Abstract

Reproductive isolation is often variable within species, a phenomenon that while largely ignored by speciation studies, can be leveraged to gain insight into the potential mechanisms driving the evolution of genetic incompatibilities. We used experimental greenhouse crosses to characterize patterns of reproductive isolation among three divergent genetic lineages of Campanulastrum americanum that occur in close geographic proximity in the Appalachian Mountains. Substantial, asymmetrical reproductive isolation for survival due to cytonuclear incompatibility was found among the lineages (up to 94% reduction). Moderate reductions in pollen viability, as well as cytoplasmic male sterility, were also found between some Mountain populations. We then compared these results to previously established patterns of reproductive isolation between these Mountain lineages and a fourth, widespread Western lineage to fully characterize reproductive isolation across the complete geographic and genetic range of C. americanum. Reproductive isolation for survival and pollen viability was consistent across studies, indicating the evolution of the underlying genetic incompatibilities is primarily determined by intrinsic factors. In contrast, reproductive isolation for germination was only found when crossing Mountain populations with the Western lineage, suggesting the underlying genetic incompatibility is likely influenced by environmental or demographic differences between the two lineages. Cytoplasmic male sterility was also limited in occurrence, being restricted to a handful of Mountain populations in a narrow geographic range. These findings illustrate the complexity of speciation by demonstrating multiple, independent genetic incompatibilities that lead to a mosaic of genetic divergence and reproductive isolation across a species range.

Highlights

  • A major goal of evolutionary biology was to understand the processes that drive differentiation of populations within a species, generating genetic incompatibility, and leading to the formation of new species (Coyne & Orr, 2004)

  • To evaluate patterns of reproductive isolation (RI) among the Mountain lineages, nine populations were identified that were distributed throughout the Mountain region of C. americanum’s range, with two, three, and four populations sampled from the Eastern, Smoky, and Appalachian clades, respectively (Table S1, Figure 1b)

  • Substantial reproductive isolation was found when crossing between the three Mountain lineages of C. americanum, with up to a 94% reduction in cumulative fitness

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Summary

| INTRODUCTION

A major goal of evolutionary biology was to understand the processes that drive differentiation of populations within a species, generating genetic incompatibility, and leading to the formation of new species (i.e., speciation) (Coyne & Orr, 2004). Much work has been done characterizing genetic incompatibilities between species, and it is fairly well established that most genetic incompatibilities fit the Bateson-­Dobzhansky-­Muller model (Bateson-­Dobzhansky-­ Muller incompatibilities; Dobzhansky, 1937; Muller, 1942; Johnson, 2002), where incompatibilities occur between, rather than within loci (Sweigart & Willis, 2012) Theory suggests these incompatibilities involve alternate divergent alleles that have arisen and become fixed within populations of a species that are isolated and evolving. Even at the earliest stages (i.e., within species), we are likely to find multiple incompatibilities of varying strength (Barnard-­Kubow, So, & Galloway, 2016; Levy, 1991; Skrede, Brochmann, Borgen, & Rieseberg, 2008) Each one of these ­incompatibilities may be influenced by different evolutionary ­processes. What traits show reproductive isolation among divergent Mountain lineages? Is reproductive isolation among Mountain lineages predicted by geographic or ­genetic distance? by combining the current and previous range-w­ ide crossing studies, does reproductive isolation vary or remain constant across the geographic and genetic range of C. americanum, and how can that inform our understanding of the underlying genetic incompatibilities?

| MATERIALS AND METHODS
| Evaluation of reproductive isolation
| DISCUSSION
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