Robin Hopkins

Pollinator-mediated selection on flower color allele drives reinforcement.

The Constant Pollinator In ecology, reinforcement is the process by which species prevent hybridization and maintain species boundaries, but the underlying genetic mechanisms are unclear. Hopkins and Rausher (p. 1090, published online 2 February) examined reinforcement between two species of a wild flowering plant called Phlox that show incomplete hybrid sterility. Down-regulation of a flavonoid gene produces red flowers and operates in concert with a color intensity locus to adjust flower color and tone. A distinct geography of flower color has emerged in which it appears that dark coloration causes less hybridization between the species because the butterfly pollinators tend to favor light-blue flower color variants. If pollinators visit flowers with similar phenotypes more frequently than flowers with dissimilar phenotypes, this will decrease gene flow between the unlike flowers. Butterfly behavior reduces hybridization and reinforces speciation among Phlox plants. Reinforcement is the process by which reduced hybrid fitness generates selection favoring the evolution of stronger prezygotic reproductive barriers between emerging species. Using common-garden field experiments, we quantified the strength of reinforcing selection in nature by demonstrating strong selection favoring an allele conferring increased pigment intensity in the plant Phlox drummondii in areas of sympatry with the closely related species Phlox cuspidata. Incomplete hybrid sterility between the two species generates selection for traits that decrease interspecies hybridization. In contrast, selection on this locus is undetectable in the absence of P. cuspidata. We demonstrate that reinforcing selection is generated by nonrandom pollinator movement, in which pollinators move less frequently between intensely pigmented P. drummondii and P. cuspidata than between lightly pigmented P. drummondii and P. cuspidata.

Identification of two genes causing reinforcement in the Texas wildflower Phlox drummondii

Species formation generates biological diversity and occurs when traits evolve that prevent gene flow between populations. Discerning the number and distribution of genes underlying these traits and, in a few cases, identifying the genes involved, has greatly enhanced our understanding over the past 15 years of species formation (reviewed by Noor and Feder and Wolf et al.). However, this work has almost exclusively focused on traits that restrict gene flow between populations that have evolved as a by-product of genetic divergence between geographically isolated populations. By contrast, little is known about the characteristics of genes associated with reinforcement, the process by which natural selection directly favours restricted gene flow during the formation of species. Here we identify changes in two genes that appear to cause a flower colour change in Phlox drummondii, which previous work has shown contributes to reinforcement. Both changes involve cis-regulatory mutations to genes in the anthocyanin biosynthetic pathway (ABP). Because one change is recessive whereas the other is dominant, hybrid offspring produce an intermediate flower colour that is visited less by pollinators, and is presumably maladaptive. Thus genetic change selected to increase prezygotic isolation also appears to result in increased postzygotic isolation.

A latitudinal cline and response to vernalization in leaf angle and morphology in Arabidopsis thaliana (Brassicaceae)

Adaptation to latitudinal patterns of environmental variation is predicted to result in clinal variation in leaf traits. Therefore, this study tested for geographic differentiation and plastic responses to vernalization in leaf angle and leaf morphology in Arabidopsis thaliana. Twenty-one European ecotypes were grown in a common growth chamber environment. Replicates of each ecotype were exposed to one of four treatments: 0, 10, 20 or 30 d of vernalization. Ecotypes from lower latitudes had more erect leaves, as predicted from functional arguments about selection to maximize photosynthesis. Lower-latitude ecotypes also had more elongated petioles as predicted by a biomechanical constraint hypothesis. In addition, extended vernalization resulted in shorter and more erect leaves. As predicted by functional and adaptive hypotheses, our results show genetically based clinal variation as well as environmentally induced variation in leaf traits.

MOLECULAR SIGNATURES OF SELECTION ON REPRODUCTIVE CHARACTER DISPLACEMENT OF FLOWER COLOR IN PHLOX DRUMMONDII

Character displacement, which arises when species diverge in sympatry to decrease competition for resources or reproductive interference, has been observed in a wide variety of plants and animals. A classic example of reproductive character displacement, presumed to be caused by reinforcing selection, is flower‐color variation in the native Texas wildflower Phlox drummondii. Here, we use population genetic analyses to investigate molecular signatures of selection on flower‐color variation in this species. First, we quantify patterns of neutral genetic variation across the range of P. drummondii to demonstrate that restricted gene flow and genetic drift cannot explain the pattern of flower‐color divergence in this species. There is evidence of extensive gene flow across populations with different flower colors, suggesting selection caused flower‐color divergence. Second, analysis of sequence variation in the genes underlying this divergence reveals a signature of a selective sweep in one of the two genes, further indicating selection is responsible for divergence in sympatry. The lack of a signature of selection at the second locus does not necessarily indicate a lack of selection on this locus but instead brings attention to the uncertainty in depending on molecular signatures to identify selection.

The Cost of Reinforcement: Selection on Flower Color in Allopatric Populations of Phlox drummondii*

Reinforcement is the process by which increased reproductive isolation between incipient species evolves due to selection against maladaptive hybrids or costly hybrid mating. Reinforcement is predicted to create a pattern of greater prezygotic reproductive isolation in regions where the two species co-occur, sympatry, than in allopatry. Although most research on reinforcement focuses on understanding the evolutionary forces acting in sympatry, here we consider what prevents the alleles conferring greater reproductive isolation from spreading into allopatry. We investigate flower color divergence in the wildflower Phlox drummondii, which is caused by reinforcement in the regions sympatric with its congener Phlox cuspidata. Specifically, we performed common garden field experiments and pollinator observations to estimate selection acting on flower color variation in allopatry. We combine our estimates of maternal and paternal fitness using simulations and predict how flower color alleles migrating from sympatry will evolve in allopatry. Our results suggest that strong pollinator preference for the ancestral flower color in allopatry can maintain divergence between allopatric and sympatric populations.

Strong Reinforcing Selection in a Texas Wildflower

Reinforcement, the process of increased reproductive isolation due to selection against hybrids, is an important mechanism by which natural selection contributes to speciation [1]. Empirical studies suggest that reinforcement has generated reproductive isolation in many taxa (reviewed in [2-4]), and theoretical work shows it can act under broad selective conditions [5-11]. However, the strength of selection driving reinforcement has never been measured in nature. Here, we quantify the strength of reinforcing selection in the Texas wildflower Phlox drummondii using a strategy that weds a population genetic model with field data. Reinforcement in this system is caused by variation in two loci that affect flower color [12]. We quantify sharp clines in flower color where this species comes into contact with its congener, Phlox cuspidata. We develop a spatially explicit population genetic model for these clines based on the known genetics of flower color. We fit our model to the data using likelihood, and we searched parameter space using Markov chain Monte Carlo methods. We find that selection on flower color genes generated by reinforcement is exceptionally strong. Our findings demonstrate that natural selection can play a decisive role in the evolution of reproductive isolation through the process of reinforcement.

Limited hybridization across an edaphic disjunction between the gabbro-endemic shrub Ceanothus roderickii (Rhamnaceae) and the soil-generalist Ceanothus cuneatus

UNLABELLED PREMISE OF THE STUDY Hybridization is thought to have played an important role in diversification of the speciose shrub genus Ceanothus; putative hybrid species have been described, and data suggest that intrinsic barriers may not exist among closely related species. However, the extent to which hybridization occurs in the wild is not known, and little is understood about how extrinsic factors such as soil chemistry may influence the process. The present research focuses on the gabbro-endemic C. roderickii and the closely related soil-generalist C. cuneatus. Though the species occur peripatrically, they remain distinct across an edaphic disjunction. • METHODS AFLP was used to quantify hybridization and introgression. Biological data and experiments were used to test for prezygotic isolation. Growth trials were used to test for local adaptation and selection against hybrids. • KEY RESULTS Ceanothus cuneatus and C. roderickii were strongly differentiated morphologically and genetically, despite a lack of evidence for prezygotic barriers. Hybrids and back-crosses were present but infrequent. Finally, there was selection against hybrids in nonnative soil. • CONCLUSIONS There is little genetic exchange between the focal species across an edaphic disjunction, despite the absence of prezygotic barriers. This result implies that soil conditions, as well as other extrinsic factors, should be considered as forces that may restrict hybridization and gene flow in Ceanothus, influencing local adaptation and speciation. Findings presented here are significant because they imply that exchange of genetic material between plants may be limited directly by the abiotic environment, rather than by the biology of the plants.

Dispensing Pollen Via Catapult: Explosive Pollen Release in Mountain Laurel (Kalmia latifolia)

The astonishing amount of floral diversity has inspired countless assumptions about the function of diverse forms and their adaptive significance, yet many of these hypothesized functions are untested. We investigated an often-repeated adaptive hypothesis about how an extreme floral form functions. In this study, we conducted four investigations to understand the adaptive function of explosive pollination in Kalmia latifolia, the mountain laurel. We first performed a kinematic analysis of anther movement. Second, we constructed a heat map of pollen trajectories in three-dimensional space. Third, we conducted field observations of pollinators and their behaviors while visiting K. latifolia. Finally, we conducted a pollination experiment to investigate the importance of pollinators for fertilization success. Our results suggest that insect visitation dramatically improves fertilization success and that bees are the primary pollinators that trigger explosive pollen release.