Oceanic Microbial Observatory SAR11 (E5/l)

1. 1.Identify spatial and temporal patterns in specific bacterioplankton/ prokaryotic populations 2. 2.Initiate experiments to investigate potential linkages between microbial processes, community structure and biogeochemical processes / events Discovery…. High throughput culturing in low nutrient media as a means to bring some of the uncultured bacterioplankton - into culture 4. Education and Outreach Oceanic Microbial Observatory Objectives

16 cruises / yr Core Measurements TemperatureSalinity Dissolved OxygenFluorescence Beam AttenuationPAR Salinity Oxygen Total CO 2 (TCO 2 )Alkalinity Nitrate Nitrite Phosphate Silicate POC/PON DOC/DON Pigments (HPLC) Bacteria Primary Production Bacteria Production Particle Fluxes Discrete Continuous Rates present

Temperature and mixed layer depth at BATS Depth (m) Temp. (°C) BATS Core

Bacterial Biomass Bacterial Production µ BP - estimated via 3H-TdR incorporation Summer max in BP and µ Composite of Bacterial biomass and production integrated throughout the Euphotic zone at BATS Steinberg, Carlson, Bates et al. 2001

BATS DOC (µM C) Hansell and Carlson

Prokaryotic Cell Abundance (cells E8 l -1 ) Depth (m)

Depth Cells E8 / l Bacterial LH-PCR 16s electropherograms BATS CDOM Cruise Aug 2001

10 m or 250 m whole water was diluted 0.22µm filtrate from surface or 250 m 10 m or 250 m whole water was diluted 0.22µm filtrate from surface or 250 m Cultures were incubated at inoculum’s in situ temperatures in the dark Cultures were incubated at inoculum’s in situ temperatures in the dark Mixing Experiments MO Objective: investigate potential linkages between microbial processes, community structure and biogeochemical patterns

DOC ( µM C) Niskin Day 1 Day 7 Day Cells E9 / l Surf / Surf 0.2 µmDeep/ deep 0.2µm Surface / surface 0.2µ treatment Deep / deep 0.2µ treatment Deep/ surf 0.2 media ba Cells E9/l DOC ( µM C) Surf / mix 0.2 µm Surface /(surf+deep) 0.2µ Deep / Surf 0.2µ HS 893 Mixing Experiment days Carlson et al. 2004

ExperimentInoc.0.2 Filtrate∆Cell ∆DOC SourceSource Cells E8/l1 week > month HS 852 (Aug-97)SurfSurf DeepSurf HS 875 (Aug-98)SurfSurf DeepSurf HS 893 (Aug-99)SurfSurf0.5-- DeepSurf Deep Deep Surfmix BATS 155 (Aug-01)SurfSurf0.5-- Deep Deep---

MLD and Integrated Prokaryotic Biomass in the Upper Mesopelagic at BATS

Identification of Terminal Restriction Fragments TRFLP Groups Fragment length observed Fragment length predicted from sequence Sequence Identification 1113, 226, , 228, 294SAR11, , , 193Unc. Gamma, Roseobacter OCS 116, , , 263SAR202, SAR324, , , 390Prasinophytes, Marine Actinobacteria SAR samples from surface samples m samples HAE III- digest 27F, 519 R

Ordination of T-RFLP Fragments ( ) Nonmetric multidimensional scaling Morris,Cho, Rappé, Vergin, Carlson, and Giovannoni, In Press

Roseobacter (E5/L) Cytophaga-Flavobacteria (E5/L) SAR 11 (E5/L) Carlson, Morris, Parsons and Giovannoni, unpublished MO objective: Identify spatial and temporal patterns in specific bacterioplankton/ prokaryotic populations FISH DATA

OSU High Throughput Cultivation Lab >2000 Strains > 80% do not grow on agar 18 strains in genome sequencing Natural Sample Dilute cells and distribute into wells of microtiter dishes Step 1: Dilution to extinction Step 2:Cell Arrays Cells arrayed on polycarbonate membranes Target species identified by multiplexed in situ Hybridization and laser scanning cytometry Step 3: High-Throughput Cell Identification MO Objective: Isolation of new organisms Courtesy of S. Giovannoni

Marine Microbial Ecology Marine Genomics MO objective: Education & Outreach

Acknowledgements BBSR Rachel Parsons Nick Bates Rod Johnson Mike Lomas Tony Michaels Dennis Hansell Debbie Steinberg Norm Nelson Tony Knap BATS Team past and present UCSB Stu Goldberg Courtney Ewart Kurt Gray Elli Wallner Aubrey Cano Meredith Meyers OSU Steve Giovannoni Kevin Vergin Bob Morris Mike Rappé Giovannoni Lab group