DEVELOPMENT OF FECAL DNA SAMPLING METHODS TO ASSESS GENETIC POPULATION STRUCTURE OF GREATER YELLOWSTONE BISON
The bison (Bison bison) of Yellowstone National Park (YNP) and Grand Teton National Park (GTNP) represent two of only three remaining populations of plains bison that have no evidence of hybridization with cattle. Therefore, these bison are an important source for ecological and genetic restoration of wild bison. Little is known regarding genetic population structure and gene flow among the Greater Yellowstone Area (GYA) bison herds. I evaluated the feasibility of fecal DNA sampling for genetic analyses of wild bison populations. I used matched blood and fecal samples from eight radio-collared bison from Hayden Valley breeding group (YNP), and multiplex polymerase chain reaction (PCR) of four microsatellite loci to assess amplification success and genotyping error rates. The amplification success rate was 92% and the genotyping error rate was 12% on average across all individuals, and loci. Exclusion of two poor quality samples from data analyses increased amplification success to 97%, and reduced the genotyping error rate to 4%. I PCR amplified a 470 bp mitochondrial DNA (mtDNA) fragment for sequencing, and successfully identified haplotypes for120 individuals. The error rate for mtDNA sequencing was 0.0005 nucleotide mis-incorporations across all samples. Sequencing and RFLP analysis of mtDNA control region from 179 fecal samples collected over two consecutive seasons was conducted to evaluate population structure among YNP breeding groups, and between GTNP and YNP bison populations. I found significant genetic distinction between YNP and GTNP bison populations (FST = 0.191, p < 0.001). The differences in haplotype frequencies between Hayden Valley and Lamar Valley breeding groups were highly significant (FST = 0.367, p < 0.001), and nearly two times greater than between GTNP and YNP thus providing evidence for at least two genetically distinct breeding groups within YNP. Differences between breeding groups remained significant even though haplotype frequencies were different between years within Hayden Valley (FST = 0.054, p < 0.05). The techniques and protocols developed have allowed high amplification success, low genotyping and sequencing error rates. This study demonstrated that non-invasive fecal DNA sampling is feasible for bison, and detected fine-scale population genetic structure in among GYA bison, suggesting female philopatry.
Advisor:Fred W. Allendorf; Gordon Luikart; Mark Hebblewhite; Richmond Clow; Rick Wallen
School:The University of Montana
School Location:USA - Montana
Source Type:Master's Thesis
Date of Publication:07/24/2007