Synoicum_adareanum_microbiome

Study Extent

Samples were collected in Anvers Island Archipelago, Gerlache Strait, Palmer Deep and LaPeyrere Bay.

Sampling

A spatial survey of Synoicum adareanum was executed in which samples were collected by SCUBA in austral fall between 23 March and 3 April 2011. Seven sampling sites (depths 24.7 - 31 m) around the region accessible by Zodiac boat from Palmer Station were selected in which we sampled in a nested design where three multi-lobed colonies were selected from each site, and three lobes per colony were sampled. In total, 63 S. adareanum lobes were sampled (9 from each site). Samples were transported to Palmer Station on ice, and frozen at -80 °C until processing at DRI and USF. Frozen S. adareanum lobes were cut longitudinally in half for parallel processing through DNA and palmerolide detection pipelines. Reference seawater data set was represented by samples that had been collected in February-March 2008 (five samples) and in August-September 2008 (nine samples) from LTER Station B near Anvers Island (east of the Bonaparte Point dive site), and at a few other locations in the region. Seawater samples were collected by a submersible pump and acid washed silicone tubing at 10 m at Station B, and using a rosette equipped with 12 L Niskin bottles for the offshore samples at 10 m and 500 m (2 samples each depth). The February-March seawater samples were processed using in-line filtration with a 2.5 um filter (Polygard, Millipore) to screen larger organisms and bacterioplankton were concentrated using a tangential flow filtration system and the cells were harvested on 25 mm 0.2 um Supor filters (Millipore). The August-September seawater samples were processed using inline filtration with a 3.0 um filter (Versapor, Millipore), and then bacterioplankton was collected onto 0.2 um Sterivex (Millipore) filters using a multichannel peristaltic pump (Masterflex). All filters were immersed in sucrose lysis buffer(68) and stored frozen at -80 °C until extraction. S. adareanum samples collected by SCUBA in 2004 and 2007 were used for cultivation. The 2004 specimen were archived in 20% glycerol at -80 °C until processing by manual homogenization using sterilized mortar and pestle prior to plating a suspension onto marine agar 2216 plates in 2006. The 2007 samples were homogenized immediately following collection using sterilized mortar and pestle, and suspensions were prepared in 1X marine broth or filter-sterilized seawater then transferred at 4 °C.

Quality Control

NA

Method steps

1. NA

Samples were collected in Anvers Island Archipelago, Gerlache Strait, Palmer Deep and LaPeyrere Bay.