1. Oceanic Microbial Observatory SAR11 (E5/l)

2. 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. 3. 3.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

3. 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 1988 - present

4. Temperature and mixed layer depth at BATS Depth (m) 0 50 100 150 200 250 300 19891990199119921993199419951996199719981999 Temp. (°C) 30 29 28 27 26 25 24 23 22 21 20 19 18 BATS Core

6. 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

7. BATS DOC (µM C) Hansell and Carlson

8. 0 50 100 150 200 250 300 12 10 8 6 4 2 0 91 92 93 94 95 96 97 98 99 00 Prokaryotic Cell Abundance (cells E8 l -1 ) Depth (m)

9. 1000 900 800 700 600 500 400 300 200 100 0 012345678910 Depth Cells E8 / l 1 10 40 60 80 100 150 250 500 1000 Bacterial LH-PCR 16s electropherograms BATS CDOM Cruise Aug 2001

10. 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

11. 50 55 60 65 70 DOC ( µM C) Niskin Day 1 Day 7 Day 37 0 0.1 0.2 0.3 0.4 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 50 55 60 65 70 0 0.1 0.2 0.3 0.4 01234567 Cells E9/l DOC ( µM C) Surf / mix 0.2 µm Surface /(surf+deep) 0.2µ Deep / Surf 0.2µ HS 893 Mixing Experiment 01234567 days Carlson et al. 2004

12. ExperimentInoc.0.2 Filtrate∆Cell ∆DOC SourceSource Cells E8/l1 week > month HS 852 (Aug-97)SurfSurf0.75-- DeepSurf2.851.93.5 HS 875 (Aug-98)SurfSurf0.75-- DeepSurf2.53.13.1 HS 893 (Aug-99)SurfSurf0.5-- DeepSurf2.13.75.1 Deep Deep0.25-- Surfmix0.9-1.3 BATS 155 (Aug-01)SurfSurf0.5-- Deep Deep---

13. MLD and Integrated Prokaryotic Biomass in the Upper Mesopelagic at BATS

14. Identification of Terminal Restriction Fragments TRFLP Groups Fragment length observed Fragment length predicted from sequence Sequence Identification 1113, 226, 290117, 228, 294SAR11, 33 2115, 193119, 193Unc. Gamma, 9 3194195Roseobacter 4302303OCS 116, 4 5155, 260158, 263SAR202, 3 6405 SAR324, 2 7303, 388304, 390Prasinophytes, 2 8328330Marine Actinobacteria 9192193SAR116 164 samples from 1992 -2002 81 - surface samples 83- 200 m samples HAE III- digest 27F, 519 R

15. Ordination of T-RFLP Fragments (1992-2002) Nonmetric multidimensional scaling Morris,Cho, Rappé, Vergin, Carlson, and Giovannoni, In Press

16. 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

17. 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

18. Marine Microbial Ecology Marine Genomics http://www.lifesci.ucsb.edu/~carlson/ MO objective: Education & Outreach

19. 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