Host-plant selection is a key factor driving the ecology and evolution of insects.While the majority of phytophagous insects is highly host specific, generalist behavior is quite widespread among bees and presumably involves physiological adaptations that remain largely unexplored. However, floral visitation patterns suggest that generalist bees do not forage randomly on all available resources. While resource availability and accessibility as well as nectar composition have been widely explored, pollen chemistry could also have an impact on the range of suitable host-plants. This study focuses on particular pollen nutrients that cannot be synthesized de novo by insects but are key compounds of cell membranes and the precursor for molting process: the sterols. We compared the sterol composition of pollen from the main hostplants of three generalist bees: Anthophora plumipes, Colletes cunicularius, and Osmia cornuta, as well as one specialist bee Andrena vaga. We also analyzed the sterols of their brood cell provisions, the tissues of larvae and nonemerged females to determine which sterols are used by the different species. Our results show that sterols are not used accordingly to foraging strategy: Both the specialist species A. vaga and the generalist species C. cunicularius might metabolize a rare C 27 sterol, while the two generalist species A. plumipes and O. cornuta might rather use a very common C 28 sterol. Our results suggest that shared sterolic compounds among plant species could facilitate the exploitation of multiple host-plants by A. plumipes and O. cornuta whereas the generalist C. cunicularius might be more constrained due to its physiological requirements of a more uncommon dietary sterol. Our findings suggest that a bee displaying a generalist foraging behavior may sometimes hide a sterol-specialized species. This evidence challenges the hypothesis that all generalist free-living bee species are all able to develop on a wide range of different pollen types. K E Y W O R D Sbee-flower interactions, generalization, insects, physiological constraints, sterols | 151 VANDERPLANCK Et AL.

Sterols are essential nutrients for bees which are thought to obtain them exclusively from pollen. It is possible that variability in pollen sterol content shapes pollinator-flower relationships, as bee species require the physiological capacity or behavioral adaptations to cope with unfavorable sterolic composition of pollen. One behavioral adaptation used by generalist bees to avoid deficiencies is the mixing of different pollen types from multiple botanical families to achieve an optimal nutritional balance. However, a possible strategy that has never been investigated is the specific addition of nutrients by adult bees to pollen during foraging trips. Here, we analyzed the pollen sterols from 48 plant species and assessed their relation with the level of bee dietary specialization. We also investigated whether sterol addition or modification might occur during pollen collection by comparing handand bee-collected pollen for nine bee species. Our results show that sterolic composition tends to be similar within the same plant family, but there was no impact of overall relatedness. For pollen sterol content and bee specialization, pollen from plants used by specialist bees displayed more uncommon sterols than pollen from plants used by generalist bees. In addition, a sterol addition behavior may occur during foraging trips and could be considered a possible strategy to balance nutrient deficiencies. Such behavior was unrelated to bee specialization as this phenomenon was observed in both the generalist Apis mellifera and specialist Dasypoda hirtipes , suggesting that sterols might constitute a physiological constraint even for specialist species.