Do flower-colonizing microbes influence floral evolution? A test with fast-cycling Brassica.

Pollinators are thought to be the main drivers of floral evolution. Flowers are also colonized by abundant communities of microbes that can affect the interaction between plants and their pollinators. Very little is known, however, about how flower-colonizing microbes influence floral evolution. Here we performed a six-generation experimental evolution study using fast-cycling Brassica rapa, in which we factorially manipulated the presence of pollinators and flower microbes to determine how pollinators and microbes interact in driving floral evolution. We measured the evolution of six morphological traits, as well as plant mating system and flower attractiveness. Only one of the six traits (flower number) evolved in response to pollinators, while microbes did not drive the evolution of any trait, nor did they interact with pollinators in driving evolution of morphological traits. Moreover, we did not find evidence that pollinators or microbes affected the evolution of flower attractiveness to pollinators. However, we found an interactive effect of pollinators and microbes on the evolution of autonomous selfing, a trait that is expected to evolve in response to pollinator limitation. Overall, we found only weak evidence that microbes mediate floral evolution. However, our ability to detect an interactive effect of pollinators and microbes might have been limited by weak pollinator-mediated selection in our experimental setting. Our results contrast with previous (similar) experimental evolution studies, highlighting the susceptibility of such experiments to drift and to experimental artefacts.

Consequences of pollen defense compounds for pollinators and antagonists in a pollen-rewarding plant.

Plants produce an array of defensive compounds with toxic or deterrent effects on insect herbivores. Pollen can contain relatively high concentrations of such defense compounds, but the causes and consequences of this enigmatic phenomenon remain mostly unknown. These compounds could potentially protect pollen against antagonists but could also reduce flower attractiveness to pollinators. We combined field observations of the pollen-rewarding Lupinus argenteus with chemical analysis and laboratory assays to test three hypotheses for the presence of pollen defense compounds: (1) these compounds are the result of spillover from adjacent tissues, (2) they protect against pollen thieves, and (3) they act as antimicrobial compounds. We also tested whether pollen defense compounds affect pollinator behavior. We found a positive relationship between alkaloid concentrations in pollen and petals, supporting the idea that pollen defense compounds partly originate from spillover. However, pollen and petals exhibited quantitatively (but not qualitatively) distinct alkaloid profiles, suggesting that plants can adjust pollen alkaloid composition independently from that of adjacent tissues. We found no relationship between pollen alkaloid concentration and the abundance of pollen thieves in Lupinus flowers. However, pollen alkaloids were negatively associated with bacterial abundance. Finally, plants with more alkaloids in their pollen received more pollinator visits, but these visits were shorter, resulting in no change in the overall number of flowers visited. We propose that pollen defense compounds are partly the result of spillover from other tissues, while they also play an antimicrobial role. The absence of negative effects of these compounds on pollinator visitation likely allows their maintenance in pollen at relatively high concentrations. Taken together, our results suggest that pollen alkaloids affect and are mediated by the interplay of multiple interactions.

Pollen chemical and mechanical defences restrict host-plant use by bees.

Plants produce an array of chemical and mechanical defences that provide protection against many herbivores and pathogens. Putatively defensive compounds and structures can even occur in floral rewards: for example, the pollen of some plant taxa contains toxic compounds or possesses conspicuous spines. Yet little is known about whether pollen defences restrict host-plant use by bees. In other words, do bees, like other insect herbivores, tolerate the defences of their specific host plants while being harmed by non-host defences? To answer this question, we compared the effects of a chemical defence from Lupinus (Fabaceae) pollen and a putative mechanical defence (pollen spines) from Asteraceae pollen on larval survival of nine bee species in the tribe Osmiini (Megachilidae) varying in their pollen-host use. We found that both types of pollen defences reduce larval survival rate in some bee species. These detrimental effects were, however, mediated by host-plant associations, with bees being more tolerant of the pollen defences of their hosts, relative to the defences of plant taxa exploited by other species. This pattern strongly suggests that bees are adapted to the pollen defences of their hosts, and that host-plant use by bees is constrained by their ability to tolerate such defences.

Earlier spring reduces potential for gene flow via reduced flowering synchrony across an elevational gradient.

PREMISE: One of the best-documented ecological responses to climate warming involves temporal shifts of phenological events. However, we lack an understanding of how phenological responses to climate change vary among populations of the same species. Such variability has the potential to affect flowering synchrony among populations and hence the potential for gene flow. METHODS: To test whether an earlier start of the growing season affects the potential for gene flow among populations, we quantified the distributions of flowering times of two spring-flowering plants (Trillium erectum and Erythronium americanum) over 6 years along an elevational gradient. We developed a novel model-based metric of potential gene flow between pairs of populations to quantify the potential for pollen-mediated gene flow based on flowering phenology. RESULTS: Earlier onset of spring led to greater separation of peak flowering dates across the elevational gradient for both species investigated, but was only associated with a reduction in potential gene flow in T. erectum, not E. americanum. CONCLUSIONS: Our study suggests that climate change could decrease gene flow via phenological separation among populations along climatic gradients. We also provide a novel method for quantifying potential pollen-mediated gene flow using data on flowering phenology, based on a quantitative, more biologically interpretable model than other available metrics.

Defence compounds in pollen: why do they occur and how do they affect the ecology and evolution of bees?

Pollen plays two important roles in angiosperm reproduction, serving as a vehicle for the plant's male gametes, but also, in many species, as a lure for pollen-feeding animals. Despite being an important food source for many pollinators, pollen often contains compounds with known deterrent or toxic properties, as documented in a growing number of studies. Here we review these studies and discuss the role of pollen defensive compounds in the coevolutionary relationship between plants and bees, the preeminent consumers of pollen. Next, we evaluate three hypotheses that may explain the existence of defensive compounds in pollen. The pleiotropy hypothesis, which proposes that defensive compounds in pollen merely reflect physiological spillover from other plant tissues, is contradicted by evidence from several species. Although plants may experience selection to defend pollen against poor-quality pollinators, we also find only partial support for the protection-against-pollen-collection-hypothesis. Finally, pollen defences might protect pollen from colonisation by antagonistic microorganisms (antimicrobial hypothesis), although data to evaluate this idea are scarce. Further research on the effects of pollen defensive compounds on pollinators, pollen thieves, and pollen-colonising microbes will be needed to understand why many plants have chemically defended pollen, and the consequences of those defences for pollen consumers.

Herbivory and pollen limitation at the upper elevational range limit of two forest understory plants of eastern North America.

Studies of species' range limits focus most often on abiotic factors, although the strength of biotic interactions might also vary along environmental gradients and have strong demographic effects. For example, pollinator abundance might decrease at range limits due to harsh environmental conditions, and reduced plant density can reduce attractiveness to pollinators and increase or decrease herbivory. We tested for variation in the strength of pollen limitation and herbivory by ungulates along a gradient leading to the upper elevational range limits of Trillium erectum (Melanthiaceae) and Erythronium americanum (Liliaceae) in Mont Mégantic National Park, Québec, Canada. In T. erectum, pollen limitation was higher at the range limit, but seed set decreased only slightly with elevation and only in one of two years. In contrast, herbivory of T. erectum increased from <10% at low elevations to >60% at the upper elevational range limit. In E. americanum, we found no evidence of pollen limitation despite a significant decrease in seed set with elevation, and herbivory was low across the entire gradient. Overall, our results demonstrate the potential for relatively strong negative interactions (herbivory) and weak positive interactions (pollination) at plant range edges, although this was clearly species specific. To the extent that these interactions have important demographic consequences-highly likely for herbivory on Trillium, based on previous studies-such interactions might play a role in determining plant species' range limits along putatively climatic gradients.