College of Arts and Sciences
Primary Subject Area
Eurycea, E. Lucifuga, E. Longicauda, E. guttolineata, Cytochrome b, polymerase chain reaction
Ecology and Evolutionary Biology | Genetics and Genomics | Laboratory and Basic Science Research
Genomic sequencing is a powerful tool that has many applications for research, one of which is in the field of taxonomy and the identification of species. This thesis discusses the mitochondrial gene cytochrome b and its utility in population genetics and identification of larval amphibians. The development of the Polymerase Chain Reaction and primers are an integral part of the modern DNA sequencing process. The Polymerase Chain Reaction is used to amplify a target DNA sequence, and the protocol for this procedure must be optimized for the specific sequence of target DNA. Primers must also be designed and modified for a selected portion of the DNA to be copied. This thesis also discusses the application of these techniques in a current study of a population of Eurycea. Three species of Eurycea, the Cave Salamander (E. lucifuga), Long-tailed Salamander (E. longicauda longicauda), and Three-lined Salamander (E. guttolineata) were discovered in an abandoned mine shaft near Riverville, Amherst County, Virginia in 1999. The population is unusual, as E. longicauda longicauda and E. lucifuga are outside their normal distribution and this was the first syntopic occurrence in Virginia of E. guttolineata and E. longicauda longicauda, the species usually indigenous to the Piedmont physiographic region. In addition, there is possible hybridization between E. guttolineata and E. longicauda. The larvae of these species are difficult to identify morphologically; so the paper discusses the use of cytochrome b sequencing for species identification among this population of Eurycea.