Eun Young Yoon

Mixotrophy in the Newly Described Phototrophic Dinoflagellate Woloszynskia cincta from Western Korean Waters: Feeding Mechanism, Prey Species and Effect of Prey Concentration

ABSTRACT. Woloszynskia species are dinoflagellates in the order Suessiales inhabiting marine or freshwater environments; their ecophysiology has not been well investigated, in particular, their trophic modes have yet to be elucidated. Previous studies have reported that all Woloszynskia species are photosynthetic, although their mixotrophic abilities have not been explored. We isolated a dinoflagellate from coastal waters in western Korea and established clonal cultures of this dinoflagellate. On the basis of morphology and analyses of the small/large subunit rRNA gene (GenBank accession number=FR690459), we identified this dinoflagellate as Woloszynskia cincta. We further established that this dinoflagellate is a mixotrophic species. We found that W. cincta fed on algal prey using a peduncle. Among the diverse prey provided, W. cincta ingested those algal species that had equivalent spherical diameters (ESDs) ≤12.6 μm, exceptions being the diatom Skeletonema costatum and the dinoflagellate Prorocentrum minimum. However, W. cincta did not feed on larger algal species that had ESDs≥15 μm. The specific growth rates for W. cincta increased continuously with increasing mean prey concentration before saturating at a concentration of ca. 134 ng C/ml (1,340 cells/ml) when Heterosigma akashiwo was used as food. The maximum specific growth rate (i.e. mixotrophic growth) of W. cincta feeding on H. akashiwo was 0.499 d−1 at 20 °C under illumination of 20 μE/m2/s on a 14:10 h light–dark cycle, whereas its growth rate (i.e. phototrophic growth) under the same light conditions without added prey was 0.040 d−1. The maximum ingestion and clearance rates of W. cincta feeding on H. akashiwo were 0.49 ng C/grazer/d (4.9 cells/grazer/d) and 1.9 μl/grazer/h, respectively. The calculated grazing coefficients for W. cincta on co‐occurring H. akashiwo were up to 1.1 d−1. The results of the present study suggest that grazing by W. cincta can have a potentially considerable impact on prey algal populations.

Mixotrophic ability of the phototrophic dinoflagellates Alexandrium andersonii, A. affine, and A. fraterculus

The dinoflagellate Alexandrium spp. have received much attention due to their harmful effects on diverse marine organisms, including commercially important species. For minimizing loss due to red tides or blooms of Alexandrium spp., it is very important to understand the eco-physiology of each Alexandrium species and to predict its population dynamics. Its trophic mode (i.e., exclusively autotrophic or mixotrophic) is one of the most critical parameters in establishing prediction models. However, among the 35 Alexandrium species so far described, only six Alexandrium species have been revealed to be mixotrophic. Thus, mixotrophic ability of the other Alexandrium species should be explored. In the present study, whether each of three Alexandrium species (A. andersonii, A. affine, and A. fraterculus) isolated from Korean waters has or lacks mixotrophic ability, was investigated. When diets of diverse algal prey, cyanobacteria, and bacteria sized micro-beads were provided, A. andersonii was able to feed on the prasinophyte Pyramimonas sp., the cryptophyte Teleaulax sp., and the dinoflagellate Heterocapsa rotundata, whereas neither A. affine nor A. fraterculus fed on any prey item. Moreover, mixotrophy elevated the growth rate of A. andersonii. The maximum mixotrophic growth rates of A. andersonii on Pyramimonas sp. under a 14:10h light/dark cycle of 20μEm-2s-1 was 0.432d-1, while the autotrophic growth rate was 0.243d-1. With increasing mean prey concentration, the ingestion rate of A. andersonii increased rapidly at prey concentrations <650ngCml-1 (ca. 16,240 cellsml-1), but became saturated at the higher prey concentrations. The maximum ingestion rate by A. andersonii of Pyramimonas sp. was 1.03ngC predator-1d-1 (25.6 cells predator-1d-1). This evidence suggests that the mixotrophic ability of A. andersonii should be taken into consideration in predicting the outbreak, persistence, and decline of its harmful algal blooms.

Determination of the Absolute Configuration of Polyhydroxy Compound Ostreol B Isolated from the Dinoflagellate Ostreopsis cf. ovata

Following isolation of the polyhydroxy compound, ostreol B, from cultivated cells of the toxic dinoflagellate Ostreopsis cf. ovata collected in South Korea, 1D and 2D NMR spectroscopy were employed to determine the planar chemical structure of this compound, which contained a tetrahydropyran ring, two terminal double bonds, and 21 hydroxyl groups. The absolute configurations of all stereogenic carbon centers in ostreol B were then determined through a combination of the J-based configuration analysis, rotating frame Overhauser effect correlations, and the modified Mosher method following cleavage of the 1,2-diol bonds. Ostreol B was also found to exhibit moderate cytotoxicity in HepG2, Neuro-2a and HCT-116 cells.

Feeding and grazing impact by the bloom-forming euglenophyte Eutreptiella eupharyngea on marine eubacteria and cyanobacteria

The phototrophic euglenophyte Eutreptiella eupharyngea often causes blooms in the coastal waters of many countries, but its mode of nutrition has not been assessed. This species has previously been considered as exclusively auxotrophic. To explore whether E. eupharyngea is a mixotrophic species, the protoplasm of E. eupharyngea cells were examined using light, epifluorescence, and transmission electron microscopy after eubacteria, the cyanobacterium Synechococcus sp., and diverse algal species were provided as potential prey. Furthermore, the ingestion rates of E. eupharyngea KR on eubacteria or Synechococcus sp. as a function of prey concentration were measured. In addition, grazing by natural populations of euglenophytes on natural populations of eubacteria in Masan Bay was investigated. This study is the first to report that E. eupharyngea is a mixotrophic species. Among the potential prey organisms offered, E. eupharyngea fed only on eubacteria and Synechococcus sp., and the maximum ingestion rates of these two organisms measured in the laboratory were 5.7 and 0.7 cells predator-1 h-1, respectively. During the field experiments, the maximum ingestion rates and grazing impacts of euglenophytes, including E. eupharyngea, on natural populations of eubacteria were 11.8 cells predator-1 h-1 and 1.228 d-1, respectively. Therefore, euglenophytes could potentially have a considerable grazing impact on marine bacterial populations.