The Genetics of Postglacial Colonization and Peripherality in Northwestern Populations of the Spring Peeper, Pseudacris crucifer

Glaciation over the Pleistocene induced dramatic range fluctuations for species across North America such that postglacial recolonization by southern refugial lineages has characterized the genetic structure of northern North American species. Based on the leading edge model of postglacial range expansion, dispersal and rapid population growth in these northern taxa is expected to produce vast areas of genetic homogeneity. Previous work on the widely distributed spring peeper (Pseudacris crucifer) revealed six distinct mitochondrial lineages that diverged between 3-11 mya, expanding and contracting with glacial cycles. Beginning 16,000 yBP, receding glaciers permitted Eastern lineage refugia residing in the southern Appalachians to migrate northward into the St. Lawrence Valley then westward through most of central Canada. Peripheral populations at the northwestern range limit of P. crucifer in central Manitoba are likely descended from this westward expanding Eastern lineage. According to the central-marginal hypothesis, founder effects from colonization as well as limited gene flow is expected to reveal genetic differentiation and lower genetic diversity in peripheral populations. The goal of my study is to further our understanding of peripheral range dynamics in peripheral Manitoba populations of P. crucifer by determining their genetic affinity and diversity relative to more central populations in Ontario and Minnesota. In this study I amplified and aligned cytochrome b sequences from sample sites across central Manitoba to reconstruct a Bayesian phylogeny for P. crucifer; additionally, microsatellite loci were genotyped to estimate genetic diversity. Results from this study affirmed Eastern lineage descent for peripheral Manitoba sites by aligning with Ontario. Initial colonization by the Interior lineage between glacial retreat and the appearance of arid vicariance events may explain the apparent introgression of non-Eastern lineages in Manitoba. However, genetic diversity measured in expected heterozygosity (H¬e) was not found to be significantly different in Manitoba genotypes. Greater isolation by distance and inbreeding relative to Ontario and Minnesota is likely the primary driver of genetic variation in these sites. Further sampling is necessary to generate a more complete genetic population structure for P. crucifer.

Maternal and paternal growth regulatory factor expression in hybrid sunfish.

Unidirectional hybridization between bluegill (Lepomis macrochirus) and pumpkinseed (L. gibbosus) sunfish enables researchers to explore the relative expression of paternal and maternal alleles in hybrids. Past studies have found that the metabolic dysfunction in bluegill-pumpkinseed hybrids may be due to incompatibilities between nuclear and mitochondrial genomes. However, the consequences of hybridization on body size and muscle growth have not been examined. This topic is particularly interesting because hybrids grow larger than parentals despite the fact that they are often sired by smaller, precociously mature bluegills. In order to improve our understanding of growth dynamics in hybrid sunfish, I conducted real-time quantitative PCR using species-specific primers on the white muscle tissue of bluegills, pumpkinseeds, and hybrids collected from Lake Opinicon, ON. Five growth factors that have been linked to muscle growth and body size demonstrated similar expression for maternal and paternal alleles. While about half of the hybrids showed the same pattern with myogenin, about half showed very low levels of mRNA for the paternal (bluegill) gene. While this did not explain the heterosis seen in hybrids, it may explain the small body phenotype of the cuckholding bluegill males. I explored the upstream genetic structure of bluegill myogenin and established that four alleles exist within the population. Furthermore, I uncovered a relationship in hybrids between the proximal promoter/ 5’ UTR of myogenin and its transcript level. I found that the hybrids demonstrating low paternal myogenin expression unfailingly possessed A3 or A4 alleles, but future studies will be needed to reveal the molecular links between the genotype and the growth phenotype. A similar genotype-phenotype association was not obvious in parentals, even those that were homozygous for these alleles. Whether this relationship can provide insight into the genetic determinants of bluegill alternative mating strategies has yet to be determined.

Genomic consequences of mating system evolution in the Pacific coastal dune endemic, Abronia umbellata (Nyctaginaceae)

The advent of next-generation sequencing has significantly reduced the cost of obtaining large-scale genetic resources, opening the door for genomic studies of non-model but ecologically interesting species. The shift in mating system, from outcrossing to selfing, has occurred thousands of times in angiosperms and is accompanied by profound changes in the population genetics and ecology of a species. A large body of work has been devoted to understanding why the shift occurs and the impact of the shift on the genetics of the resulting selfing populations, however, the causes and consequences of the transition to selfing involve a complicated interaction of genetic and demographic factors which are difficult to untangle. Abronia umbellata is a Pacific coastal dune endemic which displays a striking shift in mating system across its geographic range, with large-flowered outcrossing populations south of San Francisco and small-flowered selfing populations to the north. Abronia umbellata is an attractive model system for the study of mating system transitions because the shift appears to be recent and therefore less obscured by post-shift processes, it has a near one-dimensional geographic range which simplifies analysis and interpretation, and demographic data has been collected for many of the populations. In this study, we generated transcriptome-level data for 12 plants including individuals from both subspecies, along with a resequencing study of 48 individuals from populations across the range. The genetic analysis revealed a recent transition to selfing involving a drastic reduction in genetic diversity in the selfing lineage, potentially indicative of a recent population bottleneck and a transition to selfing due to reproductive assurance. Interestingly, the genetic structure of the populations was not coincident with the current subspecies demarcation, and two large-flowered populations were classified with the selfing subspecies, suggesting a potential need for re-evaluation of the current subspecies classification. Our finding of low diversity in selfing populations may also have implications for the conservation value of the threatened selfing subspecies.