Inheritance Of Resistance To Mammalian Herbivores and Of Plant Defensive Chemistry In A <Emphasis Type="Italic">Eucalyptus</Emphasis> Species

Hybridization in plants provides an opportunity to investigate the patterns of inheritance of hybrid resistance to herbivores, and of the plant mechanisms conferring this resistance such as plant secondary metabolites. We investigated how inter-race differences in resistance of Eucalyptus globulus to a generalist mammalian herbivore, Trichosurus vulpecula, are inherited in their F₁ hybrids. We assessed browsing damage of three-year-old trees in a common environment field trial on four hybrid types of known progeny. The progency were artificial intra-race crosses and reciprocal inter-race F₁ hybrids of two geographically distinct populations (races) of E. globulus; north-eastern Tasmania and south-eastern Tasmania. Populations of trees from north-eastern Tasmania are relatively susceptible to browsing by T. vulpecula, while populations from south-eastern Tasmania are more resistant. We assessed the preferences of these trees in a series of paired feeding trials with captive animals to test the field trial results and also investigated the patterns of inheritance of plant secondary metabolites. Our results demonstrated that the phenotypic expression of resistance of the inter-race F₁ hybrids supported the additive pattern of inheritance, as these hybrids were intermediate in resistance compared to the pure parental hybrids. The expression of plant secondary metabolites in the F₁ hybrids varied among groups of individual compounds. The most common pattern supported was dominance towards one of the parental types. Together, condensed tannins and essential oils appeared to explain the observed patterns of resistance among the four hybrid types. While both chemical groups were inherited in a dominant manner in the inter-race F₁ hybrids, the direction of dominance was opposite. Their combined concentration, however, was inherited in an additive manner, consistent with the phenotypic differences in browsing.     

Ontogeny and Environment as Determinants of the Secondary Chemistry of Three Species of White Birch

This study investigates variation in the secondary chemistry of the bark of three closely related, winter-dormant species of white birch (Betula resinifera, B. pendula, and B. platyphylla) at different ontogenetic stages by using different plant parts (top and base). The experimental birches were grown for 4 years in two growing conditions (pot and field) at different nutrient levels. There was considerable species-specific quantitative and qualitative variation in the secondary chemicals in bark, but this was also affected by fertilization and the age of the plant. In general, there was greater chemical diversity in saplings than in seedlings. The study revealed three new components, secoisolariciresinol 9-O-β-glucopyranoside and two of its derivatives, that have not been reported previously for the bark of white birches. Principal component analysis showed that the species studied had a similar chemical composition at the juvenile stage, but as the plants grew, they became more clearly differentiated, which indicates that the species of older plants can be identified by chemotaxonomy. Evidently, the secondary chemistry of birches is under genetic control, but it is affected by properties of growing conditions and ontogeny. Electronic Supplementary Material Supplementary material to this paper is available in electronic form at     

Inter- and Intraspecific Comparisons of Antiherbivore Defenses in Three Species of Rainforest Understory Shrubs

Plants defend themselves against herbivores and pathogens with a suite of morphological, phenological, biochemical, and biotic defenses, each of which is presumably costly. The best studied are allocation costs that involve trade-offs in investment of resources to defense versus other plant functions. Decreases in growth or reproductive effort are the costs most often associated with antiherbivore defenses, but trade-offs among different defenses may also occur within a single plant species. We examined trade-offs among defenses in closely related tropical rain forest shrubs (Piper cenocladum, P. imperiale, and P. melanocladum) that possess different combinations of three types of defense: ant mutualists, secondary compounds, and leaf toughness. We also examined the effectiveness of different defenses and suites of defenses against the most abundant generalist and specialist Piper herbivores. For all species examined, leaf toughness was the most effective defense, with the toughest species, P. melanocladum, receiving the lowest incidence of total herbivory, and the least tough species, P. imperiale, receiving the highest incidence. Although variation in toughness within each species was substantial, there were no intraspecific relationships between toughness and herbivory. In other Piper studies, chemical and biotic defenses had strong intraspecific negative correlations with herbivory. A wide variety of defensive mechanisms was quantified in the three Piper species studied, ranging from low concentrations of chemical defenses in P. imperiale to a complex suite of defenses in P. cenocladum that includes ant mutualists, secondary metabolites, and moderate toughness. Ecological costs were evident for the array of defensive mechanisms within these Piper species, and the differences in defensive strategies among species may represent evolutionary trade-offs between costly defenses.     

Fate of the Dietary Terpene, P-Cymene, in the Male Koala

The fate of the dietary terpene, p-cymene, is reported in a specialist Eucalyptus leaf folivore, the koala (Phascolarctos cinereus). Six male koalas were given two single oral doses of p-cymene (0.37 mmol/kg and 1.49 mmol/kg). Urine and feces were collected for two 24-hr periods after each dose and analyzed for metabolites by extraction, gas chromatography, and mass spectrometry. A total of six metabolites were detected in the urine. A novel, extensively oxidized metabolite, 4-(1,2-dihydroxy-1-methylethyl)-benzoic acid, was identified and its structure elucidated by high resolution and chemical ionization mass spectrometry. Minor amounts of two glycine conjugated metabolites were also detected. Five metabolites were present in sufficient amounts to quantify, using isolated urinary metabolites as reference standards. The mean fractional recovery of administered p-cymene was 0.77 ± 0.09 and 0.84 ± 0.12 for the low and high dose, respectively. The major metabolite excreted was the novel carboxy diol compound. No fecal metabolites were found. Thus, the koala employs a strategy in the metabolism of p-cymene that promotes the production of extensively oxidized metabolites that consequently have increased polarity and enhanced renal excretion. This strategy is compared with that employed by other Eucalyptus leaf folivores.     

Behavioral Effects and Sensory Detection of Drimane Deterrents in Myzus persicae and Aphis gossypii Nymphs

Eleven synthetic drimane compounds were tested for their deterrency to nymphs of Myzus persicae (Sulzer) and Aphis gossypii (Glover) (Homoptera: Aphididae). In general, A. gossypii nymphs were less sensitive to the drimanes than M. persicae nymphs. Warburganal (1) and polygodial (2), with a -dialdehyde configuration and double bond at C7–C8, were highly active as deterrents and/or feeding inhibitors against both species. One of the synthetic compounds (10) showed relatively high activity against A. gossypii, while it was not active at all against M. persicae, indicating that synthetic analogs of natural deterrents can be highly selective feeding deterrents. From 24-hr interval observations and ablation studies, it is concluded that nymphs of both M. persicae and A. gossypii detect polygodial and possibly the other drimanes tested with contact chemosensilla at the tips of their antennae. The ablation studies also showed that in both species no tarsal, labial, or epipharyngeal sensilla are involved in detecting polygodial within 24 hr.     

低温控温结晶法分离提纯1,8-桉叶油素的工艺

采用全结晶工艺对原料桉叶油[w(cineole)=63.24%]进行分离提纯,在无晶种添加下,降温速率4℃/h、结晶终温-30℃、发汗速率5℃/h、发汗终温-19℃时,可将原料油提纯至77.52%。添加晶种后,可有效提高操作温度,缩小结晶温度的操作范围,提高产品的纯度,在降温速率4℃/h、结晶终温-25℃、发汗速率4℃/h、发汗终温-6℃的操作条件下,将原料油提纯至89.63%。实验得到了低温控温添加晶种结晶法分离提纯1,8-桉叶油素的适宜工艺流程和操作参数,该方法相对其他分离方法具有较为明显的优势。     

Scratch resistance of automobile clearcoats: Chemistry and characterization on the micro-and nanoscale

The scratch resistance of polymeric clearcoats intended for automobile exterior surfaces was tested with scanning 3D nanoindentation and laboratory car wash simulation (AMTEC). It could be shown by using “real world” samples with comparable scratch resistance that the applied methods were suitable for discriminating even small differences in scratch resistance with high accuracy. In the area of physical characterization of scratch resistance, the results of this investigation demonstrate the ability of microscopic, single-contact testing methods to reproduce the macroscopically experienced results. By using a combination of different scratch methods, insight into the fundamentals, e.g., chemistry, that are responsible for good or bad scratch resistance, is achieved. One key for this goal is the imaging and analytical evaluation of the damage pattern after the indent, which leads to additional results and models presented in this article. It was found that an essential key to obtaining scratch resistant surfaces is a strengthening of rubber-like elasticity as well as minimizing interactions between individual polymeric chains.     

Potential aversive compounds in leafy spurge for ruminants and rats

Several wild and domestic ruminant species and horses apparently will not consume leafy spurge (Euphorbia esula) while grazing range and pasture lands. It has been demonstrated that leafy spurge can elicit conditioned food aversions in cattle and sheep, and the aversion-eliciting capacity of leafy spurge may account for why cattle seldom graze this nutritious plant and why sheep may not readily consume it at some locations. The identity of the aversive compound(s) in leafy spurge is unknown, but several different diterpenoid ingenol esters have been isolated from its tissues, and we suspect that one or more ingenol esters may be aversion-eliciting compounds in leafy spurge. The objectives of this study were to determine whether or not leafy spurge is aversive to laboratory rats and if a crude acetone extract of leafy spurge, presumably containing ingenol esters and other phytochemicals, could generate an aversive response in sheep and laboratory rats. An additional objective was to determine whether or not a particular ingenol monobenzoate, which may be similar to ingenol esters in leafy spurge, might also elicit an aversive response from rats. Rats exhibited food aversions associated with leafy spurge (P < 0.05). An acetone extract of leafy spurge induced conditioned food aversions in both sheep and rats (P < 0.01). The ingenol 3-monobenzoate also induced conditioned food aversions in rats (P < 0.01). Our interpretation of these data is that rats can be used as a model for cattle and sheep with respect to their aversion to leafy spurge ingestion. Additionally, we suggest that one or more ingenol esters may be aversion-inducing agents in leafy spurge. However, others may exist in leafy spurge that are also aversive or are the only or prime aversive chemicals.Published with the approval of the Director of the South Dakota Agricultural Research Station as publication number 2830 of the journal series.This research was supported in part by a grant from the Cooperative States Research Service (91-38300-6158). All experimental procedures with animals followed guidelines approved by animal care and use committees.     

Effects of predator chemical cues and behavioral biorhythms on foraging activity of terrestrial salamanders

Red-backed salamanders, Plethodon cinereus, show a variety of alarm responses to chemical cues from eastern garter snakes, Thamnophis sirtalis. We measured the foraging activity of red-backed salamanders exposed to water soiled by a garter snake (fed P. cinereus) or to unsoiled water. Salamanders exposed to snake-soiled water showed less foraging activity than salamanders exposed to unsoiled water; therefore, predators could have nonlethal effects on salamander populations. Our results also show additional factors influenced salamander foraging activity. Salamander foraging activity and responsiveness to chemical cues do not appear to have been affected by sex or food deprivation. Salamander foraging activity does appear to have been influenced by activity biorhythms. Foraging activity of animals in both treatments showed a bimodal periodicity that is consistent with natural activity patterns controlled by internal biorhythms. Exposure to snake-soiled water significantly reduced foraging activity during periods of peak foraging activity, but had a subtler effect0 on foraging activity during natural lulls in activity. We suggest that both activity biorhythms and exposure to chemical cues are important factors affecting salamander foraging behavior.     

Consequences of Enriched Atmospheric CO2 and Defoliation for Foliar Chemistry and Gypsy Moth Performance

Elevated concentrations of atmospheric CO₂ are likely to interact with other factors affecting plant physiology to alter plant chemical profiles and plant–herbivore interactions. We evaluated the independent and interactive effects of enriched CO₂ and artificial defoliation on foliar chemistry of quaking aspen (Populus tremuloides) and sugar maple (Acer saccharum), and the consequences of such changes for short-term performance of the gypsy moth (Lymantria dispar). We grew aspen and maple seedlings in ambient (~360 ppm) and enriched (650 ppm) CO₂ environments at the University of Wisconsin Biotron. Seven weeks after budbreak, trees in half of the rooms were subjected to 50% defoliation. Afterwards, foliage was collected for chemical analyses, and feeding trials were conducted with fourth-stadium gypsy moths. Enriched CO₂ altered foliar levels of water, nitrogen, carbohydrates, and phenolics, and responses generally differed between the two tree species. Defoliation induced chemical changes only in aspen. We found no significant interactions between CO₂ and defoliation for levels of carbon-based defenses (phenolic glycosides and tannins). CO₂ treatment altered the performance of larvae fed aspen, but not maple, whereas defoliation had little effect on performance of insects. In general, results from this experimental system do not support the hypothesis that induction of carbon-based chemical defenses, and attendant effects on insects, will be stronger in a CO₂-enriched world.     

Changes in Heartwood Chemistry of Dead Yellow-Cedar Trees that Remain Standing for 80 Years or More in Southeast Alaska

We measured the concentrations of extractable bioactive compounds in heartwood of live yellow-cedar (Chamaecyparis nootkatensis) trees and five classes of standing snags (1–5, averaging 4, 14, 26, 51, and 81 years-since-death, respectively) to determine how the concentrations changed in the slowly deteriorating snags. Three individuals from each of these six condition classes were sampled at four sites spanning a 260-km distance across southeast Alaska, and the influence of geographic location on heartwood chemistry was evaluated. Cores of heartwood were collected at breast height and cut into consecutive 5-cm segments starting at the pith. Each segment was extracted with ethyl acetate and analyzed by gas chromatography. Concentrations of carvacrol, nootkatene, nootkatol, nootkatone, nootkatin, and total extractives (a sum of 16 compounds) for the inner (0–5 cm from pith), middle (5–10 cm from pith), and surface (outer 1.1–6.0 cm of heartwood) segments from each core were compared within each tree condition class and within segments across condition classes. Heartwood of class 1 and 2 snags had the same chemical composition as live trees. The first concentration changes begin to appear in class 3 snags, which coincides with greater heartwood exposure to the external environment as decaying sapwood sloughs away, after losing the protective outer bark. Within core segments, the concentrations of all compounds, except nootkatene, decrease between snag classes 2 and 5, resulting in the heartwood of class 5 snags having the lowest quantities of bioactive compounds, although not different from the amounts in class 4 snags. This decline in chemical defense is consistent with heartwood of class 5 snags being less decay-resistant than heartwood of live trees, as observed by others. The unique heartwood chemistry of yellow cedar and the slow way it is altered after death allow dead trees to remain standing for up to a century with a profound impact on the ecology of forests in southeast Alaska where these trees are in decline.     

Ruminal metabolism of leafy spurge in sheep and goats: A potential explanation for differential foraging on spurge by sheep,goats, and cattle

Leafy spurge (Euphorbia esula) is an introduced forb that is invading western rangelands. Goats (Capra hircus) readily graze the plant, but cattle (Bos tarus) generally and sheep (Ovis aries) locally appear to develop conditioned flavor aversions to leafy spurge. They either avoid the plant entirely or graze it reluctantly. We hypothesized that: (1) a diterpene diester that can occur in leafy spurge was an aversive agent, and (2) diet selection differences among ruminant species may be partly a function of differential ruminai metabolism of aversive phytochemicals, and further that cattle and sheep may be reluctant to graze leafy spurge because their ruminai microbes do not metabolize certain leafy spurge chemicals as do ruminai microbes in goats. Sheep did not develop an aversion to a novel food when its consumption was followed by an intravenous injection of ingenol 3,20-dibenzoate (P=0.34). Sheep did develop an aversion to a novel food when its intake was followed by a dose of leafy spurge fermented with sheep ruminai digesta, but not when followed by a dose of leafy spurge fermented with goat ruminai digesta (P= 0.03). This suggests that goat ruminai microbes may modify leafy spurge such that it does not elicit an aversion in sheep.This research was supported in part by a grant from the Cooperative States Research Service (91-38300-6158). All experimental procedures with ruminants followed a protocol approved by the U.S. Sheep Experiment Station's Animal Care and Use Committee. Mention of trade names or companies does not constitute an implied warranty by USDA or the authors.     

Influence of Synthetic Oviposition Pheromone and Volatiles from Soakage Pits and Grass Infusions Upon Oviposition Site-Selection of Culex Mosquitoes in Tanzania

The response of Culex mosquitoes to (5R,6S)-6-acetoxy-5-hexadecanolide (the synthetic oviposition pheromone, SOP), emanations from soakage pit water (SPW), and grass infusions (GI) was studied in pit latrines in Muheza, Tanzania. Water treated with the synthetic oviposition pheromone received more egg rafts of Culex quinquefasciatus Say and Cx. cinereus Theobald than did the untreated water (P < 0.001). The residual activity of SOP did not decrease over a nine-day period, with the geometric mean number of egg rafts laid in the SOP-treated water varying daily between 2.5 and 8.9 as compared to 0.1–1.6 egg rafts laid in tap water. However, SOP did not attract ovipositing mosquitoes to nonbreeding sites, and both treatment and control received few eggs. SPW and GI attracted ovipositing Cx. quinquefasciatus, Cx. cinereus, and Cx. tigripes Grandpré and de Charmoy with the number of eggs rafts deposited in bowls containing SPW or GI being larger than that deposited in bowls with tap water (P < 0.05). More egg rafts of Cx. quinquefasciatus were deposited in SOP-treated water than in SPW (P < 0.05). When SOP was compared with SOP + SPW, more egg rafts of Cx. quinquefasciatus were deposited in bowls with the latter combination than in bowls containing SOP or SPW only (P < 0.05), indicating a synergistic effect between SOP and SPW. Similarly, when SOP-treated water and GI were compared with SOP + GI, more egg rafts of Cx. quinquefasciatus were laid in the latter bowl, indicating a synergistic effect between SOP and GI. The result suggest that the combined use of SOP and organically enriched water can be employed in monitoring of Cx. quinquefasciatus for control programs. This is the first record of the attraction of Cx. cinereus and Cx. tigripes to oviposition stimuli.     

Ingestion of tall larkspur by cattle

Tall larkspur (Delphinium spp.) is a palatable but toxic poisonous plant in the western United States. The toxins in tall larkspur are diterpenoid alkaloids. We examined the influences of food flavor and postingestive consequences on consumption of a 33% larkspur pellet during 30-min feeding periods for five days using esophageally fistulated cattle that were sham-fed larkspur pellets. Consumption by the sham-fed group was compared to a control group fed alfalfa pellets, and a larkspur group fed only larkspur pellets. Sham-fed cattle did not decrease (P > 0.1) feed consumption compared to controls, indicating no significant difference in food flavor. The larkspur group decreased (P < 0.05) feed consumption by 41% relative to controls and by 31% relative to sham-fed animals (P = 0.08). This reduction in feed consumption indicates the adverse postingestive consequences of tall larkspur ingestion, as the larkspur group apparently developed a conditioned taste aversion to the larkspur pellet. Even though these animals were averted to the pellets, they showed none of the classical signs of intoxication from ingestion of tall larkspur.This research was supported in part by the Utah Agricultural Experiment Station, Utah State University, Logan, Utah 84322. Approved as journal article No. 3846.     

Chemistry of male mandibular gland secretions ofPhilanthus triangulum

Males of the European and African beewolf,Philanthus triangulum, possess a sex specific mandibular gland secretion that is used for marking plant stems in mating territories. The secretion is composed of 90% (Z)-11-eicosen-1-ol plus small amounts of 10-nonadecen-2-one, nonadecenal, octadecanoic and octadecenoic acids, and eicosenal. The chemistry of this secretion differs markedly from the secretions of North AmericanPhilanthus, which consist of a larger number of components that possess different chemical functionalities and are more volatile. We postulate that the chemical differences betweenP. triangulum and its New World relatives reflect phylogenetic differences plus a possible reduced necessity for species isolating mechanisms inP. triangulum.     

Release of allelochemical agents from litter,throughfall, and topsoil in plantations ofEucalyptus globulus Labill in Spain

Natural leachates ofEucalyptus globulus (throughfall, stemflow, and soil percolates) were collected daily during rainy spells in the vegetative period (February–July), and their effects on the germination and radicle growth ofLactuca sativa were measured. Concurrently, the effects ofL. sativa of topsoil and leachates from decaying litter were determined. The results suggest that toxic allelochemicals released byEucalyptus globulus may influence the composition and structure of the understory of the plantation and that this effect is attributable mainly to the decomposition products of decaying litter rather than to aerial leachates. The soil may neutralize or dilute allelopathic agents, at least below the top few cms.     

Chemistry and functions of exocrine secretions of the antsTapinoma melanocephalum and T. erraticum

Volatile constituents produced by ant workers belonging to the speciesTapinoma melanocephalum andT. erraticum have been analyzed by gas chromatography-mass spectrometry. The pygidial (=anal) gland secretion ofT. melanocephalum is fortified with 6-methyl-5-hepten-2-one and actinidine (the mass spectrum of which is corrected in this paper). An unidentified compound was detected in cephalic extracts. The pygidial gland secretion ofT. erraticum was also dominated by 6-methyl-5-hepten-2-one, in addition to two isomers of iridodial, and iridomyrmecin. The sternal glands contained iridodial and C₁₅-C₂₀ hydrocarbons. Workers ofT. melanocephalum effectively utilize their pygidial gland secretions as an alarm-defense system during aggressive encounters with workers ofSolenopsis geminata. 6-Methyl-5-hepten-2-one is active as a releaser of alarm behavior, and actinidine is repellent to workers ofT. melanocephalum. Cephalic extracts possessed attractant and arrestant properties for workers of this species.     

Hostplant Suitability and Defensive Chemistry of the Catalpa Sphinx, Ceratomia catalpae

The growth and survival of the Catalpa sphinx, Ceratomia catalpae (Sphingidae), were measured on five different species of Catalpa: C. bignonioides, C. bungeii, C. fargeseii, C. ovata, and C. speciosa. Larval growth varied significantly among these host plant species; however, survival did not differ. Quantification of the iridoid glycoside content of larvae, pupae, adults, larval frass, and leaves of the larval host plant, C. bignonioides, by gas chromatography showed that leaves contained both catalpol and catalposide; larvae, pupae, and frass contained only catalpol; and the adults contained no detectable iridoid glycosides. Amounts were highest in the larvae and declined in the pupal stage. Very small amounts of catalpol were detected in adults of the parasitoid, Cotesia congregata, and in the silken cocoons. The hemolymph in which the parasitoid larvae grew contained over 50% dry weight catalpol. Larvae of C. catalpae often regurgitate when disturbed. This may serve as a defense against predators. A comparison of the growth of larvae pinched with forceps to induce regurgitation with those that were not so treated showed that larvae that were pinched, and usually regurgitated, grew significantly more slowly than those that were not.     

Identification of a four-component sex pheromone of the female oriental fruit moth,Grapholitha molesta (Lepidoptera: Tortricidae)

The female-emitted pheromone ofGrapholitha molesta (Busck), the Oriental fruit moth, was collected by holding females in glass flasks during calling. Flask washes were found to contain four pheromone components: (Z)-8-dodecenyl acetate and (E)-8-dodecenyl acetate in a 1007 ratio, and (Z)-8-dodecen-1-ol and dodecanol in a 10020 ratio. The ratio of (Z)-8-acetate to (Z)-8-dodecen-1-ol was approx. 10030. Approximately 0.1–0.2 ng of pheromone was recovered per female per hour of calling.Published as Journal Article No. 8598 of the Michigan State University Agricultural Experiment Station.     

Chemistry of Leichhardt's Grasshopper, Petasida ephippigera, and its Host Plants, Pityrodia jamesii, P. ternifolia, and P. pungens

Investigation of the strikingly aposematic Leichhardt's grasshopper revealed a number of host-plant-derived components. Chemical study of the three known host plants revealed a diverse array of components. Pityrodia ternifolia and P. pungens contain predominantly sesquiterpenes and sesquiterpene glycosides and P. jamesii is rich in cineole, dihydrochalcones, chalcones, and monoterpene glycosides. Feces from grasshoppers feeding on P. jamesii contain a mixture of hydroxycineoles, of which 3-hydroxycineole is the predominant isomer (68%). No evidence for previously suspected "alkaloidal" toxins was found in either the insect or host plant.