Description: Abstract: We investigated the bacterial community structure in surface waters along a 2500 km transect in the eastern Indian Ocean. Using high throughput sequencing of the 16S rRNA gene we measured a significant latitudinal increase in bacterial richness from 800 to 1400 OTUs (42% increase; r2=0.65; p〈0.001) from the tropical Timor Sea to the colder temperate waters. Total dissolved inorganic nitrogen, chl a, phytoplankton community structure and primary productivity strongly correlated with bacterial richness (all p〈0.01). Our data suggest that primary productivity drives greater bacterial richness. Because, N2-fixation accounts for up to 50% of new production in this region we tested whether higher N2-fixation rates are linked to a greater nifH diversity. The nifH diversity was dominated by heterotrophic Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria. We did not found any mechanistic links between nifH amplicon data, bacterial richness and primary productivity due to the overall low nifH evenness in this region. Scientific statement: Geographic gradients of marine microbial diversity is currently thought to be explained by two mechanisms, 1) diversity increases with increased productivity, and 2) it increases with increasing temperature. However, conclusive evidence for these mechanisms has been lacking from studies that span gradients in both, and it is unclear which organisms are responsible for the changes in diversity along these gradients. Here we present the first analysis of bacterial richness along the West Australian boundary current, the Leeuwin Current. Our analysis of bacterial richness along a latitudinal gradient in the eastern Indian Ocean shows support for the productivity mechanism rather than the temperature mechanism. Further, we show that bacterial richness increases towards the productive temperate waters are driven by productive eukaryotes (NO3- based) and heterotrophic N2-fixers.

Description: Our data, as part of the OISO (Ocean Indien Service d'Observation) campaign, contributes to a better understanding of the physical and biological factors controlling N2 fixation in the Southern Indian Ocean and the French Southern and Antarctic lands during Austral summer January and February 2017. We measured N2 and C fixation as well as NH4+ and NO3- assimilation in 3-6 replicates per station. Additionally, we measured diagnostic pigment concentrations to evaluate phtosynthetic community composition. For pigment analysis 4L water was filtered through 25mm Whatman GF/F filters (pressure drop 〈10kPa). Samples were stored at -80°C until analysis. Pigments were analysed using High Performance Liquid Chromatography (HPLC). Pigment concentration were calculated according to Kilias et al (2013, doi:10.1111/jpy.12109). N2 fixation experiments were carried out in three to six replicates for each station. Incubations were done in pre-acid washed polycarbonate bottles on deck with ambient light conditions. All polycarbonate incubation bottles were rinsed with deionized water, and seawater prior to incubation. We used the combination of the bubble approach (Montoya et al., 1996) and the dissolution method (Mohr et al., 2010, doi:10.1371/journal.pone.0012583) proposed by Klawonn et al. (2015, doi:10.3389/fmicb.2015.00769). Bottles were filled up to capacity to avoid air contamination. Incubations were initialized by adding a 10 ml 15-15N gas bubble. Bottles were gently rocked for 15 minutes. Finally, the remaining bubble was removed to avoid equilibration between gas and aqueous phase. after 24 hours a water subsample was taken to a 12 ml exetainer and preserved with 100 µl HgCl2 solution for later determination of exact 15N-15N concentration. Natural 15N2 was determined using Membrane Inlet Mass Spectrometry (MIMS; GAM200, IPI) for each station. Analysis of 15N2 incorporated was carried out by the Isotopic Laboratory at the UC Davis, California campus. We used stable isotope tracers (15N) to measure dissolved inorganic nitrogen (DIN) assimilation rates. Experiments were initiated by adding a known concentration of 0.05 of K15NO3 and 15NH4Cl for oligotrophic waters of the IO and 0.625 µmol L-1 for HNLC regions in the ACC and PF (Knap et al., 1994, Waite et al., 2007, doi:10.1016/j.dsr2.2006.12.010) to one litre polycarbonate bottles. For C assimilation experiments, we added 20 µmol L-1 of NaH13CO3 to one of each of N2 fixation, NH4+ and NO3- assimilation experiment bottles. For incubation, we followed the same procedure as for N2 fixation experiments. Findings reveal that N2 fixation occurs throughout the whole sampling area up to 55°S latitude. In addition, variations of N2 fiaxation rates between replicates were relatively high indicating a great heterogeneity of the French Southern and Antarctic waters. References: Montoya 1996: Montoya, Joseph P., et al. "A Simple, High-Precision, High-Sensitivity Tracer Assay for N (inf2) Fixation." Applied and environmental microbiology 62.3 (1996): 986-993. Knap et al 1994: Knap, A., Michaels, A., Close, A., Ducklow, H. & Dickson, A. 1994. Protocols for the Joint Global Ocean Flux Study (JGOFS) Core Measurements, JGOFS, Reprint of the IOC Manuals and Guides No. 29. UNESCO, 19, 1.

Description: Marine microbes along with micro eukaryotes are key regulators of oceanic biogeochemical pathways. Here we present a high-resolution (every 0.5° latitude) dataset describing microbial pro- and eukaryotic diversity, in the surface and just below the thermocline, along a 7000km transect from 66° S at the Antarctic ice edge to the equator in the South Pacific Ocean. The transect, conducted in Austral winter, covered key oceanographic features including crossing of the polar front (PF), the subtropical front (STF) and the equatorial upwelling region. Our data indicate that temperature does not determine patterns of marine microbial richness, complementing the global model data from Ladau, et al. (2013). Rather, NH4⁺ nanoplankton and primary productivity were the main drivers for archaeal and bacterial richness. Eukaryote richness was highest in the least productive ocean region, the tropical oligotrophic province. We also observed a novel diversity pattern in the South Pacific Ocean; a regional increase in archaeal and bacterial diversity between 10° S and the equator. Our data showed that the mean latitudinal ranges of archaea and bacteria decreased with latitude, thereby not confirming the Rapoport's rule. We show that permanent oceanographic features, such as the STF and the equatorial upwelling can have a significant influence on pro- and eukaryotic richness.

Description: During Polarstern expedition PS99.2 in July 2016 in LTER Hausgarten, particle abundances and biovolumes were acquired using an Underwater Vision Profiler 5HD. The UVP was attached to the CTD, whereas the instrument was programmed for acquisition during the descending part of the cast with a frequency of 20 Hz, resulting in 1 acquisition every 5 cm of the water column with 1 m/s lowering speed. The pixel size was 88 µm, the acquired volume ~1 L and the resolution 4 Mpx.

Description: At approximate every degree, water samples to measure C assimilation rates were taken from the clean underway flow through system (intake at 6m). Triplicate incubation bottles were inoculated with 20 µmol/L of NaH13CO3. All polycarbonate incubation bottles were acid rinsed three times, rinsed two times with deionized water, and rinsed three times with seawater directly from the sample point prior to incubation. Samples were incubated for 24 hours. C assimilation rates experiments were terminated by filtering each bottle (pressure drop 〈10 kPa) through a 25 mm precombusted GF/F filter. Natural abundance samples for particulate organic carbon, used as t-zero values, were obtained by filtering 4 L water samples onto pre-combusted GF/F filters. All filters were snap frozen in liquid N and stored at -80 °C. Back on land, the filters were acidified and dried overnight at 60 °C. Samples were analysed at the Isotopic Laboratory at UC Davis, California, with an Elementar Vario EL Cube or Micro Cube elemental analyser (Elementar Analysensysteme GmbH, Hanau, Germany), interfaced to a PDZ Europa 20-20 isotope ratio mass spectrometer (Sercon Ltd., Cheshire, UK). The external error of analyses was 0.2 per mil for d13C. C fixat assimilation ion rates (rho in nmol/L/h) were calculated following Dugdale and Goering (1967) and Knap et al. (1996).