Metagenomic Study of North East Lakes and Ponds

Conditions that favor cyanobacterial dominance in algal blooms, and thus facilitate microcystin production, are not well understood (2). To address this issue we designed an experiment to measure taxonomic diversity during different stages of algal blooms across a geographic region. Microcystin levels and bulk algal counts are reported over a fourteen week period from samples taken once weekly at five different locations in four northeastern states. In addition, we conducted a 16S survey and metagenomic sequencing of directly sampled blooms at three different time points from each of the five sites. Water samples were collected from five lakes and ponds in the northeast United States over a fourteen-week period from July 20 to October 21, 2010 by investigators at partner institutions across the North East Cyberinfrastructure Consortium (NECC) (Table 1). Samples were collected in triplicate once weekly from Lake Champlain at Highgate Springs, VT, Lake Winnipesaukee, NH, Sebasticook Lake, Maine and Trustum and Yawgoo Ponds in Rhode Island (Figure 1). Whole water was shipped overnight to the University of Vermont DNA Sequencing Facility where all further processing was carried out. A total of 204 samples were collected and processed. Microcystin levels and raw algae counts were measured for all samples. Total flourescent particle counts from flow cytometry analysis were used to select three time points representing the beginng, midpoint and end of the blooms at each location. These selected time points were further processed for DNA sequencing. Whole DNA was extracted from each of the three selected time points for all locations and subjected to 16S rRNA amplicon sequencing. Samples were barcoded, multiplexed and sequenced on a single plate of a 454 Life Sciences Genome Sequencer FLX at the Genetic Analysis Genomics Core Facility,?Huck Institutes for Life Sciences, Penn State University producing 1.1M sequence reads.