Ocean Sampling Day was initiated by the EU-funded Micro B3 (Marine Microbial Biodiversity, Bioinformatics, Biotechnology) project to obtain a snapsho

N₂ fixation should be a critical process in the nitrogen-poor surface water of the eastern Mediterranean Sea. Despite favorable conditions, diazotroph abundance and N₂ fixation rates remains low for reasons yet explained. The main goal of this study was to investigate the limiting nutrients for diazotrophy in this oligotrophic environment. Hence, we conducted dedicated bottle-microcosms with eastern Mediterranean Sea water that were supplemented with mono and polysaccharides as well as inorganic nitrogen and phosphorous. Our results indicate that the diazotrophic community expressing nifH was primarily represented by heterotrophic Proteobacteria. N₂ fixation and heterotrophic bacterial activity increased up-to tenfold following two days of dark incubations, once seawater was supplemented with organic carbon substrate in the form of glucose (monosaccharides) or gum-xanthan (polysaccharide surrogate). Furthermore, our results point that carbon-rich polysaccharides, such as transparent exopolymer particles, enhance heterotrophic N₂ fixation, by forming microenvironments of intense metabolic activity, high carbon: nitrogen ratio, and possibly low O₂ levels. The conclusions of this study indicate that diazotrophs in the eastern Mediterranean coast are primarily limited by organic carbon substrates, as possibly in many other marine regions.

Marine diazotrophs convert dinitrogen (N₂) gas into bioavailable nitrogen (N), supporting life in the global ocean. In 2012, the first version of the global oceanic diazotroph database (version 1) was published. Here, we present an updated version of the database (version 2), significantly increasing the number of in situ diazotrophic measurements from 13 565 to 55 286. Data points for N₂ fixation rates, diazotrophic cell abundance, and nifH gene copy abundance have increased by 184 %, 86 %, and 809 %, respectively. Version 2 includes two new data sheets for the nifH gene copy abundance of non-cyanobacterial diazotrophs and cell-specific N₂ fixation rates. The measurements of N₂ fixation rates approximately follow a log-normal distribution in both version 1 and version 2. However, version 2 considerably extends both the left and right tails of the distribution. Consequently, when estimating global oceanic N₂ fixation rates using the geometric means of different ocean basins, version 1 and version 2 yield similar rates (43–57 versus 45–63 Tg N yr⁻¹; ranges based on one geometric standard error). In contrast, when using arithmetic means, version 2 suggests a significantly higher rate of 223±30 Tg N yr⁻¹ (mean ± standard error; same hereafter) compared to version 1 (74±7 Tg N yr⁻¹). Specifically, substantial rate increases are estimated for the South Pacific Ocean (88±23 versus 20±2 Tg N yr⁻¹), primarily driven by measurements in the southwestern subtropics, and for the North Atlantic Ocean (40±9 versus 10±2 Tg N yr⁻¹). Moreover, version 2 estimates the N₂ fixation rate in the Indian Ocean to be 35±14 Tg N yr⁻¹, which could not be estimated using version 1 due to limited data availability. Furthermore, a comparison of N₂ fixation rates obtained through different measurement methods at the same months, locations, and depths reveals that the conventional ¹⁵N₂ bubble method yields lower rates in 69 % cases compared to the new ¹⁵N₂ dissolution method. This updated version of the database can facilitate future studies in marine ecology and biogeochemistry. The database is stored at the Figshare repository (https://doi.org/10.6084/m9.figshare.21677687; Shao et al., 2022).

Mesopelagic prokaryotes (archaea and bacteria), which are transported together with nutrient-rich intermediate-water to the surface layer by deep convection in the oceans (e.g., winter mixing, upwelling systems), can interact with surface microbial populations. This interaction can potentially affect production rates and biomass of surface microbial populations, and thus play an important role in the marine carbon cycle and oceanic carbon sequestration. The Eastern Mediterranean Sea (EMS) is one of the most oligotrophic and warm systems in the world's oceans, with usually very shallow winter mixing (<200 m) and lack of large-size spring algal blooms. In this study, we collected seawater (0–1,500 m) in 9 different cruises at the open EMS during both the stratified and the mixed seasons. We show that the EMS is a highly oligotrophic regime, resulting in low autotrophic biomass and primary productivity and relatively high heterotrophic prokaryotic biomass and production. Further, we simulated deep water mixing in on-board microcosms using Levantine surface (LSW, ~0.5 m) and intermediate (LIW, ~400 m) waters at a 9:1 ratio, respectively and examined the responses of the microbial populations to such a scenario. We hypothesized that the LIW, being nutrient-rich (e.g., N, P) and a “hot-spot” for microbial activity (due to the warm conditions that prevail in these depths), may supply the LSW with not only key-limiting nutrients but also with viable and active heterotrophic prokaryotes that can interact with the ambient surface microbial population. Indeed, we show that LIW heterotrophic prokaryotes negatively affected the surface phytoplankton populations, resulting in lower chlorophyll-a levels and primary production rates. This may be due to out-competition of phytoplankton by LIW populations for resources and/or by a phytoplankton cell lysis via viral infection. Our results suggest that phytoplankton in the EMS may not likely form blooms, even after exceptionally deep winter mixing, and therefore have a very small overall effect on the vertical flux of organic matter to the deep sea.

This study considers variability in phytoplankton and heterotrophic bacterial abundances and production rates, in one of the most oligotrophic marine regions in the world–the Levantine Basin. The temporal dynamics of these planktonic groups were studied in the coastal waters of the southeastern Mediterranean Sea approximately every two weeks for a total of two years. Heterotrophic bacteria were abundant mostly during late summer and midwinter, and were positively correlated with bacterial production and with N₂ fixation. Based on size fractionating, picophytoplankton was abundant during the summer, whereas nano-microphytoplankton predominated during the winter and early spring, which were also evident in the size-fractionated primary production rates. Autotrophic abundance and production correlated negatively with temperature, but did not correlate with inorganic nutrients. Furthermore, a comparison of our results with results from the open Levantine Basin demonstrates that autotrophic and heterotrophic production, as well as N₂ fixation rates, are considerably higher in the coastal habitat than in the open sea, while nutrient levels or cell abundance are not different. These findings have important ecological implications for food web dynamics and for biological carbon sequestration in this understudied region.