Novel Prochlorococcus ecotypes from the Anoxic Minimum Zone

Novel Prochlorococcus ecotypes from the Anoxic Minimum Zone of the Eastern Tropical South Pacific Ocean. Alvaro Munoz Plominsky

Prochlorococcus ecotypes HLII LLIV Partensky & Garczarek, 2010.

Prochlorococcus ecotypes HLII LLIV Kettler et al. , 2007.

Prochlorococcus ecotypes HLII LLIV Maximum Likelihood 16 S+ITS tree Ulloa et al. , unpublished

Prochlorococcus ecotypes HLII LLIV Maximum Likelihood 16 S+ITS tree Ulloa et al. , unpublished

Anoxic Minimum Zones (AMZs) Layer, of a hundred meters thick, where O 2 concentrations fall below the 3 n. M detection limit of the most sensible O 2 sensors. Ulloa et al. , 2012

Anoxic Minimum Zones (AMZs) Layer, of a hundred meters thick, where O 2 concentrations fall below the 3 n. M detection limit of the most sensible O 2 sensors. Ulloa et al. , 2012

Anoxic Minimum Zones (AMZs) Layer, of a hundred meters thick, where O 2 concentrations fall below the 3 n. M detection limit of the most sensible O 2 sensors. Ulloa et al. , 2012

AMZ Prochlorococcus 1 st Approach: Metagenomics (Msc. student Marcia Astorga) Characterization of 454 derived Prochlorococcus contigs. Prochlorococcus strain MIT 9313 MIT 9303 Total size (bp) 2410873 2682675 Matching contigs (bp) 831346 662306 34, 5 24, 7 88 83 Coverage (%) Number of contigs

AMZ Prochlorococcus 1 st Approach: Metagenomics (Msc. student Marcia Astorga) Characterization of 454 derived Prochlorococcus contigs. 2 nd Approach: Single-Cell Genome Sequencing

AMZ Prochlorococcus 2 nd Approach: Single-Cell Genome Sequencing

Oxygen (µmol Kg-1) 0 20 40 60 80 100 140 160 180 200 220 0 AMZ Prochlorococcus Samples were taken from the 2 nd Chl a peak at Station 1 (53 m). 20 Depth (m) Samples were enriched by flow cytometry cell sorting. 40 Cryopreserved in glycerol. 60 Sent for whole genome amplification at Bigelow Laboratory for Ocean Sciences. 80 0 1 2 3 Chl a (µg L-1) 4 5

AMZ Prochlorococcus

AMZ Prochlorococcus 1 st Approach: Metagenomics (Msc. student Marcia Astorga) Characterization of 454 derived Prochlorococcus contigs. 2 nd Approach: Single-Cell Genome Sequencing 3 rd Approach: Culture Screening by q. PCR Approx. 350 cultures (of 15 -20 m. L), from Big-RAPA Station 1, were maintained in our laboratory!!

AMZ Prochlorococcus 3 rd Approach: Culture Screening “General” primers (Synechococcus + Prochlorococcus) 101 bp

AMZ Prochlorococcus 3 rd Approach: Culture Screening LL VI 92 bp 97 bp

AMZ Prochlorococcus 3 rd Approach: Culture Screening LL VI

AMZ Prochlorococcus 3 rd Approach: Culture Screening LL VI

AMZ Prochlorococcus Single-cell genomes will be available. . . eventually. Genomic data of AMZ Prochlorococcus is also available among metagenomic datasets. Cultures are currently being scaled for future isolation of pure LL V Procholococcus. q. PCR methodology will allow screening of samples from the 2013 cruises to the ETNP and ETSP OMZs.

AMZ Prochlorococcus

Lavin et al. , 2010.