Just how insoluble are monoterpenes?

Prior generalizations about the ecological roles of monoterpenes may be misleading if based on the presumed insolubility of monoterpenes in water. We determined the aqueous solubility of 31 biologically active monoterpenes by gas chromatography. While hydrocarbons were of low solubility (< 35 ppm), oxygenated monoterpenes exhibited solubilities one or two orders of magnitude higher, with ranges of 155–6990 ppm for ketones and of 183–1360 ppm for alcohols. Many monoterpenes are phytotoxic in concentrations under 100 ppm, well below the saturated aqueous concentrations of oxygenated monoterpenes. Therefore, even dilute, unsaturated solutions of monoterpenes, occurring naturally in plant tissues and soil solutions, may act as potent biological inhibitors.     

Inhibition ofSchizachyrium scoparium (poaceae) by the allelochemical hydrocinnamic acid

Bare zones around shrubs in the Florida scrub indicate the possibility of allelopathy by shrubs controlling the distribution of grasses invading from adjacent sandhills. The allelochemical, hydrocinnamic acid, has been identified as a breakdown product of ceratiolin, which is released from the shrubCeratiola ericoides. Here, hydrocinnamic acid (HCA) was shown to have a strongly inhibitory effect on shoot and root biomass of the grassSchizachyrium scoparium in greenhouse bioassays lasting 4.5 months. Linear increases in the concentration of HCA from 0 to 200 ppm, applied biweekly, resulted in exponential decreases in root and shoot biomass at harvest. Plants grown at 200 ppm HCA had root and shoot biomasses 13% and 17% of controls, respectively. Concurrent investigation of reduced nutrient levels indicated greater inhibition by HCA in a reduced nitrogen (N) treatment and in a reduced potassium (K) treatment relative to HCA inhibition in the full nutrient treatment. The negative slopes of the regressions of log of biomass on HCA concentration were steepest in the reduced N and reduced K treatments. Root and shoot biomasses in reduced N treatments were 20–43% and 24–34% less than the respective biomasses in the full nutrient treatment. Comparable reductions in the reduced K treatment were as much as 19% and 10% for root and shoot biomasses, respectively. The effects of HCA in a reduced phosphorus (P) treatment and in a reduced P and K treatment were not significantly different from the effects of HCA in the full nutrient treatment. Extraction of the soils at harvest indicated no buildup of HCA at the end of the experiment. The sensitivity ofSchizachyrium scoparium to HCA in general and increased sensitivity under low N and low K solutions may be important in the Florida scrub community where levels of N and K are known to be low.     

Response of the ichneumonid parasiteNemeritis canescens to Kairomones from the flour moth,Ephestia kuehniella

2-Acylcyclohexane-1,3-diones, a novel class of compounds in the larval mandibular glands ofEphestia(=Anagasta) kuehniella Zeller, act as kairomones. Their relative activities in eliciting oviposition responses from the larval parasiteNemeritis(=Venturia) canescens (Grav.) are reported.     

Allelopathic potential of menthofuran monoterpenes fromCalamintha ashei

A reversed-phase HPLC analysis was used to separate and quantify five menthofuran monoterpenes inCalamintha ashei leaf soaks and washes. (+)-Evodone and desacetylcalaminthone were the major constituents of both soaks and washes. Concentrations of (+)-evodone and desacetylcalaminthone were as high as 0.66 and 0.74 mM, respectively, in leaf soaks. The highest concentration of monoterpenes in leaf washes obtained by misting was 0.021 mM. Aqueous solubilities of the menthofurans were determined to exceed concentrations required for growth inhibition. Bioassays of individualCalamintha monoterpenes demonstrated effects on germination as low as 0.05 mM for (+)-evodone. An equimolar mixture of desacetylcalaminthone and (+)-evodone reducedRudbeckia hirta germination by 17% at a combined concentration of 0.025 mM forLeptochloa dubia. Confirmation of allelopathic effects byCalamintha ashei will require long-term bioassays ofCalamintha menthofurans on the growth of native sandhill species under conditions comparable to the harsh environment of the Florida scrub.     

Preference of meadow voles (Microtus pennsylvanicus) for conifer seedlings: Chemical components and nutritional quality of bark of damaged and undamaged trees

Meadow voles (Microtus pennsylvanicus Ord.) of the 1987–1988 population density peak left several seedling plantations of Norway spruce (Picea abies) and Norway pine (Pinus resinosa) severely damaged in southern Quebec, Canada, while white spruce (Picea glauca) and white pine (Pinus strobus) suffered no damage. We compared levels of crude protein, total nonstructural carbohydrates (TNC), total phenols, and monoterpenes in the bark to detect differences between damaged and undamaged seedlings within and between plantations. There were no significant differences in levels of nutritional components between both types of seedlings within the same plantation. However, differences were noted among tree plantations for protein, TNC, and phenolics content but these differences could not be assigned to the voles' selection for particular species. Quantitative and qualitative differences also were registered in monoterpene levels between species. -Myrcene and bornyl acetate were found only in seedlings of undamaged species. Moreover, the latter contained higher levels of limonene. We conclude that the presence of specific monoterpenes is more important in defense mechanisms of conifer seedlings than relative levels of nutrients (protein or TNC) or total phenolics.     

Volatile hydrocarbons in the dufour's gland of the parasiteNemeritis canescens (Grav.) (Hymenoptera; ichneumonidae)

The Dufour's gland of the parasitic waspNemeritis (= Venturia) canescens (Grav.) was shown by gas chromatography and mass spectrometry to contain a mixture of C₂₁, C₂₃, and C₂₅ saturated and monounsaturated hydrocarbons. The main component (62%) was (Z)-10-tricosene. The biological activity of the components is discussed.     

Chemical Analysis of Squirt-gun Defense in Bursera and Counterdefense by Chrysomelid Beetles

The genus Bursera produces resin stored in canals in the leaf. When leaves are damaged, some, but not all, species release abundant resin. Species of Blepharida are specialized herbivores of Bursera, and they exhibit variation in their counterdefensive behavior. Species feeding on resin-releasing plants cut the leaf veins before feeding, which often makes them more prone to predation. They also adorn their backs with their feces and may regurgitate and release an anal secretion when attacked or disturbed by predators. Species that feed on Bursera species that release no fluids do not sever the leaf veins prior to feeding, and they do not carry their feces on their backs. Instead, they face their predators, raise their heads in a "boxing-like" display, and rapidly swing their abdomens from side to side. We performed a comparative chemical analysis of the compounds found in Bursera schlechtendalii, a species that releases abundant resins, and B. biflora, a species that does not. We also analyzed the frass, enteric discharges, and larvae of the two species of Blepharida that feed on each of these plants. The compounds found in the body, feces, and discharges of the Blepharida species that adorns itself with feces match the chemical mixture of its host plant, suggesting that this beetle species can compensate its higher risk of predation by using the compounds present in the plant for defense. The chemical mixture of B. biflora is more complex and does not match the compounds found in the body or frass of its beetle herbivore, suggesting that the defensive strategy of this insect is behavioral and does not rely on its host's constituents.     

Defensive mechanisms of loblolly and shortleaf pine against attack by southern pine beetle,Dendroctonus frontalis Zimmermann,and its fungal associate,Ceratocystis minor (Hedgecock) Hunt

Loblolly and shortleaf pine growing on a single site in the North Carolina piedmont were examined to determine similarities and differences in their defensive mechanisms against the southern pine beetle,Dendroctonus frontalis Zimmermann, and its fungal associate,Ceratocystis minor (Hedgecock) Hunt. Both species responded to wounding and fungal inoculation by forming a hypersensitive lesion around the wound site. There were significantly less soluble sugars and more monoterpenes in the lesion tissue than in unwounded inner bark. The two species were similar in resin flow rate and inner bark soluble sugar content, but the loblolly pines had thicker bark, longer hypersensitive lesions, and a higher concentration of inner bark monoterpenes. Inner bark monoterpene composition was also significantly different between the two pine species. It is hypothesized that two different defensive strategies against southern pine beetle attack may be utilized.     

Relative kairomonal activities of 2-acylcyclohexane-1,3-diones in eliciting oviposition behavior from parasiteNemeritis canescens (Grav.)

The relative activities of sixteen 2-acylcyclohexane-1,3-diones from the larval mandibular glands ofEphestia (=Anagasta) kuehniella Zeller in causing the parasiteNemeritis (=Venturia) canescens (Grav.) to make oviposition movements are reported.     

Chemical contents of stinging trichomes ofCnidoscolus texanus

Plants known as nettles bear trichomes that deliver a painful sting when touched. Studies of members of the Urticaceae have suggested that their trichomes may contain histamine, acetylcholine, and serotonin. We report investigations using TLC, HPLC, and UV-VIS spectrophotometry to analyze the hair fluids ofCnidoscolus texanus, a member of the Euphorbiaceae, along with standards of known histamine, acetylcholine, and serotonin. Of these, only serotonin was detected in hair extracts. Up to four other compounds were detected and partially characterized, but they have not been identified.     

Allelopathic potential ofPiqueria trinervia (Compositae) and piquerols A and B

Bioassays were carried out to test extracts of roots and leaves ofPiqueria trinervia and solutions of piquerol A and B, in order to appraise their allelopathic potential. Leaves seem to have greater concentrations of inhibitory compounds than roots. Both piquerol compounds show strong biological activity, but piquerol A is the more active.     

Geranium defensive agents. IV. Chemical and morphological bases of resistance

Insect-resistant and -susceptible geranium plants (Pelargonium xhortorum) were chemically and morphologically compared to assess the relative importance of resistance components. Glandular and nonglandular trichome density and quantification of trichome exudate (anacardic acids) from inbred resistant and susceptible lines were determined. Three different extraction procedures were employed to quantify the anacardic acids: (1) capillary solvent extract, (2) leaf wipe, and (3) whole leaf extract. The results support the conclusion that tall glandular trichomes and the exudate they produce are the most important factors in small pest resistance in the geranium. In addition, it was shown that tall glandular trichome densities are less and the ability of the plant to express the anacardic acids as exudate on the trichome head exterior is lacking in the susceptible lines analyzed, even though all lines possessed the capability to synthesize the anacardic acids. A plant line of intermediate resistance character was shown to possess high densities of tall glandular trichomes but resembled susceptible plants in lacking the ability to express the anacardic acids as an exudate in appreciable quantities.     

Seed-germination inhibition by volatile alcohols and other compounds associated withAmaranthus palmeri residues

Effects of 3-pentanone and eight low molecular weight aliphatic alcohols and aldehydes identifed in the mixtures of volatiles released byAmaranthus palmeri S. Wats. (AMAPA) residues were determined on germination of onion, carrot, AMAPA, and tomato seeds. Three-day exposures to these volatiles significantly inhibited germination of these assay seeds, and the inhibition was dependent upon exposure time and concentration. Based on the degree of inhibition observed in both time- and concentration-dependent assays, the following activity series was obtained: 2-heptanol > 3-methyl-1-butanol, 1-hexanol > hexanal, 1-pentanol, 3-pentanone, acetaldehyde > ethanol, 2-methyl-1-propanol. The activities of this group of compounds with that of 2-heptanone appear to be additive and related to test compound volatility and hydrophilicity. 2-Heptanol and 2-heptanone also significantly inhibited the germination of other species, including shepherdspurse, soybean, lettuce, alfalfa, common purslane, oats, and lovegrass.Name of companies or commercial products are given solely for the purpose of providing specific information; their mention does not imply recommendation or endorsement by the U.S. Department of Agriculture over others not mentioned.     

Cyanide in the Chemical Arsenal of Garlic Mustard, <Emphasis Type="Italic">Alliaria petiolata</Emphasis>

Cyanide production has been reported from over 2500 plant species, including some members of the Brassicaceae. We report that the important invasive plant, Alliaria petiolata, produces levels of cyanide in its tissues that can reach 100 ppm fresh weight (FW), a level considered toxic to many vertebrates. In a comparative study, levels of cyanide in leaves of young first-year plants were 25 times higher than in leaves of young Arabidopsis thaliana plants and over 150 times higher than in leaves of young Brassica kaber, B. rapa, and B. napus. In first-year plants, cyanide levels were highest in young leaves of seedlings and declined with leaf age on individual plants. Leaves of young plants infested with green peach aphids (Myzus persicae) produced just over half as much cyanide as leaves of healthy plants, suggesting that aphid feeding led to loss of cyanide from intact tissues before analysis, or that aphid feeding inhibited cyanide precursor production. In a developmental study, levels of cyanide in the youngest and oldest leaf of young garlic mustard plants were four times lower than in the youngest and oldest leaf of young Sorghum sudanense (cv. Cadan 97) plants, but cyanide levels did not decline in these leaves with plant age as in S. sudanense. Different populations of garlic mustard varied moderately in the constitutive and inducible expression of cyanide in leaves, but no populations studied were acyanogenic. Although cyanide production could result from breakdown products of glucosinolates, no cyanide was detected in vitro from decomposition of sinigrin, the major glucosinolate of garlic mustard. These studies indicate that cyanide produced from an as yet unidentified cyanogenic compound is a part of the battery of chemical defenses expressed by garlic mustard.     

Toxicity of allelopathic monoterpene suspensions on yeast dependence on droplet size

The toxic effects of the allelopathic nonsubstituted monoterpenes -pinene and limonene on yeast,Saccharomyces cerevisiae, were proportional to the size of the monoterpene droplets in suspension. Both the toxic effects and the size of the droplets in suspension were decreased by adding different solvents with the monoterpene as follows: dimethylsulfoxide – dimethylformamide ethanol > dioxane. Oxygen consumption was inhibited about 80% by 1 mM -pinene added in dimethylsulfoxide but less than 10% when -pinene was added in dioxane. Parallel decreases in droplet size and toxic effects of either monoterpene were also induced by hydrating the monoterpene-dimethylformamide or monoterpene-dimethylsulfoxide before addition to yeast. Molecular aggregation may be a mechanism to potentiate the allelopathic properties of monoterpenes when these associate with diverse soil components.     

In search of allelopathy in the Florida scrub: The role of terpenoids

The hypothesis was tested that allelopathic agents released from fire-sensitive plants of the Florida scrub community deter the invasion of fireprone sandhill grasses. The structures of the constituents of four endemic scrub species,Conradina canescens, Calamintha ashei, Chrysoma pauciflosculosa, andCeratiola ericiodes, were established and their phytotoxic activity against two grasses of the sandhill was examined. Effects of the secondary metabolites from the above scrub species and their degradation products upon the germination and radicle growth of little bluestem (Schizachyrium scoparium) and green sprangletop (Leptochloa dubia), two native grasses of the Florida sandhill community, were determined. The studies included determination of the water solubility and release mechanism of terpenes and other allelopathic agents from the source plants and their aqueous transport to the target species. Some of the natural products were nontoxic until activated by light and/or oxidation after release from the source plant into the environment.Dedicated to one of the pioneers in allelopathy, Cornelius H. Muller.     

Chemical Stimulants of Leaf-Trenching by Cabbage Loopers: Natural Products, Neurotransmitters, Insecticides, and Drugs

Larvae of the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae), often transect leaves with a narrow trench before eating the distal section. The trench reduces larval exposure to exudates, such as latex, during feeding. Plant species that do not emit exudate, such as Plantago lanceolata, are not trenched. However, if exudate is applied to a looper's mouth during feeding on P. lanceolata, the larva will often stop and cut a trench. Dissolved chemicals can be similarly applied and tested for effectiveness at triggering trenching. With this assay, I have documented that lactucin from lettuce latex (Lactuca sativa), myristicin from parsley oil (Petroselinum crispum), and lobeline from cardinal flower (Lobelia cardinalis) elicit trenching. These compounds are the first trenching stimulants reported. Several other constituents of lettuce and parsley, including some phenylpropanoids, monoterpenes, and furanocoumarins had little or no activity. Cucurbitacin E glycoside found in cucurbits, another plant family trenched by cabbage loopers, also was inactive. Lactucin, myristicin, and lobeline all affect the nervous system of mammals, with lobeline acting specifically as an antagonist of nicotinic acetylcholine receptors. To determine if cabbage loopers respond selectively to compounds active at acetylcholine synapses, I tested several neurotransmitters, insecticides, and drugs with known neurological activity, many of which triggered trenching. Active compounds included dopamine, serotonin, the insecticide imidacloprid, and various drugs such as ipratropium, apomorphine, buspirone, and metoclopramide. These results document that noxious plant chemicals trigger trenching, that loopers respond to different trenching stimulants in different plants, that diverse neuroactive chemicals elicit the behavior, and that feeding deterrents are not all trenching stimulants. The trenching assay offers a novel approach for identifying defensive plant compounds with potential uses in agriculture or medicine. Cabbage loopers in the lab and field routinely trench and feed on plants in the Asteraceae and Apiaceae. However, first and third instar larvae enclosed on Lobelia cardinalis (Campanulaceae) failed to develop, even though the third instar larvae attempted to trench. Trenching ability does not guarantee effective feeding on plants with canal-borne exudates. Cabbage loopers must not only recognize and respond to trenching stimulants, they must also tolerate exudates during the trenching procedure to disable canalicular defenses.     

Chemical analysis of volatiles emitted by Pinus svlvestris after induction by insect oviposition

Gas chromatography – mass spectrometry analyses of the headspace volatiles of Scots pine (Pinus sylvestris) induced by egg deposition of the sawfly Diprion pini were conducted. The odor blend of systemically oviposition-induced pine twigs, attractive for the eulophid egg parasitoid Chrysonotomyia ruforum, was compared to volatiles released by damaged pine twigs (control) that are not attractive for the parasitoid. The mechanical damage inflicted to the control twigs mimicked the damage by a sawfly female prior to egg deposition. The odor blend released by oviposition-induced pine twigs consisted of numerous mono- and sesquiterpenes, which all were also present in the headspace of the artificially damaged control twigs. A quantitative comparison of the volatiles from oviposition-induced twigs and controls revealed that only the amounts of (E)--farnesene were significantly higher in the volatile blend of the oviposition-induced twigs. Volatiles from pine twigs treated with jasmonic acid (JA) also attract the egg parasitoid. No qualitative differences were detected when comparing the composition of the headspace of JA-treated pine twigs with the volatile blend of untreated control twigs. JA-treated pine twigs released significantly higher amounts of (E)--farnesene. However, the JA treatment induced a significant increase of the amount of further terpenoid components. The release of terpenoids by pine after wounding, egg deposition, and JA treatment is discussed with special respect to (E)--farnesene.     

Effects of soil nitrogen level on ferulic acid inhibition of cucumber leaf expansion

It has been suggested that the allelopathic activity of phenolic acids should be primarily important in soils of low fertility. If this is true, then plant growth inhibition by phenolic acids may be unimportant in managed agricultural soils. The objective of this study was to determine how soil nitrogen (N) level might modify phenolic acid inhibition of growth. Cucumber seedlings (Cucumis sativus cv Early Green Cluster) grown in containers in growth chambers under varying N levels (5, 10, 15, 20, and 25 g N/g soil) in Portsmouth B,-horizon soil material were treated with ferulic acid (0 or 10 g/g soil). Nitrogen and ferulic acid (FA) were applied every other day to the soil surface. The amount of FA in the soil solution declined with depth in the containers. A more rapid disappearance of FA from the soil solution was observed for the last FA treatment (0% recovered after 10 hr on day 23) than the first treatment (44% recovered after 10 hr on day 13). Both low N (5 g N/g soil) and FA treatments reduced shoot dry weight, the mean absolute (AGR) and the mean relative (RGR) rates of leaf expansion, and increased the root-shoot ratio. High N treatments reduced shoot dry weight and the AGR. Ferulic acid inhibited cucumber seedling growth over a range of N concentrations, suggesting that the allelopathic activity of phenolic acids may be important in both nutrient limiting and nonlimiting soils for some species.Paper No. 12219 of the journal series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601. The use of tradenames in this publication does not imply endorsement by the North Carolina Agricultural Research Service of products named, nor criticism of similar ones not mentioned.     

Influence of oleoresin constituents fromPinus ponderosa andPinus jeffreyi on growth of mycangial fungi fromDendroctonus ponderosae andDendroctonus jeffreyi

Dendroctonus jeffreyi andD. ponderosae are morphologically similar sympatric species of pine bark beetles over portions of their geographic ranges; however,D. jeffreyi is monophagous onP. jeffreyi whileD. ponderosae is highly polyphagous. Both species carry a species of mycangial fungi that are also very similar in appearance. Growth of the two mycangial fungi and of the fungusLeptographium terebrantis (associated with the polyphagous and non-tree-killingDendroctonus valens) in the presence of oleoresin constituents of host and nonhost conifers was tested by placing individual chemicals on agar growth medium and by growing the cultures in saturated atmospheres of the chemicals. The fungus associated withD. jeffreyi showed greater tolerance for chemical constituents placed on the growth medium than the other two fungi, and growth after three days was enhanced by heptane, the dominant constituent ofP. jeffreyi oleoresin. Growth of all three species of fungi was reduced by the resin constituents when the chemicals were presented as saturated atmospheres. The results suggest that the influence of the tree on growth of the symbiotic fungi of the bark beetles during the initial attack process may be different than after colonization by the beetles is complete. The difference in the responses of the apparently related species of mycangial fungi may provide some new insight into the evolutionary history of these beetle/mycangial fungus/host tree systems.