1. Understanding Algal Biodiversity and Function: Comparison of in situ versus ex situ Analyses Rocky Patil Advisor: Dr. Rachael Morgan-Kiss

2. Overall Project Goal Improve the accuracy of in situ sensors of phytoplankton diversity/activity using ex situ analyses.

3. Overall Project Goal Improve the accuracy of in situ sensors of phytoplankton diversity/activity using ex situ analyses. In situ: Chlorophyll a fluorescence Ex situ: molecular phylogenetics & enzyme assays

4. Overall Project Goal Improve the accuracy of in situ sensors of phytoplankton diversity/activity using ex situ analyses. Study Sites: 1.Ohio watersheds 2.Montana Alpine Lakes 3.Antarctic Lakes *

5. McMurdo Dry Valleys Long Term Ecological Research (MCM LTER) ©2006 McMurdo Dry Valleys LTER BONNEY HOARE FRYXELL

6. GENERAL CHARACTERISTICS McMURDO DRY VALLEY LAKES Permanent ice-cover & isolation of lakes Simple microbe-dominated food web Lack of human impact & seasonal mixing Highly stratified aquatic environment More vulnerable to climate change & abiotic factors Extremely stable physical conditions 6-8 months total darkness

7. Spatial Profiling Algal Diversity with the bbe FluoroProbe

8. Photosynthetic Unit Chlorophyll-a Fluorescence as a Diagnostic Tool Chl a/b Carotenoids PhycocyaninChl a/c Phycoerythrin Chl a/c ChlorophytaCyanophytaDinophyta Haptophyta Heterokont. Cryptophyta SPECTRAL GROUPS OF MICROALGAE GREENBLUEBROWNMIXED Beutler et al. Photosynth Res 2002. 72:39-53

9. In situ measurements: FluoroProbe profiles of Lake Bonney EAST BONNEYWEST BONNEY GREEN - Chlorophyta BLUE - Cyanophyta BROWN - Haptophyta MIXED - Cryptophyta

10. Ex situ measurements: Sampling & Extraction for Phylogenetic Analyses Water samples collected at 3 meter intervals to determine microbial diversity Filtered through Whatman filters DNA extraction with FastDNA Spin Kit 18S rRNA gene amplification from universal primers (Lopez Garcia et al., 2001) –Cloning, Ligation, Transformation, & Phylogenetic Analysis

11. Phylogenetic Characterization of Lake Phytoplankton Communities Collect lake samples from major Chlorophyll-a maxima depths Concentrate Samples and Preserve 9 m 11 m 15 m Isolate Environmental DNA RFLP mapping/sequencing unique OTUs

12. Micro-Eukaryote Distribution 18S rRNA Ex situ measurements: Dry Valley Lake Phytoplankton Diversity