(Table 2) Rate of reduction in cell size (apical length) and growth rate for seven Fragilariopsis kerguelensis strains sampled during POLARSTERN cruise ANT-XXI/3

The planktonic diatom Fragilariopsis kerguelensis plays an important role in the biogeochemical cycles of the Southern Ocean, where remains of its frustules form the largest deposit of biogenic silica anywhere in the world. We assessed the genetic identity of 26 strains, from cells collected at various sites in the Southern Ocean, using three molecular markers, LSU and ITS rDNA and rbcL. The LSU sequences were identical among the tested strains, ITS sequences were highly similar, and only one base pair difference was detected among the rbcL sequences. These results, together with a large number of successful mating experiments demonstrated that the strains belong to a single biological species. We investigated the mating system and life cycle traits of F. kerguelensis. Cell size diminished gradually in clonal strains. Gamete formation only occurred when strains of opposite mating type - within a cell size range of 7-36 µm - were mixed together. Two binucleate gametes were formed in each gametangium and gamete conjugation produced a zygote that had four nuclei and was surrounded by thin siliceous scales. Two out of the four nuclei subsequently degenerated and the zygote expanded to form an auxospore surrounded by a transverse and a longitudinal perizonium. Staining with the fluorochrome PDMPO provided for the first time a clear demonstration that the longitudinal perizonium is formed after auxospore expansion is complete. Initial cells produced within the mature auxospores were 78-101 µm in length. Various authors have shown that the average valve size of F. kerguelensis varies in sediment samples collected in regions and seasons with different primary production regimes and this parameter has thus been proposed as a biological proxy for palaeo-productivity. A better understanding of the life cycle of F. kerguelensis should help the design of future investigations aimed at testing the link between cell size distribution in the natural environment and the role that environmental factors might have in the regulation of population cell size.

Occurrence of planktic foraminifera in Pliocene to Holocene sediments of DSDP Hole 81-552A in the North Atlantic (Appendix 2)

A new planktic foraminifer transfer function (GSF18) related 5 North Atlantic assemblages to winter and summer sea surface temperature. GSF18, based on recombined and simplified core top census data, preserves most environmental information and reproduces modern North Atlantic conditions with approximately the same accuracy as previous transfer functions, but can be more readily applied to faunal samples ranging in age from Pliocene to Holocene. Transfer function GSF18 has been applied to faunal data from Deep Sea Drilling Project Hole 552A to produce a 2.5 m.y. sea-surface temperature (SST) time series. Estimates show several periods between 2.3 and 4.6 Ma during which mean SST's were both several degrees warmer and several degrees cooler than modern conditions. Between 2.9 and 4.0 Ma SST was generally warmer than modern except for a 250 k.y. interval centered at 3.3 Ma. Maximum SST, with respect to modern conditions, occurred after the cool interval near 3.1 Ma when SST was approximately 3.6° C warmer than present conditions. Comparison of SST estimates with stable isotope data suggest that after peak warming at 3.1 Ma, there was an overall surface water cooling with concomitant build up of global ice volume, culminating in Northern Hemisphere glaciation. This event is also indicated by the presence of ice rafted detritus in 552A sediments at about 2.45 Ma.

Sea surface temperature estimates and alkenone C37:4 abundances in ODP Site 145-882 and 177-1090

The cold upwelling 'tongue' of the eastern equatorial Pacific is a central energetic feature of the ocean, dominating both the mean state and temporal variability of climate in the tropics and beyond. Recent evidence for the development of the modern cold tongue during the Pliocene-Pleistocene transition has been explained as the result of extratropical cooling that drove a shoaling of the thermocline. We have found that the sub-Antarctic and sub-Arctic regions underwent substantial cooling nearly synchronous to the cold tongue development, thereby providing support for this hypothesis. In addition, we show that sub-Antarctic climate changed in its response to Earth's orbital variations, from a subtropical to a subpolar pattern, as expected if cooling shrank the warm-water sphere of the ocean and thus contracted the subtropical gyres.

(Table 2) Metazoan meiofauna abundance in sediments obtained during POLARSTERN cruise ANT-XIX/3 (ANDEEP-1)

Meiofauna standing stocks and community structure are reported for the first time for abyssal soft-sediment samples in Antarctic waters. At seven stations within a depth range of 2274-5194 m a total of 128 sediment cores were retrieved with a multiple corer (MUC) on board of the R.V. Polarstern during the ANDEEP-1 cruise (ANT XIX/3). The metazoan meiofauna (defined by a lower size limit of 40 µm) was identified and counted, and one core per station was preserved for CPE, C/N, TOM and grain size analyses. Meiofauna densities are in the range of 2731 Ind./10 cm² at 2290 m depth and 75 Ind./10 cm² at 3597 m depth, with nematodes being the dominant group at all stations. Nematodes account for 84-94% followed by copepods with 2-8% of the total meiofauna. Other frequent taxa found at each station are kinorhynchs, loriciferans, tantulocarids, ostracods and tardigrades. There is a general tendency of decreasing abundances of metazoan meiofauna with increasing depth, but not all higher level taxa displayed this pattern. In addition, a tendency of decreasing higher taxon density with increasing depth was observed. Standing stocks are higher than the average found at similar depths in other oceans.

Surface Ocean CO2 Atlas (SOCAT) V1.4

A well-documented, publicly available, global data set of surface ocean carbon dioxide (CO2) parameters has been called for by international groups for nearly two decades. The Surface Ocean CO2 Atlas (SOCAT) project was initiated by the international marine carbon science community in 2007 with the aim of providing a comprehensive, publicly available, regularly updated, global data set of marine surface CO2, which had been subject to quality control (QC). Many additional CO2 data, not yet made public via the Carbon Dioxide Information Analysis Center (CDIAC), were retrieved from data originators, public websites and other data centres. All data were put in a uniform format following a strict protocol. Quality control was carried out according to clearly defined criteria. Regional specialists performed the quality control, using state-of-the-art web-based tools, specially developed for accomplishing this global team effort. SOCAT version 1.5 was made public in September 2011 and holds 6.3 million quality controlled surface CO2 data points from the global oceans and coastal seas, spanning four decades (1968-2007). Three types of data products are available: individual cruise files, a merged complete data set and gridded products. With the rapid expansion of marine CO2 data collection and the importance of quantifying net global oceanic CO2 uptake and its changes, sustained data synthesis and data access are priorities.

Biomarker in Heinrich event layers

There are controversies regarding the origin of Heinrich layer 3 (H3), the massive ice-rafting and meltwater event in the North Atlantic during the last glacial cycle spanning a time window between 29 and 30 kyr B.P. Some argue in favor of a Laurentide Ice Sheet source similar to other Heinrich layers, while a contending view argues for the European ice sheet source. Existing geochemical proxies such as 40Ar/39Ar, 206Pb/204Pb, or epsilon-Nd, etc., could not be used to distinguish among various sources of ice-rafted debris in H3 because of their low abundances, suggesting a background glacial sediment signal. In order to circumvent this problem a biomarker-based approach is used to characterize the provenance of H layers 2, 3, and 4 and other non-Heinrich layers. The presence of hopanes and steranes and their aromatic counterparts in the H layers is incompatible with Recent sediments and is attributed to the transportation of organic matter because of the glacial erosion of source rocks. The most diagnostic and useful signatures of this ancient organic matter in the H layers are the dominance of C34 hopanoids over C33 and the occurrence of isorenieratane along with palaerenieratane. Biomarkers signatures in H layers 2 and 3 of the Labrador Sea suggest no difference in their source. Hydrocarbon distributions suggest that these sediments were derived from the Middle to Late Ordovician and Silurian source rocks of the Hudson Bay of eastern Canada. Biomarker data of the H layer 4 from the northwest Atlantic reveal that the sediments of this layer have a similar source to the H layers in the Labrador Sea.

Seasonal variations in surface metabolite composition of Fucus vesiculosus and Fucus serratus sampled at outer Kiel Fjord (August 2012 - July 2013)

The chemical composition of surface associated metabolites of two Fucus species (Fucus vesiculosus and Fucus serratus) was analysed by means of gas chromatography-mass spectrometry (GC-MS) to describe temporal patterns in chemical surface composition. Method: The two perennial brown macroalgae F. vesiculosus and F. serratus were sampled monthly at Bülk, outer Kiel Fjord, Germany (54°27'21 N / 10°11'57 E) over an entire year (August 2012 - July 2013). Per month and species six non-fertile Fucus individuals were collected from mixed stands at a depth of 0.5 m under mid water level. For surface extraction approx. 50 g of the upper 5-10 cm apical thalli tips were cut off per species. The surface extraction of Fucus was performed according to the protocol of de Nys and co-workers (1998) with minor modifications (see Rickert et al. 2015). GC/EI-MS measurements were performed with a Waters GCT premier (Waters, Manchester, UK) coupled to an Agilent 6890N GC equipped with a DB-5 ms 30 m column (0.25 mm internal diameter, 0.25 mM film thickness, Agilent, USA). The inlet temperature was maintained at 250°C and samples were injected in split 10 mode. He carrier gas flow was adjusted to 1 ml min-1. Alkanes were used for referencing of retention times. For further details (GC-MS sample preparation and analysis) see the related publication (Rickert et al. submitted to PLOS ONE).

Magnetic analyses of ODP Hole 119-738B sediments

Magnetotactic bacteria intracellularly biomineralize magnetite of an ideal grain size for recording palaeomagnetic signals. However, bacterial magnetite has only been reported in a few pre-Quaternary records because progressive burial into anoxic diagenetic environments causes its dissolution. Deep-sea carbonate sequences provide optimal environments for preserving bacterial magnetite due to low rates of organic carbon burial and expanded pore-water redox zonations. Such sequences often do not become anoxic for tens to hundreds of metres below the seafloor. Nevertheless, the biogeochemical factors that control magnetotactic bacterial populations in such settings are not well known. We document the preservation of bacterial magnetite, which dominates the palaeomagnetic signal throughout Eocene pelagic carbonates from the southern Kerguelen Plateau, Southern Ocean. We provide evidence that iron fertilization, associated with increased aeolian dust flux, resulted in surface water eutrophication in the late Eocene that controlled bacterial magnetite abundance via export of organic carbon to the seafloor. Increased flux of aeolian iron-bearing phases also delivered iron to the seafloor, some of which became bioavailable through iron reduction. Our results suggest that magnetotactic bacterial populations in pelagic settings depend crucially on particulate iron and organic carbon delivery to the seafloor.

Impacts of seawater acidification on mantle gene expression patterns of the Baltic Sea blue mussel: implications for shell formation and energy metabolism, link to supplementary material

Marine organisms have to cope with increasing CO2 partial pressures and decreasing pH in the oceans. We elucidated the impacts of an 8-week acclimation period to four seawater pCO2 treatments (39, 113, 243 and 405 Pa/385, 1,120, 2,400 and 4,000 µatm) on mantle gene expression patterns in the blue mussel Mytilus edulis from the Baltic Sea. Based on the M. edulis mantle tissue transcriptome, the expression of several genes involved in metabolism, calcification and stress responses was assessed in the outer (marginal and pallial zone) and the inner mantle tissues (central zone) using quantitative real-time PCR. The expression of genes involved in energy and protein metabolism (F-ATPase, hexokinase and elongation factor alpha) was strongly affected by acclimation to moderately elevated CO2 partial pressures. Expression of a chitinase, potentially important for the calcification process, was strongly depressed (maximum ninefold), correlating with a linear decrease in shell growth observed in the experimental animals. Interestingly, shell matrix protein candidate genes were less affected by CO2 in both tissues. A compensatory process toward enhanced shell protection is indicated by a massive increase in the expression of tyrosinase, a gene involved in periostracum formation (maximum 220-fold). Using correlation matrices and a force-directed layout network graph, we were able to uncover possible underlying regulatory networks and the connections between different pathways, thereby providing a molecular basis of observed changes in animal physiology in response to ocean acidification.