Comparing mitochondrial autoradiographs of four species of Ontario turtles in order to gain a sense of the evolutionary relationship between these

species

Heather Filyk 10009402

Tuesday 4:00 Lab, Rm.2320 Samantha Lo

T.A.:

Biology 102

November 22nd, 2011

Abstract The purpose of this study was to compare mitochondrial DNA sequences of four species of Ontario turtles in order to gain a sense of the evolutionary relationship between these species. The four species of turtles are, the Clemmys guttata (spotted turtle), the Graptemys geographica (Northern map turtle), the Chelydra serpentina (snapping turtle) and, the Emydoidea blandingii (Blandings turtle). An autoradiograph of each species was examined to obtain the number of point mutations in the mitochondrial DNA. A table of differences was then set up in order to compare all the point mutations between the species of turtles. The Chelydra serpentina was found to have the most differences in its mitochondrial DNA, the Clemmys guttata had the second most differences in its mitochondrial DNA and the Graptemys geographica and the Emydoidea blandingii had the same amount of differences in their mitochondrial DNA. The Chelydra serpentina probably branched off from the common ancestor first because it has the most point mutations causing it to be the most distantly related, following this theory the Clemmys guttata would have then branched off from the common ancestor second and the Graptemys geographica and, Emydoidea blandingii would have branched off at around the same time from the common ancestor. Some factors that may have caused the species of turtles to evolve differently from each other are that they may have migrated separately to an environment and habitat that is completely different from the one they lived in. Therefore they would most likely evolve to adapt to their new surroundings by possibly changing their genetic makeup.

Introduction

Evolution, as stated by Darwin, is the process of natural selection. The four Ontario turtle species researched in this lab all have one or more changes in their nucleotide sequences in their mitochondrial DNA. These changes may be able to explain the relevant amount of divergence of the species and this in turn may be able to help create a phylogeny. This is important research because understanding relationships between organisms can lead to a better understanding of why species from one common ancestor diverge and evolve differently. Researchers typically use mitochondrial DNA to study phylogeny because it has several attributes, including lack of significant recombination due to its uniparental inheritance, and amenability to the polymerase chain reaction (PCR) (Fujita et al. 2003). PCR enables researchers to retrieve copies of DNA quickly. These qualities have made mitochondrial DNA a good phylogenic tool (Fujita et al. 2003). There have been experiments done with mitochondrial DNA one of which showed that it evolves fast, they found that this may be due its inefficient ability to repair mitochondrial DNA damage (Wesley et al. 1979). This may account for the divergence in these four different species mitochondrial DNA. The Pleistocene climate that was present during the Ice Ages caused genetic diversification of many taxa (Amato et al. 2008) possibly including the Ontario turtles of this study. The Ice Age probably caused this genetic diversification because it forced organisms to migrate to new geographical regions. A study done on a wide variety of soft shell turtles in different geographical areas of America found that turtles in the same geographical regions were more closely related to each other genetically (Weisrock and Janzen 2000). This statement proposed by Kawecki and Ebert (2004), stating that divergent selection should cause each local population to evolve traits that provide an advantage under its local environmental conditions, supports the evidence shown in the other studies. The different geographical regions in Ontario that the turtles may live in and historical climate changes, may be some of the factors associated with the point mutations in the mitochondrial DNA of the turtles that the autoradiograph had shown. I propose to contribute to current data by analyzing an autoradiograph of the four turtles mitochondrial DNA. I will analyze this by looking for point mutations that differentiate the turtles DNA from each other. With the comparison of the number of point mutations in the species, the ability to

identify how closely related the turtles are to themselves, and to the common ancestor may become apparent.

Results The results of the phylogenic tree (see Figure 1) show that the Chelydra serpentina (snapping turtle) branches off from the common ancestor first. The Clemmys guttata (spotted turtle) is the second to branch off from the common ancestor. The Emydoidea blandingii (Blandings turtle) and the Graptemys geographica (Northern map turtle) branch species the most nucleotides of mitochondrial DNA would first, thus the two most similar species to the common ancestor would branch off last. off from the common ancestor at the same point. It was determined that the of turtle that had deviations in the their branch off

Figure 1. The phylogenetic tree of the four Ontario turtle species studied: the Clemmys guttata (spotted turtle), the Graptemys geographica (Northern map turtle), the Chelydra serpentina (snapping turtle) and the Emydoidea blandingii (Blandings turtle).

Literature Cited

Amato, M. L., R. J. Brooks, and J. Fu. 2008. A phylogeographic analysis of populations of the wood turtle (Glyptemys insculpta) throughout its range. Molecular Ecology 17: 570581. Fujita, M. K., T. N. Engstrom, D. E. Starkey, and H. B. Shaffer. 2003. Turtle phylogeny: insights from a novel nuclear intron. Molecular Phylogenetics and Evolution 31: 1031-1040. Kawecki, T. J., and D. Ebert. 2004. Conceptual issues in local adaptation. Ecology Letters 7: 1225-1241. Weisrock, D. W., and F. J. Janzen. 2000. Comparative Molecular Phylogeography of North American Softshell Turtles (Apalone): Implications for Regional and Wide-Scale Historical Evolutionary Forces. Molecular Phylogenetics and Evolution 14: 152-164.

Wesley, M. B., M. George, and A. C. Wilson. 1979. Rapid evolution of animal mitochondrial DNA. Proceedings of the National Academy of Sciences of the United States of America 76: 1967-1971.