ALBERT

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.