THE BEGINNINGS OF ANTARCTIC MACROALGAL CHEMICAL ECOLOGY: DEFENSES AGAINST HERBIVORES IN A NITROGEN REPLETE,CARBON LIMITED OCEAN

Amsler, C. D.¹, Iken, K. B.¹, McClintock, J. B.¹, Furrow, F. B.², & Baker, B. J.^{2 1}Department of Biology, University of Alabama at Birmingham, Birmingham AL 35294-1170 USA; ²Department of Chemistry, Florida Institute of Technology, Melbourne, FL 32901 USA We examined palatability of Antarctic Peninsula macroalgae in feeding bioassays with three common, sympatric macroalgal-consuming omnivores (amphipod, fish, sea star). Antarctic macroalgae have low C:N ratios, high nitrogen contents, and are usually growth limited by carbon (light). The Carbon Nutrient Balance Hypothesis (CNBH) predicts that macroalgae would produce nitrogenous secondary metabolites for defense rather than high levels of non-nitrogenous defenses under these conditions. To date, feeding bioassays have been performed on fragments of thallus from 26 macroalgal species and 21 (81%) were rejected by at least one omnivore. Organic extracts from 13 macroalgal species rejected as thallus were used in feeding bioassays. At least one extract from 12 species (92%) was rejected by at least one omnivore, suggesting that chemical defenses against herbivores probably are present in at least some of the macroalgal species. We have identified a number of specific, non-nitrogenous secondary metabolites in these extracts and previous workers have also reported non-nitrogenous secondary metabolites from antarctic macroalgae. Additional extracts targeting nitrogenous metabolites from 25 species were subjected to thin layer chromatography and visualized by stains specific for nitrogenous compounds. No nitrogenous secondary metabolites were identified by this or other methods. Nitrogenous secondary metabolites are also extremely rare in macroalgae from other areas of the world. Consequently, although our bioassays suggest that chemical defenses probably do occur, our data cast doubt on the applicability of the CNBH for predicting the chemical composition of macroalgal defenses under carbon limited conditions. (NSF OPP9814538, OPP9901076)     

DIVERSITY OF EUKARYOTIC PICOPLANKTON IN COASTAL WATERS

Van Alstyne, K. L. Shannon Point Marine Center, Western Washington University, 1900 Shannon Point Road, Anacortes, WA 98221 USA Ulvoid green macroalgae, such as Enteromorpha and Ulva, can form large blooms that have deleterious impacts on the local biota. These algae are often assumed to be very palatable for most invertebrate and vertebrate herbivores because they lack obvious physical defenses, and because there have been few reports of their producing chemical defenses. However, in laboratory feeding preference assays, the ulvoid macroalgae Enteromorpha linza and Ulva fenestrata were low preference foods for green sea urchins, Strongylocentrotus droebachiensis. Both these algae and several other species of green algae produce large quantities of dimethylsulphoniopropionate (DMSP), which is enzymatically converted to dimethyl sulfide (DMS) and acrylic acid when the algae are physically damaged. In laboratory bioassays, both DMS and acrylic acid were potent feeding deterrents towards urchins at concentrations that the urchins would be likely to encounter in the field. The precursor in this system, DMSP, was a feeding attractant. Our data provide evidence that DMSP functions as a precursor in an activated defense system in marine macroalgae and suggests a similar function in phytoplankton. The presence of this activated defense system may contribute to the persistence of macroalgal blooms by making these algae unpalatable to some species of herbivores.     

The macrophytobenthos of Terra Nova Bay (Ross Sea,Antarctica): results of three Italian scientific expeditions

Abstract

The results of a study on the benthic macroalgal flora and vegetation of Terra Nova Bay (Ross Sea) carried out during the Italian expeditions to Antarctica made in the years 1987/88, 1989/90 and 1993/94, are presented. The flora is rather poor (9 Rhodophyceae, 4 Fucophyceae, 4 Chlorophyceae), but richer than that known from adjacent areas. The vegetation is on the whole characterized by a prevalence of sciaphilous species among which Iridaea cordata, Phyllophora antarctica and Clathromorphum lemoineanum dominating in the upper infralittoral, lower infralittoral and circalittoral zones, respectively. Moreover, the results of a study on biomass of a population of I. cordata as well as of a population of P. antarctica, carried out in the austral summer 1993/94, are reported too. The values of the biomass of the population of I. cordata resulted to vary inversely with respect to those of the population of P. antarctica.     

Palatability and Chemical Defenses of Macroalgae in the Antarctic Peninsula

We examined palatability of 37 species of nonencrusting macroalgae from the Antarctic Peninsula. This represents approximately 30% of the entire antarctic macroalgal flora and 75% of the 49 nonencrusting species we collected. Organic extracts from most species were also prepared and mixed into artificial foods. We examined palatability using feeding bioassays with three common, macroalga‐consuming animals (an omnivorous antarctic rockfish, Notothenia coriiceps; an omnivorous sea star, Odontaster validus; and a herbivorous amphipod, Gondogenia antarctica). Thallus pieces from 23 of 34 macroalgal species tested with the fish (68%) were rejected. Of the 23 species rejected as thallus, organic extracts of 16 were bioassayed using the fish with 9 (56%) unpalatable. Thallus pieces from 21 of 36 macroalgal species tested with the sea star (58%) were rejected. Of the 21 species rejected as thallus, organic extracts of 20 were bioassayed using the sea stars and 14 (70%) were unpalatable. Overall, 28 of the 37 species assayed as thallus (76%) were rejected by either or both the fish and sea stars. The amphipod assay was not suitable for use with thallus but was utilized with organic extracts of 23 macroalgal species that were rejected as thallus by either or both the fish and sea stars. Of these, 14 (61%) of the species' extracts were rejected by the amphipods. Unpalatability was highest among the brown algae examined with only an ephemeral, ectocarpoid species not rejected as thallus out of 10 species tested. Of the remaining nine brown algal species, six of seven tested were also unpalatable as extracts, including all the ecologically dominant, perennial species in the area. We conclude that unpalatability to herbivores is common in antarctic macroalgae and that chemical defenses may play an important role in the unpalatability of many algal species (NSF OPP9814538, OPP9901076).     

Large mobile versus small sedentary herbivores and their resistance to seaweed chemical defenses

Summary Small, relatively sedentary herbivores like amphipods and polychaetes (mesograzers) often live on the plants they consume and should therefore view plants as both foods and living sites. Large, relatively mobile herbivores like fishes commonly move among, and feed from, many plants; they should view plants primarily as foods and rarely as potential living sites. In marine communities, fishes that consume plants are also important predators on mesograzers. Since seaweeds avoided by fishes should represent safer living sites for small herbivores, mesograzers living on and consuming seaweeds that are not eaten by fishes should have higher fitness than mesograzers living on plants preferred by fishes. In previous work, we demonstrated that seaweed secondary metabolites that deterred feeding by a fish and sea urchin had no effect on feeding by a common amphipod (Hay et al. 1987a). We then hypothesized that mesograzers would, in general, be less affected by seaweed chemical defenses than larger, more mobile herbivores like fishes. In this investigation, we evaluate the generality of this hypothesis by comparing the feeding of an omnivorous fish (Lagodon rhomboides) with that of an omnivorous, tube-building polychaete (Platynereis dumerilii) to see if the mesograzer prefers seaweeds avoided by the fish and if it is less affected by seaweed chemical defense. Platynereis dumerilii fed almost exclusively on Dictyota dichotoma, the seaweed eaten least by Lagodon rhomboides. The diterpene alcohols (dictyol-E and pachydictyol-A) produced by Dictyota significantly deterred feeding by Lagodon but did not affect, or at one concentration stimulated, feeding by Platynereis. Our data support the hypothesis that small, relatively sedentary herbivores that live on plants are more resistant to chemical defenses than are large, relatively mobile herbivores that move among many plants.     

Costs of induced defenses for the invasive plant houndstongue (<Emphasis Type="Italic">Cynoglossum officinale</Emphasis> L.) and the potential importance for weed biocontrol

Inducible plant defenses—those produced in response to herbivore feeding—are thought to have evolved as a cost-saving tactic that allows plants to enact defenses only when needed. The costs of defense can be significant, and loss of plant fitness due to commitment of resources to induced defenses could affect plant populations and play a role in determining the success or failure of weed biocontrol. We used methyl jasmonate (MeJA) to experimentally induce defenses without herbivores in invasive houndstongue plants (Cynoglossum officinale L.) in the field and measured resulting growth and fitness (plant size, seed number, and seed weight). MeJA-treated plants emitted large amounts of plant volatiles and produced leaves with twice as many trichomes as untreated plants. Plants with activated defenses had fewer leaves, were smaller, and produced nutlets that weighed less than plants not investing in defenses. These data indicate that herbivore-induced defenses are costly for houndstongue plants in their invaded range and represent significant indirect costs of herbivory beyond direct feeding damage (e.g., loss of photosynthetic tissue). Notably, the magnitude of defenses elicited upon feeding varies greatly by herbivore species and a better understanding of the costs of defense could help us predict which potential biocontrol herbivores are most likely to be effective.     

Feeding preferences of the sandhopper Orchestoidea tuberculata: the importance of algal traits

Factors such as nutritional quality and the secondary metabolite content of food resources have been shown to influence the feeding behavior of herbivores in many marine habitats. For intertidal macroalgae consumers on sandy beaches, the influence of these factors on feeding behavior and the consequences on their performance is poorly understood. In this study, we evaluated the relationships of nutritional quality, chemical defenses (phlorotannins), and the structure of three macroalgal species that form the bulk of imported wrack subsidies to beaches in southern Chile, with the feeding behavior, absorption efficiency, and growth rate of the talitrid amphipod Orchestoidea tuberculata, one of the most abundant organisms in this environment. The amphipods preferred Durvillaea antarctica over Lessonia nigrescens and Macrocystis pyrifera when simultaneously offered fresh pieces of each alga. Similar results were observed when artificial food made of dry powdered algae of each species was provided, suggesting that the structure of these three algae did not influence preference. The performance of amphipods when reared on a diet of a single algal species matched feeding preferences; higher growth rates were observed in treatments with the preferred alga, D. antarctica. These results imply that D. antarctica is a superior food item for O. tuberculata when compared to L. nigrescens or M. pyrifera, and also that the alga’s intrinsic quality (i.e., not structure) may influence dietary preference in these consumers. The higher content of proteins and carbohydrates found in D. antarctica may explain why this macroalga represents better quality food for O. tuberculata. Phlorotannin content did not have obvious negative effects on diet choice or growth, as D. antarctica, the alga with greater content of these secondary metabolites, was preferred and associated with higher growth rates of O. tuberculata. However, it is necessary to emphasize that the low phlorotannins concentrations registered in the three macroalgae species examined in this study, may not have been sufficient to deter O. tuberculata. When the amphipods were fed with each alga individually, they consumed significantly higher quantities of D. antarctica, which suggests that O. tuberculata did not eat more to compensate for the lower nutritional quality of the other algal species in order to maintain growth. Nor was compensation for lower food quality achieved by increasing absorption efficiency. Our results imply that the composition of the macroalgae arriving on the beach can significantly affect the performance and subsequent life history traits of O. tuberculata and by extension other amphipod species.     

Leaf drop affects herbivory in oaks

Leaf phenology is important to herbivores, but the timing and extent of leaf drop has not played an important role in our understanding of herbivore interactions with deciduous plants. Using phylogenetic general least squares regression, we compared the phenology of leaves of 55 oak species in a common garden with the abundance of leaf miners on those trees. Mine abundance was highest on trees with an intermediate leaf retention index, i.e. trees that lost most, but not all, of their leaves for 2–3 months. The leaves of more evergreen species were more heavily sclerotized, and sclerotized leaves accumulated fewer mines in the summer. Leaves of more deciduous species also accumulated fewer mines in the summer, and this was consistent with the idea that trees reduce overwintering herbivores by shedding leaves. Trees with a later leaf set and slower leaf maturation accumulated fewer herbivores. We propose that both leaf drop and early leaf phenology strongly affect herbivore abundance and select for differences in plant defense. Leaf drop may allow trees to dispose of their herbivores so that the herbivores must recolonize in spring, but trees with the longest leaf retention also have the greatest direct defenses against herbivores.     

Effector proteins that modulate plant--insect interactions

Insect herbivores have highly diverse life cycles and feeding behaviors. They establish close interactions with their plant hosts and suppress plant defenses. Chewing herbivores evoke characteristic defense responses distinguishable from general mechanical damage. In addition, piercing-sucking hemipteran insects display typical feeding behavior that suggests active suppression of plant defense responses. Effectors that modulate plant defenses have been identified in the saliva of these insects. Tools for high-throughput effector identification and functional characterization have been developed. In addition, in some insect species it is possible to silence gene expression by RNAi. Together, this technological progress has enabled the identification of insect herbivore effectors and their targets that will lead to the development of novel strategies for pest resistances in plants.     

A Specialist Herbivore Uses Chemical Camouflage to Overcome the Defenses of an Ant-Plant Mutualism

Many plants and ants engage in mutualisms where plants provide food and shelter to the ants in exchange for protection against herbivores and competitors. Although several species of herbivores thwart ant defenses and extract resources from the plants, the mechanisms that allow these herbivores to avoid attack are poorly understood. The specialist insect herbivore, Piezogaster reclusus (Hemiptera: Coreidae), feeds on Neotropical bull-horn acacias (Vachellia collinsii) despite the presence of Pseudomyrmex spinicola ants that nest in and aggressively defend the trees. We tested three hypotheses for how P. reclusus feeds on V. collinsii while avoiding ant attack: (1) chemical camouflage via cuticular surface compounds, (2) chemical deterrence via metathoracic defense glands, and (3) behavioral traits that reduce ant detection or attack. Our results showed that compounds from both P. reclusus cuticles and metathoracic glands reduce the number of ant attacks, but only cuticular compounds appear to be essential in allowing P. reclusus to feed on bull-horn acacia trees undisturbed. In addition, we found that ant attack rates to P. reclusus increased significantly when individuals were transferred between P. spinicola ant colonies. These results are consistent with the hypothesis that chemical mimicry of colony-specific ant or host plant odors plays a key role in allowing P. reclusus to circumvent ant defenses and gain access to important resources, including food and possibly enemy-free space. This interaction between ants, acacias, and their herbivores provides an excellent example of the ability of herbivores to adapt to ant defenses of plants and suggests that herbivores may play an important role in the evolution and maintenance of mutualisms.     

Finish line plant–insect interactions mediated by insect feeding mode and plant interference: a case study of Brassica interactions with diamondback moth and turnip aphid

There are gaps in our understanding of plant responses under different insect phytophagy modes and their subsequent effects on the insect herbivores’ performance at late season. Here we compared different types of insect feeding by an aphid, Lipaphis erysimi, and a lepidopteran, Plutella xylostella, and how this affected defensive metabolites in leaves of 2 Brassica species when plants gain maturity. Thiocyanate concentrations after P. xylostella and L. erysimi feeding activities were the same. Total phenolics was higher after the phloem feeder feeding than the folivore activity. The plants compensatory responses (i.e., tolerance) to L. erysimi feeding was significantly higher than the responses to P. xylostella. This study showed that L. erysimi had higher carbon than P. xylostella whereas nitrogen in P. xylostella was 1.42 times that in L. erysimi. Population size of the phloem feeder was not affected by plant species or insect coexistence. However, there was no correlation between plant defensive metabolites and both insects’ population size and biomass. This suggests that plant root biomass and tolerance index after different insect herbivory modes are not necessarily unidirectional. Importantly, the interaction between the folivore and the phloem feeder insects is asymmetric and the phloem feeder might be a trickier problem for plants than the folivore. Moreover, as both plants’ common and special defenses decreased under interspecific interference, we suggest that specialist insect herbivores can be more challenged in ecosystems in which plants are not involved in interspecific interference.     

Chemical and Mechanical Defenses Vary among Maternal Lines and Leaf Ages in Verbascum thapsus L. (Scrophulariaceae) and Reduce Palatability to a Generalist Insect

Intra-specific variation in host-plant quality affects herbivore foraging decisions and, in turn, herbivore foraging decisions mediate plant fitness. In particular, variation in defenses against herbivores, both among and within plants, shapes herbivore behavior. If variation in defenses is genetically based, it can respond to natural selection by herbivores. We quantified intra-specific variation in iridoid glycosides, trichome length, and leaf strength in common mullein (Verbascum thapsus L, Scrophulariaceae) among maternal lines within a population and among leaves within plants, and related this variation to feeding preferences of a generalist herbivore, Trichopulsia ni Hübner. We found significant variation in all three defenses among maternal lines, with T. ni preferring plants with lower investment in chemical, but not mechanical, defense. Within plants, old leaves had lower levels of all defenses than young leaves, and were strongly preferred by T. ni. Caterpillars also preferred leaves with trichomes removed to leaves with trichomes intact. Differences among maternal lines indicate that phenotypic variation in defenses likely has a genetic basis. Furthermore, these results reveal that the feeding behaviors of T. ni map onto variation in plant defense in a predictable way. This work highlights the importance of variation in host-plant quality in driving interactions between plants and their herbivores.     

How do predators cope with chemically defended foods?

Many prey species (including plants) deter predators with defensive chemicals. These defensive chemicals act by rendering the prey's tissues noxious, toxic, or both. Here, I explore how predators cope with the presence of these chemicals in their diet. First, I describe the chemosensory mechanisms by which predators (including herbivores) detect defensive chemicals. Second, I review the mechanisms by which predators either avoid or tolerate defensive chemicals in prey. Third, I examine how effectively free-ranging predators can overcome the chemical defenses of prey. The available evidence indicates that predators have mixed success overcoming these defenses. This conclusion is based on reports of free-ranging predators rejecting unpalatable but harmless prey, or voluntarily ingesting toxic prey.     

Comparative analysis of quantitative trait loci controlling glucosinolates,myrosinase and insect resistance in Arabidopsis thaliana

Evolutionary interactions among insect herbivores and plant chemical defenses have generated systems where plant compounds have opposing fitness consequences for host plants, depending on attack by various insect herbivores. This interplay complicates understanding of fitness costs and benefits of plant chemical defenses. We are studying the role of the glucosinolate-myrosinase chemical defense system in protecting Arabidopsis thaliana from specialist and generalist insect herbivory. We used two Arabidopsis recombinant inbred populations in which we had previously mapped QTL controlling variation in the glucosinolate-myrosinase system. In this study we mapped QTL controlling resistance to specialist (Plutella xylostella) and generalist (Trichoplusia ni) herbivores. We identified a number of QTL that are specific to one herbivore or the other, as well as a single QTL that controls resistance to both insects. Comparison of QTL for herbivory, glucosinolates, and myrosinase showed that T. ni herbivory is strongly deterred by higher glucosinolate levels, faster breakdown rates, and specific chemical structures. In contrast, P. xylostella herbivory is uncorrelated with variation in the glucosinolate-myrosinase system. This agrees with evolutionary theory stating that specialist insects may overcome host plant chemical defenses, whereas generalists will be sensitive to these same defenses.     

Biogeographical variation in brown algal polyphenolics and other secondary metabolites: comparison between temperate Australasia and North America

Summary Polyphloroglucinol phenolics are the best known example of chemical deterrents against herbivores in temperate marine systems. However, most of the research on these compounds has been done in North America, where phenolic levels in algae are often low. I show here that algae in the Orders Fucales and Laminariales in temperate Australia and New Zealand typically contain very high levels of polyphenolics-much higher than species in these orders in North America. The median value for the distribution of mean phenolic levels for 25 North American species is 1.33% total phenolics (dry wt.); for 37 Australasian species, the median is 6.20%. Significant spatial, temporal, and intraplant variation in phenolic content occurs in a number of species in Australasia, but this does not significantly alter my major conclusion. Phenolic levels in drift algae (an important food source for some herbivores) detached for up to two weeks are also not significantly different from living, attached plants. Many species in the Fucales in Australasia also contain non-polyphenolic secondary metabolites that are not found in North American species. Thus herbivores in Australasia face greater amounts, and a greater range,of putative chemical defenses in brown algae than do herbivores in similar systems in North America. Any general theory for the evolution of marine plant/herbivore interactions must take into account such broad-scale biogeographical (and taxonomic) patterns.     

Resistance to Plant Invasion? A Native Specialist Herbivore Shows Preference for and Higher Fitness on an Introduced Host

The response of native herbivores to the introduction of a new plant to the community has important implications for plant invasion. Under the Enemy Release Hypothesis introduced species become invasive because of reduced enemy control in the new range, while under the New Association Hypothesis introduced species lack effective defenses against native enemies because they do not share an evolutionary history. I tested the response of a native South-American specialist herbivore Utetheisa ornatrix (Lepidoptera: Arctiidae) to a native (Crotalaria incana) and an introduced host (Crotalaria pallida) (Fabaceae: Papilionoideae). I compared seed predation rates between the two hosts in the field, and I tested preference and performance traits with common garden experiments. Utetheisa ornatrix caused much higher seed predation rates on the introduced host than on the native host. Females also preferred to oviposit on the introduced over the native host. Additionally, larvae feeding on the introduced host had higher fitness (higher pupal weight) than larvae feeding on the native host. I discuss how the response of this specialist herbivore to this introduced host plant contradicts the predictions of the Enemy Release Hypothesis and support the New Association Hypothesis. This study shows that the New Association Hypothesis can also be true for specialist herbivores.     

Lygus hesperus （Heteroptera： Miridae） tolerates high concentrations of dietary nickel

Nickel hyperaccumulator plants contain unusually elevated levels of Ni （〉 1 000 μg Ni/g）. Some insect herbivores, including Lygus hesperus （Western tarnished plant bug）, have been observed feeding on the California Ni hyperaccumulator Streptanthus polygaloides. This bug may be able to utilize S. polygaloides as a host either through its feeding behavior or by physiological tolerance of Ni. This experiment determined the Ni tolerance of L hesperus by offering insects artificial diet amended with 0, 0.4, 1, 2, 4.5, 10, 20 and 40 mmol Ni/L and recording survival. Survival varied due to Ni concentration, with diets containing 10 mmol Ni/L and greater resulting in significantly lower survival compared to the control （0 mmol Ni/L） treatment. Insects tolerated diet containing as much as 4.5 mmol Ni/L, a relatively elevated Ni concentration. I conclude that L hesperus can feed on S. polygaloides because it is Ni-tolerant, probably due to physiological mechanisms that provide it with resistance to plant chemical defenses including elemental defenses such as hyperaccumulated Ni.     

Larval feeding structure plasticity during pre-feeding stages of echinoids: Not all species respond to the same cues

Much of the work on phenotypic plasticity has focused on inducible defenses. As a result, little is known about induced phenotypes that improve the acquisition of resources (i.e. inducible offenses). Feeding larvae of several marine invertebrate species, gastropods and echinoderms, have inducible offenses, and produce larger feeding structures when given less food. To better understand inducible offenses, I investigated when in development sea urchin and sand dollar larvae can first alter their feeding morphology in response to different concentrations of food. Food induced feeding structure changes in both sea urchin and sand dollar larvae before larvae were able to ingest food. This suggests that the nervous system and a regulator gene, orthopedia, play a mechanistic role. In addition, larvae of the two species, Strongylocentrotus purpuratus and Dendraster excentricus, responded to different cues. Pre-feeding larvae of both species developed relatively shorter arms when given algal cells (i.e. chemical and physical stimuli), whereas only pre-feeding larvae of D. excentricus developed shorter arms when exposed to algal exudates (i.e. chemical stimuli). Larvae of neither species responded morphologically to the presence of polystyrene beads (i.e. physical stimuli).     

Chemotaxis and chemical defenses in seaweed susceptibility to herbivory

Recent studies have show that small marine herbivores with limited mobility (mesograzers) often feed on macroalgae chemically defended against fishes or sea-urchins. In order to verify the involved mechanisms of chemotaxis or chemical defense into this process in Brazilian littoral, two species of brown alga Dictyota menstrualis and Dictyota mertensii were studied against the limited mobility herbivores, the amphipod Parhyale hawaiensis and the crab Pachygrapsus transversus. These two species were studied in order to verify the action of their crude extracts in the defense and chemotaxis processes related to limited mobility of these herbivores. Feeding preference assays revealed that P. hawaiensis do not eaten these Dictyota species. P. transversus do not eaten D. menstrualis either, but consumed large amounts of D. mertensii. Chemical deterrent assays showed that extracts of these species act as feeding deterrent to both species of herbivores. In addition, chemotaxis assays demonstrated that both herbivores are significantly negative chemotactic probably due to the presence of complementary metabolites into artificial foods. Considering that both Dictyota species exhibit active extracts against these small herbivores, we suppose that the non-occurrence of these herbivore species in close relationship with the seaweeds D. menstrualis and D. mertensii may explain the defense action of both extracts related to these mesograzers.     

Specificity of induced plant responses to specialist herbivores of the common milkweed Asclepias syriaca

Induced plant responses to herbivory appear to be universal, yet the degree to which they are specific to sets of herbivores is poorly understood. The generalist/specialist hypothesis predicts that generalist herbivores are more often negatively affected by host plant defenses, wheras specialists may be either unaffected by or attracted to these same "plant defenses". Therefore, specialists should be less predictable than generalists in their responses to induced plant resistance traits. To better understand the variation in plant responses to herbivore attack, and the impacts these responses have on specialist herbivores, we conducted a series of experiments examining pairwise interactinos between two specialaist herbivores of the common milkweed ( Asclepias syriaca ). We damaged plants mechnically, with swamp milkweed beetles ( Labidomera clivicollis ), or with monarchs ( Danaus plexippus ), and then asessed specificity of elicitation, both by measuring a putative defensive trait (latex volume) and by challenging plants with insects of both species in bioasays. Latex production increased by 34% and 13% following beetle and monarch herbivory, respectively, but only beetles significantly elevated latex production compared to undamaged controls. While beetle growth was negatively affected by latex across all experiments, beetles were not affected by previous damage caused by conspecifies or by monarchs. In contrast, monarchs feeding on previously damaged plants were 20% smaller, and their response was the same on plants damaged mechnically or by either herbivore. Therefore, these specialist herbivores exhibit both specificity of elicitation in plant responses and specificity of effects in response to prior damage.