Antimicrobial Activity of Exocrine Glandular Secretion of Chrysomela Larvae

The exocrine glandular secretions of leaf beetle larvae of the taxon Chrysomela are well-known defensive devices used against some generalist predators. Salicylaldehyde is the major repellent component of secretions emitted by larvae of Chrysomela vigintipunctata and C. lapponica, which feed on salicin-rich Salicaceae. In this study, we examined whether salicylaldehyde is also active against the entomopathogenic fungus Metarhizium anisopliae. The germination and growth of this fungus was strongly inhibited when salicylaldehyde was applied directly onto the blastospores. The salicylaldehyde concentration of the larval secretions of the tested willow feeding Chrysomela larvae was much higher than the one necessary to display this antifungal activity. Additionally, salicylaldehyde was shown to reduce germination and growth of M. anisopliae via the gas phase over a distance of 45 mm. Further studies on the antimicrobial activity of the salicylaldehyde-containing secretions of Chrysomela larvae revealed that they act nonspecifically against prokaryotic (Escherichia coli) and eukaryotic cells (Saccharomyces cerevisiae and Trichoplusia ni). All antimicrobial and cytotoxic effects detected proved to be due to salicylaldehyde. The larval secretions of the birch-feeding allospecies of C. lapponica, that do not contain salicylaldehyde, but mainly carboxylic acids and their esters, showed no detectable effects on bacteria or fungi and no cytotoxic effects against insect cells. The results are discussed with respect to their ecological relevance.     

Toxins in chrysomelid beetles Possible evolutionary sequence from de novo synthesis to derivation from food-plant chemicals

In the Chrysomelinae, it appears that de novo synthesis of chemicals for defense is the primitive state, and the sequestration of plant chemicals for defense the derived state. The derived state evolved through both the morphological and biochemical preadaptiveness of the homologous defensive glands. In the adults, we discuss one unique case of sequestration in exocrine defensive glands of host-plant pyrrolizidine alkaloids byOreina cacaliae. However, hypericin is not sequestered either in the glands or elsewhere in the body ofChrysolina spp. feeding onHypericum, which contradicts an earlier claim. In the larvae, we examine in more detail how the phenolglucoside salicin can be used as the precursor of the salicylaldehyde present in the defensive secretion ofPhratora vitellinae andChrysomela spp. with minimal changes in the biochemical mechanisms involved in the biosynthesis of iridoid monoterpenes in related species.     

Selective predation on chemically defended chrysomelid larvae

Laboratory experiments showed that femaleTenthredo olivacea prefer to prey upon insects of a previously encountered species, instead of upon unknown ones. This has been observed when comparing two natural prey of the sawfly, the larvae ofPhratora vitellinae and those ofPlagiodera versicolora. The two species secrete copious amounts of defensive secretion, the first salicylaldehyde, and the latter a mixture of cyclopentanic monoterpenes. The predator appears less reluctant when encountering a species whose secretion has been previously experienced. A selective pressure might thus exist favoring rare secretions, which is consistant with the well-known diversity of defensive compounds among sympatric insects.     

Chemical defense secretions of some species of Malaysian rhinotermitidae (Isoptera,Rhinotermitidae)

The defense secretions of the soldiers of the Malaysian rhinotermitid,Parrhinotermes aequalis (Havilandi) andP. pygmaeus (John),Termitogeton planus (Havilandi) andSchedorhinotermes malaccensis (Holmgren) consist mainly of vinyl ketones, whereas that ofProrhinotermes flavus (Bugnion & Popoff) gives (E)-1-nitropentadecene. The chemistry of the defense secretions ofParrhinotermes andTermitogeton is documented and based on their chemical relationships;Termitogeton shows a closer affinity to the Rhinotermitinae than Heterotermitinae.     

Behavioral and growth responses of specialist herbivore,Homoeosoma electellum,to major terpenoid of its host,Helianthus SPP

The responses of a sunflower specialist,Homoeosoma electellum, the sunflower moth, to the terpenoids produced by its host plant,Helianthus, were measured. Larvae were reared on synthetic diet containing one of three concentrations of the dominant sesquiterpene lactone found in glandular trichomes ofH. maximilliani. Treatments were initiated at each of three larval ages. Pupal weight was significantly reduced, but the effect diminished as the larvae aged. Survival and development time were unaffected by various treatments. In behavioral tests, larvae showed no preference for untreated synthetic diet compared to diet containing the secondary compound at a concentration of 1% by dry weight. When the concentration was raised to 5%, all but last-instar larvae showed a significant preference for the untreated diet. A second behavioral test measured the damage done to whole florets and an array of floral parts offered to larvae of different ages. The damage pattern of floral tissues changed as the larvae grew older, demonstrating that the willingness of larvae to eat tissues contaminated with trichome contents increased with age. The value of the glandular trichome contents as a defense againstH. electellum is discussed.     

Toxicity of allelochemicals from wild insect-resistant tomatoLycopersicon hirsutum f.glabratum toCampoletis sonorensis,a parasitoid ofHeliothis zea

Greenhouse-grown plants of five tomato lines varying in their level of 2-tridecanone-mediated resistance toManduca sexta (L.) andLeptinotarsa decemlineata (Say) were found to adversely affect larvae ofCampoletis sonorensis (Cameron), a larval endoparasitoid ofHeliothis zea (Boddie), in a manner directly related to their level of resistance. The parasitoid larvae, which emerge as fifth instars from their host and construct a cocoon on the foliage of their hosts' host plant, suffered extensive mortality during cocoon spinning on highly resistant foliage. Mortality was greatest (82%) on the highly resistant plants ofLycopersicon hirsutum f.glabratum (accession PI 134417) and an F₁ backcross [(L. esculentum × PI 134417) × PI 134417] selection. Mortality was intermediate (40 and 28%, respectively) on backcross selections with moderate and low levels of resistance and least (8%) on susceptibleL. esculentum. Removal of the glandular trichomes, which contain 2-tridecanone in their tips, from the foliage eliminated differences in parasitoid mortality among plant lines.Bioassays of 2-tridecanone indicated that it is acutely toxic to fifth instarC. sonorensis larvae at the quantities associated with highly resistant foliage and produces symptoms identical to those observed on resistant foliage. 2-Undecanone, a second methyl ketone present in the glandular trichomes of resistant foliage, was also toxic toC. sonorensis larvae, but significantly less so than 2-tridecanone. The results support the hypothesis that 2-tridecanone is responsible for the observed mortality ofC. sonorensis larvae during cocoon construction on resistant foliage.     

Dermal gland secretions of tropical bont tick,Amblyomma variegatum (Acarina: Ixodidae): Biological activity on predators and pathogens

When they are mechanically disturbed, all instars of the tropical bont tickAmblyomma variegatum exude droplets of a liquid on the dorsal, lateral, and ventral cuticle. These spread out and quickly evaporate. In this study, the possible role of these secretions was investigated in relation to predators and pathogens. In laboratory bioassays, it was demonstrated that the secretions from engorged larvae, nymphs, and females have an antibiotic activity against the bacteria speciesBacillus thuringiensis andSerratia marcescens, combined with a repellent effect on a potential predator, the fire-antSolenopsis geminata.     

Similarity of Cuticular Lipids Between a Caterpillar and Its Host Plant: A Way to Make Prey Undetectable for Predatory Ants?

Ithomiine butterflies (Nymphalidae) have long-lived, aposematic, chemically protected adults. However, little is known about the defense mechanisms in larvae and other juvenile stages. We showed that larvae Mechanitis polymnia are defended from ants by a chemical similarity between their cuticular lipids and those of the host plant, Solanum tabacifolium (Solanaceae). This is a novel defense mechanism in phytophagous insects. A field survey during one season showed that larval survivorship was up to 80%, which is high when compared with other juvenile stages. In a laboratory bioassay, live larvae on their host plant were not attacked by the predatory ant Camponotus crassus (Formicidae). Two experiments showed that the similarity between the cuticular lipids of M. polymnia and S. tabacifolium protected the larvae from C. crassus: (a) when the caterpillar was switched from a host plant to a non-host plant, the predation rate increased, and (b) when a palatable larva (Spodoptera frugiperda, Noctuidae) was coated with the cuticular lipids of M. polymnia and placed on S. tabacifolium leaves, it no longer experienced a high predation rate. This defensive mechanism can be defined as chemical camouflage, and may have a double adaptive advantage, namely, protection against predation and a reduction in the cost of sequestering toxic compounds from the host plant.     

Do Caterpillars Secrete “Oral Secretions”?

The oral secretions or regurgitant of caterpillars contain potent elicitors of plant induced responses. These elicitors are recognized by host plants to differentiate between simple mechanical injury and the presence of herbivores. In some cases, this level of recognition is highly specific. Despite the in-depth chemical characterization of these elicitors, little is known about the amounts delivered in regurgitant during feeding. In this study, we use a fluorescent dye to label regurgitant in order to visualize caterpillar regurgitation during feeding. The procedure is highly sensitive and allows us to visualize nanoliter amounts of regurgitant. We examined the propensity of larval Helicoverpa zea, Heliothis virescens, Spodoptera exigua, Spodoptera frugiperda, and Manduca sexta to regurgitate on various host plants. These species were selected because they have been among the most intensely studied in terms of elicitors. Our results indicate that most larvae did not regurgitate following a brief feeding bout (∼10 min) during which they ate ca. 0.40 cm² of leaf. When larvae did regurgitate, it was typically less than 10 nl. This is several orders of magnitude less than is typically used in most studies on oral secretions. The frequency of regurgitation appears to vary depending upon the host plant. Larval H. zea are less likely to regurgitate when feeding on tomato leaves compared to corn mid-whorl tissue. Our results have importance in understanding the role of oral secretions in plant recognition of herbivory. Because caterpillars did not routinely regurgitate during feeding, it is likely that they avoid the elicitation of some plant defensive responses during most feeding bouts.     

Sequestration and metabolism of protoxic pyrrolizidine alkaloids by larvae of the leaf beetle Platyphora boucardi and their transfer via pupae into defensive secretions of adults

Several neotropical leaf-beetles of the genus Platyphora ingest and specifically metabolize plant acquired pyrrolizidine alkaloids (PAs) of the lycopsamine type (e.g., rinderine or intermedine) and enrich the processed alkaloids in their exocrine defensive secretions. In contrast to the related palaearctic leaf beetles of the genus Oreina, which absorb and store only the non-toxic alkaloid N-oxides, Platyphora sequesters PAs exclusively as protoxic tertiary amines. In this study, the ability of P. boucardi larvae to accumulate PAs was investigated. Tracer studies with [¹⁴C]rinderine and its N-oxide revealed that P. boucardi larvae, like adult beetles, utilize the two alkaloidal forms with the same efficiency, but accumulate the alkaloid as a tertiary amine exclusively. Ingested rinderine is rapidly epimerized to intermedine, which is localized in the hemolymph and all other tissues; it is also detected on the larval surface. Like adults, larvae are able to synthesize their own alkaloid esters (beetle PAs) from orally administered [¹⁴C]retronecine and endogenous aliphatic 2-hydroxy acids. These retronecine esters show the same tissue distribution as intermedine. A long-term feeding experiment lasting for almost four months revealed that retronecine esters synthesized from [¹⁴C]retronecine in the larvae are transferred from larvae via pupae into the exocrine glands of adult beetles. Pupae contain ca. 45% of the labeled retronecine originally ingested, metabolized, and stored by larvae; ca. 12% of larval radioactivity could be recovered from the defensive secretions of adults sampled successively over two and a half months. Almost all of this radioactivity is found in the insect-made retronecine esters that are highly enriched in the defensive secretions, i.e., more than 200-fold higher concentration compared to pupae.     

Sequestration of Plant-Derived Phenolglucosides by Larvae of the Leaf Beetle <Emphasis Type="Italic">Chrysomela lapponica</Emphasis>: Thioglucosides as Mechanistic Probes

Feeding larvae of Chrysomela lapponica (Coleoptera: Chrysomelidae) acquire characteristic O-glucosides from the leaves of their food plants. The glucosides are selectively channeled from the gut to the defensive gland. Subsequent enzymatic transformations generate a blend of different defensive compounds, e.g., salicylaldehyde and two series of 2-methylbutyl and isobutyryl esters. By using systematically modified and hydrolysis-resistant thioglucosides as structural mimics of the plant-derived glucosides, e.g., salicin and its o-, m-, and p-isomers 1, 2, and 3; o-, m-, and p-cresols 5, 6, 7; along with thioglucosides of 2-phenylethanol 9 and (3Z)-hexenol 10, we demonstrated that the larvae of C. lapponica are able to sequester a broad range of structurally different thioglucosides with comparable efficiency. This sharply contrasts with the sequestration habitus previously observed in Chrysomela populi and Phratora vitellinae, which secrete almost pure salicylaldehyde and posses a highly specific transport mechanism for salicin (Kuhn et al., Proc. Natl. Acad. Sci. USA 101:13808–13813, 2004). Also, neither C. lapponica nor C. populi sequester in their gland the thioglucoside of 8-hydroxygeraniol, the mimic of the glucoside specifically transported by larvae secreting iridoid monoterpenes (Phaedon cochleariae, Gastrophysa viridula). Accordingly, leaf beetle larvae possess selective membrane carriers in their gut and their defensive systems that match the orientation of the functional groups of glucosides from their food plants probably by embedding the substrate in a network of hydrogen bonds inside the membrane carriers. The synthesis and the spectroscopic properties of the test compounds along with a comparative evaluation of the transport capabilities of larvae of C. populi and C. lapponica are described. Electronic Supplementary Material Supplementary material is available at and is accessible for authorized users.     

Economics of chemical defense in chrysomelinae

Chemical defense in chrysomelid larvae (subtribe Chrysomelina and Phyllodectina) is reviewed. Most species secrete autogenous monoterpenes. The diversity of their secretion is interpreted as a mechanism to reduce adaptation by predacious arthropods. The consequences of a host plant shift to the Salicacae are explored. Salicin from these host plants is used as a precursor for the salicylaldehyde secreted by the larvae of many species. This offers several advantages. It provides the larvae with an inexpensive and efficient defense. The recovery of the glucose moiety of the salicin contributes significantly to the larval energy budget. Adults sequester salicin in the eggs at concentrations which are toxic to ants. Owing to this maternal provisioning, neonate larvae produce salicylaldehyde from hatching onwards, whereas other species secreting monoterpenes are not protected at hatching. The secretion of salicylaldehyde by different species is considered to be chemical mimicry reinforcing visual aposematic signals.     

Volicitin, An Elicitor of Maize Volatiles in Oral Secretion of Spodoptera Exigua: Isolation and Bioactivity

Plants respond to insect-inflicted injury by systemically releasing relatively large amounts of several volatile compounds, mostly terpenoids and indole. As a result, the plants become highly attractive to natural enemies of the herbivorous insects. In maize, this systemic response can be induced by the uptake via the stem of an elicitor present in the oral secretions of caterpillars. Such an elicitor was isolated from the regurgitant of Spodoptera exigua larvae, identified as N-(17-hydroxylinolenoyl)-L-glutamine, and named volicitin. Here we present details on the procedure that was used to isolate volicitin and the biosasays that demonstrate its potency as an elicitor of maize volatiles that attract parasitoids. With a series of liquid chromatography purification steps, volicitin was separated from all other inactive substances in the regurgitant of larvae of the noctuid moth S. exigua. Maize seedlings that were incubated in very low concentrations of pure natural volicitin released relatively large amounts of terpenoids and became highly attractive to the parasitoid Microplitis croceipes. The identification of this and other insect-derived elicitors should allow us to determine their precise source and function, and better understand the evolutionary history of the phenomenon of herbivore-induced volatile emissions in plants.     

Tyrian Purple Precursors in the Egg Masses of the Australian Muricid, Dicathais orbita: A Possible Defensive Role

We report a putative defensive role for the precursors of Tyrian purple in the egg masses of the Australian muricid, Dicathais orbita. The fresh egg masses contain a high proportion of tyrindoleninone, which reacts to form tyriverdin and subsequently Tyrian purple and 6-bromoisatin as the eggs develop and the larvae hatch. Antimicrobial testing revealed that tyrindoleninone is toxic to both marine and human pathogens at a concentration of 1 mg/ml. Tyriverdin inhibits the growth of two marine pathogens, as well as the yeast Candida albicans at 0.001 mg/ml and was effectively bacteriostatic at 0.0005 mg/ml against three human pathogenic bacteria. Tyriverdin did not appear to significantly lyse the microbial cells. 6-Bromoisatin has mild antimicrobial properties, whereas Tyrian purple exhibited no significant activity. The antimicrobial properties of these compounds and changes in their presence during egg development correlates with ripening in the egg masses of D. orbita. This is the first report of the chemical ripening of eggs in a marine environment.     

Microbe inhibition by Tribolium flour beetles varies with beetle species,strain, sex,and microbe group

Tribolium flour beetles produce defensive compounds, including quinones, putatively aimed at deterring predators and inhibiting microbes. Here we examine how effective the defensive secretions of Tribolium confusum and T. castaneum are at inhibiting growth of various microbes and how this varies with species, geographic strain, and sex of the beetles. We explore differences at both the kingdom and species level of common flour microbes in their susceptibility to defensive compounds. Beetle species and strains vary in their ability to inhibit microbial growth. In addition, microbes vary in their sensitivity to the beetles' defense compounds. The capability to suppress microbial growth is likely under stabilizing selection with optimum quinone production varying among populations and may be dependent on several environmental factors including temperature, humidity, and predators.     

Composition of larval secretion ofChrysomela lapponica (Coleoptera,Chrysomelidae) and its dependence on host plant

The defensive secretion ofChrysomela lapponica larvae, which is produced by nine pairs of exocrine dorsal glands, has been chemically analyzed. TheC. lapponica larvae were kept in the laboratory on leaves of either birch (Betula pendula), alder (Alnus glutinosa), or willow (Salix fragilis). Larvae developed normally on birch and willow, whereas those on alder died within a few days. GC-MS analyses of the secretion of larvae on birch and willow revealed that the composition of this secretion differs distinctly from the known ones of several otherChrysomela species feeding exclusively on Salicaceae. In the exocrine secretion of larvae on birch, 69 compounds were identified, which included the main components isobutyric acid, 2-methylbutyric acid, and esters of the two. Several of the esters have not been reported previously from nature. The alcoholic components of the esters may be hydrolysis products ofBetula glycosides. Most components of the secretion of larvae feeding on birch were also found in the secretion of larvae feeding on willow. In addition, major amounts of benzoic acid and salicylalcohol were present in the secretion of the larvae feeding on willow.C. lapponica obviously acquires salicylalcohol by hydrolysis of salicin from willow leaves. However, in contrast to otherChrysomela species,C. lapponica larvae oxidize only traces of salicylalcohol to salicylaldehyde. The repellent activity of single authentic compounds of the secretion of larvae feeding on birch and willow, respectively, was tested in laboratory bioassays with ants (Myrmica sabuleti). Biosynthetic pathways to some identified compounds are suggested and discussed under evolutionary and functional aspects.     

The Cutaneous Wound Innate Immunological Microenvironment

The skin represents the first line of defense and innate immune protection against pathogens. Skin normally provides a physical barrier to prevent infection by pathogens; however, wounds, microinjuries, and minor barrier impediments can present open avenues for invasion through the skin. Accordingly, wound repair and protection from invading pathogens are essential processes in successful skin barrier regeneration. To repair and protect wounds, skin promotes the development of a specific and complex immunological microenvironment within and surrounding the disrupted tissue. This immune microenvironment includes both innate and adaptive processes, including immune cell recruitment to the wound and secretion of extracellular factors that can act directly to promote wound closure and wound antimicrobial defense. Recent work has shown that this immune microenvironment also varies according to the specific context of the wound: the microbiome, neuroimmune signaling, environmental effects, and age play roles in altering the innate immune response to wounding. This review will focus on the role of these factors in shaping the cutaneous microenvironment and how this ultimately impacts the immune response to wounding.     

From Terpenoids to Aliphatic Acids: Further Evidence for Late-Instar Switch in Osmeterial Defense as a Characteristic Trait of Swallowtail Butterflies in the Tribe Papilionini

We compared the chemical compositions of the osmeterial secretions of fourth and fifth (last) instars of eight swallowtail species of the tribe Papilionini. Four species (Papilio demoleus, P. polytes, P. paris, and P. macilentus) are Asian Rutaceae-feeding swallowtails. The other four (Chilasa epicydes, C. agestor, P. troilus, and P. glaucus) represent more distant clades within the Papilionini and species with larval hosts in other plant families. We conducted a quantitative analysis for six species, but only qualitative analysis for P. glaucus and C. agestor. In all eight species, regardless of larval host plant, secretions of the fourth instar principally consisted of mono- and sesquiterpene hydrocarbons, whereas those of the fifth instar comprised aliphatic acids and their esters. Consistent with earlier findings, our results suggest that this “heterogeneous” pattern of osmeterial chemistry, not seen in other tribes, may characterize the Papilionini as a whole. Unlike those of most Papilio species, the fourth and fifth instars of Chilasa species resemble each other in body coloration. Thus, the heterogeneous osmeterial pattern is not necessarily associated with color change in papilionid larvae. The major terpenoids identified in fourth instar larval secretions from the six species were α-pinene, sabinene, β-myrcene, limonene, β-phellandrene, (Z)-β-ocimene, (E)-β-ocimene, p-mentha-1,4(8)-diene, β-elemene, β-caryophyllene, (E)-β-farnesene, (3Z,6E)-α-farnesene, (Z)-α-bisabolene, germacrene-A, (E)-α-bisabolene, and germacrene-B. The profiles for individual species differed both qualitatively and quantitatively from one another, and certain species also secreted methyl 3-hydroxy-n-butyrate and oxygenated sesquiterpenes in relatively large proportions. Secretions from fifth instars were composed of varying proportions of isobutyric, 2-methylbutyric, and acetic acids, and methyl and ethyl (minor) esters of both isobutyric and 2-methylbutyric acids. The heterogeneity of osmeterial chemistry in the tribe Papilionini may represent fine-tuning of chemical defense in response to shifting predation pressures as the larvae age and grow.     

Sequestration of Host Plant Glucosinolates in the Defensive Hemolymph of the Sawfly Athalia rosae

Interactions between insects and glucosinolate-containing plant species have been investigated for a long time. Although the glucosinolate–myrosinase system is believed to act as a defense mechanism against generalist herbivores and fungi, several specialist insects use these secondary metabolites for host plant finding and acceptance and can handle them physiologically. However, sequestration of glucosinolates in specialist herbivores has been less well studied. Larvae of the turnip sawfly Athalia rosae feed on several glucosinolate-containing plant species. When larvae are disturbed by antagonists, they release one or more small droplets of hemolymph from their integument. This reflex bleeding is used as a defense mechanism. Specific glucosinolate analysis, by conversion to desulfoglucosinolates and analysis of these by high-performance liquid chromatography coupled to diode array UV spectroscopy and mass spectrometry, revealed that larvae incorporate and concentrate the plant's characteristic glucosinolates from their hosts. Extracts of larvae that were reared on Sinapis alba contained sinalbin, even when the larvae were first starved for 22 hr and, thus, had empty guts. Hemolymph was analyzed from larvae that were reared on either S. alba, Brassica nigra, or Barbarea stricta. Leaves were analyzed from the same plants the larvae had fed on. Sinalbin (from S. alba), sinigrin (B. nigra), or glucobarbarin and glucobrassicin (B. stricta) were present in leaves in concentrations less than 1 mol/g fresh weight, while the same glucosinolates could be detected in the larvae's hemolymph in concentrations between 10 and 31 mol/g fresh weight, except that glucobrassicin was present only as a trace. In larval feces, only trace amounts of glucosinolates (sinalbin and sinigrin) could be detected. The glucosinolates were likewise found in freshly emerged adults, showing that the sequestered phytochemicals were transferred through the pupal stage.     

Phototoxicity ofCitrus jambhiri to fungi under enhanced UV-B radiation: Role of furanocoumarins

Extracts ofCitrus jambhiri foliage exposed to and shielded from UV-B radiation were assayed for phytochemical changes and phototoxicity against four fungal pathogens, two of which (Fusarium solani andF. oxysporum) are causative agents of root rots and two of which (Penicillium italicum andP. digitatum) are associated with fruit rots. Conidial pigment mutants of these four fungal species were assayed to determine whether pigments play a role in protecting fungi against plant photosensitizers. Exposure to 10.2 kJ/ day UV-B radiation for 95 days significantly reduced phototoxicity of leaf extracts to fungi. Although furanocoumarin levels were reduced by UV-B, analysis of covariance revealed that variation in phototoxicity of the extracts cannot be attributed entirely to variation in furanocoumarin content; thus, the possibility exists that nonfuranocoumarin phototoxic constituents, as yet unidentified, respond to UV-B exposure and contribute to overall phototoxic defense ofC. jambhiri against pathogens. Root rot fungi were substantially more sensitive to furanocoumarin phototoxicity than were fruit rot fungi, a pattern consistent with the amount of light exposure normally experienced by these fungi when associated with phototoxic plants. Although pigmented strains of all four species displayed greater resistance to phototoxicity of pure furanocoumarins, no strain differences were detected in assays of foliar extracts; this finding also suggests that nonfuranocoumarin constituents may be involved in the phototoxic defense ofC. jambhiri against pathogens.Deceased.