Genetic diversity affects the daily transcriptional oscillations of marine microbial populations

Marine microbial communities are genetically diverse but have robust synchronized daily transcriptional patterns at the genus level that are similar across a wide variety of oceanic regions. We developed a microarray-inspired gene-centric approach to resolve transcription of closely-related but distinct strains/ecotypes in high-throughput sequence data. Applying this approach to the existing metatranscriptomics datasets collected from two different oceanic regions, we found unique and variable patterns of transcription by individual taxa within the abundant picocyanobacteria Prochlorococcus and Synechococcus, the alpha Proteobacterium Pelagibacter and the eukaryotic picophytoplankton Ostreococcus. The results demonstrate that marine microbial taxa respond differentially to variability in space and time in the ocean. These intra-genus individual transcriptional patterns underlie whole microbial community responses, and the approach developed here facilitates deeper insights into microbial population dynamics

Phytoplankton transcriptomic and physiological responses to fixed nitrogen in the California current system

Marine phytoplankton are responsible for approximately half of photosynthesis on Earth. However, their ability to drive ocean productivity depends on critical nutrients, especially bioavailable nitrogen (N) which is scarce over vast areas of the ocean. Phytoplankton differ in their preferences for N substrates as well as uptake efficiencies and minimal N requirements relative to other critical nutrients, including iron (Fe) and phosphorus. In this study, we used the MicroTOOLs high-resolution environmental microarray to examine transcriptomic responses of phytoplankton communities in the California Current System (CCS) transition zone to added urea, ammonium, nitrate, and also Fe in the late summer when N depletion is common. Transcript level changes of photosynthetic, carbon fixation, and nutrient stress genes indicated relief of N limitation in many strains of Prochlorococcus, Synechococcus, and eukaryotic phytoplankton. The transcriptomic responses helped explain shifts in physiological and growth responses observed later. All three phytoplankton groups had increased transcript levels of photosynthesis and/or carbon fixation genes in response to all N substrates. However, only Prochlorococcus had decreased transcript levels of N stress genes and grew substantially, specifically after urea and ammonium additions, suggesting that Prochlorococcus outcompeted other community members in these treatments. Diatom transcript levels of carbon fixation genes increased in response to Fe but not to Fe with N which might have favored phytoplankton that were co-limited by N and Fe. Moreover, transcription patterns of closely related strains indicated variability in N utilization, including nitrate utilization by some high-light adapted Prochlorococcus. Finally, up-regulation of urea transporter genes by both Prochlorococcus and Synechococcus in response to filtered deep water suggested a regulatory mechanism other than classic control via the global N regulator NtcA. This study indicated that co-existing phytoplankton strains experience distinct nutrient stresses in the transition zone of the CCS, an understudied region where oligotrophic and coastal communities naturally mix

Evidence of nitrification associated with globally distributed pelagic jellyfish

Often considered detrimental to the environment and human activities, jellyfish blooms are increasing in several coastal regions worldwide. Yet, the overall effect of these outbreaks on ecosystem productivity and structure are not fully understood. Here we provide evidence for a so far unanticipated role of jellyfish in marine nitrogen cycling. Pelagic jellyfish release nitrogen as a metabolic waste product in form of ammonium. Yet, we observed high rates of nitrification (NH4+ → NO3−, 5.7–40.8 nM gWW−1 [wet weight] h−1) associated with the scyphomedusae Aurelia aurita, Chrysaora hysoscella, and Chrysaora pacifica and low rates of incomplete nitrification (NH4+ → NO2−, 1.0–2.8 nM gWW−1 h−1) associated with Chrysaora fulgida, C. hysoscella, and C. pacifica. These observations indicate that microbes living in association with these jellyfish thrive by oxidizing the readily available ammonia to nitrite and nitrate. The four studied species have a large geographic distribution and exhibit frequent population outbreaks. We show that, during such outbreaks, jellyfish‐associated release of nitrogen can provide more than 100% of the nitrogen required for primary production. These findings reveal a so far overlooked pathway when assessing pelagic nitrification rates that might be of particular relevance in nitrogen depleted surface waters and at high jellyfish population densities

MARTIJANEC-GAMULICA - ANALYSIS OF FINDS FROM 1950

Godine 1950. na tada još neistraženom tumulu Gamulica kod Martijanca ukopan je stup dalekovoda. Tom je prilikom mještanin Martijanca, Martin Štanglin, u tumulu prokopao jamu te otkrio tragove grobne konstrukcije i priloga. Lokalitet je nakon toga obišao prof. Stjepan Vuković, tadašnji kustos Odjela za arheologiju Gradskog muzeja Varaždin, te je pokretne nalaze prikupio, a iskop sanirao. Istraživanje pod vodstvom dr. sc. Zdenka Vinskog kao i detaljna analiza tada prikupljenih arheoloških nalaza, potvrdili su izniman značaj lokaliteta Martijanec-Gamulica, međutim, tom prilikom nisu uzeti u obzir i predmeti koje je 1950. god. prikupio S. Vuković. Dvadeset i sedam keramičkih te jedan brončani predmet čine malu, ali vrijednu skupinu nalaza čija će analiza i konačna objava zaokružiti spoznaje o ukopu u tumulu Gamulica te pridonijeti boljem poznavanju stariježeljeznodobne grupe Martijanec-Kaptol.In 1950, on the then unexcavated tumulus of Gamulica near Martijanec, a transmission line pole was set up. On that occasion Martin Štanglin, a resident of Matrijanec, found a pit in the tumulus which contained traces of a tomb with grave goods. The site was then visited by professor Stjepan Vuković who worked as a curator at the Department of Archaeology of the Varaždin City Museum and who collected movable finds and protected the tomb. Research led by Zdenko Vinski, PhD, as well as a detailed analysis of the finds, confirmed the exceptional status of the Martijanec-Gamulica site. However, the study did not include finds collected in 1950 by S. Vuković. Twenty seven ceramic and one bronze find make up a small but valuable group of finds the analysis of which will unite our knowledge of the tomb inside the Gamulica tumulus and contribute to our understanding of the Iron Age Martijanec-Kaptol group

A test in a high altitude lake of a multi-parametric rapid methodology for assessing life in liquid environments on planetary bodies: A potential new freshwater polychaete Tubeworm community

On our planet, aqueous environments such as deep sea or high-altitude aphotic lakes, subject to present or past volcanic activity and active deglaciation, may provide analogs to the aqueous environments found on such planetary bodies as Europa, Titan or Enceladus. We report here on the methodologies and technologies tested in Laguna Negra, a high altitude lake in the Central Andes, Chile, for exploring and assessing the presence of life within planetary lakes or interior oceans. We adopted a multi-parametric Rapid Ecological Assessment (REA) approach centered around collecting video imagery (by an Underwater Imaging System) and sampling benthic sediments (for sedimentological, biological and geochemical analysis) to depths of 272 m, to complement physico-chemical sampling of the water column and collection of shallow sediments for microbiological analysis (reported in separate publications). This enabled us to classify and assess the apparent status of benthic habitats, based on substrata and environmental characteristics, together with floral and faunal community characteristics and bioturbation artifacts. Video imagery showed that the lower water column was characterized by a variably intense sestonic flux of particles and debris, among which were planktonic organisms such as ostracods, copepods, and possibly cladocerans. Sediment analysis revealed at all depths abundant diatom frustules, mainly of an acidophile pennade diatom Pinnularia acidicola, amid vegetal debris likely originating from littoral macrophytes. Video imagery showed that the lakebed was partly covered by microbial mats and depositional matter and harbored an unexpectedly rich assortment of macrofauna, including sponges, tubificid worms, flatworms, bivalves and crustaceans. Various forms of bioturbation were also encountered, some with the animals in the tracks. Most notably, at the deepest site, a previously undescribed faunal feature was evident, apparently formed by a mat-like community of several layers of what appeared to be polychaete tubeworms, possibly of the family Siboglinidae. It is hypothesized that the hydrothermal activity observed in the region may supply the compounds able to support the deep-water microrganisms from which such tubeworms typically gain sustenance. Such processes could be present on other deep and aphotic liquid-water-bearing planetary bodies

Linking Hydrothermal Geochemistry to Organismal Physiology: Physiological Versatility in Riftia pachyptila from Sedimented and Basalt-hosted Vents

Much of what is known regarding Riftia pachyptila physiology is based on the wealth of studies of tubeworms living at diffuse flows along the fast-spreading, basalt-hosted East Pacific Rise (EPR). These studies have collectively suggested that Riftia pachyptila and its chemoautotrophic symbionts are physiologically specialized, highly productive associations relying on hydrogen sulfide and oxygen to generate energy for carbon fixation, and the symbiont's nitrate reduction to ammonia for energy and biosynthesis. However, Riftia also flourish in sediment-hosted vents, which are markedly different in geochemistry than basalt-hosted systems. Here we present data from shipboard physiological studies and global quantitative proteomic analyses of Riftia pachyptila trophosome tissue recovered from tubeworms residing in the EPR and the Guaymas basin, a sedimented, hydrothermal vent field. We observed marked differences in symbiont nitrogen metabolism in both the respirometric and proteomic data. The proteomic data further suggest that Riftia associations in Guaymas may utilize different sulfur compounds for energy generation, may have an increased capacity for energy storage, and may play a role in degrading exogenous organic carbon. Together these data reveal that Riftia symbionts are far more physiologically plastic than previously considered, and that -contrary to previous assertions- Riftia do assimilate reduced nitrogen in some habitats. These observations raise new hypotheses regarding adaptations to the geochemical diversity of habitats occupied by Riftia, and the degree to which the environment influences symbiont physiology and evolution

Molecular-biological sensing in aquatic environments: recent developments and emerging capabilities

Aquatic microbial communities are central to biogeochemical processes that maintain Earth’s habitability. However, there is a significant paucity of data collected from these species in their natural environment. To address this, a suite of ocean-deployable sampling and sensing instrumentation has been developed to retrieve, archive and analyse water samples and their microbial fraction using state of the art genetic assays. Recent deployments have shed new light onto the role microbes play in essential ocean processes and highlight the risks they may pose to coastal populations. Although current designs are generally too large, complex and expensive for widespread use, a host of emerging bio-analytical technologies have the potential to revolutionise this field and open new possibilities in aquatic microbial metrology

“Omics”-Enabled Microbial Sensors on Ocean Platforms

In order to assess the diversity and function of microbial communities most effectively, molecular assays need to be designed that target the phylogenetic markers and functional genes that are key to major ecological processes and microorganisms. A streamlined design process is presented here that designs probes for microarray and quantitative PCR (qPCR) assays, for application on the lab bench or on remote instrumentation. These assays can be used for DNA (genome) and RNA (gene transcription) studies. The pipeline described here establishes a database of environmental sequences, which is then used for the design of molecular probes for microarrays and to inform the design of qPCR assays, which is detailed here along with the assay optimization process. Finally, the process for the design of high-density microarrays is described. The qPCR protocol is currently used for assay optimization on the Environmental Sample Processor, a deployable robotic “genosensor” (http://www.mbari.org/esp). The protocols described here should advance applications in microbial oceanography using robotic instrumentation as well as traditional sampling methods