We study the genetic architecture of an asexual animal Habrotrocha rosa, a bdelloid rotifer that exists as a metapopulation, within the vase-shaped, rainwater-filled leaves of the carnivorous pitcher plant, Sarracenia purpurea. Our lab is conducting this study to understand how organisms interact in the S. purpurea model system. In this study we are interested in the amount of genetic variation of two enzymes, Cytochrome-oxidase-1 (cox1) and Cytochrome-oxidase-b (cob), within segments of DNA from H. rosa collected among different levels of spatial and temporal scales: 1) within a specific leaf, 2) between leaves on a specific plant, 3) between leaves from different pitchers within the same bog, and 4) between leaves of different pitcher from different bogs. We aim to answer the following question: at what level within a scale do we observe genetic variation, or are all the rotifers genetically the same? The mechanism, where H. rosa migrates between suitable habitat patches, is unknown.
Results/Conclusions:
We aseptically sampled ~5 mL of fluid from the inside of S. purpurea leaves; 45 samples were from MLB & 15 Samples were collected from CBB. In each collected sample, 3 individual H. rosa were isolated from the pitcher fluid sample; these females will become the foundation for the clone populations. Each rotifer was prepared for culture by washing them with autoclaved DI H2O to remove all organic and inorganic impurities. She then was allowed to culture a cloned population in a concave microscope slide before being transferred to larger containers. Bio-Rad DNA Extraction kits were utilized to extract and duplicate DNA segments for cox1 and cob alleles. The Polymerase chain reaction products are then analyzed through gel electrophoresis. This will allow us to determine spatial and temporal levels of genetic variation. We have approximately 100 distinct clone populations that are in process of this genomic analysis procedure.
