Pyrrolizidine alkaloids as oviposition stimulants for the cinnabar moth,Tyria jacobaeae

In choice experiments with artificial leaves, we tested related pyrrolizidine alkaloids (PAs) for their stimulatory effects on the oviposition of the cinnabar moth, a specialist on the PA-containing plant Senecio jacobaea. The PAs from S. jacobaea that we tested stimulated oviposition. Monocrotaline also stimulated oviposition although this PA is not found in plants of the genus Senecio. The moths preferred ovipositing on filter paper with a PA mixture extracted from S. jacobaea to ovipositing on filter paper with single PAs. Senkirkine, heliotrine, and retrorsine did not stimulate oviposition. The nonactive retrorsine differs only in one OH group to the active senecionine, indicating that small structural differences alter the stimulatory activity of PAs. However, a PA mixture extracted from a nonhost plant, Senecio inaequidens, that consisted of 81% of the nonactive retrorsine did stimulate oviposition. Oviposition preferences between Senecio species seem to be determined by chemical compounds other than PAs.     

Pyrrolizidine alkaloids from Senecio jacobaea affect fungal growth

We investigated the growth-reducing effects of pyrrolizidine alkaloids (PAs) from Senecio jacobaea on nine plant-associated fungi (five strains of Fusarium oxysporum, two of F. sambucinum, and two of Trichoderma sp). Fungal growth was monitored on water agar media containing different concentrations of monocrotaline, retrorsine, or a purified extract of PAs from S. jacobaea. The growth rate of six strains was inhibited by PAs at the highest test concentration (3.33 mM), with the magnitude of the inhibition (7–35%) being dependent upon the specific fungus-PA interaction. In general, the PA extract caused the largest inhibition. However, the fungi isolated from S. jacobaea were positively affected by the PA extract (7–9%). Retrorsine N oxide was as effective as retrorsine in its inhibition of mycelium growth.     

The role of leaf lipids in food selection by larvae of the tobacco hornworm,Manduca sexta

The role of leaf lipids in food plant selection by larvae ofManduca sexta was investigated by measuring preference responses in a two-choice preference test using glass fiber filter paper disks laced with extract (test) or water (control). The larvae respond to the petroleum ether extract of whole leaves of the host-plantLycopersicon esculentum (tomato) extract in a concentration-dependent manner. At natural concentration it is the most strongly stimulating extract or compound yet tested using the disk test. This response is affected by food plant experience of the larvae, suggesting stimulation by plant-specific compounds in the extract. The extract contains volatile compounds that attract the larvae. In contrast, it does not promote continued feeding on an agar-cellulose diet that incorporates the extract. Also stimulating are the extracts of leaf surfaces of two hosts,L. esculentum andSolanum pseudocapsicum, and two acceptable nonhosts,Brassicae napus andVigna sinensis, indicating the presence of nonpolar feeding stimulants at the leaf surface. However, similar leaf-surface extracts of the unacceptable plantCanna generalis were inactive, although the surface extraction process renders this plant acceptable. Leaf-surface extracts ofL. esculentum, S. pseudocapsicum, andB. napus evoke feeding responses that are qualitatively comparable to those of their corresponding leaves. However, no such parallel is found for surface extracts ofV. sinensis andC. generalis. Thus, nonpolar compounds at the leaf surface of host and some acceptable non-host-plant species strongly stimulate feeding and hence must play an important role in food selection by the tobacco hornworm.     

Species by Environment Interactions Affect Pyrrolizidine Alkaloid Expression in <Emphasis Type="Italic">Senecio jacobaea</Emphasis>, <Emphasis Type="Italic">Senecio aquaticus</Emphasis>, and Their Hybrids

We examined the effects of water and nutrient availability on the expression of the defense pyrrolizidine alkaloids (PAs) in Senecio jacobaea and S. aquaticus. Senecio jacobaea, and S. aquaticus are adapted to different natural habitats, characterized by differing abiotic conditions and different selection pressures from natural enemies. We tested if PA concentration and diversity are plastic over a range of water and nutrient treatments, and also whether such plasticity is dependent on plant species. We also tested the hypothesis that hybridization may contribute to PA diversity within plants, by comparing PA expression in parental species to that in artificially generated F₁ hybrids, and also in later generation natural hybrids between S. jacobaea and S. aquaticus. We showed that total PA concentration in roots and shoots is not dependent on species, but that species determines the pattern of PA diversification. Pyrrolizidine alkaloid diversity and concentration are both dependent on environmental factors. Hybrids produce a putatively novel PA, and this PA is conserved in natural hybrids, that are backcrossed to S. jacobaea. Natural hybrids that are backcrossed several times to S. jacobaea are with regard to PA diversity significantly different from S. jacobaea but not from S. aquaticus, while F₁ hybrids are in all cases more similar to S. jacobaea. These results collectively suggest that PA diversity is under the influence of natural selection.     

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.     

Identification and bioassay of kairomones forHelicoverpa zea

Hexane extracts of leaves of 307 accessions from 73 host plant species ofHelicoverpa zea were analyzed by gas chromatography (GC) and used forH. zea oviposition and neonate larvae orientation bioassays. The gas chromatographic (GC) retention times of compounds statistically associated with behavioral activity were identified by correlation of GC peak area with oviposition and larval orientation preferences. Although taxonomically diverse in their origin, compounds for study were purified from extracts of species of the genusLycopersicon, due to their relative abundance. The structures of eight long-chain alkanes associated with oviposition preference were assigned by mass spectrometry, and the structures of five similarly associated organic acids and a terpenoid alkene were identified by¹H and¹³C nuclear magnetic resonance spectroscopy. The structures of a number of other phytochemicals from the plant leaves were identified for comparative purposes, including a previously unknown terpene, 7-epizingiberene. Bioassays were performed on the isolated acids and on the alkane wax fractions of severalLycopersicon species, and significant differences were found in oviposition stimulation for both classes of compounds. Of the hundreds of compounds found in the extracts, none were observed to act as oviposition deterrents. The results of these bioassays may be useful in explaining the broad host range ofH. zea, as well as the process and evolution of host plant selection for oviposition.     

Chemical aspects of host-plant specificity in threeLarrea-feeding grasshoppers

The host-selection behavior of three species of grasshopper feeding on creosote bush,Larrea tridentata, in southern California was investigated. The species wereBootettix argentatus, which is monophagous;Ligurotettix coquilletti, oligophagous; andCibolacris parviceps, polyphagous. The monophagous species is stimulated to bite by nordihydroguaiaretic acid (NDGA), a compound that is characteristic of the host plant and that may comprise up to 10% of the dry weight of the leaf. Host specificity ofB. argentatus is enhanced by deterrent responses to compounds present in the surface waxes of all non-host-plant species. Both the oligophagous and polyphagous species are deterred by NDGA at naturally occurring concentrations. Their association withLarrea is probably based on tolerance of the plant chemicals rather than on dependence on specific chemicals. Factors other than the chemistry of the plant probably also contribute to the specificity ofB. argentatus andL. coquilletti.     

Ecological adaptation of an Aristolochiaceae-feeding swallowtail butterfly,Atrophaneura alcinous,to aristolochic acids

Seven analogs of aristolochic acids (I, II, III, B, C, D, and E) were isolated from the leaves ofAristolochia debilis and characterized as the larval feeding stimulants of an Aristolochiaceae-feeding swallowtail butterfly,Atrophaneura alcinous. Aristolochic acids showed synergistic activity in combination with the water-soluble components in the leaf extract. Aristolochic acids were detected in the body tissues and specialized organs throughout all life stages ofA. alcinous. Larval osmeterial fluid contained aristolochic acids as high as 2% of the secretions, the compositions of which were similar to that found in the leaf extracts. In contrast, the eggs selectively contained aristolochic acids I and II. These two acids were present both in the egg yolk and egg coating material as well as in the collateral glands (glandulae sebaceae) of the adult females. The cannibalistic activity of the larvae against the eggs and pupae seemed to be triggered by aristolochic acids present on the egg surface and pupal cuticle, suggesting a possible adaptive mechanism in this species. Aristolocic acid I deterred feeding of tree sparrows, which suggested a defensive role against vertebrate predators.     

Comparative study of allelopathy as exhibited byProsopis Juliflora swartz andProsopis cineraria (L) druce

The allelopathic effects ofProsopis juliflora were studied both in the laboratory and in nature and compared with that ofProsopis cineraria to understand the chemical nature of allelochemics. Both species occupy the same habitats butP. cineraria does not appear to have any toxic effect on other plants under its canopy.P. juliflora is highly allelopathic and does not allow the growth of any other species. Leaf extracts and leaf leachates ofP. juliflora were inhibitory. Decaying leaves were also inhibitory at early stages of decomposition. Live roots were not found to be inhibitory in cogermination and interplanting of seeds. Chemical investigation of the extracts showed the allelopathic compounds to be phenolic in nature in both the species. Slow decomposition and heavy accumulation of leaf litter belowP. juliflora may possibly result in accumulation of toxic substances in soil layers, inhibiting growth of other species.     

Protein-precipitating capacity of tannins inShorea (Dipterocarpaceae) seedling leaves

The protein-precipitating capacities of tanniferous extracts from immature and mature leaves of threeShorea spp. (Dipterocarpaceae) seedlings were measured by an adaptation of Goldstein and Swain's -glucosidase precipitation assay. Protein precipitation by the extracts was not correlated with total phenolics (Folin-Denis assay) or proanthocyanidin content (BuOH-HCl assay) as measured in an earlier study. Extracts ofS. maxwelliana mature leaves had much lower protein-precipitating capacity than those ofS. acuminata andS. leprosula, but fewer insect species feed on and cause less damage to the foliage ofS. maxwelliana compared with the other species' foliage. Immature leaf extracts ofS. leprosula andS. acuminata had substantial protein-precipitating capacities which in the latter species exceeded that of its mature leaf extracts. Leaf extracts precipitated less protein when initial protein concentration was reduced, although not limiting, but no effect or the reverse effect occurred with quebracho tannin and tannic acid. Problems in the characterization of foliage astringency and the interpretation of its role as a potential antiherbivore defense are discussed.     

Detoxification of cereal plant allelochemicals by aphids: Activity and molecular weights of glutathioneS-transferase in three species of cereal aphids

The activity and molecular weights of glutathioneS-transferase were studied in three species of cereal aphid:Metopolophium dirhodum, Sitobion avenae, andRhopalosiphum padi. The highest level of glutathioneS-transferase activity was recorded in extracts fromM. dirhodum and the lowest fromS. avenae, and extracts of larvae were more active than those from adults. The activity of this enzyme was higher in extracts ofS. avenae andR. padi previously fed on a moderately resistant wheat variety than on a susceptible variety. Gel filtration followed by SDS-PAGE revealed three protein bands in the active fractions. The first had a molecular weight of 28,500 and the second of 27,500 and were present in all three species of aphid. The third protein differed in the three species, having a molecular weight of 26,000 inS. avenae, 25,500 inM. dirhodum, and 24,000 inR. padi.     

Influence of mating on surface nutrient exchange in schistosomes

In schistosomes, the mating process influences male-female transfer and gender-specific exchange of nutrients. The paired male schistosome provides glucose to the female partner. Male-to-female intertegumental transfer of¹⁴C-labeled glucose,¹⁴C-labeled 3-O-methylglucose, [¹⁴C]2-deoxyglucose and 2-fluorodeoxyglucose has been demonstrated in schistosomes. This phenomenon has been studied extensively inSchistosoma mansoni, and confirmed inSchistosoma japonicum, as well asS. haematobium, using radioactive pulsing methods. Male schistosomes contain significantly greater quantities (nmol/mg worm water) of glucose than do females. The transfer of glucose is apparently not an energy-dependent process, but occurs along this concentration gradient. Most, if not all, of the glucose utilized by the female is transferred from the male partner via tegumentary-facilitated diffusion mechanisms, free diffusion, or some combination of these two components. Unpaired male schistosomes contain greater quantities of glycogen than do comparable paired schistosomes, indicating that the presence of a female in the gynecophoral canal depletes the reserves of the male partner; this is additional indirect evidence for male-to-female transfer of glucose. Tegumentary surface uptake of acidic amino acids has been compared in paired and separated male and female schistosomes. InS. mansoni, a saturable carrier-mediated mechanism has been defined which operates only in unpaired male and unpaired female teguments. In contrast, the uptake of aspartate and glutamate is not seen in paired worms of this species. Tegumental uptake of acidic amino acids is not observed in paired or unpaired male or femaleS. japonicum. However, inS. haematobium, significant quantities of aspartate are taken up by both paired and unpaired male schistosomes. Measurable aspartate uptake is seen in paired femaleS. haematobium, but in the separated female, there is minimal uptake of this acidic amino acid. Thus the permeability of the teguments of human schistosome species to acidic amino acids is modified in response to the paired state inS. mansoni andS. haematobium, but these characteristics are not shared byS. japonicum.     

d-(+)-Pinitol,an oviposition stimulant for the pipevine swallowtail butterfly,Battus philenor

Oviposition by females of the pipevine swallowtail butterfly,Battus philenor, was stimulated by contact with alcoholic extracts of host foliage.d-(+)-Pinitol was isolated and identified from leaf material of one host species,Aristolochia macrophylla (Aristolochiaceae). In combination with chloroform-soluble components of host leaf material, this compound was comparable to the parent extract in stimulating oviposition.     

Jasmonic Acid-Induced Changes in <Emphasis Type="BoldItalic">Brassica oleracea</Emphasis> Affect Oviposition Preference of Two Specialist Herbivores

Jasmonic acid (JA) is a key hormone involved in plant defense responses. The effect of JA treatment of cabbage plants on their acceptability for oviposition by two species of cabbage white butterflies, Pieris rapae and P. brassicae, was investigated. Both butterfly species laid fewer eggs on leaves of JA-treated plants compared to control plants. We show that this is due to processes in the plant after JA treatment rather than an effect of JA itself. The oviposition preference for control plants is adaptive, as development time from larval hatch until pupation of P. rapae caterpillars was longer on JA-treated plants. Total glucosinolate content in leaf surface extracts was similar for control and treated plants; however, two of the five glucosinolates were present in lower amounts in leaf surface extracts of JA-treated plants. When the butterflies were offered a choice between the purified glucosinolate fraction isolated from leaf surface extracts of JA-treated plants and that from control plants, they did not discriminate. Changes in leaf surface glucosinolate profile, therefore, do not seem to explain the change in oviposition preference of the butterflies after JA treatment, suggesting that as yet unknown infochemicals are involved.     

Ancestral semiochemical attraction persists for adjoining populations of siblingIps bark beetles (Coleoptera: Scolytidae)

Three species ofIps pine bark beetles in Hopping's group IX (S.L. Wood'sgrandicollis group),Ips confusus, I. lecontei, andI. paraconfusus, are parapatrically distributed in the American Southwest. They share post-Pleistocene altitudinal ecotones with their host pines. Adjacent to these areas of host overlap, we tested the hypothesis that aggregation pheromones produced by male beetles and/or host volatiles are sufficient to elicit the speciesspecific colonization behaviors typical of these threeIps species in nature. A more distantly related species,I. pini (Hopping's group IV, S.L. Wood'spini group) was used for outgroup comparison. Under the influence of pheromone, males ofI. confusus andI. paraconfusus do not discern among unin-fested log bolts of host and nonhost pine prior to bark contact. Males responding to pheromones emanating from infested bolts are similarly undiscriminating. Females ofI. confusus andI. lecontei olfactorily discern the combination of conspecific males in host pine from other possible beetlepine combinations; females ofI. paraconfusus do not. FemaleI. pini discerned conspecific pheromone from that ofI. lecontei. The bark beetle predator,Enoclerus lecontei, is attracted by, but does not discriminate among, the male-produced volatiles of these Group IXIps species. These results support a hypothesis that divergence in pheromonal responses by these group IXIps species has evolved following their speciation, having been manifested first in the female sex. Evolutionarily, the derived pheromonal messages have preceded their behavioral discrimination by these beetles. Additional speciesspecific cues may operate between the sexes in the field that may preclude heterospecific pairings.     

Host recognition and the study of a chemical basis for attraction by cuckoo bumble bees (Hymenoptera: Apidae)

Species ofPsithyrus (Hymenoptera; Apidae) are obligate bumble bee social parasites. In this study, females ofP. vestalis andP. ashtoni were presented with pentane extracts prepared from different body parts of queens of their respective host species,Bombus terrestris andB. terricola. Parasites of both species were capable of distinguishing host bees from other bumble bee species using chemical cues contained within extracts. Among extracts of several body parts presented to parasites, the abdomen produced the greatest behavioral response, with Dufour's gland and terminal tergal segments eliciting the greatest response among abdominal regions. Extracts of these two body parts obtained fromB. terrestris queens shared a number of compounds, identified by GC-MS. Among the identified compounds are a number that have been reported to be of importance in bee sociochemistry.     

Role of volatile inforchemicals emitted by feces of larvae in host-searching behavior of parasitoidCotesia rubecula (Hymenoptera: Braconidae): A behavioral and chemical study

The role of volatile infochemicals emitted by feces of larvae in the host-searching behavior of the parasitoidCotesia rubecula was evaluated during single- and dual-choice tests inside a wind tunnel. The following treatments were tested: feces produced by second and fourth instars ofPieris rapae (preferred host), second instars ofP. brassicae (inferior host), second instars ofP. napi (nonhost), and wet feces of second instars ofP. rapae. During a single-choice situation females ofC. rubecula oriented to all types of feces tested. When a preference was to be made,C. rubecula preferred feces of second instars ofP. rapae over that of fourth, feces ofP. rapae over that ofP. brassicae, feces ofP. napi over that ofP. brassicae, and wet over normal host feces. No preference was exhibited between feces of second instars ofP. napi and that of second instars ofP. rapae. The relative importance of infochemicals from host feces versus plant damage caused by host larvae to the searching behavior ofC. rubecula was also evaluated. Plant damage was more important to the searching females than host feces when feces were present in specific concentrations in relation to damage. The volatiles released by normal and wet feces of second instars ofP. rapae, wet feces of fourth instars ofP. rapae, and normal and wet feces ofP. brassicae were collected and identified. Overall, 85 chemical compounds were recorded belonging to the following chemical groups: alcohols, ketones, aldehydes, esters, isothiocyanates, sulfides, nitriles, furanoids, terpenoids and pyridines. The blend of chemicals emitted by feces of different instars ofP. rapae and different species ofPieris exhibited an instar and species specificity in both quantity and quality. Wetting of normal feces increased the amount of volatile chemicals released, and it was also responsible for the appearance of new compounds. The role of feces of larvae in the host-seeking behavior ofC. rubecula is discussed.     

Olfactory discrimination byHeteropsylla cubana (Homoptera: Psyllidae) between susceptible and resistant species ofLeucaena (Leguminosae)

In the field, adult psyllids,Heteropsylla cubana Crawford, oriented significantly more towards the caged seedlings of susceptibleLeucaena leucocephala (Lam.) de Wit than to those of the resistant tree species,Leucaena collinsii Britton & Rose, or the nonhostAmaranthus spinosus L. In a dual-choice bioassay using a still-air olfactometer, the females demonstrated a strong positive response to the hexane extract ofL. leucocephala leaves at 1×10^–3 g equivalents (g eq) of leaf material. The females did not orient to leaf extracts ofL. collinsii at high concentrations but responded positively at a lower concentration of 1×10^–4 g eq. Olfactory discrimination byH. cubana between resistant and susceptible host species should be considered in selection and breeding programs.     

Allelopathic research of subtropical vegetation in Taiwan II. Comparative exclusion of understory byPhyllostachys edulis andCryptomeria japonica

On many hillsides of Taiwan there is a unique pattern of weed exclusion byPhyllostachys edulis (bamboo) andCryptomeria japonica (conifer) in which the density, diversity, and dominance of understory species are very different. Although the physical conditions of light, soil moisture, and soil nutrients strongly favor the growth of understory in a bamboo community, the biomass of its undergrowth is significantly low, indicating that physical competition among the understory species in the bamboo and conifer communities does not cause the observed differences. However, the biochemical inhibition revealed by these two plants appeared to be an important factor. The growth ofPellionia scabra seedlings, transplanted from the study site into greenhouse pots, was evidently suppressed by the aqueous leachate of bamboo leaves but was stimulated by that of conifer leaves. The radicle growth of lettuce, rye grass, and rice plants was also clearly inhibited by the leachate and aqueous extracts of bamboo leaves but not by those of conifer leaves. Six phytotoxins,o-hydroxyphenylacetic,p-hydroxybenzoic,p-coumaric, vanillic, ferulic, and syringic acids were found in the aqueous leachate and extracts of leaves and alcoholic soil extracts ofP. edulis, while the first three compounds were absent in the extracts ofC. japonica. The phytotoxicities of extracts were correlated with the phytotoxins present in both leaves and soils. The understory species might be variously tolerant to the allelopathic compounds produced by the two plants, resulting in a differential selection of species underneath. Therefore, comparative allelopathic effects ofPhyllostachys edulis andCryptomeria japonica may play significant roles in regulating the populations of the understories.Paper No. 253 of the Scientific Journal Series of the Institute of Botany, Academia Sinica, Taipei, Taiwan. This study was supported by the National Science Council of the Republic of China.     

Influence of trichome exudates from species ofLycopersicon on oviposition behavior ofHeliothis zea (Boddie)

Cage experiments revealed that accessions of the wild tomato speciesLycopersicon hirsutum were preferred sites for oviposition byHeliothis zea. Hexane extracts from the leaves ofL. hirsutum were also preferred sites of oviposition in choice experiments among extracts from severalLycopersicon species. Extracts ofL. hirsutum were still biologically active several days after application, indicating that the phytochemical(s) involved are relatively stable and of low volatility. Gas Chromatographic analysis of leaf hexane extracts from 12 different accessions of theL. hirsutum complex and three tomato cultivars revealed substantial qualitative and quantitative variation in the chemical composition of these extracts. Comparison of these results with extract oviposition studies implicate a group of structurally related compounds as the active agents. Mass spectroscopy has tentatively identified these compounds as sesquiterpenes with the chemical formula C₁₅H₂₂O₂. These compounds are apparently synthesized and secreted from glandular trichomes on the leaf surface. These phytochemicals did not stimulate ovipositional behavior in females of the cabbage looper,Trichoplusia ni. The existence of genetic variation for the presence and amount of kairomones that serve as cues for insect orientation and oviposition could be utilized in a breeding program to develop tomato cultivars with genetically modified allelochemic profiles that would disrupt the sequential behavioral processes of insect host-plant selection.