Phytotoxic Effects of 21 Plant Secondary Metabolites on <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Germination and Root Growth

This study investigated potential phytotoxic effects on germination and root growth of 21 plant secondary metabolites (sinapinic, syringic, vanillic, ferulic, p-coumaric, chlorogenic, gallic, gentisic, protocatechuic, p-hydroxybenzoic, and trans-cinnamic acids, and eucalyptol, quercetin, vanillin, syringaldehyde, rutin, 2-benzoxazolinone, protocatechualdehyde, tyrosol, juglone, and l-mimosine) in the plant model Arabidopsis thaliana. Eleven of the 21 molecules showed significant inhibitory effects on germination, and 17 inhibited root growth. Inhibitory effects on root growth were more evident when nutrients were not added. We present dose–response curves for germination effects and IC₅₀ values for each compound, along with possible explanations of the observed inhibitory actions in terms of molecular structure.     

Altered Glucosinolate Hydrolysis in Genetically Engineered Arabidopsis thaliana and its Influence on the Larval Development of Spodoptera littoralis

The antiherbivore potential of the glucosinolate–myrosinase defense system found in plants of the order Capparales is heavily influenced by the types of hydrolysis products (e.g. isothiocyanates, nitriles) formed from the parent glucosinolates upon plant damage. However, comparison of the effects of glucosinolate hydrolysis products on insect herbivores has been hampered by the lack of suitable experimental tools for rigorous bioassays, such as intact plants differing only in the types of hydrolysis products they produce, or artificial diets that can accurately simulate glucosinolate hydrolysis. The wide array of molecular resources for Arabidopsis thaliana has facilitated the identification of several genes that play a role in glucosinolate hydrolysis. One of these encodes the epithio-specifier protein (ESP) that promotes the formation of nitriles at the expense of isothiocyanates in certain ecotypes of A. thaliana. We overexpressed the ESP cDNA from the nitrile-producing ecotype Landsberg erecta in the isothiocyanate-producing ecotype Columbia-0 to generate transgenic lines of A. thaliana that differed from wild-type plants in the type of glucosinolate hydrolysis products formed upon tissue damage, whereas parent glucosinolate profile and myrosinase activity levels, as well as plant morphology and growth habit, remained unchanged. Bioassays with the model generalist herbivore Spodoptera littoralis (Lepidoptera: Noctuidae) demonstrated that larvae reared on the nitrile-producing lines on average gained weight faster in the first larval stages than larvae that fed on isothiocyanate-producing control plants. Furthermore, larvae with medial growth rates showed a tendency to pupate earlier on the ESP-overexpressing plant lines. Together with the results of previous studies, these findings suggest that isothiocyanates are more effective defenses against insect herbivores than nitriles, and raise questions about what conditions select for nitrile formation in plants.     

Phytotoxic Effects of Leukamenin E (an ent-kaurene diterpenoid) on Root Growth and Root Hair Development in Lactuca sativa L. seedlings

Leukamenin E, an ent-kaurene diterpenoid isolated from Isodon racemosa (Hemsl) Hara, showed phytotoxic effects on root growth and root hair development of lettuce seedlings (Lactuca sativa L.). Lower concentrations (10 μM) of leukamenin E did not affect root growth, but at concentrations higher than 50 μM, the rate was inhibited. The influence of leukamenin E on root growth rate was closely correlated with alterations in the mitotic index. A low incidence of aberrant mitosis image was observed when lettuce roots were treated with higher concentrations (100 and 200 μM) of leukamenin E. This suggests that inhibition of root growth may be due to inhibition of cell division. All tested concentrations of the diterpenoid (10 μM or more) inhibited root hair development in a dose-dependent manner. At a concentration of 80 μM, leukamenin E completely blocked root hair initiation. Application of Ag⁺—an ethylene action inhibitor—to lettuce seedlings inhibited root hair elongation similar to the diterpenoid. Enhanced root hair length was stimulated by exogenous ethephon—an ethylene-releasing agent—and could be reversed by addition of leukamenin E. This suggests that leukamenin E may act as a potential ethylene action antagonist in the inhibition of lettuce root hair development. We conclude that leukamenin E may curb root hair development by interfering with ethylene action at concentrations above 10 μM and inhibits root growth via inhibition of cell division at concentrations above 50 μM. Lan Ding and Linlin Qi contributed equally to this work.     

Phenolic biosynthesis,leaf damage,and insect herbivory in birch (Betula pendula)

The effect of both caterpillar herbivory and artificial damage on phenylalanine ammonia lysase (PAL) activity of birch foliage was measured, using an intact cell assay. After artificial damage there was a small increase in PAL activity in damaged leaves but no change in adjacent undamaged ones. Insect grazing produced a larger increase in PAL activity, and the enzyme activity was also increased in adjacent undamaged leaves. Artificial damage increased the phenolic levels of the damaged leaves. Insect grazing caused a larger, longer-lasting increase in phenolic levels and also elevated phenolic levels in undamaged leaves. The possible role of these wound-induced biochemical changes in birch is discussed.     

Effects of Quantitative Variation in Allelochemicals in <Emphasis Type="Italic">Plantago lanceolata</Emphasis> on Development of a Generalist and a Specialist Herbivore and their Endoparasitoids

Studies in crop species show that the effect of plant allelochemicals is not necessarily restricted to herbivores, but can extend to (positive as well as negative) effects on performance at higher trophic levels, including the predators and parasitoids of herbivores. We examined how quantitative variation in allelochemicals (iridoid glycosides) in ribwort plantain, Plantago lanceolata, affects the development of a specialist and a generalist herbivore and their respective specialist and generalist endoparasitoids. Plants were grown from two selection lines that differed ca. 5-fold in the concentration of leaf iridoid glycosides. Development time of the specialist herbivore, Melitaea cinxia, and its solitary endoparasitoid, Hyposoter horticola, proceeded most rapidly when reared on the high iridoid line, whereas pupal mass in M. cinxia and adult mass in H. horticola were unaffected by plant line. Cotesia melitaearum, a gregarious endoparasitoid of M. cinxia, performed equally well on hosts feeding on the two lines of P. lanceolata. In contrast, the pupal mass of the generalist herbivore, Spodoptera exigua, and the emerging adult mass of its solitary endoparasitoid, C. marginiventris, were significantly lower when reared on the high line, whereas development time was unaffected. The results are discussed with regards to (1) differences between specialist and generalist herbivores and their natural enemies to quantitative variation in plant secondary chemistry, and (2) potentially differing selection pressures on plant defense.     

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

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

Effect of Phenolic Acids in Soil under and Between Rows of a Prior Sorghum (Sorghum bicolor) Crop on Germination, Emergence, and Seedling Growth of Peanut (Arachis hypogea)

Sorghum bicolor is an allelopathic crop that reduces the yield of succeeding crops. We have assessed its effect on the germination, emergence, and seedling growth of Arachis hypogea sown in soil that had had a prior sorghum cropping. A. hypogea was sown on rows and interrows of a previous sorghum crop in 1997 and 1998 in Senegal. Seedling establishment (germination rate and seedling weight) was better between rows than on rows of the previous crop. The highest concentrations of phenolic compounds occurred in the rows in 1998, while contents of row and interrow soils were similar in 1997. Vanillic acid was the main component of the six chemicals found in 1997 soils, whereas the 1998 soil samples contained mainly p-hydroxybenzoic acid, p-hydroxybenzaldehyde, vanillic, and p-coumaric acids (10 phenolics identified). The germination of peanut seeds in water (control), soil water extracts, and mixtures of pure phenolics (equivalent to those in 1997 and 1998 soil samples) was tested. All extracts inhibited germination compared to controls, but there was no significant difference among treatments, i.e., the inhibition was the same for seeds in soil solutions and those in the respective phenolic mixtures. Similarly, there were no significant differences among the germination rates in soil water extracts of rows and interrows or in the pure phenolic mixtures of rows and interrows. We propose a geometrical sowing pattern for peanuts between the rows of the previous sorghum crop to escape the latter's "allelopathic heritage."     

The Effects of Arbuscular Mycorrhizal Fungi on Direct and Indirect Defense Metabolites of Plantago lanceolata L.

Arbuscular mycorrhizal fungi can strongly influence the metabolism of their host plant, but their effect on plant defense mechanisms has not yet been thoroughly investigated. We studied how the principal direct defenses (iridoid glycosides) and indirect defenses (volatile organic compounds) of Plantago lanceolata L. are affected by insect herbivory and mechanical wounding. Volatile compounds were collected and quantified from mycorrhizal and non-mycorrhizal P. lanceolata plants that underwent three different treatments: 1) insect herbivory, 2) mechanical wounding, or 3) no damage. The iridoids aucubin and catalpol were extracted and quantified from the same plants. Emission of terpenoid volatiles was significantly higher after insect herbivory than after the other treatments. However, herbivore-damaged mycorrhizal plants emitted lower amounts of sesquiterpenes, but not monoterpenes, than herbivore-damaged non-mycorrhizal plants. In contrast, mycorrhizal infection increased the emission of the green leaf volatile (Z)-3-hexenyl acetate in untreated control plants, making it comparable to emission from mechanically wounded or herbivore-damaged plants whether or not they had mycorrhizal associates. Neither mycorrhization nor treatment had any influence on the levels of iridoid glycosides. Thus, mycorrhizal infection did not have any effect on the levels of direct defense compounds measured in P. lanceolata. However, the large decline in herbivore-induced sesquiterpene emission may have important implications for the indirect defense potential of this species.     

Sensory Thresholds of Selected Phenolic Constituents from Thyme and their Antioxidant Potential in Sunflower Oil

The odor detection thresholds of carvacrol (5-isopropyl-2-methyl-phenol), thymol (2-isopropyl-5-methyl-phenol) and p-cymene 2,3-diol (2,3-dihydroxy-4-isopropyl-1-methyl-benzene) in sunflower oil, determined by the three-alternative, forced-choice procedure, were 30.97, 124 and 794.33 mg kg⁻¹, respectively. Sunflower oil containing 13, 70, or 335 mg kg⁻¹ of carvacrol, thymol or p-cymene 2,3-diol, respectively, was judged to be similar (P < 0.01) in taste and odor to its antioxidant-free counterpart. The rate constant of sunflower oil oxidation, measured from the increase in peroxide value during storage at 25 °C, was 9.2 × 10⁻⁹ mol kg⁻¹ s⁻¹ while the rate constants were 9.3 × 10⁻⁹, 9.8 × 10⁻⁹, and 4.3 × 10⁻⁹ mol kg⁻¹ s⁻¹ in the presence of 13 mg kg⁻¹ carvacrol, 70 mg kg⁻¹ thymol, and 335 mg kg⁻¹ p-cymene 2,3-diol, respectively. At a level of 335 mg kg⁻¹, p-cymene 2,3-diol did not impart flavor taints and effected a 46.7% reduction in the rate of oxidation of sunflower oil. These findings indicate that the diphenolic p-cymene 2,3-diol could potentially replace synthetic antioxidants and is a valuable addition to the antioxidants used by the food industry in its quest to meet consumer demands for synthetic-additives-free and ‘natural’ foods.     

Herbivore-Induced Volatiles in the Perennial Shrub, <Emphasis Type="Italic">Vaccinium corymbosum</Emphasis>, and Their Role in Inter-branch Signaling

Herbivore feeding activates plant defenses at the site of damage as well as systemically. Systemic defenses can be induced internally by signals transported via phloem or xylem, or externally transmitted by volatiles emitted from the damaged tissues. We investigated the role of herbivore-induced plant volatiles (HIPVs) in activating a defense response between branches in blueberry plants. Blueberries are perennial shrubs that grow by initiating adventitious shoots from a basal crown, which produce new lateral branches. This type of growth constrains vascular connections between shoots and branches within plants. While we found that leaves within a branch were highly connected, vascular connectivity was limited between branches within shoots and absent between branches from different shoots. Larval feeding by gypsy moth, exogenous methyl jasmonate, and mechanical damage differentially induced volatile emissions in blueberry plants, and there was a positive correlation between amount of insect damage and volatile emission rates. Herbivore damage did not affect systemic defense induction when we isolated systemic branches from external exposure to HIPVs. Thus, internal signals were not capable of triggering systemic defenses among branches. However, exposure of branches to HIPVs from an adjacent branch decreased larval consumption by 70% compared to those exposed to volatiles from undamaged branches. This reduction in leaf consumption did not result in decreased volatile emissions, indicating that leaves became more responsive to herbivory (or “primed”) after being exposed to HIPVs. Chemical profiles of leaves damaged by gypsy moth caterpillars, exposed to HIPVs, or non-damaged controls revealed that HIPV-exposed leaves had greater chemical similarities to damaged leaves than to control leaves. Insect-damaged leaves and young HIPV-exposed leaves had higher amounts of endogenous cis-jasmonic acid compared to undamaged and non-exposed leaves, respectively. Our results show that exposure to HIPVs triggered systemic induction of direct defenses against gypsy moth and primed volatile emissions, which can be an indirect defense. Blueberry plants appear to rely on HIPVs as external signals for inter-branch communication.     

Spodoptera exigua Oviposition and Larval Feeding Preferences for Pigweed, Amaranthus hybridus, over Squaring Cotton, Gossypium hirsutum, and a Comparison of Free Amino Acids in Each Host Plant

The beet armyworm, Spodoptera exigua (Hübner), can utilize a number of different host plants for oviposition and larval development, but some host plants are preferred over others. This study, using cage choice tests and olfactometer assays, demonstrates the beet armyworm's preference for pigweed, Amaranthus hybridus L., over cotton, Gossypium hirsutum L. Cage and olfactometer choice assays indicated that olfaction plays an important role in host selection by ovipositing females. First instars exhibited no feeding preference, but the more mobile third instars showed a significant feeding preference for excised pigweed leaves. The higher quantities and more diverse accumulations of free amino acids in pigweed might, in conjunction with other physiochemical and mechanical attractant and deterrent factors in the two-plant species system, play roles in the tendency of the beet armyworm to select pigweed over cotton and in providing a superior array of easily absorbed amino acids as compared to cotton.     

HPLC测定金银忍冬叶、花、果实中绿原酸的含量

建立了反相高效液相色谱法测定金银忍冬叶、花和果实中的绿原酸含量的测定方法。利用超声波辅助提取金银忍冬叶、花和果实中的绿原酸,采用采用反相高效液相色谱法测定金银忍冬叶、花和果实中的绿原酸含量。结果表明:在流动相为甲醇(A)∶1.0%的冰醋酸溶液(B)=27∶73,色谱柱为Alltima C18Column(250 mm×4.6 mm,5μm),流动相流速为1.0 mL/min,检测波长为327 nm,柱温为25℃的实验条件下测定金银忍冬叶、花和果实中的绿原酸含量分别为:(31.64±2.21)mg/g,(17.45±1.05)mg/g,(5.14±1.39)mg/g,且宁夏金银忍冬植株不同部位中绿原酸类化合物的含量分布为:叶花果实。回收率为99.74%,RSD为0.23%。所建立的方法稳定,操作简单,结果可靠。     

Antagonism between jasmonate- and salicylate-mediated induced plant resistance: effects of concentration and timing of elicitation on defense-related proteins,herbivore, and pathogen performance in tomato

The jasmonate (JA) and salicylate (SA) signaling pathways in plants provide resistance to herbivorous insects and pathogens. It is known that these pathways interact, sometimes resulting in antagonism between the pathways. We tested how the timing and concentration of elicitation of each pathway influenced the interaction between the jasmonate and salicylate pathways measured in terms of five biochemical responses and biological resistance to caterpillars and bacteria. The salicylate pathway had a stronger effect on the jasmonate pathway than did the reverse. The negative signal interaction was generated by two distinct paths in the plant. A negative interaction in the biochemical expression of the two pathways was most consistent in the simultaneous elicitation experiments compared to when the elicitors were temporally separated by two days. Herbivore bioassays with Spodoptera exigua also consistently reflected an interaction between the two pathways in the simultaneous elicitation experiments. The negative signal interaction reducing biological resistance to the herbivore was also demonstrated in some temporally separated treatment combinations where attenuation of the biochemical response was not evident. Concentration of the elicitors had an effect on the pathway interaction with consistent biochemical and biological antagonism in the high concentration experiments and inconsistent antagonism in the low concentration experiments. The bacterial pathogen, Pseudomonas syringae pv. tomato (Pst), consistently showed reduced lesion development on plants with SA responses activated and, in some experiments, on JA-elicited plants. Resistance to Pst was not reduced or enhanced in dual-elicited plants. Thus, signal interaction is most consistent when elicitors are applied at the same time or when applied at high doses. Signal interaction affected the herbivore S. exigua, but not the pathogen Pst.     

Effects of secondary metabolites from marine algae on feeding by the sea urchin,Lytechinus variegatus

A bioassay was developed to test the hypothesis that secondary metabolites from marine algae affect feeding by sea urchins. During experiments measuring chemoreception and gustation, feeding by the regular sea urchin,Lytechinus variegatus (Lamarck), was inhibited by extracts from the green marine alga,Caulerpa prolifera (Forsskal) Lamouroux andCymopolia barbata (Linneaus) Lamouroux. Cymopol, a monoterpene-bromohydroquinone component ofC. barbata, was partially responsible for the inhibited feeding observed in tests of theCymopolia crude extract. Caulerpenyne, an oxygenated sesquiterpene fromC. prolifera, was responsible for all of the urchin feeding inhibition observed in tests of theC. prolifera crude extract. Feeding was not affected by: (1) extracts from several otherCaulerpa species includingC. mexicana (Sonder) J. Agardh,C. ashmaedii Harvey,C. racemosa v.macrophysa (Kutzing) Taylor, andC. racemosa v.laetevirens (Montagne) Weber-van Bosse; (2) caulerpin, an indole-containing pigment isolated from all of theCaulerpa species exceptC. mexicana; and (3) an extract from the red marine alga,Gracilaria foliifera v.angustissima (Harvey) Taylor, which has no known secondary metabolites. Feeding inhibition was independent of the test diameter which correlated with the reproductive state of the urchins. Feeding inhibition was also independent of the starvation periods between experiments, and the temperature and salinity in ranges tolerated byL. variegatus obtained from the Florida Gulf Coast. The data strongly suggest that at least one alga,Caulerpa prolifera, is chemically defended against a dominant omnivore in its community.     

Above- and Below-Ground Terpenoid Aldehyde Induction in Cotton, Gossypium herbaceum, Following Root and Leaf Injury

Studies on induced defenses have predominantly focused on foliar induction by above-ground herbivores and pathogens. However, roots are attacked by as many if not more phytophages than shoots, so in reality plants are exposed to above- and below-ground attack. Here, we report effects of foliar and/or root damage on terpenoid aldehyde accumulation in cotton (Gossypium herbaceum). Using HPLC, we analyzed concentrations of individual terpenoid aldehydes in foliage and root tissue. In undamaged plants, terpenoid aldehydes were concentrated in young immature main leaves. Concentrations in side leaves, branching from the main leaves, did not differ among leaf position. Above-ground feeding by Spodopterta exigua larvae on a mature leaf enhanced terpenoid concentrations in immature leaves but not in the damaged leaf. In particular, concentrations of hemigossypolone and the heliocides 1 and 4 were enhanced following herbivory. Root herbivory by wireworms (Agriotes lineatus) also resulted in an increase in terpenoid levels in the foliage. In contrast with foliar herbivory, both immature and mature leaves were induced. However, the level of induction after root herbivory was much lower compared to foliar herbivory. Plants exposed to root herbivory also had significantly higher levels of terpenoid aldehydes in root tissue, while no such effect was found following foliar herbivory. Plants exposed to both root and foliar herbivory appeared to induce primarily above-ground at the cost of below-ground defense. The implications for above- and below-ground Mutitrophic interactions are discussed.     

Glycine Protects Against Strong Protein-Denaturing Activity of Oleuropein, a Phenolic Compound in Privet Leaves

In previous studies, we reported the existence of a high concentration of free glycine in the digestive juice of several Lepidoptera larvae, particularly in the digestive juice of species that feed on the privet tree, Ligustrum oblusifolium. The water extract of privet leaves showed very strong protein-denaturing activity and lysine-decreasing activity, which closely resembled activity of oxidized polyphenolics. Addition of 1% glycine to the extract could completely inhibit these activities. Free glycine may be secreted into the digestive juice by larvae as an adaptive mechanism for chemical defense against its host plants. The protein-denaturing compound in privet leaves is present in the cytosol or in the vacuoles of the leaf cells. The compound does not show protein-denaturing activity without oxidation, but when mixed with intact organelles under low osmotic conditions to give an osmotic shock, a very high protein-denaturing activity is produced. Our results suggest that the privet tree is endowed with a defense mechanism in which a stable compound in the cytosol or in the vacuoles is activated into a chemically active denaturant by an enzyme present in the organelles (including chloroplasts) after the leaves are eaten by insects and the organelles are broken by osmotic shock or by digestive mechanisms. Based upon HPLC and NMR data, we conclude that the denaturing compound is oleuropein, an o-dihydroxyphenolic compound. This compound makes up 3% of the wet weight of privet leaves. The protein-denaturing activity of purified oleuropein activated by the leaf enzyme is high enough to account for all the denaturing activity in a water extract of privet leaves. The denaturing reaction is completely inhibited by free glycine. Our results suggest that the protein-denaturing activity and lysine-decreasing activity of privet leaves are caused by oxidized polyphenolics, and that some insects secrete free glycine to counter the denaturing activity of oxidized phenolics. The chemical mechanism of counteraction by glycine is also considered. Free glycine in the midgut of insects probably protects proteins from denaturation by competing with proteins for oxidized phenolics.     

Diversity of the Enzymatic Activity in the Lipoxygenase Gene Family of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Lipoxygenases (LOX) catalyze the oxygenation of polyunsaturated fatty acids, the first step in the biosynthesis of a large group of biologically active fatty acid metabolites collectively named oxylipins. In the present study we report the characterization of the enzymatic activity of the six lipoxygenases found in the genome of the model plant Arabidopsis thaliana. Recombinant expressed AtLOX-1 and AtLOX-5 had comparable oxygenase activity with either linoleic acid or linolenic acid. AtLOX-2, AtLOX-3, AtLOX-4 and AtLOX-6 displayed a selective oxygenation of linolenic acid. Analyses by high-performance liquid chromatography and gas chromatography-mass spectrometry demonstrated that AtLOX-1 and AtLOX-5 are 9S-lipoxygenases, and AtLOX-2, AtLOX-3, AtLOX-4 and AtLOX-6 are 13S-lipoxygenases. None of the enzymes had dual positional specificity. The determined activities correlated with that predicted by their phylogenetic relationship to other biochemically-characterized plant lipoxygenases. Gerard Bannenberg, Marta Martínez shared first authors.     

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.     

Evaluation of the Evolution of Increased Competitive Ability (EICA) Hypothesis: Loss of Defense Against Generalist but not Specialist Herbivores

The evolution of increased competitive ability (EICA) hypothesis predicts that invasive plant species may escape their specialized natural enemies in their introduced range and subsequently evolve with a decrease in investment in anti-herbivore chemical defenses relative to native conspecifics. We compared the chemical profile of 10 populations of US native and 20 populations of European invasive Solidago gigantea. To test for differences in inducibility between native and invasive populations, we measured secondary chemistry in both damaged and undamaged plants. We also performed bioassays with three specialist and two generalist insect herbivores from four different feeding guilds. There was no evidence that invasive populations had reduced concentrations of sesquiterpenes, diterpenes, or short-chain hydrocarbons (SCH), although significant variation among populations was detected. Sesquiterpene and diterpene concentrations were not influenced by damage to the host plant, whereas SCH concentrations were decreased by damage for both native and invasive plants. Performance of the three specialist insects was not affected by the continental origin of the host plant. However, larval mass of the generalist caterpillar Spodoptera exigua was 37% lower on native plants compared to invasive plants. The other generalist insect, a xylem-tapping spittlebug that occurs on both continents, performed equally well on native and invasive plants. These results offer partial support for the defense predictions of the EICA hypothesis: the better growth of Spodoptera caterpillars on European plants shows that some defenses have been lost in the introduced range, even though our measures of secondary chemistry did not detect differences between continents. Our results show significant variation in chemical defenses and herbivore performance across populations on both continents and emphasize the need for analysis across a broad spatial scale and the use of multiple herbivores.     

Volatile Stimuli Related to Feeding Activity of Nonprey Caterpillars, Spodoptera exigua, Affect Olfactory Response of the Predatory Mite Phytoseiulus persimilis

The effect of volatiles related to feeding activity of nonprey caterpillars, Spodoptera exigua, on the olfactory response of the predatory mites Phytoseiulus persimilis was examined in a Y-tube olfactometer. At a low caterpillar density (20 caterpillars on 10 Lima bean leaves), the predators were significantly more attracted to volatiles from infested leaves on which the caterpillars and their products were present or from infested leaves from which the caterpillars and their products had been removed when compared to volatiles from uninfested leaves. The predators, however, significantly avoided odors from 20 caterpillars and their products (mainly feces) removed from bean leaves. In contrast, at a higher caterpillar density (100 caterpillars on 10 Lima bean leaves), the predators avoided volatiles from caterpillar-infested bean leaves. Volatiles from infested leaves from which the caterpillars and their products had been removed were not preferred over volatiles from uninfested leaves. Volatiles from feces collected from 100 caterpillars were strongly avoided by the predators, while the behavior of the predatory mites was not affected by volatiles from 100 caterpillars removed from a plant. The data show that carnivorous arthropods may avoid nonprofitable herbivores. This avoidance seems to result from an interference of volatiles from herbivore products with the attraction to herbivore-induced plant volatiles.