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  1. Abstract. Eastern boundary upwelling systems (EBUS) contribute a disproportionatefraction of the global fish catch relative to their size and are especiallysusceptible to global environmental change. Here we present the evolution ofcommunities over 50 d in an in situ mesocosm 6 km offshore of Callao, Peru, andin the nearby unenclosed coastal Pacific Ocean. The communities weremonitored using multi-marker environmental DNA (eDNA) metabarcoding and flowcytometry. DNA extracted from weekly water samples were subjected toamplicon sequencing for four genetic loci: (1) the V1–V2 region of the 16SrRNA gene for photosynthetic eukaryotes (via their chloroplasts) andbacteria; (2) the V9 region of the 18S rRNA gene for exploration ofeukaryotes but targeting phytoplankton; (3) cytochrome oxidase I (COI) forexploration of eukaryotic taxa but targeting invertebrates; and (4) the 12SrRNA gene, targeting vertebrates. The multi-marker approach showed adivergence of communities (from microbes to fish) between the mesocosm andthe unenclosed ocean. Together with the environmental information, thegenetic data furthered our mechanistic understanding of the processes thatare shaping EBUS communities in a changing ocean. The unenclosed oceanexperienced significant variability over the course of the 50 d experiment,with temporal shifts in community composition, but remained dominated byorganisms that are characteristic of high-nutrient upwelling conditions(e.g., diatoms, copepods, anchovies). A large directional change was found inthe mesocosm community. The mesocosm community that developed wascharacteristic of upwelling regions when upwelling relaxes and watersstratify (e.g., dinoflagellates, nanoflagellates). The selection ofdinoflagellates under the salinity-driven experimentally stratifiedconditions in the mesocosm, as well as the warm conditions brought about bythe coastal El Niño, may be an indication of how EBUS will respond underthe global environmental changes (i.e., increases in surface temperature andfreshwater input, leading to increased stratification) forecast by the IPCC. 
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  3. Abstract

    Symbiotic relationships between vestimentiferan tubeworms and chemosynthetic Gammaproteobacteria build the foundations of many hydrothermal vent and hydrocarbon seep ecosystems in the deep sea. The association between the vent tubewormRiftia pachyptilaand its endosymbiontCandidatusEndoriftia persephone has become a model system for symbiosis research in deep‐sea vestimentiferans, while markedly fewer studies have investigated symbiotic relationships in other tubeworm species, especially at cold seeps. Here we sequenced the endosymbiont genome of the tubewormLamellibrachia barhamifrom a cold seep in the Gulf of California, using short‐ and long‐read sequencing technologies in combination with Hi‐C and Dovetail Chicago libraries. Our final assembly had a size of ~4.17 MB, a GC content of 54.54%, 137X coverage, 4153 coding sequences, and aCheckMcompleteness score of 97.19%. A single scaffold contained 99.51% of the genome. Comparative genomic analyses indicated that theL. barhamisymbiont shares a set of core genes and many metabolic pathways with other vestimentiferan symbionts, while containing 433 unique gene clusters that comprised a variety of transposases, defence‐related genes and a lineage‐specific CRISPR/Cas3 system. This assembly represents the most contiguous tubeworm symbiont genome resource to date and will be particularly valuable for future comparative genomic studies investigating structural genome evolution, physiological adaptations and host‐symbiont communication in chemosynthetic animal‐microbe symbioses.

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  4. ABSTRACT Vitamin B 1 (thiamin) is a cofactor for critical enzymatic processes and is scarce in surface oceans. Several eukaryotic marine algal species thought to rely on exogenous thiamin are now known to grow equally well on the precursor 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP), including the haptophyte Emiliania huxleyi . Because the thiamin biosynthetic capacities of the diverse and ecologically important haptophyte lineage are otherwise unknown, we investigated the pathway in transcriptomes and two genomes from 30 species representing six taxonomic orders. HMP synthase is missing in data from all studied taxa, but the pathway is otherwise complete, with some enzymatic variations. Experiments on axenic species from three orders demonstrated that equivalent growth rates were supported by 1 µM HMP or thiamin amendment. Cellular thiamin quotas were quantified in the oceanic phytoplankter E. huxleyi using the thiochrome assay. E. huxleyi exhibited luxury storage in standard algal medium [(1.16 ± 0.18) × 10 −6  pmol thiamin cell −1 ], whereas quotas in cultures grown under more environmentally relevant thiamin and HMP supplies [(2.22 ± 0.07) × 10 −7 or (1.58 ± 0.14) × 10 −7  pmol thiamin cell −1 , respectively] were significantly lower than luxury values and prior estimates. HMP and its salvage-related analog 4-amino-5-aminomethyl-2-methylpyrimidine (AmMP) supported higher growth than thiamin under environmentally relevant supply levels. These compounds also sustained growth of the stramenopile alga Pelagomonas calceolata . Together with identification of a salvage protein subfamily (TENA_E) in multiple phytoplankton, the results indicate that salvaged AmMP and exogenously acquired HMP are used by several groups for thiamin production. Our studies highlight the potential importance of thiamin pathway intermediates and their analogs in shaping phytoplankton community structure. IMPORTANCE The concept that vitamin B 1 (thiamin) availability in seawater controls the productivity and structure of eukaryotic phytoplankton communities has been discussed for half a century. We examined B 1 biosynthesis and salvage pathways in diverse phytoplankton species. These comparative genomic analyses as well as experiments show that phytoplankton thought to require exogenous B 1 not only utilize intermediate compounds to meet this need but also exhibit stronger growth on these compounds than on thiamin. Furthermore, oceanic phytoplankton have lower cellular thiamin quotas than previously reported, and salvage of intermediate compounds is likely a key mechanism for meeting B 1 requirements under environmentally relevant scenarios. Thus, several lines of evidence now suggest that availability of specific precursor molecules could be more important in structuring phytoplankton communities than the vitamin itself. This understanding of preferential compound utilization and thiamin quotas will improve biogeochemical model parameterization and highlights interaction networks among ocean microbes. 
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