International Census of Marine Microbes Explore the diversity, relative abundance, and environmental context of all microbial life forms in the oceans Archaea Eukarya Viruses Bacteria

International - Global Coverage Competitive Tag Sequencing samples 660 completed ~13.5 million tag sequences Final Data set will exceed 18 million tag sequences

Microbial Population Structure of the World’s Ocean Frisian Island Sylt Guaymas Methane Seeps Gulf of Aqaba Global Protist Survey Hood Canal Washington Hawaii Ocean Time-Series IOMM Cooperative Run Gulf of Maine LaCAR Cooperative Run Lost City Mount Hope Bay Helgoland New Zealand Sediment Ocean Drilling Project English Channel Surreptitious Algal Bacteria Station M Sediments Sponges Spatial Scaling Diversity Humboldt Marine Ecosystem Black Sea Arctic Chukchi Beaufort Amazon-Guianas Water Lau Hydrothermal Vent Anaerobic Protist Project Amundsen Sea Antarctica Azorean Shallow Vents Azores Waters Project Blanes Microbial Observatory Baltic Sea Proper Black Sea Redox Census Antarctic Marine Cariaco Basin Caribbean Coral Bacteria Deep Subseafloor Sediment Coastal Microbial Mats Coastal New England Coral Reef Sediment Deep Arctic Ocean Deep Ocean Flux Deep Sea Eukarya

Phenotypic Diversity Named species: Bacteria and Archaea: 9,000 Protists: 200,000 Microbial creatures of untold diversity dominate every corner of our biosphere. Estimates of X diversity of animals. The number of different kinds of bacteria in the oceans could eclipse 5 to 10 million

Deep Sequencing BacteriaArchaea Total Unique V6 Tag Sequences 30,1085,979 Total OTUs at 3% Difference 18,5371,931 Chao1 Estimator of Richness at 3% Difference 36,8692,754 ACE Estimator of Richness at 3% Difference 37,0382,678 Huber et al Science

Deep Sequencing Eukarya Total Unique V9 Tag Sequences 8,028 Total OTUs at 5% Difference 1,622 Chao2 Estimator of Richness at 5% Difference 2,830 (+) 100 ICE Estimator of Richness at 5% Difference 2,830 (+) 26 Amaral-Zettler et al. In prep Mt. Hope Bay

Evenness

Actinobacteria Acidobacteria Bacteroidetes Chloroflexi Cyanobacteria Deferrebacteres Firmicutes Gemmatimonadetes Nitrospira Lentisphaerae Planctomycetes Proteobacteria Verrucomicrobiae 112R 550 M °E, °N 115R 4,121 M °E, °N FS312 Axial Sea Mount - Bag City Difuse Flow FS312 Axial Sea Mount - Mkr 52 Difuse Flow Soil-Kellogg-5 species Soil-Kellogg-no plants Marine vs. Soil Evenness

How many microbes can you find in a few drops of seawater? ~ 1,000,000 Bacteria ~ 1,000,000 Archaea ~10-10,000 Protists ~10,000,000 Viruses Abundance10,000,000,000,000,000,000,000,000,000 Microbial Cells in the Ocean

Visualization

Human fecal profile Lake WaterCombined Sewage overflow Bacteroidetes Firmicutes Detection of Anthropogenic Impacts

Human fecal profile Lake Water Combined Sewage overflow Firmicutes Detection of Anthropogenic Impacts

Low abundance taxa in rank-ordered, taxon abundance curves Microbial abundance curves are “long-tail distributions” Concept of the Rare Biosphere The tail is much greater than previously known Diversity eclipses all prior estimates of Bacterial, Archaeal and Eukaryl diversity Never-before-seen populations High Abundance Populations Taxon-rank distribution curve for microbial communities Low Abundance Populations The Rare Biosphere

Explaining the Rare Biosphere Biogeography Dispersal from yet to be discovered endemic sites

Human Sponge Mouse Rio Tinto Deep subsurface Sulfides/Basalts Cariaco Salt Marsh Lau Seamount Salt Pond English Channel Amunsen Palmer LTER Global Distribution of Abundant Tags

Explaining the Rare Biosphere Biogeography Dispersal from yet to be discovered endemic sites Keystone species Always rare - possibly slow growing or dormant Persistently rare but capable of becoming abundant “Seed” organisms Genomic novelty Analogous to heterozygosity in animals or plants Products of historical ecological change? Responders to environmental shifts May serve as sentinel for global change

Limits to Knowledge Operational issues for the Rare Biosphere: Definition of microbial diversity - key data - Start and length of the long tail Existence of viral, archaeal and eukaryl Rare Biosphere Tools necessary to define limits of the rare biosphere Criteria for targeting studies of rare biosphere members Questions about the Rare Biosphere and how can we seek answers? Is the Rare Biosphere globally distributed? or does the Rare biosphere reflect dispersal from endemic sites? Does membership in the Rare Biosphere shift across different spatial/temporal scales? What mechanisms determine membership in the Rare Biosphere? Why is there a rare biosphere? Do microbes in the rare biosphere compete for niche space? Is the rare biosphere a nearly unlimited source of genetic information that can transfer between microbial populations? How important are reservoirs in protecting microbial diversity? Do rare organisms have a selective advantage? Are there specific mechanisms that allow low-abundance populations to become dominant in response to environmental shifts? How does the rare biosphere shape the environment past, present and future? How might rare organisms impact human health and well being? Can the rare biosphere serve as a sentinel for global change? Do rare microbes play a key role in preserving biodiversity?

Acknowledgements ICoMM 454 participants: Paula Aguiar Robert Andersen Felipe Artigas Stefan Bertilsson Christina Bienhold Henk Bolhuis William Brazelton David Caron D. Chandramohan Andrei Chistoserdov Marco Coolen Mark Dennett Steve D’Hondt Slava Epstein Katrina Edwards Rima Franklin Eric Gaidos Victor Ariel Gallardo Gunnar Gerdts Jack Gilbert Koji Hamasaki Julie Huber David Karl David Kirchman Connie Lovejoy Els Maas Ana Martins Alison Murray Jan Pawlowski Martin Polz Thomas Pommier Anton Post James Prosser Alban Ramette Michael Rappe Anna-Louise Reysenbach Gabrielle Rocap Juliette Rooney-Varga James Staley Thorsten Stoeck Shinichi Sunagawa Andreas Teske Michael Wagner Gordon Webster Patricia Yager ICoMM Scientific Organizing Committee: Mitchell Sogin Jan de Leeuw David Patterson Lucas Stal Gerhard Herndl Stefan Schouten ICoMM Secretariat: Linda Amaral-Zettler ICoMM IT Specialist: Phillip Neal Anne Thessen ICoMM Scientific Advisory Council: John Baross (Chair) Robert Andersen Felipe Artigas Antje Boetius D. Chandramohan Kazuhiro Kogure Carlos Pedros-Alio Francisco Rodriguez-Valera