The role of pollinator preference in the maintenance of pollen colour variation

Background and Aims Pollinators often drive the evolution of floral traits, but their capacity to influence the evolution of pollen colour remains unclear. Pollen colour in Campanula americana is variable and displays a longitudinal cline from prevalence of deep purple in western populations to white and light-purple pollen in eastern populations. While selection for thermal tolerance probably underlies darker pollen in the west, factors contributing to the predominance of light pollen in eastern populations and the maintenance of colour variation within populations throughout the range are unknown. Here we examine whether pollinators contribute to the maintenance of pollen colour variation in C. americana. Methods In a flight cage experiment, we assessed whether Bombus impatiens foragers can use pollen colour as a reward cue. We then established floral arrays that varied in the frequency of white-and purple-pollen plants in two naturally occurring eastern populations. We observed foraging patterns of wild bees, totalling >1100 individual visits. Key Results We successfully trained B. impatiens to prefer one pollen colour morph. In natural populations, the specialist pollinator, Megachile campanulae, displayed a strong and consistent preference for purple-pollen plants regardless of morph frequency. Megachile also exhibited a bias toward pollen-bearing male-phase flowers, and this bias was more pronounced for purple pollen. The other main pollinators, Bombus spp. and small bees, did not display pollen colour preference. Conclusions Previous research found that Megachile removes twice as much pollen per visit as other bees and can deplete pollen from natural populations. Taken together, these results suggest that Megachile could reduce the reproductive success of plants with purple pollen, resulting in the prevalence of light-coloured pollen in eastern populations of C. americana. Our research demonstrates that pollinator preferences may play a role in the maintenance of pollen colour variation in natural populations.