Quantitative nectar spur length governs nonrandom mating in a bee-pollinated Aquilegia species

Mating patterns in angiosperms are typically nonrandom, yet the mechanisms driving nonrandom mating remain unclear, especially regarding the effects of quantitative floral traits on plant mating success across male and female functions. In this study, we investigated how variation in spur length and flower number per plant influences mating patterns in Aquilegia rockii within a natural population. Using marker-based paternity analyses and manipulative experiments, we assessed the role of these traits in mating success across both sexual functions. We found significant variation in the mate composition between male and female function, with spur-length frequency positively associated with female outcrossing rate and mate number, but not with male outcrossing or mate number. Most mating events occurred within 10 m, and spur-length frequency positively correlated with mating distance. Regardless of selfing, there was evidence for assortative mating for spur length. Although spur length did not correlate with pollinator visitation, plants with mid-length spurs had higher seed set than those with shorter or longer spurs when autonomous selfing was excluded. Flowers number per plant was only associated with mating distance and female outcrossing rate. Our results suggest that spur length plays a key role in nonrandom mating by frequency-dependent mating, with implications for stabilizing selection and maintenance of genetic diversity. This study advances our understanding of floral diversity by dissecting the role of quantitative floral traits in plant mating through both female and male functions.