• ALGAE
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• v.38 no.1
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• pp.39-55
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• 2023

Exploring mixotrophy of dinoflagellate species is critical to understanding red-tide dynamics and dinoflagellate evolution. Some species in the dinoflagellate genus Karenia have caused harmful algal blooms. Among 10 Karenia species, the mixotrophic ability of only two species, Karenia mikimotoi and Karenia brevis, has been investigated. These species have been revealed to be mixotrophic; however, the mixotrophy of the other species should be explored. Moreover, although K. mikimotoi was previously known to be mixotrophic, only a few potential prey species have been tested. We explored the mixotrophic ability of Karenia bicuneiformis, Karenia papilionacea, and Karenia selliformis and the prey spectrum of K. mikimotoi by incubating them with 16 potential prey species, including a cyanobacterium, diatom, prymnesiophyte, prasinophyte, raphidophyte, cryptophytes, and dinoflagellates. Cells of K. bicuneiformis, K. papilionacea, and K. selliformis did not feed on any tested potential prey species, indicating a lack of mixotrophy. The present study newly discovered that K. mikimotoi was able to feed on the common cryptophyte Teleaulax amphioxeia. The phylogenetic tree based on the large subunit ribosomal DNA showed that the mixotrophic species K. mikimotoi and K. brevis belonged to the same clade, but K. bicuneiformis, K. papilionacea, and K. selliformis were divided into different clades. Therefore, the presence or lack of a mixotrophic ability in this genus may be partially related to genetic characterizations. The results of this study suggest that Karenia species are not all mixotrophic, varying from the results of previous studies.

• ALGAE
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• v.31 no.4
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• pp.351-362
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• 2016

We investigated a possible reduction in $CO_2$ uptake rate by phototrophic red tide dinoflagellates arising from mixotrophy. We measured the daily ingestion rates of Prorocentrum minimum by Prorocentrum micans over 5 days in 10 L experimental bottles, and the uptake rates of total dissolved inorganic carbon ($C_T$) by a mixture of P. micans and P. minimum(mixotrophic growth), and for the predator P. micans (phototrophic growth; control) and prey P. minimum (phototrophic growth; control) alone. To account for the effect of pH on the phototrophic growth rates of P. micans and P. minimum, measurements of $C_T$ and pH in the predator and prey control bottles were continued until the pH reached the same level (pH 9.5) as that in the experimental bottles on the final day of incubation. The measured total $C_T$ uptake rate by the mixture of P. micans and P. minimum changed from 123 to $161{\mu}mol\;C_T\;kg^{-1}\;d^{-1}$ over the course of the experiment, and was lower than the $C_T$ uptake rates shown by P. micans and P. minimum in the predator and prey control bottles, respectively, which changed from 132 to $17{\mu}mol\;C_T\;kg^{-1}\;d^{-1}$ over the course of the experiment. The reduction in total $C_T$ uptake rate arising from the mixotrophy of P. micans was 7-31% of the daily $C_T$ uptake rate seen during photosynthesis. The results suggest that red tide dinoflagellates take up less $C_T$ during mixotrophy.

• ALGAE
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• v.34 no.4
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• pp.289-301
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• 2019

Mixotrophy in marine organisms is an important aspect of ecology and evolution. The discovery of mixotrophic abilities in phototrophic dinoflagellates alters our understanding of the dynamics of red tides. In the phototrophic dinoflagellate genus Alexandrium, some species are mixotrophic, but others are exclusively autotrophic. There are differences in the ecological roles of autotrophic and mixotrophic Alexandrium in marine food webs. However, of the 34 known Alexandrium species, the mixotrophic ability of >20 species has yet to be explored. In this study, the mixotrophic capabilities of Alexandrium insuetum CCMP2082, Alexandrium mediterraneum CCMP3433, Alexandrium pacificum CCMP3434, Alexandrium tamutum ATSH1609, and Alexandrium margalefii CAWD10 were investigated by providing each species with 22 diverse prey items including bacterium-sized microbeads (1 ㎛), the cyanobacterium Synechococcus sp., algal prey species, and the ciliate Mesodinium rubrum. None of the 5 Alexandrium species fed on any of the prey items. These results increase the number of Alexandrium species lacking mixotrophic abilities to 9, compared to the 7 known mixotrophic Alexandrium species. Furthermore, the Alexandrium phylogenetic tree based on the large subunit ribosomal DNA contained 3 large clades, each of which had species with and without mixotrophic abilities. Thus, the acquisition or loss of mixotrophic abilities in Alexandrium might readily occur.

• ALGAE
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• v.26 no.1
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• pp.87-96
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• 2011

Protein profiles of a common mixotrophic dinoflagellate, Prorocentrum micans, growing autotrophically and mixotrophically (fed on the cryptophyte Rhodomonas salina) were compared using two-dimensional gel electrophoresis (2-DE) to determine if they vary in different trophic modes. Approximately 2.3% of the detected proteins were differentially expressed in the different trophic modes. Twelve proteins observed only in the mixotrophic condition had lower pI value (<5) than the fifteen proteins observed only in the autotrophic condition (>5). When the internal amino acid sequences of five selected proteins differentially expressed between autotrophic and mixotrophic conditions were analyzed using matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry, two proteins that were specifically expressed in the autotrophic condition showed homology to glyceraldehyde-3-phosphatase dehydrogenase (GAPDH) and a bacterial catalase. Three mixotrophy-specific proteins showed homology to certain hypothetical proteins from an insect and bacteria. These results suggested the presence of certain gene groups that are switched on and off according to the trophic mode of P. micans.

• ALGAE
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• v.29 no.2
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• pp.137-152
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• 2014

Mixotrophic protists play diverse roles in marine food webs as predators and prey. Thus, exploring mixotrophy in phototrophic protists has emerged as a critical step in understanding marine food webs and cycling of materials in marine ecosystem. To investigate the feeding of newly described mixotrophic dinoflagellate Ansanella granifera, we explored the feeding mechanism and the different types of species that A. granifera was able to feed on. In addition, we measured the growth and ingestion rates of A. granifera feeding on the prasinophyte Pyramimonas sp., the only algal prey, as a function of prey concentration. A. granifera was able to feed on heterotrophic bacteria and the cyanobacterium Synechococcus sp. However, among the 12 species of algal prey offered, A. granifera ingested only Pyramimonas sp. A. granifera ingested the algal prey cell by engulfment. With increasing mean prey concentration, the growth rate of A. granifera feeding on Pyramimonas sp. increased rapidly, but became saturated at a concentration of $434ngCmL^{-1}$ (10,845 cells $mL^{-1}$). The maximum specific growth rate (i.e., mixotrophic growth) of A. granifera feeding on Pyramimonas sp. was $1.426d^{-1}$, at $20^{\circ}C$ under a 14 : 10 h light-dark cycle of $20{\mu}Em^{-2}s^{-1}$, while the growth rate (i.e., phototrophic growth) under similar light conditions without added prey was $0.391d^{-1}$. With increasing mean prey concentration, the ingestion rate of A. granifera feeding on Pyramimonas sp. increased rapidly, but slightly at the concentrations ${\geq}306ngCmL^{-1}$ (7,649 cells $mL^{-1}$). The maximum ingestion rate of A. granifera feeding on Pyramimonas sp. was 0.97 ng C $predator^{-1}d^{-1}$ (24.3 cells $grazer^{-1}d^{-1}$). The calculated grazing coefficients for A. granifera feeding on co-occurring Pyramimonas sp. were up to $2.78d^{-1}$. The results of the present study suggest that A. granifera can sometimes have a considerable grazing impact on the population of Pyramimonas spp.

• Journal of Microbiology and Biotechnology
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• v.20 no.9
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• pp.1276-1282
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• 2010

In traditional mixotrophic cultures of microalgae, all the inorganic nutrients and organic carbon sources are supplied in the medium before inoculation. In this study, however, an alternative approach was adopted in Haematococcus pluvialis Flotow, a microalga capable of growing mixotrophically on sodium acetate (Na-Ac). First, the cells were grown under 75 ${\mu}Mol$ photons $m^{-2}s^{-1}$ phototrophically without Na-Ac until the stationary phase and then exposed to five different light regimes by the addition of Na-Ac (e.g., dark, 20, 40, 75, and 150 ${\mu}Mol$ photons $m^{-2}s^{-1}$). Dry weight (DW), pigments, and especially cell number in alternative mixotrophy (AM) were higher than traditional mixotrophy (TM). Cell number in AM almost doubled up from 21.7 to $42.9{\times}10^4$ cells/ml during 5-day exposure to Na-Ac, whereas the increase was only 1.2-fold in TM. Maximum cell density was reached in 75 ${\mu}Mol$ photons $m^{-2}s^{-1}$ among the light intensities tested. We propose that Na-Ac in TM of H. pluvialis can not be utilized as efficiently as in AM. With this respect, AM has several advantages against TM such as a much higher cell density in a batch culture period and minimized risk of contamination owing to the shorter exposure of cells to organic carbon sources. In consequence, this method may be used for other strains of the species, and even for the other microalgal species able to grow mixotrophically.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.1
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• pp.38-44
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• 2016

The quality and quantity of live food sources strongly influence the success of fish production in farming operations. Thus, critical studies of live forage species are a crucial element for progress in fish aquaculture. The fat content of food is an especially important determinant of growth in marine fish. Omega-3 highly unsaturated fatty acids (HUFA) are essential components of diet that determine the nutritional value of larval fish. Euglena is a protist that has potential as a forage species. These single-celled organisms have plant and animal characteristics they are motile, elliptical in shape and 15–500 μm in diameter. Their nutritional content is excellent, but most studies have focused on cells raised in autotrophic culture. We therefore examined differences in the lipid and fatty acid contents, and the growth of Euglena cells grown under autotrophic, heterotrophic, and mixotrophic conditions. Biomass production reached 15.03 g/L, 12.28 g/L, and 3.66 g/L under mixotrophy, heterotrophy, and autotrophy, respectively. The proportional n-3 HUFA content differed among culture methods: 10.04%, 5.80% and 10.01% in mixotrophic, heterotrophic and autotrophic cultures, respectively. Mixotrophy was to be the best form of cultivation for improving the growth and nutritional content of Euglena.

• ALGAE
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• v.30 no.1
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• pp.35-47
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• 2015

The marine sand-dwelling dinoflagellate Polykrikos lebourae possesses obvious gold-brown pigmented plastids as well as taeniocyst-nematocyst complex structures. Despite of the presence of the visible plastids, previous attempts to establish this species in culture all failed and thus the unavailability of cultures of this species has posed a major obstacle to further detailed exploration of ecophysiology of the dinoflagellate. Here, we isolated P. lebourae from sandy sediment of an intertidal flat on Korean western coast, successfully established it in culture, and have been maintaining the stock culture over the past 3 years. Using this stock culture, we explored phagotrophy and potential prey resources of P. lebourae, growth and grazing responses of P. lebourae to different prey organisms, the effect of prey concentration on growth and grazing rates and gross growth efficiency (GGE) of P. lebourae when fed three different prey organisms, and the growth kinetics of P. lebourae under different light regimes. P. lebourae captured prey cells using a tow filament and then phagocytized them through the posterior end. The dinoflagellate was capable of ingesting a broad range of prey species varying in size, but not all prey species tested in this study supported its sustained growth. GGE of P. lebourae was extremely high at low prey concentration and moderate or low at high prey concentrations, indicating that P. lebourae grows heterotrophically at high prey concentrations but its growth seems to be more dependent on a certain growth factor or photosynthesis of plastids derived from the prey. In the presence of prey in excess, P. lebourae grew well at moderate light intensity of $40{\mu}mol$ photons $m^{-2}s^{-1}$, but did not grow at dim and high (10 or $120{\mu}mol$ photons $m^{-2}s^{-1}$) light intensities. Our results suggest that the benthic dinoflagellate P. lebourae is an obligate mixotroph, requiring both prey and light for sustained growth and survival.

• ALGAE
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• v.38 no.2
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• pp.111-126
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• 2023

Mixotrophic dinoflagellates act as primary producers, prey, and predators in marine planktonic food webs, whereas exclusively autotrophic dinoflagellates are primary producers and prey. Species of the dinoflagellate genus Scrippsiella are commonly found in marine ecosystems and sometimes cause harmful red tides. Among the 28 formally described Scrippsiella species, S. acuminata has been found to be mixotrophic and two unidentified species have been found to be mixotrophic. To determine whether the other species in this genus are similarly mixotrophic, the mixotrophic ability of S. donghaiensis SDGJ1703, S. lachrymosa SLBS1703, S. masanensis SSMS0908, S. plana SSSH1009A, and S. ramonii VGO1053 was explored using 15 potential prey items, including 2-㎛ fluorescently labeled microspheres (FLM) and heterotrophic bacteria (FLB), the cyanobacterium Synechococcus sp., and various microalgal prey species. The ability of S. acuminata to feed on FLM and FLB was also investigated. We found that S. donghaiensis, S. lachrymosa, S. masanensis, S. plana, and S. ramonii did not feed on any potential prey tested in this study, indicating a lack of mixotrophy. However, S. acuminata fed on both FLM and FLB, confirming its mixotrophic ability. These results lowered the proportion of mixotrophic species relative to the total number of tested Scrippsiella species for mixotrophy from 100% to 29-38%. Owing to its mixotrophic ability, S. acuminata occupies an ecological niche that is distinct from that of S. donghaiensis, S. lachrymosa, S. masanensis, S. plana, and S. ramonii.

• ALGAE
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• v.35 no.3
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• pp.225-236
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• 2020

Gymnodinium smaydae is a newly described mixotrophic dinoflagellate that feeds on only Heterocapsa spp. and Scrippsiella acuminata among 19 tested algal prey. It is one of the fastest growing dinoflagellates when feeding, but does not grow well without prey. To investigate its spatial-temporal distributions in Korean waters, we quantified its abundance in water samples that were seasonally collected from 28 stations along the Korean Peninsula from April 2015 to October 2018, using quantitative real-time polymerase chain reactions. This dinoflagellate had a wide distribution, as reflected by the detection of G. smaydae cells at 23 of the sampling stations. However, this distribution had a strong seasonality; it was detected at 21 stations in the summer and only one station in winter. The abundance of G. smaydae was significantly and positively correlated with chlorophyll a concentration as well as with water temperature. However, there were no significant correlations between the abundance of G. smaydae and salinity, concentrations of nutrients, or dissolved oxygen concentration. During the study period, G. smaydae was present when water temperatures were 7.6-28.0℃, salinities were 9.6-34.1, concentrations of NO₃ were not detectable-106.0 μM, and concentrations of PO₄ were not detectable-3.4 μM. The highest abundance of G. smaydae was 18.5 cells mL^-1 in the coastal waters of Jinhae in July 2017 when the chlorophyll a concentration was 127 mg m^-3 and water temperature was 23.8℃. Therefore, the spatial-temporal distribution of G. smaydae in Korean coastal waters may be affected by chlorophyll a concentration and water temperature.