Location
D.P. Culp Center Ballroom
Start Date
4-5-2024 9:00 AM
End Date
4-5-2024 11:30 AM
Poster Number
140
Name of Project's Faculty Sponsor
Joe Bidwell
Faculty Sponsor's Department
Biological Sciences
Competition Type
Competitive
Type
Poster Presentation
Presentation Category
Science, Technology and Engineering
Abstract or Artist's Statement
Green salamanders (Plethodontidae: Aneides aeneus) are found throughout the Southern Appalachian Mountains and have been identified as a threatened species. The goal of this project is to characterize the genetic diversity in green salamanders (Aneides aeneus) at Bays Mountain Park in Kingsport, Tennessee to better understand the complex of species that make up the genus and to promote conservation efforts. There are four distinct lineages within the aeneus complex: northern, southern, hickory nut gorge, and blue ridge escarpment. Data from molecular studies investigating the genetic relationship among A. aeneus individuals are delimited making it difficult for researchers to accurately assess their conservation status. With more knowledge of the distribution of A. aeneus among the Southeast, conservation management practices can be updated and can increase the demand to recognize A. aeneus as an evolutionary significant unit (ESU). It is hypothesized that there will be no significant genetic differences between individuals in Bays; however, individuals in Bays will be more closely related to the northern lineage than the other lineages. Variation among the populations will be assessed by comparing sequenced individuals in Bays with those in the characterized northern lineage to address the presence or absence of gene flow. Comprehensive taxonomical information for A. aeneus is crucial for species identification and provides valuable insights on how to craft conservation strategies specifically tailored to the distinctive attributes and needs of green salamanders. Salamander tail tips (<5 mm) will be collected from 60 individuals in Bays Mountain Park. Locality data including the species sample number, species, state, county, latitude, longitude, elevation, and temperature will be recorded for all specimens. Correspondingly numbered plastic zip-lock bags and visible implant elastomer (VIE) tags will be used for identifying each specimen. Immediately after collection, tail tips will be preserved in 95-100% ethanol, then stored at –20°C at East Tennessee State University until extraction. Each sample will undergo DNA extraction, PCR, and gel electrophoresis. Two mitochondrial genes, cytochrome b and ND4, will be isolated and amplified using PCR. Resulting gene fragments will be visualized using gel electrophoresis to indicate successful amplification. After amplification of the targeted genes is verified, the samples will be sent for Sanger sequencing to determine the nucleotide sequence and identify genetic variants. Sequences will be cleaned, aligned, and analyzed using various programs. Variations in the genes, such as single nucleotide polymorphisms (SNPs), will be examined to assess genetic diversity and structure within and between populations. F-statistics models, such as Analysis of Molecular Variance (AMOVA) and phylogenetic models, such as Bayesian inference (BI), will be applied based on SNP allele frequencies. Genetic variation within the population in Bays is not likely but it is possible that it is distinctly different from the northern lineage. It is also possible that there is no genetic variation between those in Bays and the northern lineage.
Investigating the Genomic Variation among Green Salamanders (Aneides aeneus) in Bays Mountain Park and Planetarium
D.P. Culp Center Ballroom
Green salamanders (Plethodontidae: Aneides aeneus) are found throughout the Southern Appalachian Mountains and have been identified as a threatened species. The goal of this project is to characterize the genetic diversity in green salamanders (Aneides aeneus) at Bays Mountain Park in Kingsport, Tennessee to better understand the complex of species that make up the genus and to promote conservation efforts. There are four distinct lineages within the aeneus complex: northern, southern, hickory nut gorge, and blue ridge escarpment. Data from molecular studies investigating the genetic relationship among A. aeneus individuals are delimited making it difficult for researchers to accurately assess their conservation status. With more knowledge of the distribution of A. aeneus among the Southeast, conservation management practices can be updated and can increase the demand to recognize A. aeneus as an evolutionary significant unit (ESU). It is hypothesized that there will be no significant genetic differences between individuals in Bays; however, individuals in Bays will be more closely related to the northern lineage than the other lineages. Variation among the populations will be assessed by comparing sequenced individuals in Bays with those in the characterized northern lineage to address the presence or absence of gene flow. Comprehensive taxonomical information for A. aeneus is crucial for species identification and provides valuable insights on how to craft conservation strategies specifically tailored to the distinctive attributes and needs of green salamanders. Salamander tail tips (<5 >mm) will be collected from 60 individuals in Bays Mountain Park. Locality data including the species sample number, species, state, county, latitude, longitude, elevation, and temperature will be recorded for all specimens. Correspondingly numbered plastic zip-lock bags and visible implant elastomer (VIE) tags will be used for identifying each specimen. Immediately after collection, tail tips will be preserved in 95-100% ethanol, then stored at –20°C at East Tennessee State University until extraction. Each sample will undergo DNA extraction, PCR, and gel electrophoresis. Two mitochondrial genes, cytochrome b and ND4, will be isolated and amplified using PCR. Resulting gene fragments will be visualized using gel electrophoresis to indicate successful amplification. After amplification of the targeted genes is verified, the samples will be sent for Sanger sequencing to determine the nucleotide sequence and identify genetic variants. Sequences will be cleaned, aligned, and analyzed using various programs. Variations in the genes, such as single nucleotide polymorphisms (SNPs), will be examined to assess genetic diversity and structure within and between populations. F-statistics models, such as Analysis of Molecular Variance (AMOVA) and phylogenetic models, such as Bayesian inference (BI), will be applied based on SNP allele frequencies. Genetic variation within the population in Bays is not likely but it is possible that it is distinctly different from the northern lineage. It is also possible that there is no genetic variation between those in Bays and the northern lineage.