Oceanic Microbial ObservatoryOceanic Microbial ObservatoryQuickTime™ and a

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SAR11 (E5/l)

1. Identify spatial and temporal patterns in specific bacterioplankton/ prokaryotic populations

2. Initiate experiments to investigate potential linkages between microbial processes, community structure and biogeochemical processes / events.

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 Oceanic Microbial ObservatoryObservatory ObjectivesObjectives

16 cruises / yr16 cruises / yr

Core Measurements

TemperatureTemperature SalinitySalinityDissolved OxygenDissolved Oxygen FluorescenceFluorescenceBeam AttenuationBeam Attenuation PARPAR

SalinitySalinity Oxygen OxygenTotal COTotal CO22 (TCO (TCO22)) AlkalinityAlkalinity

NitrateNitrate Nitrite NitritePhosphate Phosphate SilicateSilicatePOC/PONPOC/PON DOC/DON DOC/DONPigments (HPLC)Pigments (HPLC) Bacteria Bacteria

Primary ProductionPrimary ProductionBacteria ProductionBacteria Production

Particle FluxesParticle Fluxes

Discrete

Continuous

Rates

1988 - present1988 - present

Temperature and mixed layer depth at BATSD

epth

(m

)0

50

100

150

200

250

3001989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999

Tem

p. (

°C)

30

29

28

27

26

25

24

23

22

21

20

19

18

BATS CoreBATS 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. 2001Steinberg, Carlson, Bates et al. 2001

BATS DOC (µM C)

Hansell and Carlson

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)D

epth

(m

)

1000

900

800

700

600

500

400

300

200

100

00 1 2 3 4 5 6 7 8 9 10

Dep

thCells E8 / l

1

10

40

60

80

100

150

250

500

1000

Bacterial LH-PCR 16s electropherograms

BATS CDOM Cruise Aug 2001

• • 10 m or 250 m whole water 10 m or 250 m whole water was diluted 0.22µm filtrate was diluted 0.22µm filtrate from surface or 250 mfrom surface or 250 m

• • Cultures were incubated at Cultures were incubated at inoculum’s in situ inoculum’s in situ temperatures in the dark temperatures in the dark

Mixing Experiments

MO Objective: investigate potential linkages between microbial processes, community structure and biogeochemical patterns

50

55

60

65

70

DO

C (

µM

C)

Niskin

Day 1

Day 7

Day 37

0

0.1

0.2

0.3

0.4C

ells

E9

/ l

Surf / Surf 0.2 µm Deep/ deep 0.2µm

Surface / surface 0.2µ

treatment

Deep / deep 0.2µ treatment

Deep/ surf 0.2 mediab a50

55

60

65

70

0

0.1

0.2

0.3

0.4

0 1 2 3 4 5 6 7

Cel

ls E

9/l

DO

C (

µM

C)

Surf / mix 0.2 µm

Surface /(surf+deep) 0.2µ

Deep / Surf 0.2µ

HS 893 Mixing ExperimentHS 893 Mixing Experiment

0 1 2 3 4 5 6 7

daysdays Carlson et al. 2004Carlson et al. 2004

Experiment Inoc. 0.2 Filtrate ∆Cell ∆DOCSource Source Cells E8/l 1 week > month

HS 852 (Aug-97) Surf Surf 0.75 - -

Deep Surf 2.85 1.9 3.5

HS 875 (Aug-98) Surf Surf 0.75 - -

Deep Surf 2.5 3.1 3.1

HS 893 (Aug-99) Surf Surf 0.5 - -

Deep Surf 2.1 3.7 5.1

Deep Deep 0.25 - -

Surf mix 0.9 - 1.3

BATS 155 (Aug-01) Surf Surf 0.5 - -

Deep Deep - - -

MLD and Integrated Prokaryotic Biomass in the Upper MLD and Integrated Prokaryotic Biomass in the Upper

Mesopelagic at BATSMesopelagic at BATS

Identification of Terminal Restriction Fragments

TRFLP

Groups

Fragment length observed

Fragment length predicted

from sequence

Sequence Identification

1 113, 226, 290 117, 228, 294 SAR11, 33

2 115, 193 119, 193 Unc. Gamma, 9

3 194 195 Roseobacter

4 302 303 OCS 116, 4

5 155, 260 158, 263 SAR202, 3

6 405 405 SAR324, 2

7 303, 388 304, 390 Prasinophytes, 2

8 328 330 Marine Actinobacteria

9 192 193 SAR116

164 samples from 1992 -2002

81 - surface samples

83- 200 m samples

HAE III- digest 27F, 519 R

Ordination of T-RFLP Fragments (1992-2002)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 agar18 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 EcologyMarine Genomics

http://www.lifesci.ucsb.edu/~carlson/

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