Specificity in Chemical Profiles of Workers,Brood and Mutualistic Fungi in <Emphasis Type="Italic">Atta</Emphasis>, <Emphasis Type="Italic">Acromyrmex,</Emphasis> and <Emphasis Type="Italic">Sericomyrmex</Emphasis> Fungus-growing Ants

Neotropical attine ants live in obligatory symbiosis with a fungus that they grow for food on a substrate of primarily plant material harvested by workers. Nestmate recognition is likely based on chemical cues as in most other social insects, but recent studies have indicated that both the ants and their mutualistic fungi may contribute to the recognition templates. To investigate the within-colony variation in chemical profiles, we extracted and identified compounds from the cuticle of workers, the postpharyngeal gland of workers, ant pupae and larvae, and the fungal symbiont of three species of higher attine ants: Atta colombica, Acromyrmex echinatior, and Sericomyrmex amabilis. The relative proportions of identified compounds were compared and represented 11 classes: n-alkanes, alkenes, branched methylalkanes, branched dimethylalkanes, trimethylalkanes, branched alkenes, aldehydes, alcohols, acetates, acids, and esters. The chemical profiles in all three species are likely to be sufficiently different to allow discrimination at the species and colony level and sufficiently similar within colonies to generate a relatively constant colony-specific chemical gestalt. The relative likelihood of individual compounds being derived from the ants, the ant brood, or the fungal symbiont are discussed. We hypothesize that hydrocarbons are particularly important as recognition cues because they appear to simultaneously allow the assessment of developmental stages and the identification of symbiont, colony, and species.     

Alkyl-Dimethylpyrazines in Mandibular Gland Secretions of Four <Emphasis Type="Italic">Odontomachus</Emphasis> Ant Species (<Emphasis Type="Italic">Formicidae: Ponerinae)</Emphasis>

Ponerine ants are known to contain mixtures of pyrazines in their mandibular glands. We analyzed the mandibular gland contents of four ponerine species (Odontomachus chelifer, O. erythrocephalus, O. ruginodis, and O. bauri) by gas chromatography coupled with mass spectrometry, and found that each species contains specific mixtures of trisubstituted alkylpyrazines among other volatiles. Attempts to identify alkylpyrazines solely by mass spectral interpretation is unrealistic because spectra of positional isomers are indistinguishable. To avoid misidentifications, we synthesized a large number of reference compounds and compared their mass spectral and gas chromatographic properties with those present in the Odontomachus species under investigation. Most of the compounds identified were 2-alkyl-3,5-dimethylpyrazines. Interestingly, when the third substituent was an isopentyl group, the two methyl groups were found to be located at the 2 and 5 ring positions. Using our data, we recognized several misidentifications in previous publications.     

Host Recognition by the Specialist Hoverfly <Emphasis Type="Italic">Microdon mutabilis</Emphasis>, a Social Parasite of the Ant <Emphasis Type="Italic">Formica lemani</Emphasis>

The larva of the hoverfly Microdon mutabilis is a specialist social parasite of the ant Formica lemani that is adapted to local groups of F. lemani colonies but mal-adapted to colonies of the same species situated only a few hundred meters away. At a study site in Ireland, F. lemani shares its habitat with four other ant species. All nest under stones, making the oviposition choice by M. mutabilis females crucial to offspring survival. In this study, we tested the hypothesis that, as an extreme specialist, M. mutabilis should respond to cues derived from its host rather than from its microenvironment, a phenomenon that has hitherto only been addressed in the context of herbivorous insects and their parasitoids. In behavioral assays, M. mutabilis females reacted to volatiles from F. lemani colonies by extending their ovipositors, presumably probing for an oviposition substrate. This behavior was not observed toward negative controls or volatiles from colonies of Myrmica scabrinodis, the host ant of the closely related Microdon myrmicae. Coupled gas chromatography-electroantennography (GC-EAG) that used antennal preparations of M. mutabilis located a single physiologically active compound within an extract of heads of F. lemani workers. Coupled GC-mass spectrometry (GC-MS) tentatively identified the compound as a methylated methylsalicylate. GC co-injection of the extract with authentic samples showed that of the four possible isomers (methyl 3-, 4-, 5-, and 6-methylsalicylate), only methyl 6-methylsalicylate co-eluted with the EAG-active peak. Furthermore, the response to methyl 6-methylsalicylate was four times higher than to those of the other isomers. Coupled GC-EAG and GC-MS also revealed physiological responses to two constituents, 3-octanone and 3-octanol, of the M. scabrinodis alarm pheromone. However, the behavioral trials did not reveal any behavior that could be attributed to these compounds. Results are discussed in the context of four phases of host location behavior, and of the characteristics, which volatile cues should provide to be useful for an extreme specialist such as M. mutabilis.     

Trade-Off Between Chemical and Biotic Antiherbivore Defense in the South East Asian Plant Genus Macaranga

The plant genus Macaranga is known for its manifold mutualistic associations with ants. The plants provide food for the ants and in turn get protection from herbivores. Depending on the strength of the plant–ant interaction, the plant's investment in ants and the biotic defense derived from them is more or less effective. We conducted a comparative study on tannin content in 12 Macaranga species that were selected based on their associations with ants (three nonmyrmecophytes and nine myrmecophytes, three of which start their ontogeny as nonmyrmecophytes). Different developmental stages were investigated in three Macaranga species. Extracts of every individual plant analyzed for tannins were also tested for their effects on larval growth employing larvae of the common cutworm (Spodoptera littoralis). The studied Macaranga species differed significantly in their tannin contents as well as in the effects of their leaf extracts on the growth of S. littoralis larvae. A correlation analysis shows a connection between tannin contents and larval growth. High tannin contents and, thus more effective chemical defense, were observed in nonmyrmecophytic Macaranga species associated only facultatively with ants as compared to obligate myrmecophytes. Our study supports the hypothesis of a trade-off between chemical and biotic defense in the genus Macaranga.     

Leaf Chemistry and Foliage Avoidance by the Thrips <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> and <Emphasis Type="Italic">Heliothrips haemorrhoidalis</Emphasis> in Glasshouse Collections

Observational studies on foliage avoidance by the polyphagous thrips species Frankliniella occidentalis (Pergande) and Heliothrips haemorrhoidalis (Bouché) (Thysanoptera: Thripidae) identified six non-host species (Allagopappus dichotomus (Asteraceae), Gardenia posoquerioides (Rubiaceae), Plectranthus aff. barbatus, Plectranthus strigosus, Plectranthus zuluensis (Lamiaceae), and Sclerochiton harveyanus (Acanthaceae) among plants growing within a major glasshouse botanical collection. The effects of sequentially obtained acetone and aqueous methanol leaf extracts on mortality in first instar Frankliniella occidentalis were assessed. The acetone leaf extract of Sclerochiton harveyanus, which had the highest activity against the thrips, yielded four new iridoids, sclerochitonosides A–C, and sclerochitonoside B 4′-methyl ether. Mortality of F. occidentalis was increased on exposure to all four iridoids, and the most active iridoid was sclerochitonoside A (8-epiloganic acid 4′-hydroxyphenylethyl ester). Choice experiments demonstrated that this compound did not significantly deter H. haemorrhoidalis from treated leaf surfaces. The significance of iridoids in the defense mechanism of plants against thrips is discussed.     

Phytotoxicity of <Emphasis Type="Italic">Phytolacca americana</Emphasis> Leaf Extracts on the Growth,and Physiological Response of <Emphasis Type="Italic">Cassia mimosoides</Emphasis>

We examined the allelochemical effects of control soil, native soil (treated soil), and leaf extracts of Phytolacca americana (pokeweed) on the germination rate and seedling growth of Cassia mimosoides var. nomame. We also studied the resulting changes in root-tip ultrastructure and peroxidase isozyme biochemistry. P. americana leaf extract inhibited seed germination, seedling growth, and biomass when compared to control and treated soil. Root and shoot growth in treated soil was stimulated relative to control soil, but root growth was inhibited by 50% in the leaf extract treatment. Biomass of C. mimosoides seedlings grown on leaf extract was reduced sevenfold when compared to the control seedlings. The amounts of total phenolic compounds in the leaf extract, treated soil, and control soil were 0.77, 0.14, and 0.03 mg l⁻¹, respectively. The root tips of C. mimsoides treated with leaf extracts of P. americana showed amyloplasts and large central vacuoles with electron-dense deposits inside them when compared to control root tips. The activity of guaiacol peroxidase (GuPOX) in whole plant, roots, and shoots of C. mimosoides increased as leaf extract increased; maximum activity was observed in extract concentrations of 75% and higher. Root GuPOX activity was three times higher than in shoots. Therefore, we conclude that inhibition of C. mimosoides growth is related to the phenolic compounds in the P. americana leaf extract and the ultrastructure changes in root-tip cells and increased GuPOX activity is a response to these allelochemicals.     

The Role of Fresh <Emphasis Type="Italic">versus</Emphasis> Old Leaf Damage in the Attraction of Parasitic Wasps to Herbivore-Induced Maize Volatiles

The odor produced by a plant under herbivore attack is often used by parasitic wasps to locate hosts. Any type of surface damage commonly causes plant leaves to release so-called green leaf volatiles, whereas blends of inducible compounds are more specific for herbivore attack and can vary considerably among plant genotypes. We compared the responses of naïve and experienced parasitoids of the species Cotesia marginiventris and Microplitis rufiventris to volatiles from maize leaves with fresh damage (mainly green leaf volatiles) vs. old damage (mainly terpenoids) in a six-arm olfactometer. These braconid wasps are both solitary endoparasitoids of lepidopteran larvae, but differ in geographical origin and host range. In choice experiments with odor blends from maize plants with fresh damage vs. blends from plants with old damage, inexperienced C. marginiventris showed a preference for the volatiles from freshly damaged leaves. No such preference was observed for inexperienced M. rufiventris. After an oviposition experience in hosts feeding on maize plants, C. marginiventris females were more attracted by a mixture of volatiles from fresh and old damage. Apparently, C. marginiventris has an innate preference for the odor of freshly damaged leaves, and this preference shifts in favor of a blend containing a mixture of green leaf volatiles plus terpenoids, after experiencing the latter blend in association with hosts. M. rufiventris responded poorly after experience and preferred fresh damage odors. Possibly, after associative learning, this species uses cues that are more directly related with the host presence, such as volatiles from host feces, which were not present in the odor sources offered in the olfactometer. The results demonstrate the complexity of the use of plant volatiles by parasitoids and show that different parasitoid species have evolved different strategies to exploit these signals.     

Changes in Monoterpene Emission Rates of <Emphasis Type="Italic">Quercus ilex</Emphasis> Infested by Aphids Tended by Native or Invasive <Emphasis Type="Italic">Lasius</Emphasis> Ant Species

The emission of volatile organic compounds (VOCs) depends on temperature and light. Other factors such as insect herbivory also may modify VOC emission. In particular, aphid feeding promotes the release of new compounds and changes the composition of plant volatile blends. Given that some aphids are tended by ants, we investigated whether ants change the emission of VOCs indirectly through attendance on aphids. The effect of Lachnus roboris aphids and two different tending ant species on terpene emission rates of 4-year-old holm oak (Quercus ilex) saplings was investigated during a field experiment. There were five treatments: saplings alone (T1), saplings infested with L. roboris aphids (T2), saplings infested with aphids tended by the local ant Lasius grandis (T3), those tended by small colonies of the invasive ant Lasius neglectus (T4), and those tended by large colonies of the same invasive ant species (T5). The infestation by L. roboris elicited the emission of Δ³-carene and increased the emission of myrcene and γ-terpinene. Terpene emissions were modified depending on the tending ant species. Attendance by the local ant L. grandis increased α and β-pinene and sabinene. Attendance by the invasive ant L. neglectus only decreased significantly the emission of myrcene, one of the major compounds of the Q. ilex blend. Aphid abundance decreased with time for all treatments, but there was no difference in aphid abundance among treatments. Total terpene emission rates were not correlated with aphid abundance. These results highlight that aphids and tending ants may change terpene emission rates, depending on the ant species.     

Allelopathic Potential of <Emphasis Type="Italic">Robinia pseudo-acacia</Emphasis> L.

Robinia pseudo-acacia L. (black locust) is a nonindigenous species currently invading the central part of Japanese grasslands. Several allelochemicals were identified and characterized from the leaf tissue. The growth of both radicle and hypocotyl in the tested species (barnyard grass, white clover, lettuce, and Chinese cabbage) was reduced when grown in soil mixed with the leaves of R. pseudo-acacia at various concentrations. Aqueous leaf extracts, when bioassayed, exhibited a significant suppression of radicle growth. Chromatographic separation of an ethanolic extract of R. pseudo-acacia leaves resulted in isolation of three compounds, identified as robinetin (1), myricetin (2), and quercetin (3) by nuclear magnetic resonance and mass spectroscopy. All inhibited root and shoot growth of lettuce. Robinetin, found in a large amount, caused 50% suppression of the root and shoot growth of lettuce at 100 ppm. The presence of these bioactive substances in leaf tissue suggests a potential role for flavonoids in R. pseudo-acacia invasion in introduced habitats.     

Sequestration of Furostanol Saponins by <Emphasis Type="Italic">Monophadnus</Emphasis> Sawfly Larvae

Sawfly larvae of the tribe Phymatocerini (Hymenoptera: Tenthredinidae), which are specialized on toxic plants in the orders Liliales and Ranunculales, exude a droplet of deterrent hemolymph upon attack by a predator. We investigated whether secondary plant metabolites from Ranunculaceae leaves are sequestered by phymatocerine Monophadnus species, i.e., Monophadnus alpicola feeding upon Pulsatilla alpina and Monophadnus monticola feeding upon Ranunculus lanuginosus. Moreover, two undescribed Monophadnus species were studied: species A collected from Helleborus foetidus and species B collected from Helleborus viridis. Comparative high-performance liquid chromatographic–photodiode array detection–electrospray ionization–mass spectrometric analyses of plant leaf and insect hemolymph extracts revealed the presence of furostanol saponins in all samples. Larvae of species A and B actively sequestered (25R)-26-[(α-l-rhamnopyranosyl)oxy]-22α-methoxyfurost-5-en-3β-yl O-β-d-glucopyranosyl-(1→3)-O-[6-acetyl-β-d-glucopyranosyl-(1→3)]-O-β-d-glucopyranoside (compound 1). This compound occurred at a 65- to 200-fold higher concentration in the hemolymph of the two species (1.6 and 17.5 μmol/g FW, respectively) than in their host plant (0.008 and 0.268 μmol/g FW, respectively). In M. monticola, compound 1 was found at a concentration (1.2 μmol/g FW) similar to that in the host plant (1.36 μmol/g FW). The compound could not be detected consistently in M. alpicola larvae where, however, a related saponin may be present. Additional furostanol saponins were found in H. foetidus and H. viridis, but not in the two Monophadnus species feeding on them, indicating that sequestration of compound 1 is a highly specific process. In laboratory bioassays, crude hemolymph of three Monophadnus species showed a significant feeding deterrent activity against a potential predator, Myrmica rubra ant workers. Isolated furostanol saponins were also active against the ants, at a concentration range similar to that found in the hemolymph. Thus, these compounds seem to play a major role for chemical defense of Monophadnus larvae, although other plant secondary metabolites (glycosylated ecdysteroids) were also detected in their hemolymph. Physiological and ecological implications of the sequestered furostanol saponins are discussed. Dedicated to the memory of Professor Ivano Morelli (1940–2005)     

Laboratory Evaluation of <Emphasis Type="Italic">Artemisia annua</Emphasis> L. Extract and Artemisinin Activity against <Emphasis Type="Italic">Epilachna paenulata</Emphasis> and <Emphasis Type="Italic">Spodoptera eridania</Emphasis>

Ethanolic extract of aerial parts of Artemisia annua L. and artemisinin were evaluated as anti-insect products. In a feeding deterrence assay on Epilachna paenulata Germ (Coleoptera: Coccinellidae) larvae, complete feeding rejection was observed at an extract concentration of 1.5 mg/cm² on pumpkin leaf tissue. The same concentration produced a feeding inhibition of 87% in Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae). In a no-choice assay, both species ate less and gained less weight when fed on leaves treated with the extract. Complete mortality in E. paenulata and 50% mortality in S. eridania were observed with extract at 1.5 mg/cm². Artemisinin exhibited a moderate antifeedant effect on E. paenulata and S. eridania at 0.03–0.375 mg/cm². However, a strong effect on survival and body weight was observed when E. paenulata larvae were forced to feed on leaves treated at 0.03 and 0.075 mg/cm². Artemisia annua ethanolic extract of aerial parts at 1.5 mg/cm² showed no phytotoxic effect on pumpkin seedlings.     

Assessment of the Allelopathic Potential of <Emphasis Type="Italic">Juniperus ashei</Emphasis> on Germination and Growth of <Emphasis Type="Italic">Bouteloua curtipendula</Emphasis>

Potential allelopathic compounds of Juniperus ashei Buchh. (Ashe juniper) and their effect on a native grass were determined in laboratory and field studies. Solid-phase microextraction and gas chromatography/mass spectrometry were used to determine if monoterpenes found in the essential oils of J. ashei are released in leaf and litter leachate, as well as volatilized from leaf tissue. Camphor, bornyl acetate, and limonene were found in leaf and fresh litter leachates; however, degraded litter did not contain any of these compounds. Camphor was the most common potentially allelopathic compound found in J. ashei leaf and litter leachate and in volatiles from leaf tissue. The effects of leaf and litter tissue on the germination of Bouteloua curtipendula (Michx.) Torr. (side-oats grama) was tested by using the “sandwich agar method”. The highest germination of B. curtipendula (29.6%) occurred in the control, which was significantly higher than fresh litter (13.2%) and degraded litter (16.2%). The lowest germination (6.2%) occurred with J. ashei leaves. In the field experiment, aboveground dry mass of B. curtipendula was evaluated in relation to position within the canopy and intercanopy of J. ashei adult trees when light and water were held constant across locations. Aboveground dry mass of B. curtipendula was significantly greater in the intercanopies of J. ashei (163.7 g m²) compared to the dry mass in the understory (44.8 g m²) and dripline (44.5 g m²), suggesting some negative influence by J. ashei. Chemical analyses indicate that monoterpenes are released through leaching and volatilization from J. ashei, and germination and field studies suggest that these compounds inhibit B. curtipendula.     

Diet-Related Modification of Cuticular Hydrocarbon Profiles of the Argentine Ant, <Emphasis Type="Italic">Linepithema humile</Emphasis>, Diminishes Intercolony Aggression

Territorial boundaries between conspecific social insect colonies are maintained through a highly developed nestmate recognition system modulated by heritable and, in some instances, nonheritable cues. Argentine ants, Linepithema humile, use both genetic and environmentally derived cues to discriminate nestmates from nonnestmates. We explored the possibility that intraspecific aggression in the Argentine ant might diminish when colonies shared a common diet. After segregating recently field-collected colony pairs into high or moderate aggression categories, we examined the effect of one of three diets: two hydrocarbon-rich insect prey, Blattella germanica and Supella longipalpa, and an artificial (insect-free) diet, on the magnitude of aggression loss. Aggression diminished between colony pairs that were initially moderately aggressive. However, initially highly aggressive colony pairs maintained high levels of injurious aggression throughout the study, independent of diet type. Each diet altered the cuticular hydrocarbon profile by contributing unique, diet-specific cues. We suggest that acquisition of common exogenous nestmate recognition cues from shared food sources may diminish aggression and promote fusion in neighboring colonies of the Argentine ant.     

Response of <Emphasis Type="Italic">Plutella xylostella</Emphasis> and its Parasitoid <Emphasis Type="Italic">Cotesia plutellae</Emphasis> to Volatile Compounds

The effects of limonene, a mixture of limonene + carvone (1:1, v/v), and methyl jasmonate (MeJA) on diamondback moth (DBM) (Plutella xylostella L.) oviposition, larval feeding, and the behavior of its larval parasitoid Cotesia plutellae (Kurdjumov) with cabbage (Brassica oleracea L. ssp. capitata, cvs. Rinda and Lennox) and broccoli (B. oleracea subsp. Italica cv Lucky) were tested. Limonene showed no deterrent effect on DBM when plants were sprayed with or exposed to limonene, although there was a cultivar difference. A mixture of limonene and carvone released from vermiculite showed a significant repellent effect, reducing the number of eggs laid on the cabbages. MeJA treatment reduced the relative growth rate (RGR) of larvae on cv Lennox leaves. In Y-tube olfactometer tests, C. plutellae preferred the odors of limonene and MeJA to filtered air. In cv Lennox, the parasitoid preferred DBM-damaged plants with limonene to such plants without limonene. C. plutellae females were repelled by the mixture of limonene + carvone. In both cultivars, exogenous MeJA induced the emission of the sesquiterpene (E,E)-α-farnesene, the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), and green leaf volatile (Z)-3-hexenyl acetate + octanal. The attractive effect of limonene and MeJA predicts that these two compounds can be used in sustainable plant protection strategies in organic farming.     

Oviposition Responses of the Mosquitoes <Emphasis Type="Italic">Aedes aegypti</Emphasis> and <Emphasis Type="Italic">Aedes albopictus</Emphasis> to Experimental Plant Infusions in Laboratory Bioassays

Attraction of the mosquitoes Aedes aegypti and Ae. albopictus to plant infusions was evaluated by using a modified sticky-screen bioassay that improved the resolution of mosquito responses to odorants. Under bioassay conditions, solid-phase microextraction-gas chromatographic analyses of the volatile marker chemical indole showed that odorants diffused from bioassay cups, forming a concentration gradient. Infusions were prepared by separately fermenting senescent leaves of eight plant species in well water. Plant infusions were evaluated over an 8-fold range of leaf biomass and/or a 28 d fermentation period. The responses of gravid females of both mosquito species varied with the plant species and biomass of plant materials used to make infusions, and with the length of the fermentation period. Infusions made from senescent bamboo (Arundinaria gigantea) and white oak (Quercus alba) leaves were significantly attractive to both mosquitoes. In general, infusions prepared by using low biomass of plant material over a 7–14 d fermentation period were most attractive to Ae. aegypti. In contrast, Ae. albopictus was attracted to infusions made using a wider range of plant biomass and over a longer fermentation period. Both mosquito species were more attracted to a non-sterile white oak leaf infusion than to white oak leaf infusion that was prepared using sterilized plant material and water, thus suggesting a role for microbial activity in the production of odorants that mediate the oviposition response of gravid mosquitoes.     

<Emphasis Type="Italic">E</Emphasis>-2-Ethylhexenal, <Emphasis Type="Italic">E</Emphasis>-2-Ethyl-2-Hexenol,Mellein, and 4-Hydroxymellein in <Emphasis Type="Italic">Camponotus</Emphasis> Species from Brunei

E-2-ethyl-2-hexen-1-ol (1), mellein (4), and 4-hydroxymellein (5) were identified as the major volatile compounds in the head and/or thorax of Camponotus quadrisectus. Neither 1 nor 5 have been previously detected in insects. Also identified were small amounts of m-cresol (2) and 6-methyl salicylic acid (3). E-2-ethylhexenal (6) and small amounts of 3 were identified in heads of Camponotus irritibilis from Kuala Belalong, Brunei. Compounds 2–4 occur in other Bornean camponotines with hypertrophied mandibular glands, and 4 is widespread in the tribe. The possibility of semiochemical parsimony (multiple functions) for these mandibular gland compounds is reviewed in the context of existing data on mandibular gland products of other camponotines, reported biological activities of the compounds, and secondary loss of metapleural glands in this ant group.     

Antimicrobial Activity of Extractable Conifer Heartwood Compounds Toward <Emphasis Type="Italic">Phytophthora ramorum</Emphasis>

Ethyl acetate extracts from heartwood of seven western conifer trees and individual volatile compounds in the extracts were tested for antimicrobial activity against Phytophthora ramorum. Extracts from incense and western redcedar exhibited the strongest activity, followed by yellow-cedar, western juniper, and Port-Orford-cedar with moderate activity, and no activity for Douglas-fir and redwood extracts. Chemical composition of the extracts varied both qualitatively and quantitatively among the species with a total of 37 compounds identified by mass spectrometry. Of the 13 individual heartwood compounds bioassayed, three showed strong activity with a Log₁₀ EC₅₀ less than or equal to 1.0 ppm (hinokitiol, thymoquinone, and nootkatin), three expressed moderate activity ranging from 1.0–2.0 ppm (nootkatol, carvacrol, and valencene-11,12-diol), four compounds had weak activity at 2.0–3.0 ppm [α-terpineol, valencene-13-ol, (+)-β-cedrene, (−)-thujopsene], and three had no activity [(+)-cedrol, δ-cadinene, and methyl carvacrol]. All of the most active compounds contained a free hydroxyl group, except thymoquinone. The importance of a free hydroxyl was demonstrated by the tremendous difference in activity between carvacrol (Log₁₀ EC₅₀ 1.81 ± 0.08 ppm) and methyl carvacrol (Log₁₀ EC₅₀ >3.0 ppm). A field trial in California, showed that heartwood chips from redcedar placed on the forest floor for 4 months under Umbellularia californica (California bay laurel) with symptoms of P. ramorum leaf blight significantly limited the accumulation of P. ramorum DNA in the litter layer, compared with heartwood chips from redwood.     

Spectrum of Cyanide Toxicity and Allocation in <Emphasis Type="Italic">Heliconius erato</Emphasis> and <Emphasis Type="Italic">Passiflora</Emphasis> Host Plants

The larvae of three races of Heliconius erato were fed various species of Passiflora containing varying levels of cyanoglucosides. The mortality rate of larvae and pupae rose when larvae were fed species of Passiflora capable of releasing larger quantities of cyanide. When larvae were fed species of Passiflora with these properties, the resulting adult butterflies also released higher levels of cyanide. This may serve as a defense mechanism. The compounds responsible for the release of cyanide were not evenly distributed throughout the adult butterfly’s body. The thorax contained the highest concentration of cyanogenic substances, followed by the head, wings, and abdomen. The younger tissues of Passiflora plants had higher levels of cyanide-releasing compounds than stems and mature leaves. Cyanogenic glycoside distribution within the plants is consistent with optimal allocation theory. The levels of cyanide-releasing substances in plants varied depending on the season.     

<Emphasis Type="Italic">N</Emphasis>-Acylated Bacteriohopanehexol-Mannosamides from the Thermophilic Bacterium <Emphasis Type="Italic">Alicyclobacillus acidoterrestris</Emphasis>

Identification of molecular species of various N-acylated bacteriohopanehexol-mannosamides from the thermophilic bacterium Alicyclobacillus acidoterrestris by semipreparative HPLC and by RP-HPLC with ESI is described. We used triple-quadrupole type mass spectrometer, ¹H and ¹³C NMR for analyzing this complex lipid. CD spectra of two compounds (model compound—7-deoxy-d-glycero-d-allo-heptitol obtained by stereospecific synthesis, and an isolated derivative of hopane) were also measured and the absolute configuration of both compounds was determined. On the basis of all the above methods, we identified the full structure of a new class of bacteriohopanes, represented by various N-acylated bacteriohopanehexol-mannosamides.     

Male-Produced Aggregation Pheromones of the Cerambycid Beetles <Emphasis Type="Italic">Xylotrechus colonus</Emphasis> and <Emphasis Type="Italic">Sarosesthes fulminans</Emphasis>

Adults of both sexes of the cerambycid beetles Xylotrechus colonus (F.) and Sarosesthes fulminans (F.) were attracted to odors produced by male conspecifics in olfactometer bioassays. Analyses of headspace volatiles from adults revealed that male X. colonus produced a blend of (R)- and (S)-3-hydroxyhexan-2-one and (2 S,3 S)- and (2R,3R)-2,3-hexanediol, whereas male S. fulminans produced (R)-3-hydroxyhexan-2-one and (2 S,3R)-2,3-hexanediol. All of these compounds were absent in the headspace of females. Two field bioassays were conducted to confirm the biological activity of the synthesized pheromones: (1) enantiomerically enriched pheromone components were tested singly and in species-specific blends and (2) four-component mixture of racemic 3-hydroxyhexan-2-one plus racemic 2-hydroxyhexan-3-one and the four-component blend of the stereoisomers of 2,3-hexanediols were tested separately and as a combined eight-component blend. In these experiments, adult male and female X. colonus were captured in greatest numbers in traps baited with the reconstructed blend of components produced by males, although significant numbers were also captured in traps baited with (R)-3-hydroxyhexan-2-one alone or in blends with other compounds. Too few adult S. fulminans were captured for a statistical comparison among treatments, but all were caught in traps baited with lures containing (R)-3-hydroxyhexan-2-one. In addition to these two species, adults of two other species of cerambycid beetles, for which pheromones had previously been identified, were caught: Neoclytus a. acuminatus (F.) and its congener Neoclytus m. mucronatus (F.). Cross-attraction of beetles to pheromone blends of other species, and to individual pheromone components that are shared by two or more sympatric species, may facilitate location of larval hosts by species that compete for the same host species.