Evolution of resistance to single and combined floral phytochemicals by a bumble bee parasite.

Repeated exposure to inhibitory compounds can drive the evolution of resistance, which weakens chemical defence against antagonists. Floral phytochemicals in nectar and pollen have antimicrobial properties that can ameliorate infection in pollinators, but evolved resistance among parasites could diminish the medicinal efficacy of phytochemicals. However, multicompound blends, which occur in nectar and pollen, present simultaneous chemical challenges that may slow resistance evolution. We assessed evolution of resistance by the common bumble bee gut parasite Crithidia bombi to two floral phytochemicals, singly and combined, over 6 weeks (~100 generations) of chronic exposure. Resistance of C. bombi increased under single and combined phytochemical exposure, without any associated costs of reduced growth under phytochemical-free conditions. After 6 weeks' exposure, phytochemical concentrations that initially inhibited growth by > 50%, and exceeded concentrations in floral nectar, had minimal effects on evolved parasite lines. Unexpectedly, the phytochemical combination did not impede resistance evolution compared to single compounds. These results demonstrate that repeated phytochemical exposure, which could occur in homogeneous floral landscapes or with therapeutic phytochemical treatment of managed hives, can cause rapid evolution of resistance in pollinator parasites. We discuss possible explanations for submaximal phytochemical resistance in natural populations. Evolved resistance could diminish the antiparasitic value of phytochemical ingestion, weakening an important natural defence against infection.

Conspecific and Heterospecific Aboveground Herbivory Both Reduce Preference by a Belowground Herbivore.

Insect herbivores damage plants both above- and belowground, and interactions in each realm can influence the other via shared hosts. While effects of leaf damage on aboveground interactions have been well-documented, studies examining leaf damage effects on belowground interactions are limited, and mechanisms for these indirect interactions are poorly understood. We examined how leaf herbivory affects preference of root-feeding larvae [Acalymma vittatum F. (Coleoptera: Chrysomelidae)] in cucumber (Cucumis sativus L.). We manipulated leaf herbivory using conspecific adult A. vittatum and heterospecific larval Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) herbivores in the greenhouse and the conspecific only in the field, allowing larvae to choose between roots of damaged and undamaged plants. We also examined whether leaf herbivory induced changes in defensive cucurbitacin C in leaves and roots. We hypothesized that induced changes in roots would deter larvae, and that effects would be stronger for damage by conspecifics than the unrelated caterpillar because the aboveground damage could be a cue to plants indicating future root damage by the same species. In both the greenhouse and field, plants with damaged leaves recruited significantly fewer larvae to their roots than undamaged plants. Effects of conspecific and heterospecific damage did not differ. Leaf damage did not induce changes in leaf or root cucurbitacin C, but did reduce root biomass. While past work has suggested that systemic induction by aboveground herbivory increases resistance in roots, our results suggest that decreased preference by belowground herbivores in this system may be because of reduced root growth.

Indirect effects on mutualisms: parasitism of bumble bees and pollination service to plants.

Researchers increasingly recognize the important role of mutualisms in structuring communities and view positive interactions in a community context rather than as simple pairwise interactions. Indirect effects, such as those that predators have on lower trophic levels, are a key process in community ecology. However, such top-down indirect effects have rarely been extended to mutualisms. Antagonists of one mutualist have the potential to negatively affect the second mutualist through negative effects on their partner, and the magnitude of such effects should vary with mutualism strength. Bumble bees are ecologically and economically important pollinators that are an ideal system to determine if such indirect effects play an important role in mutualisms. Bumble bees are attacked by an array of parasites and predators, and they interact with a range of plants that vary in their dependence on bumble bees for reproduction. We tested whether variation in parasitism rates by Nosema bombi, Crithidia bombi, and conopid flies correlated with reproduction of greenhouse-raised plants placed in the field. At multiple sites over two years, we studied four plant species that varied in reliance on bumble bees as pollinators. We found a consistent negative relationship between Nosema parasitism and measures of pollination for Trifolium pratense and Solanum carolinense, plant species with high bumble bee visitation, whereas Rudbeckia hirta and Daucus carota, plant species with generalized pollination, experienced no impacts of Nosema. However, both Crithidia and conopids showed inconsistent relationships with pollination service. Although these patterns are correlational, they provide evidence that parasites of bumble bees may have negative indirect effects on plants, and that mutualism strength can moderate the magnitude of such effects.

Comparison of perimeter trap crop varieties: effects on herbivory, pollination, and yield in butternut squash.

Perimeter trap cropping (PTC) is a method of integrated pest management (IPM) in which the main crop is surrounded with a perimeter trap crop that is more attractive to pests. Blue Hubbard (Cucurbita maxima Duch.) is a highly effective trap crop for butternut squash (C. moschata Duch. ex Poir) attacked by striped cucumber beetles (Acalymma vittatum Fabricius), but its limited marketability may reduce adoption of PTC by growers. Research comparing border crop varieties is necessary to provide options for growers. Furthermore, pollinators are critical for cucurbit yield, and the effect of PTC on pollination to main crops is unknown. We examined the effect of five border treatments on herbivory, pollination, and yield in butternut squash and manipulated herbivory and pollination to compare their importance for main crop yield. Blue Hubbard, buttercup squash (C. maxima Duch.), and zucchini (C. pepo L.) were equally attractive to cucumber beetles. Border treatments did not affect butternut leaf damage, but butternut flowers had the fewest beetles when surrounded by Blue Hubbard or buttercup squash. Yield was highest in the Blue Hubbard and buttercup treatments, but this effect was not statistically significant. Native bees accounted for 87% of pollinator visits, and pollination did not limit yield. There was no evidence that border crops competed with the main crop for pollinators. Our results suggest that both buttercup squash and zucchini may be viable alternatives to Blue Hubbard as borders for the main crop of butternut squash. Thus, growers may have multiple border options that reduce pesticide use, effectively manage pests, and do not disturb mutualist interactions with pollinators.

Using trap crops for control of Acalymma vittatum (Coleoptera: Chrysomelidae) reduces insecticide use in butternut squash.

Striped cucumber beetle, Acalymma vittatum F., is the primary insect pest of cucurbit crops in the northeastern United States. Adult beetles colonize squash crops from field borders, causing feeding damage at the seedling stage and transmitting bacterial wilt Erwinia tracheiphila Hauben et al. 1999. Conventional control methods rely on insecticide applications to the entire field, but surrounding main crops with a more attractive perimeter could reduce reliance on insecticides. A. cittatum shows a marked preference for Blue Hubbard squash (Cucurbita maxima Duchesne) over butternut squash (C. moschata Poir). Given this preference, Blue Hubbard squash has the potential to be an effective perimeter trap crop. We evaluated this system in commercial butternut fields in 2003 and 2004, comparing fields using perimeter trap cropping with Blue Hubbard to conventionally managed fields. In 2003, we used a foliar insecticide to control beetles in the trap crop borders, and in 2004, we compared systemic and foliar insecticide treatments for the trap crop borders. We found that using a trap crop system reduced or eliminated the need to spray the main crop area, reducing insecticide use by up to 94% compared with conventional control methods, with no increase in herbivory or beetle numbers. We surveyed the growers who participated in these experiments and found a high level of satisfaction with the effectiveness and simplicity of the system. These results suggest that this method of pest control is both effective and simple enough in its implementation to have high potential for adoption among growers.

Transfer of quinolizidine alkaloids from hosts to hemiparasites in two Castilleja-Lupinus associations: analysis of floral and vegetative tissues.

Many hemiparasites, including several members of the Castilleja genus (Scrophulariaceae), obtain secondary compounds from their host plants. Both Castilleja miniata in subalpine Colorado and C. indivisa in central Texas have reduced herbivory when obtaining alkaloids from the hosts Lupinus argenteus and L. texensis (Fabaceae), respectively. However, pollinators were not deterred from visiting Castilleja parasitizing alkaloid-containing hosts. To determine if alkaloids are present in all tissues of plants parasitizing lupins, we analyzed floral tissue as well as leaves of both Castilleja species. Leaves, bracts, calices, corollas, gynoecium and nectar of both Castilleja species were examined for quinolizidine alkaloid presence using a Dragendorff reagent, and alkaloids were identified in vegetative tissue and nectar by capillary GLC and GLC-MS. Lupanine and alpha-isolupanine were the principal alkaloids in C. indivisa parasitizing L. texensis, while principal alkaloids of C. miniata parasitizing L. argenteus were 5,6-iso-dehydrolupanine, alpha-isolupanine, thermopsine, and 17-oxolupanine. Except for 17-oxolupanine, which was probably synthesized by biotransformation in the parasite, all other alkaloids correspond to those present in the host plants. Alkaloids were present in the leaves of both Castilleja species, and in the bracts, calices and gynoecium of some plants, but never in the corollas. Alkaloids from L. texensis and L. argenteus were not detected in nectar of either Castilleja species. The presence of alkaloids in leaves and outer floral tissue of both Castilleja species, but not nectar, may explain why alkaloid uptake and storage affected herbivores but not pollinators.