Appendix A. Photograph of experimental mesocosms.

Photograph of experimental mesocosms

Morphological Divergence Driven by Predation Environment within and between Species of <i>Brachyrhaphis</i> Fishes

<div><p>Natural selection often results in profound differences in body shape among populations from divergent selective environments. Predation is a well-studied driver of divergence, with predators having a strong effect on the evolution of prey body shape, especially for traits related to escape behavior. Comparative studies, both at the population level and between species, show that the presence or absence of predators can alter prey morphology. Although this pattern is well documented in various species or population pairs, few studies have tested for similar patterns of body shape evolution at multiple stages of divergence within a taxonomic group. Here, we examine morphological divergence associated with predation environment in the livebearing fish genus <i>Brachyrhaphis</i>. We compare differences in body shape between populations of <i>B. rhabdophora</i> from different predation environments to differences in body shape between <i>B. roseni</i> and <i>B. terrabensis</i> (sister species) from predator and predator free habitats, respectively. We found that in each lineage, shape differed between predation environments, consistent with the hypothesis that locomotor function is optimized for either steady swimming (predator free) or escape behavior (predator). Although differences in body shape were greatest between <i>B. roseni</i> and <i>B. terrabensis</i>, we found that much of the total morphological diversification between these species had already been achieved within <i>B. rhabdophora</i> (29% in females and 47% in males). Interestingly, at both levels of divergence we found that early in ontogenetic development, females differed in shape between predation environments; however, as females matured, their body shapes converged on a similar phenotype, likely due to the constraints of pregnancy. Finally, we found that body shape varies with body size in a similar way, regardless of predation environment, in each lineage. Our findings are important because they provide evidence that the same source of selection can drive similar phenotypic divergence independently at multiple divergence levels.</p></div

Map of collection sites for <i>Brachyrhaphis terrabensis</i>, <i>B. roseni</i>, and <i>B. rhabdophora</i> used in this study.

<p><i>Brachyrhaphis terrabensis</i> (open circles) occur at higher elevations in streams that are void of fish predators. <i>Brachyrhaphis roseni</i> (closed circles) occur at lower elevations in streams that have abundant predators. <i>Brachyrhaphis rhabdophora</i> occur at sites that are both predator (closed squares) and predation free (open squares).</p

Results of mixed-repeated-measures MANOVA testing for interactions between combinations of species-group, predation-environment, size and index-variable.

<p>DF  =  degrees of freedom, f =  females, m =  males.</p

Results of mixed-repeated-measures MANOVA examining shape variation and sexual dimorphism in <i>Brachyrhaphis</i>.

<p>DF  =  degrees of freedom.</p

Statistical assessment of differences in trajectory size/ direction among trajectories characterizing sexual dimorphism in <i>Brachyrhaphis</i>.

<p><i>MD</i>_1,2 =  trajectory size, <i>θ</i>_1,2 =  trajectory direction, Taxa codes: 1 =  <i>Brachyrhaphis roseni</i>, 2 =  <i>B. terrabensis</i>, 3 =  <i>B. rhabdophora</i> from predator environments, and 4 =  <i>B. rhabdophora</i> from predator free environments. Significant differences generated empirically from 1,000 permutations are indicated in bold.</p