Distribution and Conservation Genetics of the Cow Knob Salamander, Plethodon punctatus Highton (Caudata: Plethodontidae) Thesis submitted to The Graduate College of Marshall University In partial fulfillment of the Requirements for the degree Master of Science Biological Sciences by Matthew R. Graham Thomas K. Pauley, Committee Chairman Victor Fet, Committee Member Guo-Zhang Zhu, Committee Member April 29, 2007 ii Distribution and Conservation Genetics of the Cow Knob Salamander, Plethodon punctatus Highton (Caudata: Plethodontidae) MATTHEW R. GRAHAM Department of Biological Sciences, Marshall University Huntington, West Virginia 25755-2510, USA email:
[email protected] Summary Being lungless, plethodontid salamanders respire through their skin and are especially sensitive to environmental disturbances. Habitat fragmentation, low abundance, extreme habitat requirements, and a narrow distribution of less than 70 miles in length, makes one such salamander, Plethodon punctatus, a species of concern (S1) in West Virginia. To better understand this sensitive species, day and night survey hikes were conducted through ideal habitat and coordinate data as well as tail tips (10 to 20 mm in length) were collected. DNA was extracted from the tail tips and polymerase chain reaction (PCR) was used to amplify mitochondrial 16S rRNA gene fragments. Maximum parsimony, neighbor-joining, and UPGMA algorithms were used to produce phylogenetic haplotype trees, rooted with P. wehrlei. Based on our DNA sequence data, four disparate management units are designated. Surveys revealed new records on Jack Mountain, a disjunct population that expands the known distribution of the species 10 miles west. In addition, surveys by Flint verified a population on Nathaniel Mountain, WV and revealed new records on Elliot Knob, extending the known range several miles south.