1. A Summer of Research Annie Roessler Centre College Brown Fellows Program
2. Project Schedule Centre College Prolactin and Nest Attachment University of Cincinnati DNA-based species Delimitation Sierra Club Alpine Beginner Backpacking Course
8. DNA Types Mitochondrial DNA Inherited from mother only Not highly conserved Rapid mutation rate Form single clades quickly Easy to amplify Sometimes does not provide monophyletic clades (paraphyly) Nuclear DNA Inherited from both parents Highly conserved Fewer mutations Form single clades slowly More difficult to amplify Monophyletic info
9. Ravinia Limited morphological characters Agriculturally important 17 species living in N. America (12 present in this study)
10. Subjects Twelve species of North American Ravinia Multiple individuals Multiple populations Additional samples collected in Burnet Woods
12. Experiment Design Ravinia DNA Digestion Plasmid Extraction Vector Digestion Ligation Vector and Insert Transformation of E. coli Sequencing of Inserts
13. SUMR-UC Schedule 40 hour work week Hands on laboratory work Subject collections Literature reviews
14. 1 of 12 samples Sequence length: 710 base pairs Not similar to anything in database Not vector or human Results
15. Conclusions and Implications Newly sequenced nuclear segment Provide phylogenetic information Create a gene tree with mitochondrial info Create a species tree with nuclear info Understanding of evolutionary systems Conservation of protected species Other fields Agriculture Ecology
16. Generate primers off of acquired DNA Testing generated primers on multiple species of Ravinia Creating species and gene trees from sequence data Testing species boundaries and limits Infer species status of questionable Raviniataxa Future Directions
17. Terminology Lab techniques Lack of chemistry background Communication Literature reading Career advice SUMR-UC Challenges and Strengths
19. Field Research Laboratory research Prolactin and Nest Attachment Ongoing research Ravinia and Gene Trees 8 weeks in summer 2011 Field research Natural habitat Free-living, wild animals Avoid changing/altering behavior
20. Famous Field Researchers Jane Goodall Anthropology 120 Thomas Defler American Primatology Convention speaker Lived and worked in Colombia
21. My Interests Animal behavior Ecology The Green Movement Psychology Conservation Animal Welfare
22. The Sierra Club To explore, enjoy, and protect the wild places of the earth; To practice and promote the responsible use of the earth's ecosystems and resources; To educate and enlist humanity to protect and restore the quality of the natural and human environment; and to use all lawful means to carry out these objectives.
24. Alpine Beginner Backpack Design Slow, steady approach Rocky Mountain National Park Continental Divide Trail Six day, 28 mile loop Variety of terrain Gain and lose 4,000 feet in elevation Summit Hallet’s Peak: 12,700 ft
25. Environment Subalpine and Alpine Regions Forest Tundra Snow fields Boulder fields High winds Meadows Glacial Lakes
26. Equipment Medium weight, waterproof hiking boots Backpack Sleeping system Rain gear Lightweight tent
28. Results Fauna Flora Elk Pika Marmots Mule deer Big horn sheep Moose Bluebell Mountain Daisy Elephant Head Indian Paintbrush Columbine Berries Strawberries Rasberries Blueberries Whortleberries Deadly Nightshade
29. Conclusions and Implications Immersion in and response to the elements “Leave No Trace” Policy Outdoor experience Aligned with my interests Development of field research techniques Note-taking behavior Identification Foster a sense of health and pride Concern with the Mountain Pine Beetle
For the past two years I have been investigating the role of the hormone prolactin in nest attachment in ring neck dove under the Centre College BNS department. This project is my focus during the academic year and for the first several weeks of this summer I was continuing with the work. Those of you who have seen me present before should be familiar with the project and so for the sake of time, this is not going to be a large part of my presentation, although I credit the Brown Foundation for the recognition and reputation that allowed me the opportunity to do this work. If you’re curious about it, I would be pleased to speak with you after the program or during the break. This summer, the majority of my Brown Foundation stipend went toward were an internship at UC and a Sierra Club backpacking trip. I’ll cover both in the next 15 minutes.
There is currently a debate going on in the field of phylogeny (the division of science that determines relationships between organisms) because multiple definitions of a species are circulating. Many of these definitions contain similar characteristics or are variations of the same general idea. The ambiguity surrounding the definition of a species makes it difficult to determine the boundary between one species and the next. To a general audience, identifying what a species is can appear to be a simple task, however, understanding what makes up a species can be challenging, even to professional biologists. In the midst of this debate, any tool that can help better define a species will have broad implications for multiple fields including agriculture, biology, zoology, ecology, and more.
The traditional method of determining whether an individual belongs to an existing or new species begins by identifying the morphological characteristics, for example, species-typical markings on the abdominal. However, morphological identification techniques are prone to human error, making DNA sequencing a useful supplement. DNA exists in two forms, mitochondrial and nuclear, both of which can potentially be used in species identification experiments.
DNA sequence data can help to generate phylogenetic trees, such as this hypothetic one. I would like to point out a few things about this diagram: the highlighted species are monophyletic, meaning that they have one most recent common ancestor with other species in the tree.
The highlighted species, on the other hand, form non-monophyletic clades, meaning that they have more than one most recent common ancestor. This is one of the problems of using DNA to determine species.
The two types of DNA differ in several ways. Mitochondrial DNA is inherited from the mother. Nuclear DNA , located in the nucleus, is inherited from both parents. Mitochondrial DNA is prone to mutations and nuclear DNA tends to be more conserved, and as a result, each provides us which different phylogenetic info.
Ravinia is the genus that we use in our laboratory for genetic experiments. Ravinia is a genus of fly, and is a valuable organism in the breakdown of animal waste. Of the 12 species in the US, we tried to incorporate as many as possible in our experiments.
Because of the problems seen from the mitochondrial DNA we wanted to use nuclear DNA, and since the genome isn’t available for this genus, we obtained nuclear data by shearing random fragments of Ravinia DNA to create a genomic DNA library using a plasmid vector to obtain numerous fragments of cloned DNA.
The is the design of the experiment. I will go into more detail on each segment later in the presentation. As an overview, we needed to insert sections of ravinia DNA into e. coli DNA, allow the e. coli to amplify and clone the ravinia DNA, and then send the cloned ravinia DNA to be identified.
As a student research, this translated to approximately a 40 hour work weed composed primarily of working alongside my student mentor in the laboratory. Additionally, the program included field collections of potential subjects and literature reviews of similar published studies.
One of our 12 samples provided us with useable genetic information.
In conclusion, from this experiment we have obtained a nuclear segment that had not been sequenced before. The phylogenetic information obtained from this segment allows us to create a gene tree and a species tree, helping us to understand evolutionary systems. In addition, techniques similar to what we used in this experiment might help us identify and conserve protected species, which has implications in other fields such as agriculture and ecology.
In the future, we will attempt to generate primers from the DNA that we have acquired, and we will test the generated primers on multiple species of Ravinia. We will also create species and gene trees and test the boundaries and limitations of different species, allowing us to clarify some of the ambiguity of the raviniataxa.
From what you can see, in my academic career I have built up a strong and competitive background in laboratory research. However, I lack experience in field research, which differs in laboratory research in many respects.
My first introduction to field work was in a class I took at Centre…. My interest was also sparked by Thomas Defler, who….
It was becoming apparent to me that field research appropriately tied into many of my interests, and that in order to be a successful field researcher I needed to prepare myself and acquire some type of basic outdoor skills
As my interests evolved, I found myself becoming more and more involved with the Sierra Club, of which I am a member. The purpose of the club, which you see on the screen, aligns with my own values. In addition to service and education, the Sierra club is famous for the numerous outings that it holds across the United States and abroad. Trips typically last between 4 and 10 days. This time frame coordinated perfectly with my SUMR-UC internship, which lasted 8 weeks. I chose to spend the majority of my stipend on an outing sponsored by The Sierra Club.
The Alpine Beginner Backpacking trip seemed appropriate for my needs. It would teach me…, expose me to climate that I was unfamiliar and uncomfortable with, and allow me to take field notes on….
The alpine environment is harsh and unforgiving. Much of our trip took place above the tree line, where the air was scarce and the weather was cold and windy.
I had to extensively research the equipment appropriate for the alpine environment, the most notable components included… I also had to train with my alpine equipment in the 90 degree heat of Cincinnati
Training was composed of …
I took detailed sketches and notes on all of the flora and fauna that I spotted. Here is a sampling of the fauna and flora. I became quite skilled at identifying not only animals, but also prints and scat. A typical field note entry includes a sketch, dimensions, colors, proximity and location, and if an animal, whatever it was doing at the time of identification. The exercise of keeping a journal of field notes became a routine from day to day.
From these picture you can see the damage that the beetle has done so far. Some people believe that global warming is to blame- it is not getting cold enough, long enough to control the larvae. Whatever the cause, it is quite a sobering sight and something that we should be concerned about in Kentucky as well as Colorado.
I would hope to continue doing research in my future. Whether it be field research or animal research, after this summer I have a stronger arsenal of skills that I would hope to offer a company in need of an intern in a lab. On the more recreational side, I enjoyed the immense physical challenge that this trip offered me and would hope to test my limits again in some way. I will certainly continue backpacking, and would not have been able to get into the sport without the support of The Brown Foundation, and for that I am forever grateful.
