Molecular basis ofMorinda citrifolia (L.): Toxicity on drosophila

The ripe fruit ofMorinda citrifolia, host plant forDrosophila sechellia is highly toxic for three closely related species (D. melanogaster, D. simulans, andD. mauritiana). Green and rotten fruits are not toxic for all species tested. Short fatty acids were found to be present in large quantities in the extract of the ripe fruit. The most abundant (octanoic acid) was tested pure for its toxicity in a dose-dependent manner;D. sechellia is five to six times more resistant thanD. melanogaster to octanoic acid. Octanoic acid alone seems to be sufficient to explain the toxic effect of the pulp. It is less abundant in the rotten fruit and absent in the green fruit.     

Fatty Acid Amides,Previously Identified in Caterpillars,Found in the Cricket <Emphasis Type="Italic">Teleogryllus taiwanemma</Emphasis> and Fruit Fly <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> Larvae

Fatty acid amides (FAAs) are known elicitors that induce plants to release volatile compounds that, in turn, attract foraging parasitoids. Since the discovery of volicitin [N-(17-hydroxylinolenoyl)-l-glutamine] in the regurgitant of larval Spodoptera exigua, a series of related FAAs have been identified in several other species of lepidopteran caterpillars. We screened 13 non-lepidopteran insects for the presence of FAAs and found that these compounds were present in adults of two closely related cricket species, Teleogryllus taiwanemma and T. emma (Orthoptera: Gryllidae), and larvae of the fruit fly, Drosophila melanogaster (Diptera: Drosophilidae). When analyzed by liquid chromatography/mass spectrometry-ion trap-time-of-flight (LCMS-IT-TOF), the gut contents of both crickets had nearly identical FAA composition, the major FAAs comprising N-linolenoyl-l-glutamic acid and N-linoleoyl-l-glutamic acid. There were also two previously uncharacterized FAAs that were thought to be hydroxylated derivatives of these glutamic acid conjugates, based on their observed fragmentation patterns. In addition to these four FAAs containing glutamic acid, N-linolenoyl-l-glutamine and a small amount of volicitin were detected. In D. melanogaster, N-linolenoyl-l-glutamic acid and N-linoleoyl-l-glutamic acid were the major FAAs found in larval extracts, while hydroxylated glutamic acid conjugates, volicitin and N-linolenoyl-l-glutamine, were detected as trace components. Although these FAAs were not found in ten of the insects studied here, their identification in two additional orders of insects suggests that FAAs are more common than previously reported and may have physiological roles in a wide range of insects besides caterpillars.     

Aggregation pheromones ofDrosophila immigrans,D. phalerata,andD. subobscura

Aggregation pheromones ofDrosophila immigrans, D. phalerata andD. subobscura were demonstrated by testing attraction of adult flies to hexane extracts of the flies in a windtunnel bioassay. Extracts of adult males of all species attracted conspecific males and females. However,D. subobscura flies were attracted only when the extract (cVA) in the extracts of adult maleD. immigrans andD. phalerata. Both species were attracted to synthetic cVA. Male and femaleD. phalerata. Both species were attracted to synthetic cVA. Male and femaleD. subobscura produced 5,9-pentacosadiene, 5-pentacosene, 2-methylhexacosene and 5,9-heptacosadiene, while only maleD. subobscura produced (Z)-5-tricosene and minor amounts of cVA.     

Roquefortine/Oxaline Biosynthesis Pathway Metabolites in <Emphasis Type="Italic">Penicillium</Emphasis> ser. <Emphasis Type="Italic">Corymbifera</Emphasis>: <Emphasis Type="Italic">In Planta</Emphasis> Production and Implications for Competitive Fitness

Three strains of each of the seven taxa comprising the Penicillium series Corymbifera were surveyed by direct injection mass spectrometry (MS) and liquid chromatography–MS for the production of terrestric acid and roquefortine/oxaline biosynthesis pathway metabolites when cultured upon macerated tissue agars prepared from Allium cepa, Zingiber officinale, and Tulipa gesneriana, and on the defined medium Czapek yeast autolysate agar (CYA). A novel solid-phase extraction methodology was applied for the rapid purification of roquefortine metabolites from a complex matrix. Penicillium hordei and P. venetum produced roquefortine D and C, whereas P. hirsutum produced roquefortine D and C and glandicolines A and B. P. albocoremium, P. allii, and P. radicicola carried the pathway through to meleagrin, producing roquefortine D and C, glandicolines A and B, and meleagrin. P. tulipae produced all previously mentioned metabolites yet carried the pathway through to an end product recognized as epi-neoxaline, prompting the proposal of a roquefortine/epi-neoxaline biogenesis pathway. Terrestric acid production was stimulated by all Corymbifera strains on plant-derived media compared to CYA controls. In planta, production of terrestric acid, roquefortine C, glandicolines A and B, meleagrin, epi-neoxaline, and several other species-related secondary metabolites were confirmed from A. cepa bulbs infected with Corymbifera strains. The deposition of roquefortine/oxaline pathway metabolites as an extracellular nitrogen reserve for uptake and metabolism into growing mycelia and the synergistic role of terrestric acid and other Corymbifera secondary metabolites in enhancing the competitive fitness of Corymbifera species in planta are proposed.     

Odorant binding protein homologues of the malaria mosquito Anopheles gambiae; possible orthologues of the OS-E and OS-F OBPs OF Drosophila melanogaster

Twenty-nine Anopheles gambiae candidate Odorant Binding Proteins (OBPs) were characterized for similarity to OBPs of Drosophila melanogaster and other insects. Twenty-five of these sequences were identified by BLAST searching the A. gambiae genome database. Several A. gambiae sequences were significantly similar to the D. melanogaster OBPs OS-E/OS-F, LUSH and PBPRP2/PBPRP5. Exon boundary comparisons suggests that two A. gambiae genes are orthologues of OS-E and OS-F, justifying the names AgamOS-E (EAA01090, AF437886) and AgamOS-F (EAA14641, AF437884). If these are orthologues, then the gene duplication establishing the OS-E and OS-F lineages predated the divergence of mosquitoes and flies. The identification of orthologous OBPs and other chemosensory genes between D. melanogaster and A. gambiae should accelerate the transfer of physiological and behavioral information between these two species.     

Sexual Isolation and Cuticular Hydrocarbon Differences between <Emphasis Type="Italic">Drosophila santomea</Emphasis> and <Emphasis Type="Italic">Drosophila yakuba</Emphasis>

Drosophila santomea and Drosophila yakuba are two sister species inhabiting Saõ Tomé island. Previous studies showed that both species display strong reproductive isolation, although they can produce a few viable hybrids. Our study tried to understand the mechanism of this ethological isolation between two allopatric strains. A strong sexual isolation was confirmed, with a marked asymmetry. Comparisons of latency times to either courtship or copulation suggest that males do not discriminate females, whereas D. yakuba females, but not D. santomea females, accept their homospecifics more quickly. Cuticular hydrocarbon compositions of both species and sexes were also established with gas chromatography (GC) and GC/mass spectrometry analysis. All have (Z)-7-tricosene as their major compound. There are several quantitative differences between species for few minor compounds. The largest difference concerns n-heneicosane, which is more abundant in D. santomea than in D. yakuba flies (up to seven times more between males). A similar quantitative difference was also found in a pair of sympatric strains. Furthermore, D. yakuba males artificially perfumed with n-heneicosane were discriminated negatively by D. yakuba females, suggesting a role for this compound in the sexual isolation between these two species.     

Performance of Generalist and Specialist Herbivores and their Endoparasitoids Differs on Cultivated and Wild <Emphasis Type="Italic">Brassica</Emphasis> Populations

Through artificial selection, domesticated plants often contain modified levels of primary and secondary metabolites compared to their wild progenitors. It is hypothesized that the changed chemistry of cultivated plants will affect the performance of insects associated with these plants. In this paper, the development of several specialist and generalist herbivores and their endoparasitoids were compared when reared on a wild and cultivated population of cabbage, Brassica oleracea, and a recently established feral Brassica species. Irrespective of insect species or the degree of dietary specialization, herbivores and parasitoids developed most poorly on the wild population. For the specialists, plant population influenced only development time and adult body mass, whereas for the generalists, plant populations also affected egg-to-adult survival. Two parasitoid species, a generalist (Diadegma fenestrale) and a specialist (D. semiclausum), were reared from the same host (Plutella xylostella). Performance of D. semiclausum was closely linked to that of its host, whereas the correlation between survival of D. fenestrale and host performance was less clear. Plants in the Brassicaceae characteristically produce defense-related glucosinolates (GS). Levels of GS in leaves of undamaged plants were significantly higher in plants from the wild population than from the domesticated populations. Moreover, total GS concentrations increased significantly in wild plants after herbivory, but not in domesticated or feral plants. The results of this study reveal that a cabbage cultivar and plants from a wild cabbage population exhibit significant differences in quality in terms of their effects on the growth and development of insect herbivores and their natural enemies. Although cultivated plants have proved to be model systems in agroecology, we argue that some caution should be applied to evolutionary explanations derived from studies on domesticated plants, unless some knowledge exists on the history of the system under investigation.     

A Comparison of Semiochemically Mediated Interactions Involving Specialist and Generalist <Emphasis Type="Italic">Brassica</Emphasis>-feeding Aphids and the Braconid Parasitoid <Emphasis Type="Italic">Diaeretiella rapae</Emphasis>

Diaeretiella rapae, a parasitoid that predominately specializes in the parasitism of Brassica-feeding aphids, attacks Lipaphis erysimi, a specialist feeding aphid of the Brassicaceae and other families in the Capparales, at a greater rate than the generalist-feeding aphid, Myzus persicae. In this study, we investigated the orientation behavior of D. rapae to the volatile chemicals produced when these two aphid species feed on turnip (Brassica rapa var rapifera). We showed no significant preference orientation behavior to either aphid/turnip complex over the other. Isothiocyanates are among the compounds emitted by plants of the Brassicaceae in response to insect feeding damage, including by aphids. We assessed parasitoid orientation behavior in response to laboratory-formulated isothiocyanates. We tested two formulations and discovered significant orientation toward 3-butenyl isothiocyanate. We also assessed plant and aphid glucosinolate content, and showed large levels of glucosinolate concentration in L. erysimi, whereas there was little change in plant content in response to aphid feeding. Our results suggest that during the process of host location, similar cues may be utilized for locating L. erysimi and M. persicae, whereas the acceptance of hosts and their suitability may involve aspects of nonvolatile aphid chemistry.     

cis-Vaccenyl acetate as an aggregation pheromone inDrosophila melanogaster

Pentane extracts of matureDrosophila melanogaster males substantially increased the attractiveness of food odors to both males and females in a wind-tunnel olfactometer. Extracts of females caused no such increase. An active component of the extract was isolated and identified as (Z)-11-octadecenyl acetate (cis-vaccenyl acetate, cVA), and synthetic cVA was active in bioassay. Hydrolysis of the ester linkage or movement of the double bond to the 9 position destroyed the activity. Mature virgin males released cVA into their feeding vials, and amounts of synthetic CVA equal to that released per male caused significant bioassay responses. Females, which were known to receive cVA from males during copulation, were found to emit relatively large amounts of the ester into their feeding vials within 6 hr after mating. cVA had been demonstrated previously to be a close-range pheromone inD. melanogaster, discouraging males from courting other males or recently mated females; it now appears to have a longer-range function as well.     

Fatty Acid Composition and Oil Yield in Fruits of Five <Emphasis Type="Italic">Arecaceae</Emphasis> Species Grown in Cuba

The present study targeted the whole-fruit oil yield and fatty acid composition from five of the most abundant Arecaceae species grown in Cuba. The oil yields (% dry weight), determined by the Soxhlet extraction technique with hexane, were 25.5, 5.3, 6.9, 5.4, and 6.4% for Roystonea regia, Colpothrinax wrightii, Sabal maritima, Sabal palmetto and Thrinax radiata, respectively. The free fatty acid (FFA) content varied from 2.7 to 6.8%. Fatty acid (FA) profiles of the oils indicated that lauric acid (13.7–44.4%), myristic acid (9.4–22.4%) and palmitic acid (9.2–17.1%) as major saturated FA; whereas oleic acid (9.6–42.7%) and linoleic acid (9.3–17.0%) as major unsaturated FA. R. regia fruit seemed the most promising among Arecaceae grown in Cuba because of its high oil yield and low oil FFA content.     

Precise characterization of cuticular compounds in youngDrosophila by mass spectrometry

Cuticular hydrocarbons of youngDrosophila flies are singular with very long chains and complex diene mixtures. A precise characterization of these substances was carried out by epoxidation and analysis of the products by GC-MS with negative chemical ionization. InD. melanogaster, double bonds of dienes are more probable at carbon positions 11 or 13 and 21 or 23. InD. simulans, double bonds are shifted more towards the interior of the chain. Such a difference is also found among monoenes of both species. The analyses of monoenes and dienes confirm the similarity of cuticular compounds of young flies of both sexes in both species. A main cuticular compound ofD. erecta females, 9, 23-tritriacontadiene, is also presented.     

Forest Successional Stage Affects the Cortical Secondary Chemistry of Three Old Forest Lichens

Three epiphytic old forest lichens (Usnea longissima, Pseudocyphellaria crocata, and Lobaria pulmonaria) were transplanted along a natural shade–sun gradient comprising three successional stages in boreal spruce forests (dense young forest, open old forest, and clear-cut) for one summer. After harvest, extractable secondary compounds were analyzed by high-performance liquid chromatography, and the brown pigmentation in melanic species was quantified by reflectance measurements. Cortical compounds in all species increased from shady young forests to exposed clear-cuts. Usnic acid, the major cortical, secondary compound in U. longissima, showed consistently higher concentration in the clear-cut than in the two forested stands. Pseudocyphellaria crocata and L. pulmonaria, lacking extractable secondary compounds in the cortex, significantly increased their amounts of cortical melanins in well-lit stands. The medullary compounds showed more complex responses. Many were not influenced by environmental conditions during the transplantation, whereas the majority of those that responded showed the lowest concentration in clear-cut transplants. Only a few medullary compounds showed the highest concentration in the clear-cut, and at a low level of significance. The synthesis of UV-B-absorbing usnic acid and melanins seems to be part of an acclimation to increased light exposure. The medullary compounds in studied species barely function as solar screens despite their strong UV-B absorbance.     

Effects of phenolic acids on germination and early growth of herbaceous woodland plants

Four herbaceous plant species from woodland (clearings),Deschampsia flexuosa, Scrophularia nodosa, Senecio sylvaticus, andChamaenerion angustifolium, were tested for their sensitivity to phenolic acids. Seven commonly occurring phenolic compounds were used in a germination experiment in concentrations ranging from 0.01 to 10 mM, i.e., salicylic,p-hydroxybenzoic, syringic, caffeic, vanillic,p-coumaric, and ferulic acids. Germination was delayed rather than inhibited. Radicle elongation was strongly affected; at lower concentrations stimulatory effects were observed, whereas at high concentrations radicle elongation was severely reduced. Salicylic acid was the most effective phenolic compound, whereas caffeic acid caused no effects. Early growth was studied in more detail in a second experiment withDeschampsia flexuosa andSenecio sylvaticus and the phenolic acids, ferulic and p-coumaric acid. Primary root length, number and length of secondary roots, and dry weight were stimulated at 0.01 mM but were inhibited at 10 mM of both compounds. The results are discussed in view of the allelopathic relations between trees and herbaceous understory vegetation.     

Host selection behavior of bark beetles (Coleoptera: Scolytidae) attackingPinus ponderosa,with special emphasis on the western pine beetle,Dendroctonus brevicomis

Detection of weakened hosts from a distance by bark beetles through olfaction was investigated in field experiments. No significant numbers of Scolytidae were attracted to anaerobically treated pine bolts, stem disks, or sugar and ponderosa pine bark including phloem. Treatment of living trees with cacodylic acid induced attacks byDendroctonus brevicomis, D. ponderosae, Ips latidens, Gnathotrichus retusus, andPityophthorus scalptor, beginning two weeks after treatment. There was no significant difference between landing rates ofD. brevicomis andD. ponderosae on screened treated trees and screened controls. There was a significant increase in landing rates ofG. retusus andI. latidens, because both species had penetrated the screen and produced pheromones. Tree frilling alone did not increase the landing rate of bark beetles. Freezing of the lower trunk with dry ice did not increase significantly the landing rate ofD. brevicomis, D. ponderosae, G. retusus, orI. latidens on screened trees, whereas unscreened frozen trees were attacked by all four species. There was no significantly higher landing rate byD. brevicomis, D. ponderosae, I. paraconfusus, I. latidens, G. retusus, orHylurgops subcostulatus on screened trees evidencing symptoms of severe infection by the root pathogenVerticicladiella wagenerii, than on symptornless trees. These experiments show thatD. brevicomis, D. ponderosae, I. paraconfusus, I. latidens, andG. retusus land, apparently indiscriminately, on healthy and stressed hosts. Thus, in these species host discrimination must occur after landing and prior to sustained feeding.These studies were supported in part by the U.S.D.A. Forest Service, Cooperative State Research Services (2598-RRF.W-1 10) and the National Science Foundation and Environmental Protection Agency through a grant (NSF GB-34719/BM575-04223) to the University of California; and by the Canada Department of the Environment. The findings, opinions and recommendations are not necessarily those of the University of California or the funding agencies.From a thesis submitted by H.A. Moeck to the University of California, Berkeley, in partial fulfillment of the requirements for the degree Doctor of Philosophy in Entomology.     

Phytotoxic and Antifungal Compounds from Two <Emphasis Type="Italic">Apiaceae</Emphasis> Species, <Emphasis Type="Italic">Lomatium californicum</Emphasis> and <Emphasis Type="Italic">Ligusticum hultenii</Emphasis>, Rich Sources of Z-ligustilide and Apiol,Respectively

The seeds of two Apiaceae species, Ligusticum hultenii and Lomatium californicum, were investigated. Preliminary bioassays indicated that methylene chloride extracts of seeds of both species contained selective phytotoxic activity against monocots and antifungal activity against Colletotrichum fragariae. Active constituents were isolated by bioassay-guided fractionation, and the structures were elucidated by NMR and GC-MS as apiol and Z-ligustilide, isolated from L. hultenii and L. californicum, respectively. Apiol and Z-ligustilide had I₅₀ values of about 80 and 600 μM, respectively, for inhibition of the growth of Lemna paucicostata. The methylene chloride (CH₂Cl₂) extracts of the seeds and the isolated and purified compounds were tested against the 2-methylisoborneol-producing cyanobacterium (blue-green alga) Oscillatoria perornata, and the green alga Selenastrum capricornutum. The CH₂Cl₂ extracts of both Apiaceae species and apiol were weakly toxic to both species of phytoplankton, while Z-ligustilide was toxic to both with a lowest complete inhibitory concentration (LCIC) of 53 μM. Seeds of L. californicum and L. hultenii were found to be rich sources of Z-ligustilide (97 mg/g of dry seed) and apiol (40 mg/g of dry seed), respectively.     

Comparison of Age-dependent Quantitative Changes in the Male Labial Gland Secretion of <Emphasis Type="Italic">Bombus Terrestris</Emphasis> and <Emphasis Type="Italic">Bombus Lucorum</Emphasis>

Age-related changes of antennal-active components of male labial gland extracts were studied in two closely related bumblebee species, Bombus terrestris and B. lucorum. In B. terrestris, compounds eliciting electroantennogram (EAG) responses of virgin queens were ethyl dodecanoate, 2,3-dihydrofarnesal, 2,3-dihydrofarnesol, hexadecan-1-ol, octadeca-9,12,15-trien-1-ol, and geranylcitronellol. Compounds that elicited EAG responses from queens of B. lucorum were ethyl dodecanoate, ethyl tetradec-7-enoate, ethyl tetradec-9-enoate, ethyl hexadec-9-enoate, hexadecan-1-ol, hexadec-7-enal, octadeca-9,12-dien-1-ol, octadeca-9,12,15-trien-1-ol, and octadecan-1-ol. Quantities of these compounds in the labial glands changed significantly over the lifetime of the respective males of the two species. In both species, concentrations of the respective compounds reached their maximum within seven days after eclosion. Subsequently, a rapid decrease in the amount of EAG-active compounds occurred in B. terrestris, whereas in B. lucorum the amount of active compounds stayed approximately constant or decreased at a slow rate. Microscopy showed that in B. terrestris secretory cells of the labial glands undergo apoptosis from the fifth to the tenth day of life, whilst in B. lucorum labial gland cells remain unchanged throughout the life of the males.     

Sequestration of lichen compounds by three species of terrestrial snails

Three species of lichen-grazing snails,Balea perversa, Chondria clienta, andHelicigona lapicida, all from the Swedish island of Öland, were found to sequester lichen compounds when feeding on the crustous lichen speciesAspicila calcarea, Caloplaca flavovirescens, Lecanora muralis, Physcia adscendens, Tephromela atra, andXanthoria parietina. The lichen compounds detected in the soft bodies of the snail species analyzed included the anthraquinone parietin, the depside atranorin, as well as a presumable degradation product of the latter. Other lichen compounds such as (+)-usnic acid or -collatolic acid were not found in the soft bodies but were only detected in the feces, suggesting selective uptake of lichen compounds by the snails. In individuals ofC. clienta initially fed on the lichenX. parietina, the amount of sequestered parietin decreased over time on a parietin-free diet but was still detectable in the soft bodies after 28 days. In the ovoviviparous land snail,B. perversa, sequestered parietin was transferred from the mother to the eggs in the reproductive tract.Dedicated to Prof. Dr. F.-C. Czygan on the occasion of his 60th birthday.     

Isolation of Three Diterpenoid Acids from Sunflowers,as Oviposition Stimulants for the Banded Sunflower Moth, <Emphasis Type="Italic">Cochylis hospes</Emphasis>

The banded sunflower moth (BSFM), Cochylis hospes Walshingham (Lepidoptera: Cochylidae) is a specialist insect, the larvae of which feed on sunflowers, Helianthus spp., and a few other species of Compositae. It is one of the most important pests of sunflower in the USA. Previous work on H. annuus, the cultivated sunflower, revealed two diterpenoids that function as oviposition stimulants for female BSFM, and that other, more polar compounds also stimulated oviposition. Using a bioassay-guided approach, we isolated three additional diterpenoids, grandifloric acid (1), 15β-hydroxy-ent-trachyloban-19-oic acid (2), and 17-hydroxy-16α-ent-kauran-19-oic acid (3), from polar fractions of pre-bloom sunflower head extracts. In laboratory bioassays, purified natural samples of each of these compounds stimulated oviposition by female BSFM. Structure–activity relationships of the five diterpenoids known to stimulate oviposition by female BSFM are discussed.     

The Effect Of Pollination On Floral Fragrance in Thistles

We investigated postpollination changes in fragrance composition and emission rates, as well as pollinator discrimination in hand-pollinated flower heads of two thistle species: Canada thistle (Cirsium arvense) and sandhill thistle (C. repandum). Following pollination, neither species emitted any novel compounds that could function as repellents. Scent emission rates declined in pollinated plants of both species by approximately 89% within 48 hr. This decline was evident in all 13 scent components of C. arvense. Apis mellifera, the dominant pollinator in the study population of C. arvense, was nearly three times more likely to visit an unpollinated rather than a pollinated flower head. A more complex pattern was observed for C. repandum, whose scent comprised 42 compounds. Quantities of aromatic and sesquiterpenoid volatiles declined after pollination, whereas two classes of scent compounds, fatty acid derivatives and monoterperpenoids, continued to be emitted. In C. repandum, discrimination against pollinated flower heads by Papilio palamedes (its primary pollinator) was not as marked. Unpollinated control plants of both species maintained moderate levels of scent production throughout this experiment, demonstrating that senescence and floral advertisement may be delayed until pollination has occurred. We expect postpollination changes in floral scent contribute to communication between plants with generalized pollinator spectra and their floral visitors. This study provides the first field study of such a phenomenon outside of orchids.     

Epicuticular Hydrocarbon Variation in Drosophila mojavensis Cluster Species

Epicuticular hydrocarbon variation was investigated among the three species of the Drosophila mojavensis cluster (D. mojavensis, D. arizonae, and D. navojoa) within the large D. repleta group. Because these hydrocarbons serve as contact pheromones in adult D. mojavensis, the chemical characteristics and differences in hydrocarbon profiles in populations of these three sibling species were further investigated. Twenty-seven hydrocarbon components with chain lengths ranging from C₂₈ to C₄₀, including n-alkanes, methyl-branched alkanes, n-alkenes, methyl-branched alkenes, and alkadienes were observed. Hydrocarbon profiles among the three species reared on different cactus hosts were easily aligned with previously identified components in D. mojavensis. Male and female D. navojoa possessed a 31-methyldotricont-6-ene absent in both D. arizonae and D. mojavensis, while lacking the 8,24-tritricontadiene present in these two species. D. navojoa adults had far less 2-methyloctacosane than these sibling species, but the significance of this difference was obscured by the degree of variation among populations in amounts of this hydrocarbon. Mainland and Baja California populations of D. mojavensis were fixed for differences in the amounts 8,24-tritricontadiene, 9,25-pentatricontadiene, and 9,27-heptatricontadiene, consistent with all previous studies. Amounts of 18 of the 27 hydrocarbon components were greater in flies reared on Opuntia cactus. Canonical discriminant function analysis resolved all three species into distinct, nonoverlapping groups, suggesting that epicuticular hydrocarbon profiles are species-specific in the D. mojavensis cluster. Based on the amounts of interpopulation variation in hydrocarbon profiles in these three species, we hypothesize that epicuticular hydrocarbon differences may evolve early during the formation of new species.