Iridoid glycoside sequestration byThessalia leanira (Lepidoptera: Nymphalidae) feeding onCastilleja integra (Scrophulariaceae)

A small population of a polyvoltine checkerspot butterfly,Thessalia leanira fulvia (also known asChlosyne leanira ssp.fulvia), was found to useCastilleja integra as a larval food plant at a localized site (Burnt Mill) southwest of Pueblo, Colorado. Field-captured adult butterflies contained the major iridoid glycosides (catalpol and macfadienoside) of theCastilleja. The content of a third iridoid glycoside, methyl shanzhiside, was also relatively high in the collected butterflies even though most individualCastilleja plants at Burnt Mill contained little or no methyl shanzhiside. Only a few plants, restricted to a small area, did contain appreciable methyl shanzhiside. Most of the plants that lacked the ester methyl shanzhiside contained shanzhiside, the corresponding free carboxylic acid.Thessalia larvae did not normally methylate the acid to produce methyl shanzhiside. Larvae that stopped feeding at an early instar, but yet survived several weeks, did contain major amounts of methyl shanzhiside. It is suggested that only larvae that overwinter or otherwise enter diapause convert shanzhiside to methyl shanzhiside. TheCastilleja food plant also contained iridoids other than catalpol and macfadienoside, sometimes in major amounts, but these were never found in larvae, pupae, or butterflies.Paper 27 in the series Chemistry of the Scrophulariaceae. Paper 26: Stermitz, F.R., Foderaro, T.A., and Li, Y.-X., 1993.Phytochemistry 32:1151     

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

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

Defensive steroids from a carrion beetle (Silpha novaboracensis)

Two novel steroids, 3,7-dihydroxy-14-pregn-4-en-15,20-dione (1) and 3,7, 20-trihydroxy-14-pregn-4-en-15-one (4), have been characterized from the rectal gland ofSilpha novaboracensis (Coleoptera: Silphidae). Judging from the function of comparable pregnanes in another species ofSilpha, the compounds may play an antipredator role.Paper No. 81 of the seriesDefense Mechanisms of Arthropods. Paper No. 80 is Eisner, T., Deyrup, M., Jacobs, R., and Meinwald, J.,J. Chem. Ecol. 12:1407–1415.     

Leaf surface chemicals fromNicotiana affecting germination ofPeronospora tabacina (adam) sporangia

A bioassay was used to evaluate the effects of cuticular leaf components, isolated fromN. tabacum, N. glutinosa (accessions 24 and 24a), and 23other Nicotiana species, on germinationof P. tabacina (blue mold). The leaf surface compounds included- and-4,8,13,-duvatriene-l,3-diols (DVT-diols), (13-E)-labda-13-ene-8-,15-diol (labdenediol), (12-Z)-labda-12,14-diene-8-ol (cis-abienol), (13-R)-labda-8,14-diene-13-ol (manool), 2-hydroxymanool, a mixture of (13-R)-labda-14-ene-8,13-diol (sclareol), and (13-S)-labda-14-ene-8,13-diol (episclareol), and various glucose and/or sucrose ester isolates. The above in acetone were applied onto leaf disks of the blue moldsusceptibleN. tabacum cv. TI 1406, which was then inoculated with blue mold sporangia. Estimated IC₅₀ values (inhibitory concentration) were 3.0g/cm² for-DVT-diol, 2.9/cm² for-DVT-diol, 0.4g/cm² for labdenediol and 4.7g/cm² for the sclareol mixture. Manool, 2-hydroxymanool, andcis-abienol at application rates up to 30g/cm² had little or no effect on sporangium germination. Glucose and/or sucrose ester isolates from the cuticular leaf extracts of 23Nicotiana species and three different fractions fromN. bigelovii were also evaluated for antimicrobial activity at a concentration of 30g/cm². Germination was inhibited by >20% when exposed to sugar esters isolated fromN. acuminata, N. benthamiana, N. attenuata, N. clevelandii, andN. miersii, and accessions 10 and 12 ofN. bigelovii. These results imply that a number of compounds may influence resistance to blue mold in tobacco.     

Defensive secretions of three species ofAcilius (Coleoptera: Dytiscidae) and their seasonal variations as determined by high-pressure liquid chromatography

A rapid, sensitive, stable, and quantitative high-pressure liquid chromatographic technique was developed for the analysis of defensive secretions obtained from the pygidial and prothoracic glands of dytiscids. Methods were developed for both normal phase (Porasil) and reverse phase (Bondapak C₁₈) columns. The applicability of this technique was demonstrated when defensive compounds ofAcilius semiculcatus, A. sylvanus andA. mediatus were isolated, identified, and quantitated. No major differences were found in the composition of the defensive secretions between the three species. The seasonal defensive titer ofA. semisulcatus was determined from June through October 1977. The pygidial defensive agents (benzoic acid,p-hydroxybenzaldehyde, and methylp-hydroxybenzoate varied from 9.0 to 67.8 g and exhibited a maximum in July, and a steroid from the prothoracic gland varied from 7.1 to 33.2 g and was maximum in October.     

Iridoid glycoside content ofEuphydryas anicia (Lepidoptera: Nymphalidae) and its major hostplant,Besseya plantaginea (Scrophulariaceae), at a high plains colorado site

The checkerspot butterfly,Euphydryas anicia, utilizes mainlyBesseya plantaginea and only occasionallyCastilleja integra as a larval hostplant at Michigan Hill, a few kilometers from a site whereC. integra is used by over 90% of the butterflies. TheB. plantaginea leaves that are consumed contain 9–22% iridoid glycosides, composed mainly of catalpol and catalpol esters, while larvae from the same plants contain 6–18% iridoids, mainly catalpol and no esters. Field-collected adult butterflies contain 0.5–4.3% iridoids. Laboratory-reared adults secrete iridoids in the meconium upon eclosion and retain similar amounts. The adult and meconium iridoid content is considerably lower than in the larvae, and metabolism in the pupal stage may be occurring.This work was supported by grant CHE-8521382 from the National Science Foundation. Paper 13 in the series Chemistry of the Scrophulariaceae. Paper 12: G.N. Belofsky and F.R. Stermitz,J. Nat. Prod. 51:614–616, 1988.     

Iridoid glycoside metabolism and sequestration byPoladryas minuta (Lepidoptera: Nymphalidae) feeding onPenstemon virgatus (Scrophulariaceae)

A bivoltine checkerspot butterfly,Poladryas minuta, is aPenstemon specialist, not known to utilize any other plant genus for oviposition and larval feeding. At several intermontane plains sites of central Colorado, the butterfly utilizesPenstemon virgatus as its sole host plant. Analysis of the host plant showed it to contain three cinnamyl-type catalpol esters (scutellarioside-II, globularin, globularicisin) and catalpol. The host plant contained an average of 10% dry weight iridoids, but some variation among individual plants and leaves within plants was noted. Field-collected butterflies contained 2.1–8.7% dry weight catalpol, but no other iridoids. Adults from larvae fedP. virgatus in the lab contained 4.2–9.0% dry weight catalpol and excreted large amounts of catalpol in the meconium. No catalpol was found in the larval frass. Larvae did not consume three alternate iridoid-containing host-plant species, and most eventually died rather than feed on the alternate plants. Larvae did consume small amounts of artificial diets containing the alternate species andP. virgatus, but most went into diapause and some died. Survival was good on artificial diet containing 10% dry weight of the iridoid esters fromP. virgatus. Only catalpol was found in pupae and adults, but it was absent from the larval frass. The cinnamic-type acids expected from larval hydrolysis of the esters were not found in larval frass, pupae, or adults. These results are contrasted with those found for another checkerspot,Euphydryas anicia, which consumes a different host-plant species but was present at one of the same sites withPoladryas minuta.Paper 15 in the series Chemistry of the Scrophulariaceae. Paper 14 Boros, C.M., Stermitz, F.R., and Harris, G.H. 1990.J. Nat. Prod. 5372–80.     

Sex pheromone of aPlanotortrix excessana sibling species and reinvestigation of related species

The sex pheromone of aPlanotortrix excessana sibling species was investigated. Females were found to produce eight potential pheromone components: dodecyl acetate, tetradecyl acetate (14OAc). (Z)-5-tetradecenyl acetate (Z5-14OAc), (Z)-7-tetradecenyl acetate (Z7-14OAc), (Z)-9-tetradecenyl acetate, hexadecyl acetate, (Z)-7-hexadecenyl acetate, and (Z)-9-hexadecenyl acetate. When these compounds were bioassayed using field-trapping and wind-tunnel techniques, only 14OAc,Z5-14OAc, andZ7-14OAc were found to be behaviorally active. The sex pheromone glands of females of other species including,Planotortrix MBS,Planotortrix M,P. notophaea, Ctenopseustis servana, and aC. obliquana sibling species, were also found to containZ5-14OAc orZ7-14OAc, singly or in combination. In the case ofPlanotortrix M, the addition ofZ7-14OAc to the previously identified sex pheromone blend ofZ5-14OAc and 14OAc was found to increase trap captures of male moths of this species. Thus in these New Zealand species (and in some Australian species),Z5-14OAc andZ7-14OAc appear to be utilized in combination in pheromonal communication just as (Z)-11-tetradecenyl acetate and (E)-11-tetradecenyl acetate are used by many species of Holarctic Tortricidae in the tribe Archipini.Lepidoptera: Tortricidae: Tortricinae.     

Selective Phytotoxic Activity of Withanolides from Iochroma australe to Crop and Weed Species

Phytotoxic effects of Iochroma australe extract and 4,7,20-oxowithanolides were examined in Petri dish bioassays at 10, 100, and 1000 ppm. The extract and the major constituent (17S,20R,22R)-4,7,20-trihydroxy-1oxowitha-2,5,24-trienolide (2) reduced growth of the radicle of the weeds Sorghum halepense (Monoct.) and Chenopodium album (Dicot.). Neither the extract nor withanolides had significant effect on germination or radicle length of the commercial crop species Lactuca sativa. From our experimental data we conclude that the withanolides tested here are natural products with selective herbicidal activity against weed species.     

Euphydryas anicia (Lepidoptera: Nymphalidae) utilization of iridoid glycosides fromCastilleja andBesseya (Scrophulariaceae) host plants

Iridoid glycosides were found to be sequestered by natural populations ofEuphydryas anicia after ingestion from the host plantsBesseya alpina, B. plantaginea, andCastilleja integra. Both major iridoids ofB. alpina, cataipol and aucubin, were found in butterfly populations where this was the only host plant. The catalpol-aucubin ratio was higher in the butterflies than in the host plant. AnE. anicia population which uses bothB. plantaginea andC. integra as host plants was found to sequester cataipol as well as another iridoid, macfadienoside. Macfadienoside was the major iridoid ofC. integra, while catalpol esters were the major iridoids ofB. plantaginea. Although it was a major sequestered iridoid, catalpol was a minor constituent in both host plants. The macfadienoside-catalpol ratio in the butterflies from this population was highly variable, and there appeared to be both sex and individual variation in host plant and/or iridoid glucoside utilization byE. anicia. Although other iridoids were present in the host plants, none was sequestered in more than trace amounts.This work was supported by grant CHE-8213714 to FRS, in part by grant DEB-06961 to PRE from the National Science Foundation, and a grant from the Koret Foundation of San Francisco to PRE. Paper 5 in the series Chemistry of the Scrophulariaceae. Paper 4: Roby, M.R. and Stermitz, F.R. 1984.J. Nat. Prod. 47:854–857.     

Light-induced variation in phenolic levels in foliage of rain-forest plants

Levels of phenolic secondary metabolites in the leaves of four west African rain-forest plants,Acacia pennata, Cynometra leonensis, Diopyros thomasii, andTrema guineensis, were correlated with incident light intensity at both the inter- and intraindividual level. Enhanced phenolic levels under high light intensity appeared to be due to production of both polyphenolics (condensed and hydrolyzable tannins) and simple phenolics. InTrema guineensis, where it is possible to separate leaves in terms of both their age and the light incident upon them, condensed tannin production progressed differently during the development of sun and shade leaves, suggesting continuing production of new oligomers in the former but not in the latter. The results of this study suggest that the production of phenolics in relation to variation in incident light is a finely tuned process, which must be explained in terms of plant physiology and intermediate metabolism rather than in terms of resource allocation or a direct response to herbivory.     

Chemical constituents and inhibitory activities of essential oils fromCyperus brevifolius andC. kyllingia

Cyperus brevifolius andC. kyllingia are two common weeds in Hawaii. The underground parts, including the rhizomes and roots, contain allelopathic essential oils, which may contribute to the aggressive spread of these weeds in grasslands and lawns. Although morphologically similar,C. kyllingia contains more essential oils thanC. brevifolius. Also, the former is rich in terpenes, including -cyperone, -selinene, and -humulene, while the latter is rich in C₁₇ to C₂₅ n-paraffins. These chemical differences agree with their inhibitory activities against indicator species and the general observation thatC. kyllingia is the more prevalent weed of the two.On leave from the Department of Agricultural Chemistry, Kinki University, Osaka, Japan.     

Sex pheromone chemistry of the western spruce budwormChoristoneura occidentalis Free

SCOT capillary Chromatographic and SCOT capillary chromatographic-mass spectrometic analyses of gland washes and effluvia of virgin femaleChoristoneura occidentalis Free, have been conducted with both a diapausing and nondiapausing strain of this insect. The following compounds were identified in gland washes and effluvia in both strains:E andZ11–14Ald,E andZ11–14Ac,E andZ11–14OH and 14Ald, 14Ac, and 14OH. The average aldehyde: acetate: alcohol ratio found by analysis of single glands by virgin females (nondiapausing strain) was 170.73. Analysis of virgin female effluvia gave this ratio as 1038 (diapausing strain: %Z=8, 11, 15, respectively) and 1036 (nondiapausing strain: %Z=8, 11, 12, respectively). The saturated components were generally 1–2% of theE isomer in each case. Comparisons of EAG responses of bothC. occidentalis andC. fumiferana toE11–14Ald,E11–14Ac andE11–14OH were made. Correlations with both laboratory and field data previously published were also made betweenC. fumiferana andC. occidentalis.Lepidoptera: Tortricidae.     

Iridoid glycoside sequestration by two aposematicPenstemon-feeding geometrid larvae

The iridoid glycoside catalpol was found to be sequestered by larvae ofMeris alticola feeding onPenstemon virgatus and by larvae ofNeoterpes graefiaria which utilizeP. barbatus. The strikingly similar larval patterns of these two ennomine geometrids were previously considered to be disruptive, but predator-based Mullerian mimicry is equally likely to be involved. The cryptic adult moths generally contain only small amounts of catalpol, having left most of the bitter iridoid in the pupal case and in the meconium excreted after eclosion. OneNeoterpes female did contain considerable catalpol in the abdomen, presumably in the eggs.This work was supported by grant BSR-8506064 from the National Science Foundation. Paper 11 in the series Chemistry of the Scrophulariaceae. Paper 10: F.R. Stermitz and G.H. Harris, 1987.J. Chem. Ecol. 13:1917–1925.     

Inhibition of β-Glucosidase and Esterase by Tannins from Betula, Salix, and Pinus Species

The reaction of tannin fractions isolated from the bark of Betula, Salix, and Pinus species with two enzymes, -glucosidase and esterase, was investigated. The influence of precipitation to the hydrolytic capacity of -glucosidase also was studied. All tannins studied precipitated -glucosidase and esterase, and moderate differences in the precipitating capacities of the tannins were observed. Interestingly, complex formation between -glucosidase and tannin did not markedly affect the activity of the enzyme. Therefore, complex formation during the insect/herbivore feeding does not necessarily change the defense activity of phenolic glycosides or decrease activity of digestive enzymes.     

Defensive secretion of chrysomelid larvaeGastrophysa atrocyanea Motschulsky andPhaedon brassicae Baly

The active substance of the larval defensive secretion of the Japanese chrysomelid beetles,Gastrophysa atrocyanea Motschulsky andPhaedon brassicae Baly was identified as chrysomelidial.This is report No. 2 of the Defensive Secretion of Chrysomelid Beetles, Report No. 1 is Sugawara et al. (1978).     

Chemical nature of larval osmeterial secretions of papilionid butterflies in the generaParnassius,Sericinus andPachliopta

Volatile components of the larval osmeterial secretion ofParnassius glacialis (Parnassiinae, Parnassiini) consisted of isobutyric acid, 2-methylbutyric acid, and their methyl esters. In contrast, the osmeterial exudate ofSericinus montela (Parnassiinae, Zerynthiini) was characterized as monoterpene hydrocarbons comprising-myrcene (Major),-pinene, sabinene, limonene, and-phellandrene, whereas that ofPachliopta aristolochiae (Papilioninae, Troidini) was composed of numerous sesquiterpene hydrocarbons, including-himachalene,-amorphene, and germacrene-A, and a few oxygenated sesquiterpenoids. In these three species, the chemical nature of the secretions of the last and the penultimate instars was essentially of similar quality, suggesting that the three genera,Parnassius, Sericinus, andPachliopta, are assigned to homogeneous types.     

Occurrence and Correlations of Monoterpene Hydrocarbon Enantiomers in Pinus sylvestris and Picea abies

The relative amounts and enantiomeric compositions of monoterpene hydrocarbons in branch and trunk xylem, in needles, and in resin from apical buds in 18 Pinus sylvestris trees have been determined and compared with the terpene content in branch xylem and needles of Picea abies. Besides the high amount of (+)-3-carene, an excess of (+)--pinene has been found in P. sylvestris, whereas in P. abies (–)--pinene dominates over (+)--pinene. In P. sylvestris, clear positive correlations were found between (+)--pinene and (+)-camphene in the four tissues analyzed. Good positive correlations were also observed between (–)--pinene and (–)-camphene in the two types of xylem, between (+)--pinene and (+)--pinene in the resin, and between tricyclene and (–)-camphene in resin and needles. In P. abies, positive correlations were found between (+)--pinene and (+)-camphene in the branch xylem and between tricyclene and (–)-camphene as well as between (–)--pinene and (–)-camphene in the needles. Complex relationships between (–)--pinene and (–)--pinene were found both in the P. abies and in the P. sylvestris tissues. The importance of the enantiomeric composition of -pinene for the host selection of Ips typographus, Tomicus piniperda, and Hylobius abietis is discussed.     

Response on the ground of bark beetle and weevil species colonizing conifer stumps and roots to terpenes and ethanol

Responses of threeHylastes species,Dryocoetes autographus, and twoHylobius species to terpenes and ethanol were studied in field experiments on clear-cut forest sites in Sweden using baited ground traps.-Pinene alone did not attract any of the six species. A terpene blend (spruce turpentine consisting mainly of-pinene,-pinene, and 3-carene) attractedHylastes cunicularius, H. brunneus, andHylobius abietis in some experiments, but not in others. The attractiveness of ethanol also varied; the only species consistently attracted wasH. abietis. Baits containing both terpenes and ethanol, particularly the combination of spruce turpentine and ethanol, were attractive to all species exceptHylobius pinastri. InH. abietis, the terpene plus ethanol/ ethanol catch ratios increased during early summer. Seasonal differences in catch levels were observed inH. cunicularius andH. abietis. The addition of-pinene reduced the attractiveness of the combination of spruce turpentine and ethanol toH. cunicularius, H. opacus, andD. autographus. The differences in response to the volatiles between species are probably related to differences in reproductive behavior and host preferences.     

The influence of dietaryβ-carboline alkaloids on growth rate,food consumption,and food utilization of larvae ofSpodoptera exigua (Hubner)

-Carboline alkaloids are found worldwide in many plant families. Harman, harmine, and other simple -carboline alkaloids were tested for activity against a generalist phytophagous insect, the beet army worm [Spodoptera exigua (Hubner)]. Chronic dietary exposure tests (neonate to pupa) reveal potent antifeedant and possible toxic effects. Acute dietary exposure tests on fifth-instar larvae also demonstrate antifeedant activity.