum, irrespective of their population origin.     

Venom Alkaloid and Cuticular Hydrocarbon Profiles Are Associated with Social Organization,Queen Fertility Status,and Queen Genotype in the Fire Ant <Emphasis Type="Italic">Solenopsis invicta</Emphasis>

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.     

Biologically active secondary metabolites of barley. II. Phytotoxicity of barley allelochemicals

The release of alkaloids by barley was quantified by HPLC. Hordenine was released from the roots of barley in a hydroponic system for up to 60 days. The amount reached a maximum, 2g/plant/day, at 36 days, then declined. Effects on white mustard by hordenine and gramine included reduction of radicle length and apparent reduction in health and vigor of radicle tips. Transmission electron microscopic examination of white mustard radicle tips exposed to hordenine and gramine showed damage to cell walls, increase in both size and number of vacuoles, autophagy, and disorganization of organelles. The evidence of the morphological and primary effects of barley allelochemicals at the levels released by living plants indicates that the biologically active secondary metabolites of barley may lead to a significant role in selfdefense by the crop.     

Cuticular lipid profiles of queens,workers, and males of social waspPolistes metricus say are colony-specific

The cuticular lipids ofPolistes metricus queens, workers and males from seven laboratory-maintained colonies were extracted and analyzed by combined gas chromatography-mass spectrometry. Males had higher proportions of alkenes (20.5%) in their cuticular lipids than did queens (2.3%) or workers (7.7%). Discriminant analyses of the cuticular lipid profiles of the adult wasps showed that males group separately from females. Additional analyses showed that queens group with their respective workers by colony and that queens group even more closely with males by colony. The most distinct groupings occurred with workers only by colony and with males only by colony. Stepwise discriminant analyses showed that each type of grouping was dependent upon a different combination of cuticular lipids.     

Cuticular Hydrocarbon Composition Reflects Genetic Relationship Among Colonies of the Introduced Termite Reticulitermes santonensis Feytaud

Nestmate recognition plays a key role in kin selection to maintain colony integrity in social insects. Previous studies have demonstrated that nestmate recognition is dependent on detection of cuticular hydrocarbons. However, the absence of intraspecific aggression between some colonies of Isoptera and social Hymenoptera questions whether kin recognition must occur in social insects. The purpose of this study was to determine if cuticular hydrocarbon similarity and high genetic relatedness could explain the lack of intraspecific aggression among and within colonies of the introduced subterranean termite Reticulitermes santonensis. We performed both GC analysis of cuticular hydrocarbons and genotyping by using 10 DNA microsatellite loci on the same 10 workers from each of 14 parisian colonies. Multivariate analyses demonstrated correspondence between cuticular hydrocarbon patterns and genetic variation. By using a redundancy analysis combining chemical and genetic data, we found that a few hydrocarbons (mainly short vs. long chains; saturated vs. unsaturated alkanes) were associated with most genetic variation. We also found a strong positive correlation between chemical and genetic distances between colonies, thus providing evidence of a genetic basis for cuticular hydrocarbon variation. However, genetic distance did not account for all chemical variation, thus suggesting that some hydrocarbon variation was environmentally derived. Investigation at the intracolony level indicated that cuticular hydrocarbons did not depend on colony social structure. Based on our findings, we speculate that the absence of intraspecific aggression in R. santonensis may result from a loss of diversity in genetically derived recognition compounds in this species that presumably descended from R. flavipes populations imported from North America.     

Temporal and Spatial Variation in Alkaloid Levels in Achnatherum robustum, a Native Grass Infected with the Endophyte Neotyphodium

The native North American perennial grass Achnatherum robustum (Vasey) Barkworth [= Stipa robusta (Vasey) Scribn.] or sleepygrass is toxic and narcotic to livestock. The causative agents are alkaloidal mycotoxins produced from infections by a systemic and asexual Neotyphodium endophyte. Recent studies suggest that toxicity is limited across the range of sleepygrass in the Southwest USA. We sampled 17 populations of sleepygrass with varying distance from one focal population known for its high toxicity levels near Cloudcroft, NM, USA. For some, we sampled individual plants twice within the same growing season and over successive years (2001–2004). We also determined infection levels in each population. In general, all populations were highly infected, but infection levels were more variable near the focal population. Only infected plants within populations near the Cloudcroft area produced alkaloids. The ergot alkaloid, ergonovine, comprised the bulk of the alkaloids, with lesser amounts of lysergic and isolysergic acid amides and ergonovinine alkaloids. Levels of all alkaloids were positively correlated among individual plants within and between growing seasons. Infected plants that produced no alkaloids in 1 yr did not produce any alkaloids within the same growing season or in other years. Levels of alkaloids in sleepygrass populations declined with distance from the Cloudcroft population, although infection levels increased. Infected plants in populations in northern New Mexico and southern Colorado produced no alkaloids at all despite 100% infectivity. Our results suggest that only specific Neotyphodium haplotypes or specific Neotyphodium–grass combinations produce ergot alkaloids in sleepygrass. The Neotyphodium haplotype or host–endophyte combination that produces toxic levels of alkaloids appears restricted to one locality across the range of sleepygrass. Because of the wide variation in alkaloid levels among populations, interactions between the endophyte and host, and consequences for herbivores, competitors, and pathogens and other components of the community, are likely to vary widely across the geographic range of this native grass.     

Variation of Brominated Indoles and Terpenoids Within Single and Different Colonies of the Marine Bryozoan Flustra foliacea

The variation of the brominated indole and diterpenoid content within single and different colonies of the bryozoan Flustra foliacea was investigated. The secondary metabolite profile and concentrations of individual components of F. foliacea samples were established using GC-MS. Samples from 17 different collecting sites were analyzed. The alkaloid and diterpene composition of F. foliacea varied greatly depending upon the site of collection. Investigation of F. foliacea samples from a single site (Helgoland, North Sea) over a period of time showed that the alkaloid and diterpenoid profile remained constant, however concentrations of individual components varied significantly. The alkaloid and diterpenoid composition of different segments of a single colony was found to be constant. Only small differences could be detected in the essential oil composition of different colonies and segments of single colonies of F. foliacea. Two of the F. foliacea alkaloids were found in the gastropods Hydrobia ulva and Gibbula cinerea, and one alkaloid in the common starfish Asteria rubens, all collected from the surface of the bryozoan.     

Intercaste,intercolony, and temporal variation in cuticular hydrocarbons ofCopotermes formosanus shiraki (Isoptera: Rhinotermitidae)

We characterized the variation in cuticular hydrocarbon mixtures between seven colonies of the Formosan subterranean termite,Coptotermes formosanus Shiraki, from the same population. We report differences between castes, between colonies, and within the population over time to assess seasonality. Colonies ofC. formosanus from Oahu, Hawaii, were sampled for 25 months. Each month, one sample each of 200 workers, 50 soldiers, nymphs, or alates from each colony was subjected to gas chromatography-mass spectrometry (GC-MS) analysis of the cuticular hydrocarbons. We resolved 39 individual peaks and identified 52 individual or isomeric mixtures of hydrocarbons. Onlyn-alkanes and methyl-branched alkanes occur; no olefins were found. Internally branched monomethylalkanes were the most abundant class of hydrocarbons, representing 45% to 50% of the total 9-;11-;13-Methyl-heptacosane accounted for over 30% of the total hydrocarbon for all castes. 2-Methyl- and 3-methylalkanes comprise approximately 30% of the total. Internally branched dimethylalkanes constitute 15% to 20% of the total cuticular hydrocarbon. Only one trimethylalkane, 13,15,17-trimethylnonacosane, was found in small amounts. The hydrocarbon mixtures of all four castes were similar. Quantitative differences in hydrocarbon mixtures among the castes were easily displayed using canonical discriminant analysis. Soldiers and workers are significantly different from one another and from nymphs and alates. Nineteen peaks are statistically significant between workers and soldiers. Nymphs and alates were not statistically different. We detected statistically significant quantitative differences between colonies in 18 peaks for workers and 12 peaks for soldiers. Each of the colonies ofC. formosanus can be separated from the others by the proportions of their hydrocarbon components. We detected statistically significant differences between months of the year for 12 peaks for workers and four peaks for soldiers; two peaks each for workers and soldiers showed distinct, seasonal trends. This seasonal shift in proportions of hydrocarbons correlates with the production of alates.     

Temporal changes in colony cuticular hydrocarbon patterns ofSolenopsis invicta

Heritable cuticular hydrocarbon patterns ofSolenopsis invicta workers are consistent within colonies for a given sampling time but vary sufficiently from colony to colony to distinguish the colonies from each other. In addition, cuticular hydrocarbon patterns change within colonies over time. Nestmate recognition cues found on the individual's cuticle, can be from heritable or environmental sources, and are a subset of colony odor. The cuticular hydrocarbons can be used as a model for heritable nestmate recognition cues. We propose that because potential nestmate recognition cues, both environmental and genetic, are dynamic in nature rather than static, during its lifetime a worker must continually update its perception (template) of colony odor and nestmate recognition cues.     

Ergovaline and peramine production in endophyte-infected tall fescue: Independent regulation and effects of plant and endophyte genotype

Peramine and ergovaline have ecological and economical significance because they possess insect and/or mammalian toxicity properties. The relationship between these endophytically derived alkaloids in tall fescue (Festuca arundinacea Schreb.) is unknown. Seasonal and plant tissue effects on the concentration of peramine and ergovaline was investigated in field and greenhouse experiments. The relationship between the alkaloids and the regulatory effects of endophyte and plant on their content were investigated among progeny of reciprocal crosses between high- and low-ergovaline and peramine plant-endophyte combinations. Variation in peramine concentration ranged from 750 to 1742 µg/kg in greenhouse-grown plants, and there was no seasonal trend in peramine content. There was a correlation (r=0.69) between peramine and ergovaline content among leaf tissues of field-grown plants, but there was no correlation between the alkaloids in the culm (r=0.20) or panicle (r=0.17) tissues. Mean leaf ergovaline concentration of progeny derived from the low-ergovaline parent (163 µg/kg) was less than the midparent value (228 µg/kg), but mean of progeny from the high-ergovaline parent was not different from the mid-parent value. Ranges within each progeny set were approximately double their mean. Mean leaf peramine concentrations of the progeny sets were similar to the mid-parent value (3354 µg/kg) but ranges were from 1716 to 8753 µg/kg. There was no correlation between leaf peramine and ergovaline (r=0.01). These data suggest that endophytically produced alkaloids are independently regulated and are controlled by both plant and endophyte genotype.     

Colony Fusion in Argentine Ants is Guided by Worker and Queen Cuticular Hydrocarbon Profile Similarity

Introduced populations of the Argentine ant, Linepithema humile, have experienced moderate to severe losses of genetic diversity, which may have affected nestmate recognition to various degrees. We hypothesized that cuticular hydrocarbons (CHC) serve as nestmate recognition cues, and facilitate colony fusion of unrelated L. humile colonies that share similar CHC profiles. In this study, we paired six southeastern U.S. L. humile colonies in a 6-month laboratory fusion assay, and determined if worker and queen CHC profile similarity between colonies was associated with colony fusion and intercolony genetic similarity. We also compared worker and queen CHC profiles between fused colony pairs and unpaired controls to determine if worker and queen chemical profiles changed after fusion. We found that colony fusion correlated with the CHC similarity of workers and queens, with the frequency of fusion increasing with greater CHC profile similarity between colonies. Worker and queen CHC profile similarity between colonies also was associated with genetic similarity between colonies. Queen CHC profiles in fused colonies appeared to be a mix of the two colony phenotypes. In contrast, when only one of the paired colonies survived, the CHC profile of the surviving queens did not diverge from that of the colony of origin. Similarly, workers in non-fused colonies maintained their colony-specific CHC, whereas in fused colonies the worker CHC profiles were intermediate between those of the two colonies. These results suggest a role for CHC in regulating interactions among mutually aggressive L. humile colonies, and demonstrate that colony fusion correlates with both genetic and CHC similarities. Further, changes in worker and queen chemical profiles in fused colonies suggest that CHC plasticity may sustain the cohesion of unrelated L. humile colonies that had fused.     

Nestmate recognition cues in laboratory and field colonies ofSolenopsis invicta buren (Hymenoptera: Formicidae)

Laboratory-rearedSolenopsis invicta workers were tested for the ability to discriminate nestmates from nonnestmate conspecifics. Postcontact aggressive response to workers from local field colonies was significantly greater than the response to lab-reared workers, even when the latter were selected from colonies originating hundreds of miles away. Behavioral observations support the conclusion that lab-reared ants were less distinctive than field-collected ants with respect to recognition cues detectable on the cuticle. Potential environmental factors affecting colony odor are discussed. In addition, gas-liquid Chromatographic and statistical analyses of the majorS. invicta cuticular hydrocarbons indicate that cuticular hydrocarbon pattern was a poor predictor of laboratory colony response to field colony workers.     

Changes in Chemical Signature and Host Specificity from Larval Retrieval to Full Social Integration in the Myrmecophilous Butterfly Maculinea rebeli

The ant social parasite, Maculinea rebeli shows high levels of host specificity at a regional scale. While 68-88% of caterpillars in the field are adopted by nonhost Myrmica ants, 95-100% of the butterflies emerge from the natural host M. schencki the following year. While retrieval of preadoption caterpillars is specific to the genus Myrmica, it does not explain differential survival with different Myrmica species. We present survival data with host and nonhost Myrmica species suggesting that, with nonhosts (M. sabuleti and M. rubra), survival depends on the physiological state of the colony. We also compared the similarities of the epicuticular surface hydrocarbon signatures of caterpillars that were reared by host and nonhost Myrmica for 3 weeks with those from tending workers. Counterintuitively, the hydrocarbons of postadoption caterpillars were more similar (78%, 73%) to the ant colony profiles of the nonhost species than were caterpillars reared in colonies of M. schencki (42% similarity). However, caterpillars from M. schencki nests that were then isolated for 4 additional days showed unchanged chemical profiles, whereas the similarities of those from nonhost colonies fell to 52 and 56%, respectively. Six compounds, presumably newly synthesized, were detected on the isolated caterpillars that could not have been acquired from M. sabuleti and M. rubra (nor occurred on preadoption caterpillars), five of which were found on the natural host M. schencki. These new compounds may relate to the high rank the caterpillars attain within the hierarchy of M. schencki societies. The same compounds would identify the caterpillars as intruders in non-schencki colonies, where their synthesis appeared to be largely suppressed. The ability to synthesize or suppress additional compounds once adopted explains the pattern of mortalities found among fully integrated caterpillars in Myrmica colonies of different species and physiological states.     

Interpopulational variation in emitted pheromone blend of cabbage looper moth,Trichoplusia ni

Female cabbage looper moths,Trichoplusia ni, from laboratory colonies initiated from three locations across the United States emitted similar quantities and blend ratios of the six known pheromone components. In contrast, females from a long-established laboratory colony emitted a greater proportion of four of the five minor components relative to the major component, (Z)-7-dodecenyl acetate; only the relative proportion of 11-dodecenyl acetate was similar in all of the populations sampled. Females from this population emitted (Z)-7-dodecenyl acetate at a rate similar to that from females from field-collected colonies. Within each population there were highly significant correlations among the quantities of pheromone components of similar molecular weights. Correlations between components of different molecular weights were not as great, but often were significant. Similarities of blend ratios among field populations may indicate that the chemical signal in this species is conservative. The difference of the blend ratios in our laboratory population from the other populations may indicate a decrease in the intensity of selection pressure that usually would maintain these values.     

Damage-induced alkaloids in tobacco: Pot-bound plants are not inducible

Field-grown wild tobacco plants (Nicotiana sylvestris) were subjected to a defoliation regime designed to mimic the rate and amount of leaf mass removed by one tobacco hornworm per plant. Undamaged leaves on these plants undergo a dramatic (457% for leaf position 5, 410% for leaf position 8) increase in total leaf alkaloids compared to same-age and positioned control leaves on undamaged control plants. However, potted greenhouse-grown plants fail to exhibit the same damage-induced increase in alkaloid content. The greenhouse environment differs from the field environment in factors known to influence leaf alkaloid content, particularly soil N, P, K, near-UV radiation, and relative humidity. However, altering these environmental factors does not make potted plants able to increase their leaf alkaloid levels in response to defoliation. Transplanting plants into larger pots with more soil does allow the plants to respond to defoliation. Thirty days after transplanting, the plants are again unresponsive to damage, probably as a result of becoming pot-bound. This result suggests a mechanism for the induction response, specifically that leaf damage triggers synthesis of these alkaloids in the roots, and offers a potentially valuable experimental tool for the study of induced-plant defenses in tobacco and other plants that synthesize alkaloids in their root tissues.     

Discriminant analysis of cuticular hydrocarbons of social waspPolistes exclamans viereck and surface hydrocarbons of its nest paper and pedicel

The articular lipids were analyzed from individualPolistes exclamans workers collected from 10 nests. Surface lipids were also recovered from the paper and pedicels of these nests. Twenty-two hydrocarbons were identified in these lipid fractions. The major components of the wasp cuticular lipids weren-heptacosane,n-nonacosane, methylhentriacontane, and methyltritriacontane. Discriminant analysis of the hydrocarbon profiles of the adult wasps showed that the wasps group together according to their respective colonies. Several colonies from the same geographical location clustered more closely together than colonies from diverse locations. The nest papers and pedicels did not group with the wasps from their respective colonies, but the nest papers clustered together separately, as did the nest pedicels.     

Caffeine Does Not Protect Coffee Against the Leaf Miner Perileucoptera coffeella

The larvae of Perileucoptera coffeella, a leaf miner, is one of the main pests attacking coffee plantations. Caffeine is the major alkaloid in this crop and may have a role in insect resistance. Since there is genetic variability in both resistance to the coffee leaf miner and in caffeine content among different coffee species, we investigated the role of this alkaloid as an antiherbivory compound. Coffee plants containing different levels of caffeine were exposed to oviposition of the insect, and the caffeine content and damaged leaf area were evaluated. The same procedure was carried out with interspecific hybrids between C. arabica and C. racemosa, varying in resistance against the leaf miner. In addition, plants were exposed to the insect, but one leaf of each pair was protected from oviposition with paper bags. In another experiment, leaf disks from plants with known susceptibility to attack by the leaf miner were infiltrated with 0–2% aqueous caffeine solutions and exposed to oviposition. When one leaf of a pair was protected from the insect, there was an increase in caffeine in the infested leaves, particularly in younger leaves. None of the experiments established a significant correlation between reduction of leaf damage and caffeine content of the tissue. The results indicate that P. coffeella is well adapted to coffee, and it has evolved a mechanism to tolerate the potentially toxic effects of caffeine.     

Surface lipids of social waspPolistes melricus say and its nest and nest pedicel and their relation to nestmate recognition

In eight replicate laboratory tests wherePolistes metricus adults were allowed to choose between their own nest, a second nest, and neither nest, they selected their own nest 66% of the observed time. When the surface hydrocarbons had been extracted from the nests, the wasps chose their own nest only 8% of the time, but after the hydrocarbons were reapplied to the surface of the respective nests, they selected their own nest 47% of the time. These changes are significant. The cuticular lipids were analyzed from individualP. metricus adult females collected from 13 colonies. Surface lipids were recovered from the paper and pedicels of their nests. Eighteen hydrocarbons were identified in these lipid fractions. The major components of the wasp cuticular lipids weren-heptacosane,n-nonacosane, methylhentriacontane, and methyltritriacontane. Factor analysis revealed that extracts of pedicels are all similar in composition, while cuticle and paper extracts vary, sometimes similarly according to colony identity.     

Cuticular hydrocarbons of the paper wasp,Polistes fuscatus: A search for recognition pheromones

The cuticular chemicals of 124 individual wasps (foundresses and workers) from 23 colonies ofPolistes fuscatus were analyzed. The compounds identified, all of which were hydrocarbons, were similar to those of other vespid wasps in that the bulk of the hydrocarbons were 23–33 carbons in chain length. However, the hydrocarbon profile ofP. fuscatus differed from those of its congeners in its proportions of straight-chain alkanes, methylalkanes, and alkenes. Three of the 20 identified hydrocarbons, 13- and 15-MeC₃₁, 11,15- and 13,17-diMeC₃₁, and 13-, 15-, and 17-MeC₃₃, had properties postulated for recognition pheromones: colony specificity, efficacy in assigning wasps to the appropriate colony, heritability, lack of differences between foundresses and workers, and distinctive stereochemistry.     

Fate of quinolizidine alkaloids through three trophic levels:Laburnum anagyroides (Leguminosae) and associated organisms

The quinolizidine alkaloids (QA) of golden rain,Laburnum anagyroides, and those of phytophagous insects associated with the plant, as well as of parasitoids of the latter, were analyzed by capillary GLC and GLC-MS. The alkaloid content in samples of vegetative plant parts was high at the beginning of the season, then decreased, while that of reproductive organs was high throughout flowering, pod formation, and maturation. The analyses showed that the QA of the plant passed through two higher trophic levels (herbivorous insects and their parasitoids) and that the alkaloid pattern changed little during the passage. The alkaloids were present in two phytophagous insect species associated with golden rain: the predispersal seed predator,Bruchidius villosus [5–13g/g fresh weight (fw)], andAphis cytisorum (182–1012g/g fw), an aphid that feeds on shoots, leaves, and inflorescences. Braconid and chalcidoid parasitoids emerging from the bruchid host also contained alkaloids (1.3–3g/g fw), as did three foraging ant species,Lasius niger, Formica rufibarbis, andF. cunicularia (45g/g fw), that visited the aphid colonies or honeydew-covered leaves of aphid-infested plants. The hypothesis that developing bruchid larvae and/or the plant manipulate QA supply to infested seeds was not supported, because QA content of leftover endosperm in seeds after bruchid development was similar to that of uninfested seeds. The frass of developing bruchid larvae was rich in QA (31g/ g dry weight). While aphids sequestered, the bruchid larvae took up and eliminated QA with the frass without chemical transformation.