Milkweed Asclepias Syriaca Research Articles

Temporal matches between monarch butterfly and milkweed population changes over the past 25,000 years

In intimate ecological interactions, the interdependency of species may result in correlated demographic histories. For species of conservation concern, understanding the long-term dynamics of such interactions may shed light on the drivers of population decline. Here, we address the demographic history of the monarch butterfly, Danaus plexippus, and its dominant host plant, the common milkweed Asclepias syriaca (A.syriaca), using broad-scale sampling and genomic inference. Because genetic resources for milkweed have lagged behind those for monarchs, we first release a chromosome-level genome assembly and annotation for common milkweed. Next, we show that despite its enormous geographic range across eastern North America, A.syriaca is best characterized as a single, roughly panmictic population. Using approximate Bayesian computation with random forests (ABC-RF), a machine learning method for reconstructing demographic histories, we show that both monarchs and milkweed experienced population expansion during the most recent recession of North American glaciers 10,000-20,000 years ago. Our data also identify concurrent population expansions in both species during the large-scale clearing of eastern forests (∼200 years ago). Finally, we find no evidence that either species experienced a reduction in effective population size over the past 75 years. Thus, the well-documented decline of monarch abundance over the past 40 years is not visible in our genomic dataset, reflecting a possible mismatch of the overwintering census population to effective population size in this species.

Compound-Specific Behavioral and Enzymatic Resistance to Toxic Milkweed Cardenolides in a Generalist Bumblebee Pollinator.

Plant secondary metabolites that defend leaves from herbivores also occur in floral nectar. While specialist herbivores often have adaptations providing resistance to these compounds in leaves, many social insect pollinators are generalists, and therefore are not expected to be as resistant to such compounds. The milkweeds, Asclepias spp., contain toxic cardenolides in all tissues including floral nectar. We compared the concentrations and identities of cardenolides between tissues of the North American common milkweed Asclepias syriaca, and then studied the effect of the predominant cardenolide in nectar, glycosylated aspecioside, on an abundant pollinator. We show that a generalist bumblebee, Bombus impatiens, a common pollinator in eastern North America, consumes less nectar with experimental addition of ouabain (a standard cardenolide derived from Apocynacid plants native to east Africa) but not with addition of glycosylated aspecioside from milkweeds. At a concentration matching that of the maximum in the natural range, both cardenolides reduced activity levels of bees after four days of consumption, demonstrating toxicity despite variation in behavioral deterrence (i.e., consumption). In vitro enzymatic assays of Na+/K+-ATPase, the target site of cardenolides, showed lower toxicity of the milkweed cardenolide than ouabain for B. impatiens, indicating that the lower deterrence may be due to greater tolerance to glycosylated aspecioside. In contrast, there was no difference between the two cardenolides in toxicity to the Na+/K+-ATPase from a control insect, the fruit fly Drosophila melanogaster. Accordingly, this work reveals that even generalist pollinators such as B. impatiens may have adaptations to reduce the toxicity of specific plant secondary metabolites that occur in nectar, despite visiting flowers from a wide variety of plants over the colony's lifespan.

Decline in common milkweed along roadsides around Ottawa, Canada

ABSTRACT Over the past two decades, monarch butterfly populations have been declining. This decline has been partly attributed to the extensive loss of breeding habitat through the reduction of common milkweed (Asclepias syriaca, the larval host plant) through herbicides. While the decline of milkweed has been well documented in the US, less is known about its decline in Canada. To take a first step in quantifying its potential decline, we compared roadside milkweed abundance at sites around Ottawa, Ontario, Canada in 2018, to a county-wide estimate of milkweed abundance in 1943 and 1944. We also evaluated the effect of roadside characteristics on milkweed abundance at these sites. Current milkweed density in the Ottawa region is 33–86% lower than it was 75 years ago. We found milkweed at 67% of our 100 sites and found less milkweed at sites with a higher number of lanes in the road adjacent to the roadside. Interestingly, mowing indices were not significant predictors of milkweed abundance. Here we document the first quantitative evidence for milkweed decline over the past 75 years in Canada, which has likely contributed to the decline of breeding monarchs in Canada.

Experimental field evidence shows milkweed contaminated with a common neonicotinoid decreases larval survival of monarch butterflies.

Neonicotinoid insecticides are the most widely used class of insecticides in the world and can have both lethal and sub-lethal effects on non-target organisms in agricultural areas. Monarch butterflies Danaus plexippus have experienced dramatic declines in recent decades and, given that a large proportion of milkweed on the landscape grows in agricultural areas, there is concern about the negative effects of neonicotinoids on this non-target insect. In the field, we exposed common milkweed Asclepias syriaca, an obligate host plant of monarch butterflies, to agriculturally realistic levels of clothianidin, a widely used neonicotinoid insecticide. We tested whether this treatment influenced the number of eggs laid and larval survival over 2years. Milkweeds were transplanted into 60 experimental plots alongside a corn crop planted with a clothianidin seed coat and 60 control plots alongside an untreated corn crop. The number of eggs, larvae at each stage (first to fifth instar), and the presence of other arthropods were recorded weekly from June to the end of August and survival from egg to fifth instar was estimated using a Bayesian state-space statistical model. We counted more eggs in treated plots compared to control plots, suggesting a preference for treated milkweed. The number of plots with arthropods did not differ between treatments, but within treated plots, there was a greater decrease in the number of arthropods throughout the season. There was no evidence that monarchs selected plots with fewer arthropods for oviposition. Larval survival was lower in clothianidin-treated plots compared to control plots. Our results suggest milkweed near clothianidin-treated crops can reduce larval survival of monarch butterflies. While we provide some evidence that clothianidin could also act as an ecological trap for this species, further work is needed to identify additional components of fitness, including individual egg-laying rates and survival beyond the pupal stage. Our findings add to a growing body of evidence that neonicotinoids can negatively affect non-target organisms. ​.

Survival and regeneration ability of clonal common milkweed (Asclepias syriaca L.) after a single herbicide treatment in natural open sand grasslands

Invasive species are a major threat to biodiversity, human health, and economies worldwide. Clonal growth is a common ability of most invasive plants. The clonal common milkweed Asclepias syriaca L. is the most widespread invasive species in Pannonic sand grasslands. Despite of being an invader in disturbed semi-natural vegetation, this plant prefers agricultural fields or plantations. Herbicide treatment could be one of the most cost-effective and efficient methods for controlling the extended stands of milkweed in both agricultural and protected areas. The invasion of milkweed stand was monitored from 2011 to 2017 in a strictly protected UNESCO biosphere reserve in Hungary, and a single herbicide treatment was applied in May 2014. This single treatment was successful only in a short-term but not in a long-term period, as the number of milkweed shoots decreased following herbicide treatment. The herbicide translocation by rhizomatic roots induced the damage of dormant bud banks. The surviving buds developing shoots, growth of the milkweed stand showed a slow regeneration for a longer-term period. We concluded that the successful control of milkweed after herbicide treatment depends on repeated management of treated areas to suppress further spreading during subsequent seasons.

Distribution Modeling ofAsclepiasspp. Predicts Potential Conservation Benefits of Marginal Ecosystems as Nature Reserves in Northwest Ohio

Due to grassland and prairie conversion for agriculture, milkweed species (Asclepias spp.) are now confined to small natural areas, agricultural fields, and marginal ecosystems (e.g., ditches and field margins). Monarch butterflies (Danaus plexippus) rely exclusively on milkweed and are in decline in part due to declining milkweed populations. We conducted a species distribution modeling exercise using MaxEnt to model milkweed species and common milkweed (Asclepias syriaca) presence in natural areas and ditches throughout northwest Ohio in order to identify which local or regional landscape attributes most strongly influence milkweed presence. We found that while natural areas and ditches have similar presence probability ranges (milkweed: natural areas 3–98% and ditches 1–98%; common milkweed: natural areas and ditches 0–98%), natural areas have a higher average probability of presence than ditches (milkweed: natural areas = 30 ± 26% and ditches = 7 ± 7%; common milkweed: natural areas = 22 ± 26% and ditches = 5 ± 6%). However, due to the large area covered by ditches and field margins in our study system, ditches and field margins with a probability of milkweed occurrence greater than the average probability of occurrence in natural areas encompass a larger total area than natural areas. We also observed that the presence of milkweed is positively correlated with the presence of shrub and C4 grasses. We conclude that marginal ecosystems in ditches and field margins are an important component of regional milkweed and, thusly, monarch conservation by providing potential habitat across a diffuse network.

Allelopathic effect of invasive plants (Eriochloa villosa, Asclepias syriaca, Fallopia x bohemica, Solidago gigantea) on seed germination

The aim of this study was to determine the allelopathic potential of invasive species woolly cupgrass (Eriochloa villosa), common milkweed (Asclepias syriaca), bohemian knotweed (Fallopia x bohemica), and giant goldenrod (Solidago gigantea Ait.) on germination crop (Lepidium sativum L.). Experiments were conducted under laboratory conditions to determine effect of water extracts in petri dish bioassay. Water extracts from fresh biomass (leaves and stem) of invasive weeds in concentrations of 4 and 8 g/100 ml were investigated. All invasive plants showed allelopathic effect on germination. In giant goldenrod stem water extract experiment, allelopathic effect was less pronounced.
 The cress germination was greatly suppressed with the woolly cupgrass, common milkweed and the giant goldenrod. The experiment showed that the seed germination depended on the concentrations and the plant material used (leaves and stem).

Changing oviposition times of the crab spider Misumena vatia (Thomisidae) correlate with climate change

The crab spider Misumena vatia (Clerck, 1757) (Thomisidae) is an important sit-and-wait predator at flowers visited by nectar or pollen-seeking insects. Typically, female M. vatia molt into their adult stage when many insect-attracting flowers come into bloom, and the spiders quickly gain weight leading up to oviposition. Between 1979 and 2010, the first spider ovipositions shifted one month earlier, from late July to late June, at my study site in coastal Maine, USA, in accordance with a concurrent temperature increase of ca. 0.44°C and a lengthening growing season. Flowering times of the spiders' most important hunting site, common milkweed Asclepias syriaca, as well as recruitment dates of their most important prey, bumblebees Bombus spp., to flowering milkweed, advanced as well. The shift in spider oviposition times increased the feasibility of second broods, though I found no successful second broods in the field, and the success of such broods would be problematic because of heavy overwintering losses. Differing rates of change of spider, milkweed and bumblebee activity indicated decreasing synchrony among these species; in particular, lessening future hunting opportunities for the spiders.

A dual infection of two microscopic fungi on common milkweed (Asclepias syriaca) in Hajdúság region (East-Hungary)

Common milkweed (Asclepias syriaca L.) is one of the most noxious and invasive weed species in Hungary. A. syriaca invades arable lands, horticultural and forestry plantations, natural and semi-natural habitats too. In cases of field crops it can cause considerable yield losses mostly in maize (2–10%), soybean (12–32%) and sorghum (4–29%), but only with high rate of coverage. It can also increase these problems that the common milkweed can be serve as reservoir and host for viruses, other pathogens and pests.Because of the importance of common milkweed and in spite of demand to develop effective biological control, until now has not been developed a proper control program against A. syriaca. The aim of our research was to identify the necrotrophic fungal pathogens, which were involved in notable disease occurrence on this weed in different parts of Hajdúság region of Eastern-Hungary in 2016.To the isolation of fungi from leaves and their identification were based on morphological colony characters on potato dextrose agar (PDA) and Sabouraud dextrose agar (SDA). To the description of conidia features were used PDA for Alternaria and synthetic low-nutrient agar (SNA) for Fusarium species, respectively. The examination of axenic cultures revealed that the fungi isolated from the leaves of common milkweed were Fusarium sporotrichioides and Alternaria alternata.

Examination of dogbane beetle (Chrysochus auratus) feeding and phenology on spreading dogbane, and considerations for biological control

We carried out experiments that considered the feeding, phenology, and biocontrol potential of dogbane beetle, Chrysochus auratus, on spreading dogbane, Apocynum androsaemifolium, a native perennial weed in lowbush blueberry (Vaccinium angustifolium). In no-choice host-feeding experiments, adult beetles did not feed upon common milkweed (Asclepias syriaca), periwinkle (Vinca minor), wild raisin (Viburnum cassenoides), and lowbush blueberry, all plants related to spreading dogbane or found around lowbush blueberry fields. In a field experiment, significant decreases in spreading dogbane total and foliar weight occurred at a density of 16 beetles per ramet, but not at lower beetle densities. In our Nova Scotia (NS) field sites, beetles were present for 8–12 weeks, beginning in late June or early July (225–335 growing degree days, GDD). Beetle abundance peaked at 4–7 beetles/m2 and occurred at 357–577 GDD, which temporally coincides with the incidence of mature spreading dogbane plants in the field. The results suggest that although inundations of C. auratus could cause significant defoliation of spreading dogbane, natural populations of the beetle probably could not satisfactorily suppress development of this weed as a stand-alone control tactic. Conservation and augmentation of C. auratus populations should nonetheless be encouraged in integrated management programs for spreading dogbane.

Assessment and prediction of the invasiveness of some alien plants in conditions of climate change in the steppe Dnieper region

The flora of the steppe Dnieper region is characterized by an abundance of naturalized alien species, some of which colonised over the last decade. Climate change, associated primarily with increasing temperature, became clearly manifested in this period. We tested the hypothesis that there is an association between climate change and the initiation of invasiveness of some alien plant species in the steppe Dnieper region. For this purpose, comparative studies of the distribution boundaries of naturalized alien trees, shrubs and herbaceous plants were conducted. Along the research route numerous 5–10-year-old broadleaf linden trees ( Tilia platyphyllos Scop.) were found in the man-made plantation communities of Dnipro city in areas with moist soil; seeded undergrowth was located at a significant distance from the adult linden plants. Numerous groups of young 7–10-year-old plants of the smoke trees ( Cotinus coggygria Scop.), which had a seed origin, were found in the shelterbelt and urban recreational plantations. Young 10–12-year-old virginal and generative plants of the black cherry ( Padus serotina Ehrh.) were found in large numbers in both the semi-natural and artificial plant communities at great distances from the adult trees. The alien plant species common hackberry ( Celtis occidentalis L.) showed the ability to form fairly sparse seminal seedlings, which was presented by the plants at the age of 4–7 years in both the natural and urban plant communities. The perennial herbaceous plant common milkweed ( Asclepias syriaca L.) was found in the course of the research in ruderal habitats, urban plant communities, and also in the agrocoenoses. The common milkweed reached its greatest abundance in fields of winter crops, where the spread of this species was accompanied by a sharp decrease in the number of other species of segetal plants. Our study results confirm that the extension of the distribution boundaries of alien species over the last decade was not related to the ground conditions of the steppe Dnieper region. At the same time, changes in climatic conditions were favourable for some naturalized alien species because they have created the opportunity for seed reproduction of species away from the maternal plants. Alien species C. coggigria , P. serotina and A. syriaca were also the most sensitive to the influence of the climate changes. Consequently, these species have the greatest potential for increasing their level of invasiveness and endangering the biodiversity in the steppe Dnieper region under conditions of climate change. We suggest that a simultaneous initiation of invasiveness of these several alien species leads to an increase in the degree of threat to the diversity of natural plants in the region. The study results confirm the urgent need for analysis and forecasting of the consequences of introduction of alien species, in order to prevent the undesirable effects that this would bring for the region’s native vegetation.

New localities and habitat preferences of common milkweed Asclepias syriaca L. in Toruń (Central Poland)

The paper presents the location and general description of habitats and plant communities with common milkweed occurring within the city limits of Torun. The data set comprised new localities of this species as well as those where Asclepias syriaca has been observed for several years but the sites have not been described in the ecological literature. Releves made at these sites were compared with those described by Puchalka et al. (2013) from three sites of Asclepias syriaca in Torun. A larger set of new sites included in the analysis allows for a more accurate description of the impact exerted by some ecological factors, e.g. afforestation, on the growth of the studied species. As evidenced by the new sites of Asclepias syriaca found recently in Torun and the surrounding area, it is necessary to monitor the dynamics of the population and to undertake the research on the impact of common milkweed on plant communities.

Population growth and sequestration of plant toxins along a gradient of specialization in four aphid species on the common milkweed Asclepias syriaca

Summary Dietary specialization in insect herbivores has long been hypothesized to predict tolerance of plant defences, with more specialized herbivores being highly tolerant of and sometimes sequestering plant secondary compounds. Plant variation in secondary compounds should thus play an important and predictable role in shaping the performance and distribution of insect communities. We compared the performance of four naturally co‐occurring aphid species on twenty genotypes of the common milkweed Asclepias syriaca. Genotypes of milkweed consistently differed in functional traits, including concentrations of toxic cardenolides, while the diet breadths of the four aphids ranged from broadly generalized to monophagous. The two more generalized species had the highest population growth rate overall, while growth rates decreased with increasing specialization. In contrast, honeydew exudation as a measure of phloem consumption increased with specialization; thus, resource‐use efficiency was lower in specialist aphids. The two more generalized aphids grew best on genotypes with the highest plant growth rate (as an approximation for resource availability), while specialist aphids were not affected by plant growth. All four species contained apolar cardenolides in their bodies and excreted polar cardenolides, but only the most specialized aphid Myzocallis asclepiadis was negatively affected by increasing cardenolide concentrations of the host plant. Sequestration of cardenolides increased with diet specialization, with M. asclepiadis accumulating twice as much as any other species, perhaps explaining its susceptibility to plant cardenolides. Heritable plant traits differentially impacted co‐occurring insect herbivores within the same guild. Generalist aphids were susceptible to variation in plant vigour but not defensive compounds. Increased host specialization resulted in lower resource‐use efficiency, increased phloem throughput and ultimately higher cardenolide sequestration. Variation in these traits is thus likely to determine the relative distribution of generalist and specialist herbivores on plants in natural communities.

Evolution of plant growth and defense in a continental introduction.

Substantial research has addressed adaptation of nonnative biota to novel environments, yet surprisingly little work has integrated population genetic structure and the mechanisms underlying phenotypic differentiation in ecologically important traits. We report on studies of the common milkweed Asclepias syriaca, which was introduced from North America to Europe over the past 400 years and which lacks most of its specialized herbivores in the introduced range. Using 10 populations from each continent grown in a common environment, we identified several growth and defense traits that have diverged, despite low neutral genetic differentiation between continents. We next developed a Bayesian modeling approach to account for relationships between molecular and phenotypic differences, confirming that continental trait differentiation was greater than expected from neutral genetic differentiation. We found evidence that growth-related traits adaptively diverged within and between continents. Inducible defenses triggered by monarch butterfly herbivory were substantially reduced in European populations, and this reduction in inducibility was concordant with altered phytohormonal dynamics, reduced plant growth, and a trade-off with constitutive investment. Freedom from the community of native and specialized herbivores may have favored constitutive over induced defense. Our replicated analysis of plant growth and defense, including phenotypically plastic traits, suggests adaptive evolution following a continental introduction.

Growth–defense tradeoffs for two major anti‐herbivore traits of the common milkweed Asclepias syriaca

Costs of plant defense are a key assumption in evolutionary ecology, yet their detection has remained challenging. Here we introduce a novel method for quantifying plant growth using the common milkweed Asclepias syriaca and repeated non‐destructive size measurements to experimentally test for costs of defensive traits. We estimated mechanistic components of plant growth (relative growth rate, net assimilation rate, specific leaf area and leaf‐mass ratio) at two levels of fertilization (high and low), and related them to production of toxic cardenolides and exudation of sticky latex. We found negative genetic correlations between cardenolides and growth (most strongly with net assimilation rate) at both nutrient levels. Additionally, plants varied in their cardenolide response to low nutrients, and genetic families maintaining higher cardenolide production at low nutrient availability suffered proportionally larger reductions in growth. In contrast, the amount of latex was positively correlated with plant growth. Because latex is instantly deployed from a plant‐wide system of pressurized laticifers, larger plants may simply exude proportionally more latex when damaged and thus plant size is likely to mask potential costs of latex synthesis. Unbiased quantification of mechanistic growth processes, coupled with the manipulation of nutrient or stress levels, is thus an effective approach to demonstrate allocation to defense and tradeoffs with growth, especially in long‐lived plant species.

Tests of the coupled expression of latex and cardenolide plant defense in common milkweed (Asclepias syriaca)

The coexpression of plant resistance traits suggests the hypothesis that they may have complementary functions in defense against herbivory. To address the extent to which defensive traits are necessarily coupled in plants grown under various conditions, we focused on latex and cardenolides, two potent defenses of milkweeds. We measured defenses across ontogenetic stages, different biotic and abiotic environments, and across genetic families of the common milkweed Asclepias syriaca. We first addressed the extent to which foliar cardenolides are derived from latex because latex actively flows through canals in leaves. We rinsed latex out of shredded leaves, which had no impact on foliar cardenolides, suggesting cardenolides are allocated to leaves independently of latex. Accordingly, there is potential for independent expression of the two traits. We next followed a cohort of plants from germination over three years; expression of both latex exudation and cardenolides increased annually, with the exception of a second year dip in cardenolides. Damage by monarch caterpillars induced ≈50% increases of both latex and cardenolides, with the former occurring rapidly within a day and the latter taking five days of herbivory; these responses were preceded by an earlier peak of the signaling hormones jasmonic acid and abscisic acid. Endogenous jasmonic acid showed an instantaneous positive correlation with latex exudation and foliar cardenolides. Under drought stress, latex and cardenolide expression were reversed, with water stress suppressing latex exudation, but nearly doubling cardenolide concentrations. These drought effects were not driven by phytohormones in the expected manner, as jasmonic acid was unaffected, salicylic acid was strongly suppressed, and abscisic acid tripled in response to drought. Finally, a meta‐analysis of four previously published field studies representing 85 genetic families of A. syriaca revealed no evidence for a genetic correlation between latex exudation and foliar cardenolide concentrations. The same lack of a correlation was observed across 22 populations of A. syriaca when grown in a common environment. Thus, the two most important defensive traits of milkweeds, although often coexpressed, can become uncoupled during some ontogenetic stages, under some biotic and abiotic conditions, and there is no evidence that they evolve together.

Reciprocal interactions between native and introduced populations of common milkweed, Asclepias syriaca, and the specialist aphid, Aphis nerii

Following its introduction into Europe (EU), the common milkweed (Asclepias syriaca) has been free of most specialist herbivores that are present in its native North American (NA) range, except for the oleander aphid Aphis nerii. We compared EU and NA populations of A. nerii on EU and NA milkweed populations to test the hypothesis that plant–insect interactions differ on the two continents. First, we tested if herbivore performance is higher on EU plants than on NA plants, because the former have escaped most of their herbivores and have perhaps been selected for lower defence levels following introduction. Second, we compared two A. nerii lines (one from each continent) to test whether genotypic differences in the herbivore may influence species interactions in plant–herbivore communities in the context of species introductions. The NA population of A. nerii developed faster, had higher fecundity and attained higher population growth rates than the EU population. There was no overall significant continental difference in aphid resistance between the plants. However, milkweed plants from EU supported higher population growth rates and faster development of the NA line of A. nerii than plants from NA. In contrast, EU aphids showed similar (low) performance across plant populations from both continents. In a second experiment, we examined how chewing herbivores indirectly mediate interactions between milkweeds and aphids, and induced A. syriaca plants from each continent by monarch caterpillars (Danaus plexippus) to compare the resulting changes in plant quality on EU aphid performance. As specialist chewing herbivores of A. syriaca are only present in NA, we expected that plants from the two continents may affect aphid growth in different ways when they are challenged by a specialist chewing herbivore. Caterpillar induction decreased aphid developmental times on NA plants, but not on EU plants, whereas fecundity and population growth rates were unaffected by induction on both plant populations. The results show that genetic variation in the plants as well as in the herbivores can determine the outcome of plant–herbivore interactions.

Specific impacts of two root herbivores and soil nutrients on plant performance and insect–insect interactions

Soil‐dwelling insects commonly co‐occur and feed simultaneously on belowground plant parts, yet patterns of damage and consequences for plant and insect performance remain poorly characterized. We tested how two species of root‐feeding insects affect the performance of a perennial plant and the mass and survival of both conspecific and heterospecific insects. Because root damage is expected to impair roots’ ability to take up nutrients, we also evaluated how soil fertility alters belowground plant–insect and insect–insect interactions. Specifically, we grew common milkweed Asclepias syriaca in low or high nutrient soil and added seven densities of milkweed beetles Tetraopes tetraophthalmus, wireworms (mainly Hypnoides abbreviatus), or both species. The location and severity of root damage was species‐specific: Tetraopes caused 59% more damage to main roots than wireworms, and wireworms caused almost seven times more damage to fine roots than Tetraopes. Tetraopes damage decreased shoot, main root and fine root biomass, however substantial damage by wireworms did not decrease any component of plant biomass. With the addition of soil nutrients, main root biomass increased three times more, and fine root biomass increased five times more when wireworms were present than when Tetraopes were present. We detected an interactive effect of insect identity and nutrient availability on insect mass. Under high nutrients, wireworm mass decreased 19% overall and was unaffected by the presence of Tetraopes. In contrast, Tetraopes mass increased 114% overall and was significantly higher when wireworms were also present. Survival of wireworms decreased in the presence of Tetraopes, and both species’ survival was negatively correlated with conspecific density. We conclude that insect identity, density and soil nutrients are important in mediating the patterns and consequences of root damage, and suggest that these factors may account for some of the contradictory plant responses to belowground herbivory reported in the literature.

Allocation of resources away from sites of herbivory under simultaneous attack by aboveground and belowground herbivores in the common milkweed, Asclepias syriaca

Following herbivory, plants can preferentially allocate newly acquired resources away from attacked sites as an important mechanism conferring tolerance. Although reported previously for both aboveground and belowground herbivores, it remains unclear whether plants can simultaneously allocate resources away from both kinds of herbivore attack, and whether they have interactive effects on plant resource allocation. In the current study, we used dual-isotopic techniques to compare the allocation of newly acquired carbon (C) and nitrogen (N) by the common milkweed Asclepias syriaca following attack by an aboveground herbivore, the monarch caterpillar Danaus plexippus and a belowground herbivore, larvae of the red milkweed beetle Tetraopes tetraophthalmus. Both species induced significant changes in the allocation of C and N in A. syriaca. Specifically, A. syriaca increased allocation of new N to stems at the expense of allocation to damaged tissues (i.e., leaf or root). When under simultaneous attack, the allocation of resources to stems was greater than that induced by either herbivore alone, suggesting that (1) the herbivores have additive effects on allocation patterns by A. syriaca and (2) A. syriaca was able to mitigate the effects of future attack by both herbivore species simultaneously.

The Effect of <I>BT</I>-Maize on Butterflies – Reckoning the Risk

Insect-resistant genetically-modified (GM) crops have been developed to provide protection against target pests by the introduction of a Bacillus thuringiensis (Bt) gene encoding various insecticidal Cry proteins. Such is the pervasiveness of this technology that the global area planted commercially with these genetically-modified Bt crops exceeded 50 million hectares in 2010. Bt maize crops engineered for toxicity to the larvae of lepidopteran pests such as the European corn borer, Ostrinia nubilalis, present a potential risk to the larvae of non-target butterflies that are not pests, but that feed in the vicinity of the crop and that might therefore be affected by toxic maize pollen. The hazard is from contact with the Bt protein by ingestion of maize pollen adhering to host plants. In North America, such a potential hazard was highlighted by a laboratory assay of Monarch butterfly larvae (Danaus plexippus) that consumed Bt maize pollen on milkweed (Asclepias syriaca) and led to extensive exposure assessments in the field in the USA. However, these found that that the amounts of maize pollen potentially ingested by monarch larvae on their host-plants in and around Bt-maize fields are unlikely to adversely affect a significant proportion of this species. However, extrapolating the monarch butterfly results to other species is problematic because of between-species variability in acute sensitivity to specific Cry1 proteins, differences in host-plants and other ecological factors. Within Europe, although lethal and sub-lethal effects of Bt-maize pollen consumption by lepidopteran larvae have now been reported for several non-target lepidopteran species under laboratory conditions, there have been few intensive field-based studies of exposure on prominent non-target butterflies of conservation concern. Only a minority of lepidopteran species live in habitats inside arable maize ecosystems, and the larvae of those that do are often found in greater numbers on host-plants in field margins, rather than in the crop itself, because host-plant densities are usually much greater in margins than in the crop. However, the intensification of agriculture over the last 50 years has been associated with substantial losses of biodiversity in arable ecosystems. Butterflies are popular indicators of biodiversity and are in decline in arable ecosystems throughout Europe, partly as a result of farming practice. In addition, genetic modification remains a controversial area for environmental policy in Europe, where public acceptance of genetically-engineered plants is much weaker than in North America. For example, in January 2005 Hungary invoked an emergency 'safeguard clause' to attempt to ban the cultivation of the Bt-maize event MON810 (which expresses the Cry1Ab protein). Despite its lack of success, this was followed in 2008 and 2009 by other EU states, including France and Germany; these safeguard clauses currently remain in place but are being challenged legally. MON810 is the only GM crop currently cultivated on a wide scale (c. 105 ha) in Europe, mainly in Spain, but also in the Czech Republic, Poland, Portugal, Romania and Slovakia. Obtaining direct data on the mortality of non-target Lepidoptera is difficult because there are many species potentially at risk; these have different host-plants; exposure studies require good estimates of population densities of both lepidopteran species and host-plant, in both crop, margin and alternative habitats; the species at risk may be rare and have a low population density; larvae may be difficult to find in the field; and the period over which pollen is shed is relatively short. Such difficulties were discussed by Aviron et al. (2009), who calculated that to detect a change in abundance of 20% with adequate statistical power would require more than 100 pairs of fields or field margins. Furthermore, there are restrictions due to limited resources and time limitations for largescale agro-ecological experiments (Perry et al., 2003). Additional problems include refusals to grant permission for trials in many EU member states and the risk of damage by activists