Variation In Chemical Defenses Research Articles

Circum‐Arctic distribution of chemical anti‐herbivore compounds suggests biome‐wide trade‐off in defence strategies in Arctic shrubs

Spatial variation in plant chemical defence towards herbivores can help us understand variation in herbivore top–down control of shrubs in the Arctic and possibly also shrub responses to global warming. Less defended, non‐resinous shrubs could be more influenced by herbivores than more defended, resinous shrubs. However, sparse field measurements limit our current understanding of how much of the circum‐Arctic variation in defence compounds is explained by taxa or defence functional groups (resinous/non‐resinous). We measured circum‐Arctic chemical defence and leaf digestibility in resinous (Betula glandulosa, B. nana ssp. exilis) and non‐resinous (B. nana ssp. nana, B. pumila) shrub birches to see how they vary among and within taxa and functional groups. Using liquid chromatography–mass spectrometry (LC–MS) metabolomic analyses and in vitro leaf digestibility via incubation in cattle rumen fluid, we analysed defence composition and leaf digestibility in 128 samples from 44 tundra locations.We found biogeographical patterns in anti‐herbivore defence where mean leaf triterpene concentrations and twig resin gland density were greater in resinous taxa and mean concentrations of condensing tannins were greater in non‐resinous taxa. This indicates a biome‐wide trade‐off between triterpene‐ or tannin‐dominated defences. However, we also found variations in chemical defence composition and resin gland density both within and among functional groups (resinous/non‐resinous) and taxa, suggesting these categorisations only partly predict chemical herbivore defence. Complex tannins were the only defence compounds negatively related to in vitro digestibility, identifying this previously neglected tannin group as having a potential key role in birch anti‐herbivore defence.We conclude that circum‐Arctic variation in birch anti‐herbivore defence can be partly derived from biogeographical distributions of birch taxa, although our detailed mapping of plant defence provides more information on this variation and can be used for better predictions of herbivore effects on Arctic vegetation.

Multimodal Aposematic Defenses Through the Predation Sequence

Aposematic organisms warn predators of their unprofitability using a combination of defenses, including visual warning signals, startling sounds, noxious odors, or aversive tastes. Using multiple lines of defense can help prey avoid predators by stimulating multiple senses and/or by acting at different stages of predation. We tested the efficacy of three lines of defense (color, smell, taste) during the predation sequence of aposematic wood tiger moths (Arctia plantaginis) using blue tit (Cyanistes caeruleus) predators. Moths with two hindwing phenotypes (genotypes: WW/Wy = white, yy = yellow) were manipulated to have defense fluid with aversive smell (methoxypyrazines), body tissues with aversive taste (pyrrolizidine alkaloids) or both. In early predation stages, moth color and smell had additive effects on bird approach latency and dropping the prey, with the strongest effect for moths of the white morph with defense fluids. Pyrrolizidine alkaloid sequestration was detrimental in early attack stages, suggesting a trade-off between pyrrolizidine alkaloid sequestration and investment in other defenses. In addition, pyrrolizidine alkaloid taste alone did not deter bird predators. Birds could only effectively discriminate toxic moths from non-toxic moths when neck fluids containing methoxypyrazines were present, at which point they abandoned attack at the consumption stage. As a result, moths of the white morph with an aversive methoxypyrazine smell and moths in the treatment with both chemical defenses had the greatest chance of survival. We suggest that methoxypyrazines act as context setting signals for warning colors and as attention alerting or “go-slow” signals for distasteful toxins, thereby mediating the relationship between warning signal and toxicity. Furthermore, we found that moths that were heterozygous for hindwing coloration had more effective defense fluids compared to other genotypes in terms of delaying approach and reducing the latency to drop the moth, suggesting a genetic link between coloration and defense that could help to explain the color polymorphism. Conclusively, these results indicate that color, smell, and taste constitute a multimodal warning signal that impedes predator attack and improves prey survival. This work highlights the importance of understanding the separate roles of color, smell and taste through the predation sequence and also within-species variation in chemical defenses.

Toxin variation among salamander populations: discussing potential causes and future directions.

Amphibians produce defensive chemicals which provide protection against both predators and infections. Within species, populations can differ considerably in the composition and amount of these chemical defenses. Studying intraspecific variation in toxins and linking it to environmental variables may help us to identify the selective drivers of toxin evolution, such as predation pressure and infection risk. Recently, there has been a renewed interest in the unique toxins produced by salamanders from the genus Salamandra: the samandarines. Despite this attention, intraspecific variation has largely been ignored within Salamandra-species. The aim of this study was to investigate whether geographic variation in profiles of samandarines exists, by sampling 4 populations of Salamandra atra over its range in the Dinaric Alps. In addition, we preliminary explored whether potential variation could be explained by predation (counting the number of snake species) and infection risk (cultivation and genomic analyses of collected soil samples). Salamanders from the 4 populations differed in toxin composition and in the size of their poison glands, although not in overall toxin quantity. Nor predation nor infection risk could explain this variation, as populations barely differed in these variables. Sampling over a much broader geographic range, using better estimators for predation and infection risk, will contribute to an improved understanding of how environment may shape variation in chemical defenses. Nevertheless, as the 4 populations of S. atra did differ in their toxin profiles, we propose that this species provides an interesting opportunity for further ecological and evolutionary studies on amphibian toxins.

Defense against predators incurs high reproductive costs for the aposematic moth Arctia plantaginis.

To understand how variation in warning displays evolves and is maintained, we need to understand not only how perceivers of these traits select color and toxicity but also the sources of the genetic and phenotypic variation exposed to selection by them. We studied these aspects in the wood tiger moth Arctia plantaginis, which has two locally co-occurring male color morphs in Europe: yellow and white. When threatened, both morphs produce defensive secretions from their abdomen and from thoracic glands. Abdominal fluid has shown to be more important against invertebrate predators than avian predators, and the defensive secretion of the yellow morph is more effective against ants. Here, we focused on the morph-linked reproductive costs of secretion of the abdominal fluid and quantified the proportion of phenotypic and genetic variation in it. We hypothesized that, if yellow males pay higher reproductive costs for their more effective aposematic display, the subsequent higher mating success of white males could offer one explanation for the maintenance of the polymorphism. We first found that the heritable variation in the quantity of abdominal secretion was very low (h2 = 0.006) and the quantity of defensive secretion was not dependent on the male morph. Second, deploying the abdominal defensive secretion decreased the reproductive output of both color morphs equally. This suggests that potential costs of pigment production and chemical defense against invertebrates are not linked in A. plantaginis. Furthermore, our results indicate that environmentally induced variation in chemical defense can alter an individual’s fitness significantly.

Critical Phenological Events Affect Chemical Defense of Plant Tissues: Iridoid Glycosides in a Woody Shrub.

Plants are chemically-complex organisms; each individual contains diverse tissue-types, has the ability to differentially allocate secondary metabolites to these tissues and can change this allocation through time. The interaction of variation in chemical defense of different tissue types and variation in chemical defense through time, however, is rarely examined and has not been studied for iridoid glycoside-producing woody plants. In this study, we quantified allocation of iridoid glycosides (IGs) to the leaves, flowers, fruits, and seeds of 25 individuals of a long-lived shrub (Lonicera x bella Zabel, Caprifoliaceae), at five important phenological timepoints (leaf-out, flowering, fruit appearance, fruit ripening, and fruit dispersal) throughout a growing season. We found that leaves had 2x higher IG concentrations during flowering and fruiting than earlier in the season (after leaf-out), and later in the season (after fruit dispersal). The individual IG driving this increase in leaves during reproduction, secologanin, was also the most abundant IG in semiripe fruits. Flowers and seeds were composed of different proportions of individual IGs than fruits or leaves, but did not change across time and had overall low concentrations of IGs. In L. x bella, phenological events such as flowering and fruiting lead to an increase in leaf chemical defense that is likely to influence interactions with leaf-feeders. Our results stress the importance of considering phenology when sampling plants for the quantification of chemical defenses.

Divergent host plant utilization by adults and offspring is related to intra-plant variation in chemical defences.

Adult and juvenile herbivores of the same species can use divergent feeding strategies, and thus may inhabit and consume different parts of the plant. Because the expression of chemical defences often differs between host plant tissues, this variation may result in disparate performance outcomes for adult and juvenile conspecifics that feed on distinct dietary substrates. The goal of this study was to evaluate how host range may differ between adults and juveniles in a generalist herbivore. We addressed the impacts of among- and within-plant defence variation using the wood-feeding Asian longhorned beetle (Anoplophora glabripennis) and three host plants having a range of putative resistance. Impacts of host plants on adult and offspring performance were assessed using a series of controlled bioassays. We evaluated adult-feeding and egg-laying behaviours in choice and no-choice experiments using the different hosts, and subsequent offspring establishment. We then evaluated host plant chemical composition related to nutrition and defence. Different plants had strong impacts on adult performance, but these patterns did not extend to effects on offspring. Females were capable of developing eggs when provided Acer rubrum, but not Populus deltoides or Populus tomentosa. Females that produced eggs by feeding on A.rubrum, however, deposited eggs into all three plant species. Larvae hatched and consumed tissues in all three hosts. The differences between adult and juvenile utilization of Populus spp. were reflected in markedly higher salicinoid phenolic concentrations in bark (>2% dw), while wood had trace quantities. Our results demonstrate that plant resistance mechanisms can differentially act upon adult and juvenile life stages of a polyphagous herbivore when there is differential expression of chemical defences among plant tissue types. Anoplophora glabripennis has been a globally successful invader due in part to its broad host range, and our results suggest a mechanism that permits the beetle to exploit marginally resistant plants. This study has implications for how host range differs between insect feeding stages, which is particularly important for invasive, polyphagous species encountering novel food sources.

Physical, but not chemical, antiherbivore defense expression is related to the clustered spatial distribution of tropical trees in an Amazonian forest.

The conspecific negative density dependence hypothesis states that mortality of young trees (seedlings and saplings) is higher near conspecific adults due to mechanisms such as allelopathy, intraspecific competition, and pest facilitation, explaining why in the tropics, most of plant species tend to be rare and live dispersed. However, there are some tree species that defy this expectation and grow in large clusters of conspecific juveniles and adults. We hypothesize that conspecifics living in clusters show higher and/or more variable defensive profiles than conspecifics with dispersed distributions.We evaluated our hypothesis by assessing the expression of physical leaf traits (thickness, and the resistance to punch, tear and shear) and leaf chemical defenses for six clustered and six non‐clustered tree species in Yasuní National Park, Ecuadorian Amazon. We ask ourselves whether (a) clustered species have leaves with higher physical resistance to damage and more chemical defenses variability than non‐clustered species; (b) saplings of clustered species may show higher physical resistance to damage and higher variation on chemical leaf defenses than their conspecific adults, and (c) saplings of non‐clustered species show lower resistance to physical damage and lower variation in chemical defenses compared to conspecific adults.Overall, our study did not support any of our hypotheses. Remarkably, we found that soluble metabolites were significantly species‐specific.Our study suggests that plants physical but not chemical leaf antiherbivore defenses may be a crucial strategy for explaining survivorship of clustered species. Trees in Yasuní may also fall along a suite of tolerance/escape/defense strategies based on limitations of each species physiological constraints for survival and establishment. We conclude that other mechanisms, such as those related to indirect defenses, soil nutrient exploitation efficiency, volatile organic compounds, delayed leaf‐greening, and seed dispersal mechanisms, shall be evaluated to understand conspecific coexistence in this forest.

Divergence in Glucosinolate Profiles between High- and Low-Elevation Populations of Arabidopsis halleri Correspond to Variation in Field Herbivory and Herbivore Behavioral Preferences.

Variation in local herbivore pressure along elevation gradients is predicted to drive variation in plant defense traits. Yet, the extent of intraspecific variation in defense investment along elevation gradients, and its effects on both herbivore preference and performance, remain relatively unexplored. Using populations of Arabidopsis halleri (Brassicaceae) occurring at different elevations in the Alps, we tested for associations between elevation, herbivore damage in the field, and constitutive chemical defense traits (glucosinolates) assayed under common-garden conditions. Additionally, we examined the feeding preferences and performance of a specialist herbivore, the butterfly Pieris brassicae, on plants from different elevations in the Alps. Although we found no effect of elevation on the overall levels of constitutive glucosinolates in leaves, relative amounts of indole glucosinolates increased significantly with elevation and were negatively correlated with herbivore damage in the field. In oviposition preference assays, P. brassicae females laid fewer eggs on plants from high-elevation populations, although larval performance was similar on populations from different elevations. Taken together, these results support the prediction that species distributed along elevation gradients exhibit genetic variation in chemical defenses, which can have consequences for interactions with herbivores in the field.

Testing for latitudinal gradients in defense at the macroevolutionary scale.

Plant defenses against herbivores are predicted to evolve to be greater in warmer climates, such as lower latitudes where herbivore pressure is also thought to be higher. Instead, recent findings are often inconsistent with this expectation, suggesting alternative hypotheses are needed. We tested for latitudinal gradients in plant defense evolution at the macroevolutionary scale by characterizing plant chemical defenses across 80 species of the evening primroses, spanning both North and South America. We quantified phenolics in leaves, flowers, and fruits, using advanced analytical chemistry techniques. Dominant individual ellagitannin compounds, total concentrations of ellagitannins, flavonoids, total phenolics, and compound diversity were quantified. Variation in these compounds was predicted with latitude, temperature, precipitation, and continent using phylogenetic generalized least squares (PGLS) multiple regression models. Latitude did not strongly explain variation in chemical defenses. Instead, fruit total ellagitannins, oenothein A, and total phenolics were greater in species inhabiting regions with colder climates. Using analytical chemistry and 80 species in two continents, we show that contrary to classic predictions, concentrations of secondary metabolites are not greater at lower latitudes or in warmer regions. We propose higher herbivore pressure in colder climates and gradients in resource availability as potential drivers of the observed patterns in Oenothera.

Testing the optimal defense hypothesis in nature: Variation for glucosinolate profiles within plants.

Plants employ highly variable chemical defenses against a broad community of herbivores, which vary in their susceptibilities to specific compounds. Variation in chemical defenses within the plant has been found in many species; the ecological and evolutionary influences on this variation, however, are less well-understood. One central theory describing the allocation of defenses in the plant is the Optimal Defense Hypothesis (ODH), which predicts that defenses will be concentrated in tissues that are of high fitness value to the plant. Although the ODH has been repeatedly supported within vegetative tissues, few studies have compared vegetative and reproductive tissues, and the results have not been conclusive. We quantified variation in glucosinolate profile and tissue value between vegetative and reproductive tissues in Boechera stricta, a close relative of Arabidopsis. B. stricta manufactures glucosinolates, a set of defensive compounds that vary genetically and are straightforward to quantify. Genetic diversity in glucosinolate profile has been previously demonstrated to be important to both herbivory and fitness in B. stricta; however, the importance of glucosinolate variation among tissues has not. Here, we investigate whether allocation of glucosinolates within the plant is consistent with the ODH. We used both clipping experiments on endogenous plants and ambient herbivory in a large-scale transplant experiment at three sites to quantify fitness effects of loss of rosette leaves, cauline leaves, and flowers and fruits. We measured glucosinolate concentration in leaves and fruits in the transplant experiment, and asked whether more valuable tissues were more defended. We also investigated within-plant variation in other aspects of the glucosinolate profile. Our results indicated that damage to fruits had a significantly larger effect on overall fitness than damage to leaves, and that fruits had much higher concentrations of glucosinolates, supporting the ODH. This is, to the best of our knowledge, the first study to explicitly compare both tissue value and chemical defense concentrations between vegetative and reproductive tissues under natural conditions.

Variation in Chemical Defense Among Natural Populations of Common Toad, Bufo bufo, Tadpoles: the Role of Environmental Factors.

Defensive toxins are widespread in nature, yet we know little about how various environmental factors shape the evolution of chemical defense, especially in vertebrates. In this study we investigated the natural variation in the amount and composition of bufadienolide toxins, and the relative importance of ecological factors in predicting that variation, in larvae of the common toad, Bufo bufo, an amphibian that produces toxins de novo. We found that tadpoles' toxin content varied markedly among populations, and the number of compounds per tadpole also differed between two geographical regions. The most consistent predictor of toxicity was the strength of competition, indicating that tadpoles produced more compounds and larger amounts of toxins when coexisting with more competitors. Additionally, tadpoles tended to contain larger concentrations of bufadienolides in ponds that were less prone to desiccation, suggesting that the costs of toxin production can only be afforded by tadpoles that do not need to drastically speed up their development. Interestingly, this trade-off was not alleviated by higher food abundance, as periphyton biomass had negligible effect on chemical defense. Even more surprisingly, we found no evidence that higher predation risk enhances chemical defenses, suggesting that low predictability of predation risk and high mortality cost of low toxicity might select for constitutive expression of chemical defense irrespective of the actual level of predation risk. Our findings highlight that the variation in chemical defense may be influenced by environmental heterogeneity in both the need for, and constraints on, toxicity as predicted by optimal defense theory.

Photoperiod-Induced Geographic Variation in Plant Defense Chemistry

Spatial variation in chemical defense of plants can be caused by genetic, biotic, and abiotic factors. For example, many plants exhibit a latitudinal cline in chemical defense, potentially due to latitudinal variation in abiotic environmental factors such as the light regime during the growing season. In the worldwide distributed Plantago lanceolata, the levels of deterrent iridoid glycosides (IGs), aucubin and catalpol, vary geographically, including latitudinally. To examine whether latitudinal variation in photoperiod can explain part of this geographic variation, plants from the Netherlands and Finland were exposed to two different photoperiods, simulating the Dutch (middle European) and Finnish (northern European) light period during the growing season. The experiment showed that although most variation in IG content was genetic, plants from both Dutch and Finnish origin produce relatively more catalpol under a northern European than under a middle European photoperiod. Our results confirm that latitudinal effects on photoperiod can contribute to geographic variation in plant defense chemistry, which should be considered when studying latitudinal clines in plant-enemy interactions.

Host plant-specific remodeling of midgut physiology in the generalist insect herbivore Trichoplusia ni

Species diversity in terrestrial ecosystems is influenced by plant defense compounds that alter the behavior, physiology, and host preference of insect herbivores. Although it is established that insects evolved the ability to detoxify specific allelochemicals, the mechanisms by which polyphagous insects cope with toxic compounds in diverse host plants are not well understood. Here, we used defended and non-defended plant genotypes to study how variation in chemical defense affects midgut responses of the lepidopteran herbivore Trichoplusia ni, which is a pest of a wide variety of native and cultivated plants. The genome-wide midgut transcriptional response of T. ni larvae to glucosinolate-based defenses in the crucifer Arabidopsis thaliana was characterized by strong induction of genes encoding Phase I and II detoxification enzymes. In contrast, the response of T. ni to proteinase inhibitors and other jasmonate-regulated defenses in tomato (Solanum lycopersicum) was dominated by changes in the expression of digestive enzymes and, strikingly, concomitant repression of transcripts encoding detoxification enzymes. Unbiased proteomic analyses of T. ni feces demonstrated that tomato defenses remodel the complement of T.ni digestive enzymes, which was associated with increased amounts of serine proteases and decreased lipase protein abundance upon encountering tomato defense chemistry. These collective results indicate that T. ni adjusts its gut physiology to the presence of host plant-specific chemical defenses, and further suggest that plants may exploit this digestive flexibility as a defensive strategy to suppress the production of enzymes that detoxify allelochemicals.

Chemical Defense Across Three Trophic Levels: Catalpa bignonioides, the Caterpillar Ceratomia catalpae, and its Endoparasitoid Cotesia congregata

Plant secondary chemistry can vary among plant tissues, individuals, and populations, and this variation has population-level consequences for upper trophic levels. In this study, we examined the multi-trophic consequences of variation in iridoid glycosides, which are a component of plant defense against generalist herbivores and also contribute to the unpalatability of sequestering herbivores to both vertebrate and invertebrate predators. Several populations of Catalpa bignonioides were located and examined for the presence of the Catalpa Sphinx, Ceratomia catalpae, a specialist herbivore of Catalpa. We quantified iridoid glycoside content in Catalpa Sphinx caterpillars and in damaged and undamaged C. bignonioides leaves. Overall, leaves of C. bignonioides that were damaged by Catalpa Sphinx caterpillars contained lower concentrations of two major iridoid glycosides, catalpol and catalposide, than leaves of undamaged trees from naturally occurring populations. Catalpa Sphinx caterpillars sequester only catalpol, and increasing catalpol and catalposide concentrations in leaves were associated with increased catalpol sequestration by caterpillars. The parasitoid Cotesia congregata develops successfully inside catalpol-sequestering Catalpa Sphinx caterpillars, and we examined parasitoid larvae for the presence of catalpol. Parasitoid larvae dissected from caterpillars contained catalpol, but at lower concentrations than their host caterpillars. The variation in chemical defense documented here has rarely been documented over multiple trophic levels, but such resolved systems are ideal for examining competing hypotheses about the effects of plant secondary metabolites on higher trophic levels.

Iridoid Glycoside Variation in the Invasive Plant Dalmatian Toadflax, Linaria dalmatica (Plantaginaceae), and Sequestration by the Biological Control Agent, Calophasia lunula

Invasive plant species can have significant ecological and economic impacts. Although numerous hypotheses highlight the importance of the chemical defenses of invasive plant species, the chemical ecology of many invasive plants has not yet been investigated. In this study, we provide the first quantitative investigation of variation in iridoid glycoside concentrations of the invasive plant Dalmatian toadflax (Linaria dalmatica). We examined variation in chemical defenses at three levels: (1) variation within and among populations; (2) variation due to phenology and/or seasonal differences; and (3) variation among plant parts (leaves, flowers, and stems). Further, we examined two biological control agents introduced to control L. dalmatica for the ability to sequester iridoid glycosides from this invasive plant. Results indicate that L. dalmatica plants can contain high concentrations of iridoid glycosides (up to 17.4% dry weight of leaves; mean = 6.28 ± 0.5 SE). We found significant variation in iridoid glycoside concentrations both within and among plant populations, over the course of the growing season, and among plant parts. We also found that one biological control agent, Calophasia lunula (Lepidoptera: Noctuidae), was capable of sequestering antirrhinoside, an iridoid glycoside found in L. dalmatica, at levels ranging from 2.7 to 7.5% dry weight. A second biological control agent, Mecinus janthinus (Coleoptera: Curculionidae), a stem-mining weevil, did not sequester iridoid glycosides. The demonstrated variation in L. dalmatica chemical defenses may have implications for understanding variation in the degree of invasiveness of different populations as well as variation in the efficacy of biological control efforts.

Feeding preferences and host associations of specialist marine herbivores align with quantitative variation in seaweed secondary metabolites

Consequences of congeneric quantitative variation in secondary metabolites of seaweeds on diet specificity and host association in specialist marine herbivores have received little attention. We investigated quantitative variation in caulerpenyne and oxytoxin 1 in 7 species of green seaweeds from the genus Caulerpa, along with the feeding preferences and host associations of 4 co-occurring sacoglossan molluscs. C. taxifolia and C. sertularioides contained high concentrations of metabolites and were preferred least by all herbivores. Algae with intermediate metabolite concentrations (C. racemosa, C. serrulata, and C. cupressoides) were preferred by Elysia tomentosa and Lobiger viridis. Oxynoe viridis and Stiliger smaragdinus had strong preferences for different low concentration Caulerpa species (C. racemosa var. laetevirens and C. lentillifera), suggesting not all feeding preferences are based exclusively on the major metabolites. In situ host associations of L. viridis and S. smaragdinus mirrored their feeding preferences, but this was not the case for E. tomentosa. Furthermore, those algal species with the highest and lowest metabolite concentrations had the lowest overall densities of sacoglossans. The results imply that the direct influence of quantitative variation in Caulerpa chemistry may only be limited to host associations in some sacoglossans. However, feeding pressure from multiple herbivore species with unique preferences could still contribute to variation in chemical defence amongst congeneric algae.

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.

Secondary metabolite and inorganic contents in Cystodytes sp. (Ascidiacea): temporal patterns and association with reproduction and growth

Temporal variation is one of the least known components of defence production in marine organisms. Here we examined whether there is a predictable temporal pattern in the production of chemical and structural defences by a marine invertebrate. To assess the seasonal variation in chemical defence, we measured ascididemin, the main pyridoacridine alkaloid in the blue Mediterranean morph of the ascidian Cystodytes sp. Structural defence variability was assessed on the basis of colony ash content, as it contains mainly spicules. Ascididemin concentration and the colony ash content displayed an annual cycle, reaching a minimum in spring and peaking in summer. Cross-correlation analyses with existing data on growth and reproduction suggested a significant trade-off between investment in reproduction and the other biological parameters considered (growth, inorganic content and ascididemin concentration). Our results suggest that optimization of resource allocation, probably influenced by biotic interactions and physical factors, shaped the temporal trends observed in secondary metabolite concentration and inorganic content.

Phenological Variation in Chemical Defense of the Pipevine Swallowtail, Battus philenor

Larvae of the pipevine swallowtail, Battus philenor, feed on plants in the genus Aristolochia, which contains aristolochic acids, toxic alkaloids unique to the Aristolochiaceae. Pipevine swallowtails sequester these compounds and, as a consequence, are chemically defended against many natural enemies. In California, the primary aristolochic acid present in the butterfly is aristolochic acid I. Newly eclosed adult females possess greater amounts of these sequestered toxins compared to males. However, over the course of the flight season, the aristolochic acid content of females in the population declines, whereas male aristolochic acid content remains relatively constant. Transference of sequestered aristolochic acids to eggs by females might explain the decline of these sequestered chemical defenses observed over time. We found no evidence that males transfer aristolochic acids to females via the spermatophore. The possibility that females at the end of the flight season may be automimics of males is discussed. Temporal variation in the aristolochic acid defenses exhibited by this pipevine swallowtail population is both age- and sex-dependent.

Within-Plant Variation In Glucosinolate Concentrations of Raphanus sativus Across Multiple Scales

Variation in chemical defenses remains underexplored. In particular, little is known about patterns of variation at small scales within leaves and spatial variation of induction. I examined variation in glucosinolate concentrations in the leaves of Raphanus sativus at several different spatial scales in two related experiments. I used samples equivalent in area to the amount an intermediate-sized caterpillar might eat in 1 d, a smaller scale than used in most previous studies. I examined variation due to induction and leaf age and small-scale spatial variation within leaves. The mean and variance of glucosinolate concentrations were higher in induced plants, young leaves, and the proximal half of leaves. Higher glucosinolate concentrations in the proximal half of leaves are previously unreported. Small-scale variation was extreme, accounting for 57% of the total random variation, and spatially random. There was no spatial autocorrelation found at scales as small as 1-2 cm. The high degree of small-scale, spatially random variation in glucosinolate concentrations in leaves is previously unreported. This small-scale variation and the variation caused by induction may have significant effects on herbivores and could be an important component of plant defense.