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This study investigates the phytoplankton community in the Columbia River and its coastal zone across seasons, utilizing 18S rRNA clone libraries to identify changes and interactions with bacteria and archaea. The significance of phytoplankton as primary producers and base of the aquatic food chain is explored through genetic fingerprinting and sequencing analysis. Traditional methods like flow cytometry and microscopy are employed to study phylogenetic variations and DNA/RNA from water samples collected onsite. By comparing different locations and seasons, unexpected changes in phytoplankton populations are monitored to assess ecosystem health.
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18S rRNA clone library of phytoplankton in the Columbia River and its coastal zone By Pete Kahn Mentors: Lydie Herfort and Peter Zuber http://www.mapwatch.com/news-blog/images/phytoplankton.jpg
Significance of phytoplankton • Major producers: O2: 90% CO2=>C3: 50% • Base of aquatic food chain
Interactions with other Domains BacterioplanktonColonization : POC=>DOC Competition for NO32- and NH3? http://www.fossilmuseum.net/Tree_of_Life/Domains_Archaea_Bacteria/Domains_of_life.jpg
Significance: Why is CMOP interested in Phytoplankton? • Base of aquatic food chain • Major producers: O2: 90%; CO2=>C3: 50% • Interaction with other two Domains -Crenarchaeota: “competition” -Bacterioplankton: colonization -POC=>DOC Marine Phytoplankton bloom off the coast of Argentina http://www.mapwatch.com/news-blog/images/phytoplankton.jpg
Traditional Methods Flow cytometry Microscopy: Pediastrium • Microscopy • Flow Cytometry • Chlorophyll a • Photosynthesis: O2 production C14 uptake http://upload.wikimedia.org/wikipedia/commons/c/c7/Picoplancton_cytometrie.jpg http://www.keweenawalgae.mtu.edu/ALGAL_IMAGES/chlorophyceans/Pediastrium_n16_dollartow3_402_c.jpg Filtering for chlorophyll a
Objectives • 18S rRNA clone library for different seasons and different locations: 0 PSU, 30 PSU for spring and summer; compare to winter samples • Compare with Bacteria & Archaea clone libraries from Dan Murphy • Identify unexpected changes • Monitor health of ecosystem Pediastrium http://www.keweenawalgae.mtu.edu/ALGAL_IMAGES/chlorophyceans/Pediastrium_n16_dollartow3_402_c.jpg
Methods Overview Understand Phytoplankton community in estuary Phylogenetic analysis Sequence analysis w/ BLAST Yan, Q. Y., Yu, Y. H. & Feng, W. S. (2006). Genetic fingerprinting of plankton community provides new insights into aquatic ecology. Progress in Natural Science16, 893-898.
Methods Overview Water sample => Sterivex on site Total nucleic acid extraction from sterivex in lab Amplification through PCR
Variations in nucleic acid in environmental samples Oct 2006 Feb 2007 • Between seasons • Between locations April Wecoma & Forerunner: Freshwater << Surface Coastal DNA RNA