Successful DNA Extractions from South Dakota Only Leedy’S Roseroot Population
Location
Village Square
Start Date
28-4-2023 11:20 AM
Department
Biology
Abstract
Leedy’s roseroot (Rhodiola integrifolia ssp. leedyi) is a U.S. federally threatened plant species known in only seven populations. There are four populations in Minnesota, two populations in New York, and a single population with 218 individuals in South Dakota. The South Dakota population has been recognized as Leedy’s roseroot for less than 10 years and is located in the Black Elk Wilderness in the Black Hills National Forest. The population’s small size and isolation suggest that it might harbor unique alleles and that it might be experiencing inbreeding depression. We collected leaf tissue from 25 plants in August 2022 under Recovery Permit number ES86044B with an overall goal of describing the population’s genetic variability. A first challenge in studying the population’s genetic variability is that under our permit we could collect only approximately 200 mg of leaf tissue, and that like other succulents, Rhodiola species are rich in mucilaginous polysaccharides that can interfere with DNA extractions. Thus, our technical goal in this early phase of the project is to extract enough DNA (at least 100 ng) of a quality that we can use in Polymerase Chain Reaction (PCR) analyses. We used Qiagen DNeasy Plant Pro Kits to extract and purify DNA from 25 samples. DNA concentrations and purity of the extracts were measured using a nanodrop spectrophotometer. We checked the compatibility of the extracts with our PCR protocols by testing for amplification of the Internal Transcribed Spacer region from the extracts using agarose gel electrophoresis. The mean yield of DNA was 445 ng (SE = 45). Gel electrophoresis detected successful PCR amplification in 24 of the 25 samples tested. Our findings suggest that we can successfully move forward with our larger goal of describing the population’s genetic variability using microsatellite markers, which are non-coding, simple sequence repeats in DNA used for measuring genetic diversity within populations or among closely related species. Our future work will help estimate the South Dakota Leedy’s roseroot population’s potential adaptability to the environment, its relationships with the Minnesota and New York Leedy’s roseroot populations and will inform conservation practices for the population.
Faculty Sponsor
Joel Olfelt , Northeastern Illinois University
Successful DNA Extractions from South Dakota Only Leedy’S Roseroot Population
Village Square
Leedy’s roseroot (Rhodiola integrifolia ssp. leedyi) is a U.S. federally threatened plant species known in only seven populations. There are four populations in Minnesota, two populations in New York, and a single population with 218 individuals in South Dakota. The South Dakota population has been recognized as Leedy’s roseroot for less than 10 years and is located in the Black Elk Wilderness in the Black Hills National Forest. The population’s small size and isolation suggest that it might harbor unique alleles and that it might be experiencing inbreeding depression. We collected leaf tissue from 25 plants in August 2022 under Recovery Permit number ES86044B with an overall goal of describing the population’s genetic variability. A first challenge in studying the population’s genetic variability is that under our permit we could collect only approximately 200 mg of leaf tissue, and that like other succulents, Rhodiola species are rich in mucilaginous polysaccharides that can interfere with DNA extractions. Thus, our technical goal in this early phase of the project is to extract enough DNA (at least 100 ng) of a quality that we can use in Polymerase Chain Reaction (PCR) analyses. We used Qiagen DNeasy Plant Pro Kits to extract and purify DNA from 25 samples. DNA concentrations and purity of the extracts were measured using a nanodrop spectrophotometer. We checked the compatibility of the extracts with our PCR protocols by testing for amplification of the Internal Transcribed Spacer region from the extracts using agarose gel electrophoresis. The mean yield of DNA was 445 ng (SE = 45). Gel electrophoresis detected successful PCR amplification in 24 of the 25 samples tested. Our findings suggest that we can successfully move forward with our larger goal of describing the population’s genetic variability using microsatellite markers, which are non-coding, simple sequence repeats in DNA used for measuring genetic diversity within populations or among closely related species. Our future work will help estimate the South Dakota Leedy’s roseroot population’s potential adaptability to the environment, its relationships with the Minnesota and New York Leedy’s roseroot populations and will inform conservation practices for the population.