Sex-biased oviposition by a nursery pollinator on a gynodioecious host plant: Implications for breeding system evolution and evolution of mutualism

Dioecy, a breeding system where individual plants are exclusively male or female, has evolved repeatedly. Extensive theory describes when dioecy should arise from hermaphroditism, frequently through gynodioecy, where females and hermaphrodites coexist, and when gynodioecy should be stable. Both pollinators and herbivores often prefer the pollen-bearing sex, with sex-specific fitness effects that can affect breeding system evolution. Nursery pollination, where adult insects pollinate flowers but their larvae feed on plant reproductive tissues, is a model for understanding mutualism evolution but could also yield insights into plant breeding system evolution. We studied a recently established nursery pollination interaction between native Hadena ectypa moths and introduced gynodioecious Silene vulgaris plants in North America to assess whether oviposition was biased toward females or hermaphrodites, which traits were associated with oviposition, and the effect of oviposition on host plant fitness. Oviposition was hermaphrodite-biased and associated with deeper flowers and more stems. Sexual dimorphism in flower depth, a trait also associated with oviposition on the native host plant (Silene stellata), explained the hermaphrodite bias. Egg-receiving plants experienced more fruit predation than plants that received no eggs, but relatively few fruits were lost, and egg receipt did not significantly alter total fruit production at the plant level. Oviposition did not enhance pollination; egg-receiving flowers usually failed to expand and produce seeds. Together, our results suggest that H. ectypa oviposition does not exert a large fitness cost on host plants, sex-biased interactions can emerge from preferences developed on a hermaphroditic host species, and new nursery pollination interactions can arise as negative or neutral rather than as mutualistic for the plant

Variable Effects of Nicotine, Anabasine, and their Interactions on Parasitized Bumble Bees

Secondary metabolites in floral nectar have been shown to reduce parasite load in two common bumble bee species. Previous studies on the effects of nectar secondary metabolites on parasitizedbees have focused on single compounds in isolation; however, in nature, bees are simultaneously exposed to multiple compounds. We tested for interactions between the effects of two alkaloids found in the nectar of Nicotiana spp. plants, nicotine and anabasine, on parasite load and mortality in bumble bees (Bombus impatiens) infected with the intestinal parasite Crithidia bombi. Adult worker bees inoculated with C. bombi were fed nicotine and anabasine diet treatments in a factorial design, resulting in four nectar treatment combinations: 2 ppm nicotine, 5 ppm anabasine, 2ppm nicotine and 5 ppm anabasine together, or a control alkaloid-free solution. We conducted the experiment twice: first, with bees incubated under variable environmental conditions (‘Variable’; temperatures varied from 10-35°C with ambient lighting); and second, under carefully controlled environmental conditions (‘Stable’; 27°C incubator, constant darkness). In ‘Variable’, each alkaloid alone significantly decreased parasite loads, but this effect was not realized with the alkaloids in combination, suggesting an antagonistic interaction. Nicotine but not anabasine significantly increased mortality, and the two compounds had no interactive effects on mortality. In ‘Stable’, nicotine significantly increased parasite loads, the opposite of its effect in ‘Variable’. While not significant, the relationship between anabasine and parasite loads was also positive. Interactive effects between the two alkaloids on parasite load were non-significant, but the pattern of antagonistic interaction was similar to that in the variable experiment. Neither alkaloid, nor their interaction, significantly affected mortality under controlled conditions. Our results do not indicate synergy between Nicotiana nectar alkaloids; however, they do suggest a complex interaction between secondary metabolites, parasites, and environmental variables, in which secondary metabolites can be either toxic or medicinal depending on context

Parasite Removal, but Not Herbivory, Deters Future Parasite Attachment on Tomato

Plants face many antagonistic interactions that occur sequentially. Often, plants employ defense strategies in response to the initial damage that are highly specific and can affect interactions with subsequent antagonists. In addition to herbivores and pathogens, plants face attacks by parasitic plants, but we know little about how prior herbivory compared to prior parasite attachment affects subsequent host interactions. If host plants can respond adaptively to these different damage types, we predict that prior parasitism would have a greater deterrent effect on subsequent parasites than would prior herbivory. To test the effects of prior parasitism and prior herbivory on subsequent parasitic dodder (Cuscuta spp.) preference, we conducted two separate greenhouse studies with tomato hosts (Solanum lycopersicum). In the first experiment, we tested the effects of previous dodder attachment on subsequent dodder preference on tomato hosts using three treatments: control plants that had no previous dodder attachment; dodder-removed plants that had an initial dodder seedling attached, removed and left in the same pot to simulate parasite death; and dodder-continuous plants with an initial dodder seedling that remained attached. In the second experiment, we tested the effects of previous caterpillar damage (Spodoptera exigua) and mechanical damage on future dodder attachment on tomato hosts. Dodder attached most slowly to tomato hosts that had dodder plants previously attached and then removed, compared to control plants or plants with continuous dodder attachment. In contrast, herbivory did not affect subsequent dodder attachment rate. These results indicate that dodder preference depended on the identity and the outcome of the initial attack, suggesting that early-season interactions have the potential for profound impacts on subsequent community dynamics

The Effect of Larval Diet and Sex on Nectar Nicotine Feeding Preferences in Manduca Sexta (Lepidoptera: Sphingidae)

The article offers information on the study conducted regarding the implication of larval diet and nectar nicotine feeding to the sexual behavior of Manduca Sexta. Researchers found that many of these lepidoptera interact with the same plant species, both herbivorous larvae and nectar-feeding adults. Moreover, they found that the said behavior paves the way to influence both pollination and herbivory in floral and foliar tissue

Pollen from multiple sunflower cultivars and species reduces a common bumblebee gut pathogen

Pathogens are one of the factors driving pollinator declines. Diet can play an important role in mediating pollinator health and resistance to pathogens. Sunflower pollen (Helianthus annuus) dramatically reduced a gut pathogen (Crithidia bombi) of Bombus impatiens previously, but the breadth of this effect was unknown. We tested whether pollen from nine H. annuus cultivars, four wild H. annuus populations, H. petiolarus, H. argophyllus and two Solidago spp., reduced Crithidia in B. impatiens compared to mixed wildflower pollen and buckwheat pollen (Fagopyrum esculentum) as controls. We also compared hand- and honeybee-collected pollen (which contains nectar) to assess whether diet effects on pathogens were due to pollen or nectar. All Helianthus and Solidago pollen reduced Crithidia by 20–40-fold compared to buckwheat pollen, and all but three taxa reduced Crithidia compared to wildflower pollen. We found no consistent differences between hand- and bee-collected pollen, suggesting that pollen alone can reduce Crithidia infection. Our results indicate an important role of pollen diet for bee health and potentially broad options within the Asteraceae for pollinator plantings to manage bee disease

First Records of \u3cem\u3ePerdita bequaerti\u3c/em\u3e (Hymenopertera: Andrenidae) from New England

We document the first New England records of the bee Perdita bequaerti Viereck. Perdita bequaerti is a specialist on Asteraceae flowers, especially Helianthus, and is native to much of eastern North America, where it is often associated with predominantly sandy habitats. Previous records indicated P. bequaerti reached the northeastern extent of its range in the state of New York, but in August 2019, during a survey of bees on Helianthus, we collected five specimens of P. bequaerti at two sites in western Massachusetts. Both sites were located on small-scale farms with abundant non-native Helianthus, on soils characteristic of sandy outwash plains and silty floodplains. These records constitute an extension of P. bequaerti\u27s known range into New England

Testing Dose-Dependent Effects of the Nectar Alkaloid Anabasine on Trypanosome Parasite Loads in Adult Bumble Bees

The impact of consuming biologically active compounds is often dose-dependent, where small quantities can be medicinal while larger doses are toxic. The consumption of plant secondary compounds can be toxic to herbivores in large doses, but can also improve survival in parasitized herbivores. In addition, recent studies have found that consuming nectar secondary compounds may decrease parasite loads in pollinators. However, the effect of compound dose on bee survival and parasite loads has not been assessed. To determine how secondary compound consumption affects survival and pathogen load in Bombus impatiens, we manipulated the presence of a common gut parasite, Crithidia bombi, and dietary concentration of anabasine, a nectar alkaloid produced by Nicotiana spp. using four concentrations naturally observed in floral nectar. We hypothesized that increased consumption of secondary compounds at concentrations found in nature would decrease survival of uninfected bees, but improve survival and ameliorate parasite loads in infected bees. We found medicinal effects of anabasine in infected bees; the high-anabasine diet decreased parasite loads and increased the probability of clearing the infection entirely. However, survival time was not affected by any level of anabasine concentration, or by interactive effects of anabasine concentration and infection. Crithidia infection reduced survival time by more than two days, but this effect was not significant. Our results support a medicinal role for anabasine at the highest concentration; moreover, we found no evidence for a survival-related cost of anabasine consumption across the concentration range found in nectar. Our results suggest that consuming anabasine at the higher levels of the natural range could reduce or clear pathogen loads without incurring costs for healthy bees

Possible Synergistic Effects of Thymol and Nicotine Against Crithidia Bombi Parasitism in Bumble Bees

Floral nectar contains secondary compounds with antimicrobial properties that can affect not only plant-pollinator interactions, but also interactions between pollinators and their parasites. Although recent work has shown that consumption of plant secondary compounds can reduce pollinator parasite loads, little is known about the effects of dosage or compound combinations. We used the generalist pollinator Bombus impatiens and its obligate gut parasite Crithidia bombi to study the effects of nectar chemistry on host-parasite interactions. In two experiments we tested (1) whether the secondary compounds thymol and nicotine act synergistically to reduce parasitism, and (2) whether dietary thymol concentration affects parasite resistance. In both experiments, uninfected Bombus impatiens were inoculated with Crithidia and then fed particular diet treatments for 7 days, after which infection levels were assessed. In the synergism experiment, thymol and nicotine alone and in combination did not significantly affect parasite load or host mortality. However, the thymol-nicotine combination treatment reduced log-transformed parasite counts by 30% relative to the control group (P = 0.08). For the experiment in which we manipulated thymol concentration, we found no significant effect of any thymol concentration on Crithidia load, but moderate (2 ppm) thymol concentrations incurred a near-significant increase in mortality (P = 0.054). Our results tentatively suggest the value of a mixed diet for host immunity, yet contrast with research on the antimicrobial activity of dietary thymol and nicotine in vertebrate and other invertebrate systems. We suggest that future research evaluate genetic variation in Crithidia virulence, multi-strain competition, and Crithidia interactions with the gut microbe community that may mediate antimicrobial activities of secondary compounds