21Diapause is a physiological arrest of development ahead of adverse environmental conditions and is 22 a critical phase of the life cycle of many insects. In bees, diapause has been reported in species from 23 all seven taxonomic families. However, they exhibit a variety of diapause strategies. These different 24 strategies are of particular interest since shifts in the phase of the insect life cycle in which diapause 25 occurs has been hypothesized to promote the evolution of sociality. Here we provide a comprehensive 26 evaluation of this hypothesis with phylogenetic analysis and ancestral state reconstruction of the 27 ecological and evolutionary factors associated with diapause phase. We find that social lifestyle, 28 latitude, and voltinism are significant predictors of the life stage in which diapause occurs. Ancestral 29 state reconstruction revealed that the most recent common ancestor of all bees likely exhibited 30 developmental diapause and shifts to adult or reproductive diapause have occurred in the ancestors 31 of lineages in which social behavior has evolved. These results provide fresh insight regarding the 32 role of diapause as a prerequisite for the evolution of sociality in bees.33 34 35 36 37 38 39 40 41 42 45 this phase of dormancy in insects, including diapause, adult diapause, reproductive diapause, 46 hibernation, adult-wintering and overwintering. A defining feature of all these terms is an arrest in 47 development or activity that is hormonally programmed in advance of environmental adversities such 48 as harsh winter, dry seasons, or food restriction [1,2]. Diapause may occur at any stage of life: egg, 49 larval, pupal or adult [1,3], and metabolic suppression varies from a decrease in activity (diapause in 50 adult phase) to complete developmental arrest (diapause during development) [4]. Diapause may also 51 be obligatory or facultative. Most obligatory diapausers live at high latitudes and produce only one 52 generation per year. Conversely, in warmer regions, there are multiple active generations before 53 winter and only one will pass through diapause (facultative diapause), and in tropical regions without 54 extreme seasonal variation in resources, many insects forego diapause [5].
55Bees are a diverse group of holometabolous insects in the Order Hymenoptera encompassing 56 more than 20,000 species in seven taxonomic families [6]. Diapause has been reported in species of 57 each family, with a great deal of variation in strategies. Although diapause in bees may occur in any 58 developmental phase, the vast majority of species diapause during the last larval instar, immediately 59 prior to metamorphosis, called the prepupal phase [7,8] or in the adult phase after mating and before 60 the foundation of a new nest [9,10]. Also, a considerable number of species diapause as 61 reproductively active adults, which is also known as reproductive diapause [11,12]. Some bee species 62 are active year round and do not diapause [13,14].
63The diversity of diapause strategies among bees i...