Performance of potential European biological control agents of Vincetoxicum spp. with notes on their distribution

Exploratory surveys were conducted in Europe to locate potential biological control agents of Vincetoxicum nigrum and Vincetoxicum rossicum and to collect known specialist herbivores of Vincetoxicum hirundinaria (Abrostola asclepiadis, Chrysolina a. asclepiadis, Eumolpus asclepiadeus and Euphranta connexa). Development of the most abundant herbivores found attacking leaves, roots, and developing seeds was evaluated in the laboratory on three Vincetoxicum spp. Field surveys revealed that the highest diversity of herbivores was associated with the abundant and widespread V. hirundinaria. No new herbivores were reported from V. hirundinaria or V. nigrum, but the leaf‐feeding noctuid Hypena opulenta was recorded for the first time attacking V. rossicum and V. scandens in Ukraine. Based upon larval survival and development and adult fecundity, the leaf feeders A. asclepiadis, C. a. asclepiadis, and H. opulenta, perform better on V. nigrum and V. rossicum than on V. hirundinaria. Larval performance of the root feeder E. asclepiadeus followed a similar pattern, but adult fecundity of this insect did not vary among host plant species. Immature development time of the pre‐dispersal seed feeder E. connexa is similar among hosts, but larvae grow larger on V. nigrum and V. hirundinaria than on V. rossicum. All herbivores are promising biological control agents of Vincetoxicum in North America and we have prioritized host range testing with H. opulenta and E. asclepiadeus.     

Impact of herbivory on performance of <Emphasis Type="Italic">Vincetoxicum</Emphasis> spp., invasive weeds in North America

The alien invasive vines Vincetoxicum rossicum and Vincetoxicum nigrum (swallow-wort) are of major concern in eastern North America, where both species invade forested landscapes and threaten faunal and plant diversity. Among the few native natural enemies reported in Eurasia, the specialist chrysomelid, Chrysochus (Eumolpus) asclepiadeus (Coleoptera; Chrysomelidae), feeds on Vincetoxicum both above ground (as adults) and below ground (as larvae). The goal of our study was to assess the potential for using this beetle to manage invasive Vincetoxicum spp. in North America by quantifying the impact of herbivory by C. asclepiadeus on Vincetoxicum and determining whether this effect was influenced by plant density (“Allee effect”). Experimental work was carried out using a split plot design in the field in southern France. Pots of V. nigrum and V. hirundinaria, a substitute for V. rossicum, were planted at high (255 plants/m²), medium (127 plants/m²), and low (32 plants/m²) plant densities, and received treatments of 0 (control), 2 or 4 C. asclepiadeus adult beetles/pot. Leaf damage, root and shoot biomass, and quantity of seeds were measured after 4 weeks of adult feeding. Densities of 2 and 4 beetles/pot caused similar damage, with significant reductions in plant biomass at low plant density. While V. hirundinaria increased allocation of resources to roots in response to herbivory, V. nigrum did not. Seed production was greatest for both species grown at low plant densities, but only V. nigrum produced fewer seeds in response to herbivory. Our results, based on the effects of herbivory by C. asclepiadeus adults, suggest that if this beetle were to be introduced into North America for the management of Vincetoxicum spp. such as V. rossicum, reductions in plant biomass and spread would be greatest if beetles were released on edges or in newly-established satellite populations at low plant densities. In the case of V. nigrum, beetles could be released irrespective of plant density as reproductive output and seed dispersal would be reduced similarly.     

Oviposition preference and larval performance of North American monarch butterflies on four Asclepias species

Monarch butterflies, Danaus plexippus L. (Lepidoptera: Nymphalidae), occur world‐wide and are specialist herbivores of plants in the milkweed family (Asclepiadaceae). In North America, two monarch populations breed east and west of the continental divide in areas populated by different host plant species. To examine the population variation in monarch responses to different Asclepias species, we measured oviposition preference and larval performance among captive progeny reared from adult butterflies collected in eastern and western North America. Host plant use was evaluated using two milkweed species widely distributed in eastern North America (A. incarnata and A. syriaca), and two species common to western North America (A. fascicularis and A. speciosa). We predicted that exposure to different host plant species in their respective breeding ranges could select for divergent host use traits, so that monarchs should preferentially lay more eggs on, and larvae should perform better on, milkweed species common to their native habitats. Results showed that across all adult female butterflies, oviposition preferences were highest for A. incarnata and lowest for A. fascicularis, but mean preferences did not differ significantly between eastern and western monarch populations. Larvae from both populations experienced the highest survival and growth rates on A. incarnata and A. fascicularis, and we again found no significant interactions between monarch source population and milkweed species. Moreover, the average rank order of larval performance did not correspond directly to mean female oviposition preferences, suggesting that additional factors beyond larval performance influence monarch oviposition behavior. Finally, significant family level variation was observed for both preference and performance responses within populations, suggesting an underlying genetic variation or maternal effects governing these traits.     

Honey bees are the dominant diurnal pollinator of native milkweed in a large urban park

In eastern North America, the field milkweed, Asclepias syriaca L. (Asclepiadaceae), is used in planting schemes to promote biodiversity conservation for numerous insects including the endangered monarch butterfly, Danaus plexippus (Linnaeus) (Nymphalidae). Less is known about its pollinators, and especially in urban habitats where it is planted often despite being under increasing pressure from invasive plant species, such as the related milkweed, the dog‐strangling vine (DSV), Vincetoxicum rossicum (Kleopow) Barbar. (Asclepiadaceae). During the A. syriaca flowering period in July 2016, we surveyed bees in open habitats along a DSV invasion gradient and inspected 433 individuals of 25 bee species in 12 genera for pollinia: these were affixed to bees that visited A. syriaca for nectar and contain pollen packets that are vectored (e.g., transferred) between flowers. Of all bees sampled, pollinia were found only on the nonindigenous honeybee, Apis mellifera (43% of all bees identified), as well as one individual bumblebee, Bombus impatiens Cresson. Pollinia were recorded from 45.2% of all honeybees collected. We found no relationship between biomass of DSV and biomass of A. syriaca per site. There was a significant positive correlation between A. syriaca biomass and the number of pollinia, and the proportion vectored. No relationship with DSV biomass was detected for the number of pollinia collected by bees but the proportion of vectored pollinia declined with increasing DSV biomass. Although we find no evidence of DSV flowers attracting potential pollinators away from A. syriaca and other flowering plants, the impacts on native plant–pollinator mutualisms relate to its ability to outcompete native plants. As wild bees do not appear to visit DSV flowers, it could be altering the landscape to one which honeybees are more tolerant than native wild bees.     

A comparison of the host preference of monarch butterflies (Danaus plexippus) for milkweed (Asclepias syriaca) over dog-strangler vine (Vincetoxicum rossicum)

Observations in the field indicate that monarch butterflies will oviposit on dog‐strangler vine, an invasive introduced species in the same family as milkweed (Asclepias spp.), the principal larval host of monarchs. The potential impact of this behaviour depends on the strength of the preference of monarch adults to oviposit on these two hosts and the relative ability of larvae to survive on each. We determined the preference for milkweed vs. dog‐strangler vine of ovipositing adults and first instar larvae in choice and no‐choice tests. We also compared the ability of larvae to consume, develop, and survive on either host. In the presence of both hosts, adults exhibited a strong preference to oviposit on milkweed over dog‐strangler vine (mean 80.7 eggs compared to 0.4 eggs over 48 h, respectively). In the absence of milkweed, adults ceased oviposition (mean 0.9 eggs in 48 h), but resumed oviposition when the dog‐strangler vine was replaced with milkweed (mean 99.1 eggs in 48 h). Given a choice between hosts over 24 h, 92% of larvae moved to milkweed leaves and consumed 3.94 cm² of milkweed leaves compared to 2% of larvae that moved to dog‐strangler vine and consumed negligible amounts of leaf material (0.01 cm²). Without a choice, larvae on dog‐strangler vine never consumed more than mean 0.02 cm² larva^?1 in a 24‐h period, did not develop beyond the first instar, and died within 96 h. We obtained no data in support of an effect of the presence of dog‐strangler vine on monarch butterfly populations.     

The effect of an invasive alien vine, <Emphasis Type="Italic">Vincetoxicum rossicum</Emphasis> (Asclepiadaceae), on arthropod populations in Ontario old fields

Vincetoxicum rossicum is an invasive alien vine introduced from Europe in the late 1800s that is now an emerging pest in upstate New York and eastern Ontario. The plant can form dense, monotypic stands in woodlots and old fields, and may be displacing native vegetation. As a consequence, V. rossicum may be displacing arthropod fauna associated with native vegetation. In June and August 2002, we sampled V. rossicum and three other old field plants (Asclepias syriaca, Solidago altissima, and mixed graminoids) for arthropods using pitfall traps and by sampling individual plants. A total of 7868 arthropods were counted on plants and 18,195 individuals were trapped; these were sorted by feeding guild. Overall, stands of V. rossicum supported the lowest abundance of both stem- and ground-dwelling individuals, as well as the lowest number of arthropods in most phytophagous guilds. Some feeding guilds are entirely absent: V. rossicum stands are completely devoid of gall-makers and miners, and support few pollinators. This study suggests that arthropod diversity will decline if V. rossicum displaces native old-field plants.     

Host specificity of Asian <Emphasis Type="Italic">Chrysochus</Emphasis> Chevr. in Dej. (Coleoptera,Chrysomelidae: Eumolpinae) and their potential use for biological control of invasive <Emphasis Type="Italic">Vincetoxicum</Emphasis> species

Three Asian leaf beetles of the genus Chrysochus were investigated as potential biological control agents of Vincetoxicum rossicum and V. nigrum, invasive weeds in northeastern North America. Chrysochus chinensis and Ch. globicollis were collected from a field host in a different genus and subtribe. Preliminary no-choice laboratory tests with Ch. goniostoma showed that its physiological host range is too broad. Based on these data, we are not considering these three species as potential biological control agents of invasive Vincetoxicum species.     

Development of microsatellite markers for the endangered grassland perennial herb Vincetoxicum atratum (Apocynaceae–Asclepiadoideae)

Eight microsatellite markers were developed for the endangered grassland perennial herb Vincetoxicum atratum. The number of alleles ranged from 4 to 14, and the expected heterozygosities were from 0.575 to 0.933 in a population of V. atratum. Five of the eight loci did not significantly deviated from the Hardy–Weinberg equilibrium. All eight loci were tested for cross-species amplification in five other species of Vincetoxicum in Japan. These microsatellite loci will be useful for conservation genetics of V. atratum and other species of Vincetoxicum.     

Monarch butterfly oviposition on swallow-worts (Vincetoxicum spp.)

As black swallow-wort, Vincetoxicum nigrum L. Moench, and pale swallow-wort, V. rossicum (Kleopow) Barbar., spread throughout the northeastern United States and southern Canada, there is concern about the impact of these invasive plants on populations of the native North American monarch butterfly Danaus plexippus L. Recent laboratory studies in New York and Ontario, Canada, have found little or no oviposition by monarch butterflies on the two Vincetoxicum species. In Rhode Island, we found 10.5-21.7% oviposition on Vincetoxicum species relative to common milkweed Asclepias syriaca L. in choice tests and 11.9-20.3% in no-choice tests in 2 yr of laboratory testing. These results were supported by field cage trials where monarchs given a choice between V. nigrum and A. syriaca laid 24.5% of their eggs on V. nigrum. In surveys of three pasture fields in Rhode Island where relative coverage of A. syriaca exceeded that of V. nigrum by a 0.77:0.23 ratio, 15.4% of monarch eggs were found on V. nigrum plants. In V. nigrum stands with very little A. syriaca (6.25 stems/ha), monarch egg density on V. nigrum was found to be over five times greater than in the three mixed pasture fields. In none of our laboratory or field evaluations was there any survival of monarch larvae on Vincetoxicum species. It seems that in Rhode Island, Vincetoxicum species serve as an oviposition sink for monarch butterflies. These findings suggest that East Coast butterflies may differ in host selection from those in central New York and southern Ontario, Canada.     

Effects of leaf and root herbivory by potential insect biological control agents on the performance of invasive Vincetoxicum spp.

The European leaf-feeding moth Abrostola asclepiadis and root-feeding beetle Eumolpus asclepiadeus are promising biological control agents for two European swallow-worts (Vincetoxicum rossicum and Vincetoxicum nigrum) in North America, however, their impact on plant performance is uncertain. Densities of each herbivore were manipulated in a common garden to determine whether leaf and root herbivory affect the performance of these plants. During the second year of the experiment, V. rossicum and V. nigrum unexpectedly became infected with the fungal pathogens Ascochyta sp. and Cercospora sp. (Ascomycota), respectively. Although pathogen infection mainly reduced shoot height and delayed reproduction, herbivore effects on plant growth were still evident. Leaf herbivory by A. asclepiadis had no effect on plant growth 1 year after defoliation. Root herbivory by E. asclepiadeus reduced shoot height and plant biomass and decreased the ability of plants to compensate for pathogen attack. Pathogen infection prevented detection of herbivore effect on reproduction. Due to its substantial impact on plant biomass, E. asclepiadeus should be further evaluated as a biological control agent against Vincetoxicum spp. populations invading open habitats in North America. Further research is needed to evaluate the impact of A. asclepiadis in combination with E. asclepiadeus and plant competition under high and low light conditions.     

Establishment of the invasive perennial <Emphasis Type="Italic">Vincetoxicum rossicum</Emphasis> across a disturbance gradient in New York State,USA

Vincetoxicum rossicum (pale swallow-wort) is a non-native, perennial, herbaceous vine in the Apocynaceae. The species’ abundance is steadily increasing in the northeastern United States and southeastern Canada. Little is known about Vincetoxicum species recruitment and growth. Therefore, we conducted a field experiment in New York State to address this knowledge gap. We determined the establishment, survival, and growth of V. rossicum during the first 2 years after sowing in two old fields subjected to four disturbance regimens. We hypothesized that establishment and survival would be higher in treatments with greater disturbance. At the better-drained location, overall establishment was 15 ± 1% [mean ± standard error] and did not differ among treatments. At the poorly drained location, establishment varied by treatment; mowed and control plots had greater establishment [10 ± 2%] than herbicide + tillage and herbicide-only plots [1.6 ± 0.5%]. Of those seedlings that emerged, overall survival was high at both locations (70–84%). Similarly, total (above + belowground) biomass was greater in herbicide + tillage and herbicide-only plots than in mowed and control plots at both locations. Thus, V. rossicum was successful in establishing and surviving across a range of disturbance regimens particularly relative to other old field species, but growth was greater in more disturbed treatments. The relatively high-establishment rates in old field habitats help explain the invasiveness of this Vincetoxicum species in the northeastern U.S. and southeastern Canada.     

Response of Danaus plexippus to pollen of two new Bt corn events via laboratory bioassay

Laboratory bioassays were conducted to evaluate the response of first instar larvae of the monarch butterfly, Danaus plexippus L. (Lepidoptera: Danaidae), a non‐target species, to pollen from corn, Zea mays L. (Commelinales: Poaceae), from two new corn hybrids genetically modified to express different types of insecticidal proteins derived from the bacterium Bacillus thuringiensis Berliner (Bacillales: Bacillaceae) (Bt). One hybrid expresses both Cry1Ab and Cry2Ab2 proteins (MON 810 × MON 84006), active against lepidopteran pests, and the other expresses Cry3Bb1 protein (MON 863), targeted against coleopteran pests. First instar larvae were placed on milkweed leaves (Asclepias syriaca L.) (Gentianales: Asclepiadaceae) dusted with doses of either Bt pollen or its nonexpressing (isoline) pollen counterpart ranging from 50 to 3200 grains cm^?2 of milkweed leaves, or no pollen at all. Larvae were exposed to pollen for 4 days, then moved to pollen‐free leaves and observed for another 6 days. Survival was observed after 2, 4, and 10 days. Weight gain was estimated after 4 and 10 days, leaf consumption after 2 and 4 days, and larval development after 10 days. Exposure to pollen of the Cry1Ab/Cry2Ab2‐Bt expressing hybrid reduced larval survival approximately 7.5–23.5% at the dose ranges tested relative to a no pollen control. Larval weight gain and consumption were reduced for larvae exposed to pollen of this hybrid and a small minority of larvae (3.1%) never developed past the third instar after 10 days of observation. Exposure to pollen of the Cry3Bb1‐Bt expressing hybrid had no negative effects on larval mortality, weight gain, consumption, or development relative to the consumption of Bt‐free corn pollen. The relevance of these findings to the risk that these Bt corn hybrids pose to monarch populations is discussed.     

Plant invasion alters trait composition and diversity across habitats

Increased globalization has accelerated the movement of species around the world. Many of these nonnative species have the potential to profoundly alter ecosystems. The mechanisms underpinning this impact are often poorly understood, and traits are often overlooked when trying to understand and predict the impacts of species invasions on communities. We conducted an observational field experiment in Canada's first National Urban Park, where we collected trait data for seven different functional traits (height, stem width, specific leaf area, leaf percent nitrogen, and leaf percent carbon) across an abundance gradient of the invasive Vincetoxicum rossicum in open meadow and understory habitats. We assessed invasion impacts on communities, and associated mechanisms, by examining three complementary functional trait measures: community‐weighted mean, range of trait values, and species’ distances to the invader in trait space. We found that V. rossicum invasion significantly altered the functional structure of herbaceous plant communities. In both habitats V. rossicum changed the community‐weighted means, causing invaded communities to become increasingly similar in their functional structure. In addition, V. rossicum also reduced the trait ranges for a majority of traits indicating that species are being deterministically excluded in invaded communities. Further, we observed different trends in the meadow and understory habitats: In the understory, resident species that were more similar to V. rossicum in multivariate trait space were excluded more, however this was not the case in the meadow habitat. This suggests that V. rossicum alters communities uniquely in each habitat, in part by creating a filter in which only certain resident species are able to persist. This filtering process causes a nonrandom reduction in species' abundances, which in turn would be expected to alter how the invaded ecosystems function. Using trait‐based frameworks leads to better understanding and prediction of invasion impacts. This novel framework can also be used in restoration practices to understand how invasion impacts communities and to reassemble communities after invasive species management.     

Maintenance of narrow diet breadth in the monarch butterfly caterpillar: response to various plant species and chemicals

In order to better understand the maintenance of a fairly narrow diet breadth in monarch butterfly larvae, Danaus plexippus L. (Lepidoptera: Nymphalidae: Danainae), we measured feeding preference and survival on host and non-host plant species, and sensitivity to host and non-host plant chemicals. For the plant species tested, a hierarchy of feeding preferences was observed; only plants from the Asclepiadaceae were more or equally preferred to Asclepias curassavica, the common control. The feeding preferences among plant species within the Asclepiadaceae are similar to published mean cardenolide concentrations. However, since cardenolide data were not collected from individual plants tested, definitive conclusions regarding cardenolide concentrations and plant acceptability cannot be made. Although several non-Asclepiadaceae were eaten in small quantities, all were less preferred to A. curassavica. Additionally, these non-Asclepiadaceae do not support continued feeding, development, and survival of first and fifth-instar larvae. Preference for a host versus a non-host (A. curassavica versus Vinca rosea) increased for A. curassavica reared larvae as compared to diet-reared larvae suggesting plasticity in larval food preferences. Furthermore, host species were significantly preferred over non-host plant species in bioassays using a host plant or sucrose as a common control. Larval responses to pure chemicals were examined in order to determine if host and non-host chemicals stimulate or deter feeding in monarch larvae. We found that larvae were stimulated to feed by some ubiquitous plant chemicals, such as sucrose, inositol, and rutin. In contrast, several non-host plant chemicals deterred feeding: caffeine, apocynin, gossypol, tomatine, atropine, quercitrin, and sinigrin. Additionally the cardenolides digitoxin and ouabain, which are not in milkweed plants, were neutral in their influence on feeding. Another non-milkweed cardenolide, cymarin, significantly deterred feeding. Extracts of A. curassavica leaves were tested in bioassays to determine which components of the leaf stimulate feeding. Both an ethanol extract of whole leaves and a hexane leaf-surface extract are phagostimulatory, suggesting the involvement of both polar and non-polar plant compounds. These data suggest that the host range of D. plexippus larvae is maintained by both feeding stimulatory and deterrent chemicals in host and non-host plants.     

Plant water stress and previous herbivore damage affect insect performance

1. Short‐term changes in plant resistance traits can be affected by abiotic factors or damage by herbivores, although how the combined effects of abiotic factors and previous damage affect subsequent insect larval development is not well understood. 2. Complementary glasshouse and field experiments were conducted to evaluate whether plant water stress and previous herbivore damage influenced monarch (Danaus plexippus) larval development on common milkweed, Asclepias syriaca. 3. In the glasshouse, water stress altered a suite of A. syriaca functional traits but did not affect nutrient content, whereas herbivore damage increased leaf nitrogen (N) and reduced the carbon:nitrogen (C:N) ratio. A bioassay experiment showed that monarch larval survival was lower on well‐watered plants that were previously damaged by monarch larva than on damaged and drought‐stressed plants. Bioassay larvae consumed less leaf tissue of previously damaged plants, whereas monarch larval mass was affected additively by water stress and previous damage, after correcting for the amount of leaf tissue consumed. 4. In a 2‐year field experiment, monarch larval performance was higher on previously damaged A. syriaca plants that received experimentally reduced rainfall, relative to plants receiving ambient rainfall. 5. Collectively, these results from glasshouse and field experiments suggest that insect performance was highest on previously damaged plants under water stress and highlight the additive and interactive roles of abiotic and biotic factors on herbivore performance.     

Variability of microsatellite loci in <Emphasis Type="Italic">Vincetoxicum</Emphasis> Wolf species in southeastern Ukraine

Genetic diversity of 13 species of the genus Vincetoxicum Wolf found in Ukraine with the use of four of eight nuclear microsatellite markers previously developed for Vincetoxicum atratum from Japan was studied. The number of alleles in studied loci varied in the range from 8 to 25. The expected heterozygosity was 0.690–0.938; the observed heterozygosity varied in the range from 0.205 to 0.806. The total rate of genetic variability of studied species was found to be comparable to the rate of variability of Vincetoxicum atratum from Japan. Microsatellite loci Vinc5, Vinc104, Vinc123, and Vinc124 can be successfully used for estimating the intra- and interspecific polymorphism of the species of genus Vincetoxicum Wolf in Ukraine.     

Effects of food plant and group size on predator defence: differences between two co-occurring aposematic Lygaeinae bugs

1. The role of food plant and aggregation on the defensive properties of two aposematic sympatrically occurring seed bugs, Tropidothorax leucopterus and Lygaeus equestris (Lygaeinae, Heteroptera), was investigated. Larvae reared on seeds either of their natural host plant Vincetoxicum hirundinaria (Asclepiadaceae) or of sunflower Helianthus annuus were subjected to predation by chicks. 2. The two species differ in their dependency on the host for their defence. Lygaeus equestris was better defended on its natural host plant than on the alternative food, as indicated by fewer attacks, lower mortality, and predator avoidance after experience. No such effect of food plant could be found for T. leucopterus, suggesting the existence of alternative defences in this species. 3. The number of attacks was lower when host plant‐fed larvae of both species were presented in groups. 4. The discussion concerns how major components of an aposematic syndrome, such as host plant chemistry, insect colouration, and aggregation, are integrated with other life‐history traits to form alternative lifestyles in L. equestris and T. leucopterus.     

Colonization and usage of eight milkweed (<Emphasis Type="Italic">Asclepias</Emphasis>) species by monarch butterflies and bees in urban garden settings

Planting milkweeds on public and private lands has emerged as a central conservation strategy for restoring declining North American migratory populations of the monarch butterfly (Danaus plexippus). Nearly all actionable science on this issue has focused on restoring common milkweed (Asclepias syriaca L.) in rural land types. The aim of this study was to develop recommendations for the best milkweeds for managed gardens intended to support both monarch butterflies and bees. Eight milkweed (Asclepias) species varying in height, form, and leaf shape were grown in a common-garden experiment at a public arboretum. We measured milkweed growth, tillering, and bloom periods, conducted bi-weekly counts of eggs and larvae to assess colonization by wild monarchs, and evaluated suitability for growth of monarch larvae. We also quantified bee visitation and compared the bee assemblages associated with six of the eight species, augmented with additional collections from other sites. Monarchs rapidly colonized the gardens, but did not equally use all of the milkweed species. More eggs and larvae were found on taller, broad-leaved milkweeds, but there was relatively little difference in larval performance, suggesting ovipositional preference for more apparent plants. Asclepias tuberosa and A. fascicularis attracted the greatest number of bees, whereas bee genus diversity was greatest on A. verticillata, A. fascicularis, and A. tuberosa. Milkweeds that do not spread extensively by tillering may be best suited for managed gardens. Combining milkweeds that are preferred by ovipositing monarchs with ones that are particularly attractive to bees may enhance conservation value of small urban gardens.