Responses of three mouse species to deterrent chemicals in the monarch butterfly. II. Taste tests using intact monarchs

Summary Peromyscus melanotis is the only one of three mouse species that eats monarch butterflies at their overwintering sites in Mexico. I tested two hypotheses: 1)P. aztecus avoids monarchs because of a bitter taste aversion to cardiac glycosides (CGs) and an inability to reject CG-rich body parts; 2)Reithrodontomys sumichrasti avoids monarchs principally because of a bitter taste aversion to the CGs. None of the species are sensitive to the toxic effects of ingested CGs. Feeding responses of laboratory-reared mice of each species to monarchs with low, medium and high CG concentrations were compared. BothP. aztecus andR. sumichrasti ate significantly fewer of all three types of monarchs thanP. melanotis. ForP. aztecus andR. sumichrasti, the number of monarchs eaten decreased with increasing CG concentration, whereas forP. melanotis, the number remained constant.Peromyscus melanotis andR. sumichrasti developed a feeding technique for rejecting the CG-laden cuticular material, which reduced the bitterness of ingested monarch material. However,R. sumichrasti displayed the technique significantly less often thanP. melanotis; andP. aztecus never developed it. I conclude that high taste sensitivity to CGs and less versatile food handling preventP. aztecus andR. sumichrasti from overcoming the monarch's chemical defenses.     

Sequestration of plant pyrrolizidine alkaloids by chrysomelid beetles and selective transfer into the defensive secretions

Summary Oreina cacaliae andO. speciosissima (Coleoptera, Chrysomelidae) sequester in their elytral and pronotal defensive secretions pyrrolizidine alkaloids (PAs) as Noxides (PA N-oxides). The PA N-oxide patterns found in the beetles and their host plants were evaluated qualitatively and quantitatively by capillary gas chromatography/mass spectrometry (GC-MS). Of the three host plantsAdenostyles alliariae (Asteraceae) is the exclusive source for PA N-oxide sequestration in the defensive secretions of the beetles. With the exception of O-acetylseneciphylline the N-oxides of all PAs ofA. alliariae, i.e. senecionine, seneciphylline, spartioidine, integerrimine, platyphylline and neoplatyphylline were identified in the secretion. PA N-oxides typical ofSenecio fuchsii (Asteraceae) were detected in the bodies of the beetles but not in their secretion. No PAs were found in the leaves of the third host plant,Petasites paradoxus (Asteraceae). The results suggest the existence of two distinctive storage compartments for PA N-oxides in the beetle: (1) the defensive secretion, containing specifically PA N-oxides acquired fromA. alliariae; (2) the body of the beetle, sequestering additionally but less selectively PA N-oxides from other sources,e.g. S. fuchsii or monocrotaline N-oxide fed in the laboratory. The concentration of PA N-oxides in the defensive secretion is in the range of 0.1 to 0.3 mol/1, which is more than 2.5 orders of magnitude higher than that found in the body of the beetle. No significant differences exist in the ability of the two species of beetles to sequester PA N-oxides fromA. alliariae, althoughO. speciosissima, but notO. cacaliae, produces autogenous cardenolides. A negative correlation seems to exist between the concentrations of plant-derived PA N-oxides andde novo synthesized cardenolides in the defensive secretion ofO. speciosissima.     

Pyrrolizidine alkaloids on three trophic levels – evidence for toxic and deterrent effects on phytophages and predators

Summary. Pyrrolizidine alkaloids (PAs) present a model system in the investigation of tritrophic interactions mediated by plant secondary compounds. However, their toxicity for insect herbivores has never been experimentally proven. Here, we demonstrate the toxic effects of a PA on growth and survival of the eri silk moth Philosamia ricini. In a feeding experiment, larvae of this generalist herbivore fed with an artificial PA diet gained weight significantly slower than control animals, and died as pupae. We suggest that derivatives of the ingested PA N-oxide damage developmental functions during metamorphosis. A tracer test with [¹⁴C]senecionine N-oxide revealed that the caterpillars lack adaptations that would prevent conversion of the chemical into the pro-toxic free base. In contrast, the PA adapted leaf beetle Longitarsus anchusae accumulates PAs as N-oxides. We tested the purpose of sequestration in this species as defence against predators. Through a series of prey choice experiments with three carabid predator species, chemically non-protected bark beetle pupae were chosen almost uniformly over L. anchusae pupae. In a following choice test with one of these predators, artificially PA-treated mealworm segments deterred the predator from feeding. Overall the study corroborates the immediate toxic effect of PAs on non-adapted herbivores and the protective effect that adapted insects may gain by sequestering them. It thereby underlines the potential for PAs to play a central role in multitrophic interactions between plants, phytophages and their predators.     

Variation of pyrrolizidine alkaloids in Ithomiinae: A comparative study between species feeding on Apocynaceae and Solanaceae

Summary The primitive, Apocynaceae-feeding Ithomiine,Tithorea harmonia, incorporates dehydropyrrolizidine alkaloids (PAs) from its larval foodplant (Prestonia acutifolia), rarely visiting PA sources pharmacophagously in the adult; females show higher concentrations of PAs than males, with similar variance. The close relativeAeria olena (feeding onP. coalita, without PAs) shows similar PA concentrations in both sexes and greater variation in males, like more advanced Solanaceae-feeding Ithomiine such asMechanitis polymnia, which likeA. olena obtain PAs by pharmacophagy in the adult (mainly males). This difference is due to the dynamics of PA incorporation in these species. Little variation in PA content was found among allopatric populations of the same species, but variation in available PA sources in different months was correlated with different average storage levels in the butterflies.     

Sequestration of pyrrolizidine alkaloids by larvae ofTellervo zoilus (Lepidoptera: Ithomiinae) and their role in the chemical protection of adults against the spiderNephila maculata (Araneidae)

Summary Life stages of the primitive Australian ithomiine butterflyTellervo zoilus and its larval hostplant, the apocynaceous vineParsonsia straminea, were quantitatively assayed for pyrrolizidine alkaloids (PAs). PAs were found in all stages, mainly as N-oxides, being most concentrated in larvae and freshly-emerged adults. Although adults feed at various confirmed PA sources this probably does not compensate for losses, as wild-caught adults had considerably lower concentrations of PAs. The main alkaloid present in both freshly-emerged adults and in leaves of the host-plant was lycopsamine (1b), stored by butterflies in the N-oxide form. Its presence in higher proportion, in relation to intermedine (1a), in larvae, pupae and adults ofTellervo in relation to the host-plants suggests the inversion of intermedine to lycopsamine by the insects. No 14-member ring macrocyclic PAs were detected in either food-plant or butterflies. Several other PAs were found in wild-caught adults reflecting visits to other PA sources. PAs were also found in high concentrations in freshly-emerged individuals of the danaineEuploea core bred onParsonsia straminea. Wild-caughtDanaus affinis had high PA levels acquired from adult feeding. Freshly emergedEuploea raised onIschnocarpus frutescens andDanaus raised onIschnostemma carnosum (both PA-free) were preyed on by the orb weaving spiderNephila maculata, and showed no PAs. In all cases where PAs were present, most butterflies were liberated, usually cut out of the web unharmed, byNephila. The spider's response was not closely linked to PA concentration, however, and may also depend on hunger levels and previous experience with PA-containing butterflies. All control and other non-PA containing butterflies were consumed although rejection of some body parts of freshly-emergedDanaus affinis suggests that compounds other than PAs may be involved.     

<Emphasis Type="Italic">Ceroplastes albolineatus</Emphasis>, the first scale insect shown to sequester pyrrolizidine alkaloids from its host-plant <Emphasis Type="Italic">Pittocaulon praecox</Emphasis>

Summary. The occurrence of pyrrolizidine alkaloids (PAs) in Pittocaulon (ex Senecio) praecox (Asteraceae) a species endemic to Mexico was established. The aboveground plant organs contain the 1,2-saturated monoester 7-angeloyl-dihydroxyheliotridane together with a small proportion of its 9-angeloyl isomer as major alkaloid. The monoesters are accompanied by the macrocyclic otonecine derivative senkirkine. Roots contain only related macrocyclic PAs with senecionine, senkirkine and platyphylline as major components; monoesters are absent. The broom-like succulent stems of P. praecox are infested by the scale insect Ceroplastes albolineatus conspicuously visible by its huge wax cover. All life-history stages, i.e. females, eggs, first instar nymphs (crawlers) and the wax cover were found associated with PAs. The measured PA concentrations clearly indicate sequestration. The highest PA concentrations (mg / g dry weight) reached are: mature females, 0.44; eggs, 0.58; crawlers, 0.37; wax cover, 0.08. The host plant as well as in the infesting scale insect contain the PAs exclusively as free bases. As a phloem-feeder C. albolineatus must acquire the PAs with the ingested phloem sap. This appears plausible since in Senecio species PA are transmitted and circulated through the phloem path. It is suggested that PAs may protect particularly the crawlers as the most endangered stage in the life-cycle of the scale insect.     

Phenological fate of plant-acquired pyrrolizidine alkaloids in the polyphagous arctiid Estigmene acrea

Summary. The alkaloid profiles of the life history stages of the highly polyphagous arctiid Estigmene acrea were established. As larvae individuals had free choice between a plain diet (alkaloid-free) and a diet that was supplemented with Crotalaria-pumila powder with a known content and composition of pyrrolizidine alkaloids (PAs). Idiosyncratic retronecine esters (insect PAs) accounted for approximately half of the PAs recovered from the larvae. These alkaloids were synthesized by the larvae through esterification of dietary supinidine yielding the estigmines, and esterification of retronecine yielding the creatonotines. The retronecine is derived from insect-mediated degradation of the sequestered pumilines (macrocyclic PAs of the monocrotaline type). With one exception, the PA profiles established for larvae were found almost unaltered in all life-stages as well as larval exuviae and pupal cocoons. The exception is the males, which in comparison to pupae and adult females, showed a significantly decreased quantity of the creatonotines and pumilines. These data support the idea that the creatonotines are direct precursors of the PA-derived male courtship pheromone, hydroxydanaidal. Crosses of PA-free males with PA-containing females and vice versa confirmed an efficient trans-mission of PAs from males to females and then from females to eggs. In single cases a male bestowed almost his total PA load to the female, and a female her total load to the eggs. The results are discussed with respect to pheromone formation, PA transmission between life-stages, and the defensive role of PAs against predators and parasitoids     

Some polyphagous Homoptera gain protection from their natural enemies by feeding on the toxic plantsSpartium junceum andErythrina corallodendrum (Leguminosae)

Summary Interactions were studied among alkaloid-containing legumes (Erythrina corallodendrum andSpartium junceum) and non-toxic plants (Citrus sinensis, Cucurbita moschata andEuphorbia tirucalli), several polyphagous homopterans,Aphis craccivora (Aphididae),Icerya purchasi, I. aegyptiaca (Margarodidae),Lepidosaphes ulmi (Diaspididae) andPlanococcus citri (Pseudococcidae), and some major natural enemies of these homopterans. Significant reductions in survival due to negative effects of alkaloid containing as compared with non-alkaloidal plants were recorded for the predatorsRodolia cardinalis andChilocorus bipustulatus, but not forCryptolaemus montrouzieri (Coleoptera: Coccinellidae),Chrysoperla carnea (Neuroptera: Chrysopidae) andSympherobius sanctus (Neuroptera: Sympherobiidae). The development time of the larvae or pupae ofR. cardinalis, C. carnea andS. sanctus was longer on the toxic plants than on the non-toxic ones. The percentage of parasitism ofA. craccivora collected from the non-alkaloidal plantsVicia palaestina andMelilotus albus was much higher than that onS. junceum. The parasitoid complexes ofA. craccivora differed between both plant groups. The nutritive value of honeydew ofI. purchasi andA. craccivora, as expressed by the life span ofEncyrtus infelix (Hymenoptera: Encyrtidae) adults, was also investigated. Life spans were significantly longer when the wasps fed on honeydew produced on non-alkaloidal plants (C. sinensis andPittosporum tobira) than on alkaloid containing plants whenI. purchasi — but notA. craccivora — was the producer. It is suggested that the chemical defense ofE. corallodendrum andS. junceum is exploited by polyphagous phytophages to reduce predation. In nature, population growth and density of four of the investigated homopterans are conspicuously high when they developed on the alkaloid containing plant species, and very low on non-alkaloid plants. The efficiency of their natural enemies may be reduced by sequestration of alkaloids (or other toxic plant compounds) or their transfer into excreted honeydew. Therefore it is assumed that a generalist phytophagous homopteran may be protected from its natural enemies, although at different rates of efficiency, if it can safely sequester the host allelochemical when it develops on toxic species within its host range.     

Toxicity and attraction effects produced byMorinda citrifolia fruits on theDrosophila melanogaster complex of species

Summary The ripe fruit of the Indian mulberry,Morinda citrifolia, is the host plant forDrosophila sechellia but is highly toxic for three closely related species (D. melanogaster, D. simulans, D. mauritiana). A simple bioassay is described with which a clear dose response to the fruit was found for these species. Significant differences in reactivity to the ripe fruit were found among species. Tested strains ofD. simulans andD. mauritiana adults were more sensitive to the toxic properties of the fruit thanD. melanogaster. A marked intraspecific variability was shown inD. melanogaster. Reciprocal interspecific hybridizations betweenD. sechellia andD. mauritiana suggested an autosomal dominant control of resistance. MoreoverD. melanogaster intraspecific crossings suggested the influence of an additional X-linked factor. Responses of flies toMorinda fruit in different states were tested in a T olfactometer. The less resistant strains ofDrosophila generally showed less preference for the ripe fruit.     

Pyrrolizidine alkaloids between plants and insects: A new chapter of an old story

Summary Among alkaloids the pyrrolizidine alkaloids (PAs) play a unique role in the interactions between plants and adapted insects. InSenecio spp. (Asteraceae) PAs are synthesized in the roots as alkaloidN-oxides which are specifically translocated into shootsvia the phloem-path and channeled to the preferred sites of storage (e.g. inflorescences) where they are stored in the cell vacuoles. In differentSenecio spp. senecionineN-oxide is produced as the common product of biosynthesis, which subsequentlyvia a number of simple but specific reactions is transformed into typical speciesspecific PA-patterns. Insects from diverse taxa sequester PAs for their own defense. Lepidopterans (e.g. arctiids such asTyria jacobaeae andCreatonotos transiens) may hydrolyze plant acquired ester-PAs and convert the resulting necine base into insect-specific PAs by esterification with an acid of their own metabolism. Adapted arctiids and the grasshopperZonocerus take up PAs in the state of the tertiary amine.N-Oxides are reduced in the guts prior to uptake. In the bodies the tertiary PAs are rapidlyN-oxidized by a specific mixed-function oxigenase and are maintained in theN-oxide state. The importance of the reversible interconversion of the nontoxicN-oxide (pro-toxine) into the toxic tertiary alkaloid is discussed as the specific feature of PAs in plant-insect interactions.     

Tropane and pyrrolizidine alkaloids in the ithomiinesPlacidula euryanassa andMiraleria cymothoe (Lepidoptera: Nymphalidae)

Summary Larvae of the ithomiine butterflyPlacidula euryanassa sequester tropane alkaloids (TAs) from the host plantBrugmansia suaveolens and pass them through the pupae to freshly emerged adults. Wild caught adults also show in their tissues, variable amounts of pyrrolidizine alkaloids (PAs), probably sequestered from variable plant sources and subject to dynamics of incorporation, accumulation and utilization of PAs by ithomiine butterflies. The ratio TAs/PAs is also variable between different populations.Miraleria cymothoe, another ithomiine that feeds onB. suaveolens as larvae, does not sequester TAs from the host-plant, but sequesters PAs from plant sources visited by the adult butterflies. The main alkaloid found in both butterflies is lycopsamine, which also is the principal PA found in all genera of Ithomiinae.     

Uptake and metabolism of [<Superscript>14</Superscript>C]rinderine and [<Superscript>14</Superscript>C]retronecine in leaf-beetles of the genus <Emphasis Type="Italic"> Platyphora</Emphasis> and alkaloid accumulation in the exocrine defensive secretions

Summary. Sequestration and processing of pyrrolizidine alkaloids (PAs) by leaf beetles of the genus Platyphora were investigated. Tracer experiments with labeled alkaloids were performed with P. eucosma feeding on Koanophyllon panamense (Asteraceae, tribe Eupatorieae). P. eucosma catalyzes the same reactions previously demonstrated for P. boucardi specialized to Prestonia portobellensis (Apocynaceae): (i) epimerization of rinderine to intermedine; (ii) esterification of retronecine yielding insect-specific PAs; (iii) efficient transport of the PAs as free bases into the defensive secretions. P. bella feeding on Tournefortia cuspidata (Boraginaceae) shows the same sequestration behavior and ability to synthesize the specific retronecine esters. P. ligata, a species phylogenetically closely related to the PA adapted species and clustering in the same clade, but feeding on a host plant devoid of PAs, feeds easily on PA treated host-plant leaves, but does not sequester or metabolize PAs. P. kollari a species clustering outside the PA clade refused to feed on its food-plant leaves painted with PAs. The results are discussed in relation to host-plant selection of the PA adapted species and the role of PAs in chemical defense. Received 20 September 2002; accepted 18 November 2002.     

Phototransformation and ecotoxicity of the drug Naproxen-Na

The phototransformation of naproxen Na in aqueous medium has been investigated. Irradiation of the drug in drinking water affords seven photoproducts. Three of them are dimeric photoproducts isolated for the first time. The compounds, isolated by chromatographic processes, have been identified by spectroscopic means. The toxicity of the photoproducts and the parent drug has been assayed on Daphnia magna and Vibrio fischeri. The results indicate that some photoproducts are more toxic than naproxen.     

Oviposition stimulants and deterrents control acceptance ofAlliaria petiolata byPieris rapae andP. napi oleracea

Summary Differential acceptance of garlic mustard,Alliaria petiolata byPieris rapae L. andP. napi oleracea is explained by their differential sensitivities to oviposition stimulants and deterrents in the plant. Fractions containing the stimulants and deterrents were isolated by solvent partitioning between water and n-butanol and by open-column chromatography followed by HPLC.P. napi oleracea showed no preference when offered a choice ofA. petiolata or cabbage, but was strongly stimulated to oviposit by post-butanol water extracts ofA. petiolata. The most abundant glucosinolate in this extract was identified as sinigrin, which could explain the high degree of stimulatory activity.P. rapae preferred cabbage plants overA. petiolata, and the relatively low stimulatory activity was also associated with the glucosinolate-containing aqueous extract. However, this species was strongly stimulated by a fraction that contained small amounts of glucotropaeolin along with unknown compounds. Deterrents to both species were found in the butanol extract fromA. petiolata, andP. napi oleracea was more sensitive thanP. rapae to these deterrents. Some HPLC fractions from the BuOH extract were strongly deterrent toP. napi oleracea, but were inactive toP. rapae. The ecological significance of these behavioral differences between the twoPieris species is discussed.     

Pyrrolizidine alkaloids and pentacyclic triterpene saponins in the defensive secretions of Platyphora leaf beetles

Summary. Field collected exocrine defensive secretions of nine neotropical Platyphora species were analyzed for the presence of plant acquired pyrrolizidine alkaloids (PAs) and pentacyclic triterpene saponins. All species secrete saponins. In addition, five species feeding on Tournefortia (Boraginaceae), Koanophyllon (Asteraceae, tribe Eupatorieae) and Prestonia (Apocynaceae) were shown to sequester PAs of the lycopsamine type, which are characteristic for species of the three plant families. The PA sequestering species commonly store intermedine, lycopsamine and their O^3′-acetyl or propionyl esters as well as O⁷- and O⁹-hydroxyisovaleryl esters of retronecine. The latter as well as the O^3′-acyl esters were not found in the beetles’ host plants, suggesting the ability of the beetles to esterify plant derived retronecine and intermedine or its stereoisomers. Despite the conformity of the beetles’ PA patterns, considerable inconsistencies exist regarding the PA patterns of the respective host plants. One host plant was devoid of PAs, while another contained only simple necines. Since the previous history of the field collected beetles was unknown this discrepancy remains obscure. In contrast to the Palearctic chrysomeline leaf beetles, e.g. some Oreina species which ingest and store PAs as their non-toxic N-oxides, Platyphora leaf beetles absorb and store PAs as the toxic free base (tertiary PA), but apparently avoid to accumulate PAs in the haemolymph. This suggests that Chrysolina and Platyphora leaf beetles developed different lines of adaptations in their parallel evolution of PA mediated chemical defense. Received 30 November 2000; accepted 5 February 2001     

Autotoxic effects of essential oils on photosynthesis in parsley, parsnip, and rough lemon

Summary. Many plant species contain essential oils with allelochemical properties, yet the extent to which these same chemicals can be autotoxic is unclear. In this study, we tested the toxicity of several essential oil components to three species that produce them—Pastinaca sativa and Petroselinum crispum (Apiaceae), and Citrus jambhiri (Rutaceae). The effects of exogenous application of small amounts of essential oil components to the surface of foliage, followed by a pinprick to allow entry into the leaf, were monitored by chlorophyll fluorescence imaging. Rapid and spatially extensive declines in photosynthetic capacity were detected within 200 s. The most toxic constituents were monoterpenes. Two sesquiterpenes, caryophyllene and farnesene, and the phenylpropanoid myristicin, by comparison, were not toxic. Autotoxicity of endogenous essential oil was investigated by slicing through containment structures (glands or tubes); significant toxicity, manifested by reduced photosynthetic activity, was observed in all three species but was most pronounced in P. sativa and P. crispum, both of which possess oil tubes.     

Detoxification of pyrrolizidine alkaloids by the harvestman <Emphasis Type="Italic">Mitopus morio</Emphasis> (Phalangidae) a predator of alkaloid defended leaf beetles

Summary. The harvestman Mitopus morio (Phalangidae) is a generalist predator. It is known to prey on larvae of the chrysomelid leaf beetle Oreina cacaliae defended by plant acquired pyrrolizidine alkaloids (PAs). Tracer feeding experiments were performed to determine how harvestmen tolerate protoxic PAs. Minced meat containing either [¹⁴C]senecionine or [¹⁴C]senecionine N-oxide was fed to M. morio and subsequently feces and bodies were analyzed. Labeled alkaloid N-oxide remained stable and was eliminated almost unaltered with the feces; only 10% was recovered as tertiary PA. In contrast, approximately 80% of labeled tertiary alkaloid (senecionine) ingested with the diet was N-oxidized and eliminated; the remaining 20% consisted of unchanged senecionine and a polar metabolite of unknown structure. Harvestmen process their diet by excreting digestive juice, indicated by bleaching of the meat color. Analysis of the processed diet revealed some N-oxidation of [¹⁴C]senecionine, suggesting the gut as the site of Noxidation. Analysis of the bodies of harvestmen 80 hours after the tracer feeding pulse revealed only trace amounts of the polar metabolite. Neither senecionine nor its N-oxide could be detected in the body extracts. The results are discussed in relation to the strategies of PA adapted insects to avoid accumulation of tertiary PAs in living tissues.     

Toxicity assays of ladybirds using natural predators

Nestling blue titsParus caeruleus L. were given diets containing homogenized ladybirds, to assess the effects of their chemical defences. The 2spot, 10spot and water ladybirds produced no apparent toxic effects when small numbers were given at regular intervals.The pine and kidney-spot ladybirds slightly inhibit growth and may be toxic to very young nestlings. The Water ladybird is extremely distasteful to this predator, despite its lack of toxicity. The results are discussed with reference to the proposed Müllerian and Batesian mimetic relationships between the ladybird species.     

Resistance of the generalist moth Trichoplusia ni (Noctuidae) to a novel chemical defense in the invasive plant Conium maculatum

Summary. Conium maculatum is an apiaceous species native to Eurasia that is highly toxic to vertebrates due to the presence of piperidine alkaloids, including coniine and γ-coniceine. More than 200 years after invading the United States this species remains mostly free from generalist insect herbivores. The presence of novel chemical defenses in the introduced range could provide invasive species with a competitive advantage relative to native plants. The cabbage looper (Trichoplusia ni) is a generalist lepidopteran found throughout the US that occasionally feeds on C. maculatum. We evaluated the toxicity of piperidine alkaloids to T. ni and determined putative resistance mechanisms, both behavioral and physiological, that allows this insect to develop successfully on C. maculatum foliage. T. ni larvae raised on diets enriched with coniine and γ-coniceine showed a decrease in consumption and longer development time, but no effects on growth were found at any alkaloid concentration. In a diet choice experiment T. ni larvae showed no avoidance of alkaloid-enriched diets, suggesting that the deterrence produced by alkaloids was related to a post-ingestive metabolic response. The ability of T. ni to consume diets high in alkaloid content could be due to at least three different mechanisms: 1) a decreased consumption rate, 2) efficient excretion of at least 1/3 of ingested alkaloids unmetabolized in frass, and 3) partial detoxification of alkaloids by cytochrome P450 s, as shown by the decreased larval growth in the presence of piperonyl butoxide, a P450 inhibitor. Even though T. ni tolerates C. maculatum alkaloids, the use of this species as a host plant could be ecologically disadvantageous due to prolonged larval growth and thus increased exposure to predators. Novel plant secondary compounds do not guarantee increased resistance to generalist herbivores.     

Comparative electrophysiological analysis of plant odor perception in females of threePapilio species

Summary Antennae of femalePapilio butterflies perceive many volatile plant constituents with widely differing, constituent-specific sensitivities. We compared the responses of threePapilio species to volatiles from host and non-host plants to assess species-specificity and the degree of evolutionary conservatism in olfactory responses.Since previous studies had demonstrated that the polar constituents in odor fromDaucus carota stimulate oviposition behavior inPapilio polyxenes, we collected headspace volatiles fromD. carota, Pastinaca sativa (both Apiaceae) andArtemisia dracunculus (Asteraceae) and separated the polar fraction of these volatiles by gas chromatography. GC-coupled electroantennograms (GC-EAG) were recorded from the speciesPapilio polyxenes, P. machaon hippocrates andP. troilus. In addition, the responses of the three species to five compounds known as generally occurring constituents of plant odor were recorded. The relative sensitivities for these compounds were nearly identical in all threePapilio species. The response spectra to the separated plant volatiles also showed considerable similarities among the species.From the limited set of GC peaks evoking a response in one of the species, 64% (D. carota), 44% (P. sativa) and 29% (A. dracunculus) also evoked a response in both of the other species. The responses of the two closely related Apiaceae feeders (P. polyxenes, P. m. hippocrates) to volatiles fromD. carota were more similar to each other than was either to the response ofP. troilus, which feeds on Lauraceae. However, this was not true for the responses to volatiles fromP. sativa. The least congruence among the three species was found in the responses to volatiles fromA. dracunculus, a non-host for all of them. The differences and similarities found in the response profiles of the threePapilio species are discussed with respect to evolutionary adaptation to host odor versus evolutionary conservatism in adaptation of olfactory receptors.