A Summer of Research

Annie Roessler

Centre College

Brown Fellows Program

Project Schedule

• Centre College

– Prolactin and Nest Attachment

• University of Cincinnati

– DNA-based species Delimitation

• Sierra Club

– Alpine Beginner Backpacking Course

DNA-based Species Delimitation in the Agriculturally Important

Genus, Ravinia

The Species Debate

• Multiple species concepts

– Wiens, J. J. (2007)

• Boundary Limitations

• Broad Implications

Species Delimitation

• Morphological characteristics

• DNA sequence data

– Mitochondrial (mtDNA)

– Nuclear

• Monophyly

• One most recent common ancestor

Phylogenetic Trees

• Non-monophyletic

• More than one most recent common ancestor

Phylogenetic Trees

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

Ravinia

• Limited morphological characters

• Agriculturally important

• 17 species living in N. America (12 present in this study)

Subjects

• Twelve species of North American Ravinia

– Multiple individuals

– Multiple populations

• Additional samples collected in Burnet Woods

Purpose

1. Establish species trees generated from gene trees comprised of mitochondrial and nuclear loci

2.Create a genomic DNA library of Ravinia using a plasmid vector to obtain numerous fragments of cloned DNA

Experiment Design

1. Ravinia DNA Digestion

2. Plasmid Extraction

3. Vector Digestion

4. Ligation Vector and Insert

5. Transformation of E. coli

6. Sequencing of Inserts

SUMR-UC Schedule

• 40 hour work week

• Hands on laboratory work

• Subject collections

• Literature reviews

• 1 of 12 samples

• Sequence length: 710 base pairs

• Not similar to anything in database

• Not vector or human

Results

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

• 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 Ravinia taxa

Future Directions

• Terminology

• Lab techniques

• Lack of chemistry background

• Communication

• Literature reading

• Career advice

SUMR-UC Challenges and Strengths

Walking Along the Spine of the Rockies: A Week in the Mountains

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

Famous Field Researchers

• Jane Goodall

– Anthropology 120

• Thomas Defler

– American Primatology Convention speaker

– Lived and worked in Colombia

My Interests

• Animal behavior

• Ecology

• The Green Movement

• Psychology

• Conservation

• Animal Welfare

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.

Purpose

1. Learn and develop basic outdoor survival skills under the guidance of an experienced instructor

2. Practice field research techniques, such as observing wildlife and taking detailed field notes

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

Environment

• Subalpine and Alpine Regions

• Forest

• Tundra

– Snow fields

– Boulder fields

– High winds

• Meadows

• Glacial Lakes

Equipment

• Medium weight, waterproof hiking boots

• Backpack

• Sleeping system

• Rain gear

• Lightweight tent

Training

• Cardiovascular– Running

– Spinning

• Weightlifting– Walking with 45

lb pack

– Unusual muscle groups

• Flexibility– Yoga

Results

Fauna

• Elk

• Pika

• Marmots

• Mule deer

• Big horn sheep

• Moose

Flora• Bluebell

• Mountain Daisy

• Elephant Head

• Indian Paintbrush

• Columbine

• Berries– Strawberries

– Rasberries

– Blueberries

– Whortleberries

– Deadly Nightshade

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

Mountain Pine Beetle

Future Directions

• Continuation of Research

– Current Projects

– Potential Internship

• Next Challenge

– To be determined

Thank You!

• The Brown Foundation

• Centre College

• The University of Louisville

• Clarence Wyatt

• Melissa Burns-Cusato

• The University of Cincinnati Graduate School

• SUMR-UC/ GSUM program

• Evan Wong• Ron DeBry• Michael Baldwin

• And many more!