Description: The genesis of phytoplankton blooms and the fate of their biomass in iron-limited, high-nutrient−low-chlorophyll regions can be studied under natural conditions with ocean iron fertilization (OIF) experiments. The Indo-German OIF experiment LOHAFEX was carried out over 40 d in late summer 2009 within the cold core of a mesoscale eddy in the productive southwest Atlantic sector of the Southern Ocean. Silicate concentrations were very low, and phytoplankton biomass was dominated by autotrophic nanoflagellates (ANF) in the size range 3−10 μm. As in all previous OIF experiments, the phytoplankton responded to iron fertilization by increasing the maximum quantum yield (Fv/Fm) and cellular chlorophyll levels. Within 3 wk, chlorophyll levels tripled and ANF biomass doubled. With the exception of some diatoms and dinoflagellates, the biomass levels of all other groups of the phyto- and protozooplankton (heterotrophic nanoflagellates, dinoflagellates and ciliates) remained remarkably stable throughout the experiment both inside and outside the fertilized patch. We attribute the unusually high biomass attained and maintained by ANF to the absence of their grazers, the salps, and to constraints on protozooplankton grazers by heavy predation exerted by the large copepod stock. The resistance to change of the ecosystem structure over 38 d after fertilization, indicated by homogeneity at regional and temporal scales, suggests that it was locked into a stable, mature state that had evolved in the course of the seasonal cycle. The LOHAFEX bloom provides a case study of a resistant/robust dynamic equilibrium between auto- and heterotrophic ecosystem components resulting in low vertical flux both inside and outside the patch despite high biomass levels.

Description: Most of the iron fertilization experiments conducted in the Southern Ocean during the past two decades were carried out in high silicic acid waters and have induced phytoplankton blooms, dominated by diatoms. The iron fertilization experiment LOHAFEX, performed during the RV Polarstern cruise ANT XXV/3 from January to March 2009, was however carried out in a silicic acid depleted mesoscale eddy in the Atlantic sector of the Southern Ocean. The low silicic acid concentrations (〈2 µM) limited diatom growth and the phytoplankton assemblage was instead dominated by nano- and picoeukaryotes. In the present study we used molecular methods to investigate the composition and succession of small phytoplankton (0.2-5µm) during LOHAFEX. This involves on the one hand ARISA (automated intergenic spacer analysis) and on the other hand 454 next generation sequencing. The ARISA approach is based on the heterogeneity of the region between the 18S and the 28S rRNA gene and delivers a quick community structure overview. The 454 sequencing is a high throughput approach and provides high resolution information on the phytoplankton diversity, including the rare biosphere. During LOHAFEX the 0.2-5µm phytoplankton fraction shows a high diversity. The most prominent classes are the Prasinophyceae (dominated by Micromonas pusilla), the Haptophyceae (dominated by Phaeocystis antarctica) and the Dinophyceae (dominated by Syndiniales). The fertilized and non-fertilized samples show a similar community structure and no significant differences concerning the abundance of the dominant species. In all samples there are a large number of sequences belonging to the rare biosphere. The results support the general notion that the diversity of the picoplankton community was highly underestimated in the past. There is still a vast number of unknown organisms, hiding in the rare biosphere, to discover. According to other studies, our outcome shows that in the northern part of the Southern Ocean the picoplankton assemblage is dominated by Micromonas pusilla and Phaeocystis antarctica. Measurements during the experiment revealed an increase in total biomass, attributed to the addition of iron. In our samples there is no species or group in the picoplankton community that is favored by the iron addition and the relative abundances remain constant. In conclusion the iron addition has increased picoplankton biomass but the assemblage composition did not change significantly.

Description: The taxonomic composition and types of particles comprising the downward particle flux were examined during the mesoscale artificial iron fertilisation experiment LOHAFEX. The experiment was conducted in low-silicate waters of the Atlantic Sector of the Southern Ocean during austral summer (January–March 2009), and induced a bloom dominated by small flagellates. Downward particle flux was low throughout the experiment, and not enhanced by addition of iron; neutrally buoyant sediment traps contained mostly faecal pellets and faecal material apparently reprocessed by mesozooplankton. TEP fluxes were low, ≤5 mg GX eq. m−2 d−1, and a few phytodetrital aggregates were found in the sediment traps. Only a few per cent of the POC flux was found in the traps consisting of intact protist plankton, although remains of taxa with hard body parts (diatoms, tintinnids, thecate dinoflagellates and foraminifera) were numerous, far more so than intact specimens of these taxa. Nevertheless, many small flagellates and coccoid cells, belonging to the pico- and nanoplankton, were found in the traps, and these small, soft-bodied cells probably contributed the majority of downward POC flux via mesozooplankton grazing and faecal pellet export. TEP likely played an important role by aggregating these small cells, and making them more readily available to mesozooplankton grazers.

Description: The iron fertilization experiment LOHAFEX was conducted in a cold-core eddy in the Southern Atlantic Ocean during austral summer. Within a few days after fertilization, a phytoplankton bloom developed dominated by nano- and picoplankton groups. Unlike previously reported for other iron fertilization experiments, a diatom bloom was prevented by iron and silicate co-limitation. We used 18S rRNA gene tag pyrosequencing to investigate the diversity of these morphologically similar cell types within the nano- and picoplankton and microscopically enumerated dominant clades after catalyzed reported deposition fluorescence in situ hybridization (CARD-FISH) with specific oligonucleotide probes. In addition to Phaeocystis, members of Syndiniales group II, clade 10–11, and the Micromonas clades ABC and E made up a major fraction of the tag sequences of the nano- and picoplankton community within the fertilized patch. However, the same clades were also dominant before the bloom and outside the fertilized patch. Furthermore, only little changes in diversity could be observed over the course of the experiment. These results were corroborated by CARD-FISH analysis which confirmed the presence of a stable nano- and picoplankton community dominated by Phaeocystis and Micromonas during the entire course of the experiment. Interestingly, although Syndiniales dominated the tag sequences, they could hardly be detected by CARD-FISH, possibly due to the intracellular parasitic life style of this clade. The remarkable stability of the nano- and picoplankton community points to a tight coupling of the different trophic levels within the microbial food web during LOHAFEX.

Description: The taxonomic composition and types of particles comprising the downward particle flux were examined during the mesoscale artificial iron fertilisation experiment LOHAFEX. The experiment was conducted in low-silicate waters of the Atlantic Sector of the Southern Ocean during austral summer (January-March 2009), and induced a bloom dominated by small flagellates. Downward particle flux was low throughout the experiment, and not enhanced by addition of iron; neutrally buoyant sediment traps contained mostly faecal pellets and faecal material apparently reprocessed by mesozooplankton. TEP fluxes were low, 〈5 mg GX eq/m**2/day, and a few phytodetrital aggregates were found in the sediment traps. Only a few per cent of the POC flux was found in the traps consisting of intact protist plankton, although remains of taxa with hard body parts (diatoms, tintinnids, thecate dinoflagellates and foraminifera) were numerous, far more so than intact specimens of these taxa. Nevertheless, many small flagellates and coccoid cells, belonging to the pico- and nanoplankton, were found in the traps, and these small, soft-bodied cells probably contributed the majority of downward POC flux via mesozooplankton grazing and faecal pellet export. TEP likely played an important role by aggregating these small cells, and making them more readily available to mesozooplankton grazers.