Fate of green plastids in Dinophysis caudata following ingestion of the benthic ciliate Mesodinium coatsi: Ultrastructure and psbA gene

Phototrophic Dinophysis species are known to acquire plastids of the cryptophyte Teleaulax amphioxeia through feeding on the ciliate Mesodinium rubrum or M. cf. rubrum. In addition, several molecular studies have detected plastid encoding genes of various algal taxa within field populations of Dinophysis species. The trophic pathway by which Dinophysis species acquire plastids from algae other than the cryptophyte genus Teleaulax, however, is unknown. In this study, we examined the fate of prey organelles and plastid genes obtained by Dinophysis caudata through ingestion of Mesodinium coatsi, a benthic ciliate that retains green plastids of Chroomonas sp. Transmission electron microscopy and molecular analysis revealed relatively rapid digestion of prey-derived plastids. Following digestion of M. coatsi, however, photodamaged D. caudata cells having olive-green rather than reddish-brown plastids were able to recover some of their original reddish-brown pigmentation. Results further suggest that plastid genes of various algal taxa detected in field populations of Dinophysis species may reflect prey diversity rather than sequestration of multiple plastid types. Ingestion and digestion of prey other than M. rubrum or M. cf. rubrum may also provide nutritional requirements needed to repair and perhaps maintain sequestered T. amphioxeia plastids.     

The Effect of Starvation on Plastid Number and Photosynthetic Performance in the Kleptoplastidic Dinoflagellate Amylax triacantha

The dinoflagellate Amylax triacantha is known to retain plastids of cryptophyte origin by engulfing the mixotrophic ciliate Mesodinium rubrum, itself a consumer of cryptophytes. However, there is no information on the fate of the prey's organelles and the photosynthetic performance of the newly retained plastids in A. triacantha. In this study, we conducted a starvation experiment to observe the intracellular organization of the prey's organelles and temporal changes in the photosynthetic efficiency of acquired plastids in A. triacantha. The ultrastructural observations revealed that while the chloroplast‐mitochondria complexes and nucleus of cryptophyte were retained by A. triacantha, other ciliate organelles were digested in food vacuoles. Acquired plastids were retained in A. triacantha for about 1 mo and showed photosynthetic activities for about 18 d when measured by a pulse‐amplitude modulation fluorometer.     

Preferential Plastid Retention by the Acquired Phototroph Mesodinium chamaeleon

The ciliate genus Mesodinium contains species that rely to varying degrees on photosynthetic machinery stolen from cryptophyte algal prey. Prey specificity appears to scales inversely with this reliance: The predominantly phototrophic M. major/rubrum species complex exhibits high prey specificity, while the heterotrophic lineages M. pulex and pupula are generalists. Here, we test the hypothesis that the recently described mixotroph M. chamaeleon, which is phylogenetically intermediate between M. major/rubrum and M. pulex/pupula, exhibits intermediate prey preferences. Using a series of feeding and starvation experiments, we demonstrate that M. chamaeleon grazes and retains plastids at rates which often exceed those observed in M. rubrum, and retains plastids from at least five genera of cryptophyte algae. Despite this relative generality, M. chamaeleon exhibits distinct prey preferences, with higher plastid retention, mixotrophic growth rates and efficiencies, and starvation tolerance when offered Storeatula major, a cryptophyte that M. rubrum does not appear to ingest. These results suggest that niche partitioning between the two acquired phototrophs may be mediated by prey identity. M. chamaeleon appears to represent an intermediate step in the transition to strict reliance on acquired phototrophy, indicating that prey specificity may evolve alongside degree of phototrophy.     

psbA based molecular analysis of cross-feeding experiments suggests that Dinophysis acuta does not harbour permanent plastids

Toxic marine dinoflagellate species of the genus Dinophysis Ehrenberg are obligate mixotrophs that require feeding on the ciliate Mesodinium rubrum and light to achieve growth. It is now well known that they harbour plastids of cryptophyte origin, particularly of the genus Teleaulax, Plagioselmis or Geminigera group (TPG clade). Nevertheless, whether these plastids are permanent, or periodically acquired from M. rubrum prey, need additional studies in different phototrophic Dinophysis species. The origin of plastids from Dinophysis acuta Ehrenberg, one of the main agents of diarrhetic shellfish poisoning (DSP) outbreaks in Western Europe, was investigated here. Cross feeding-starvation experiments were carried out with cultures of D. acuta using M. rubrum as prey, the latter fed with two cryptophyte species, Teleaulax amphioxeia Hill and Teleaulax gracilis, belonging to the TPG clade in addition to Falcomonas sp. and Hemiselmis sp. The fate of cryptophyte plastids transferred to D. acuta through its ciliate prey was investigated using the plastid psbA gene as a tracer.     

Harmful effects of Dinophysis to the ciliate Mesodinium rubrum: Implications for prey capture

Toxigenic Dinophysis spp. are obligate mixotrophic dinoflagellates that require a constant supply of prey—Mesodinium rubrum—to achieve long-term growth by means of kleptoplasty. Mesodinium rubrum is, however, a fast moving, jumping ciliate exhibiting an effective escape response from suspensivorous predators. In the present study, a series of laboratory experiments evaluating the motility and survival of M. rubrum in the presence of Dinophysis cells and/or substances contained in their culture medium was designed, in order to assess the mechanisms involved in prey capture by Dinophysis spp. Cell abundance of M. rubrum decreased in the presence of Dinophysis cf. ovum cells producing okadaic acid (OA; up to 7.94 ± 2.67 pg cell⁻¹) and smaller amounts of dinophysistoxin-1 (DTX-1) and pectenotoxin-2 (PTX-2). Prey capture was often observed after the ciliate had been attached to adhesive “mucus traps”, which only appeared in the presence of Dinophysis cells. Before being attached to the mucus traps, M. rubrum cells reduced significantly their swimming frequency (from ∼41 to 19 ± 3 jumps min⁻¹) after only 4 h of initial contact with D. cf. ovum cells. M. rubrum survival was not affected in contact with purified OA, DTX-1 and PTX-2 solutions, but decreased significantly when the ciliate was exposed to cell-free or filtered culture medium from both D. cf. ovum and D. caudata, the latter containing moderate concentrations of free eicosapentaenoic acid and docosahexaenoic acid. The results thus indicate that Dinophysis combines the release of toxic compounds other than shellfish toxins, possibly free PUFAs, and a “mucus trap” to enhance its prey capture success by immobilizing and subsequently arresting M. rubrum cells.     

PHOTOACCLIMATION IN THE PHOTOTROPHIC MARINE CILIATE MESODINIUM RUBRUM (CILIOPHORA)1

Mesodinium rubrum (=Myrionecta rubra), a marine ciliate, acquires plastids, mitochondria, and nuclei from cryptophyte algae. Using a strain of M. rubrum isolated from McMurdo Sound, Antarctica, we investigated the photoacclimation potential of this trophically unique organism at a range of low irradiance levels. The compensation growth irradiance for M. rubrum was 0.5 μmol quanta · m⁻² · s⁻¹, and growth rate saturated at ∼20 μmol quanta · m⁻² · s⁻¹. The strain displayed trends in photosynthetic efficiency and pigment content characteristic of marine phototrophs. Maximum chl a–specific photosynthetic rates were an order of magnitude slower than temperate strains, while growth rates were half as large, suggesting that a thermal limit to enzyme kinetics produces a fundamental limit to cell function. M. rubrum acclimates to light‐ and temperature‐limited polar conditions and closely regulates photosynthesis in its cryptophyte organelles. By acquiring and maintaining physiologically viable, plastic plastids, M. rubrum establishes a selective advantage over purely heterotrophic ciliates but reduces competition with other phototrophs by exploiting a very low‐light niche.     

Acquired phototrophy in Mesodinium and Dinophysis – A review of cellular organization,prey selectivity,nutrient uptake and bioenergetics

Acquired phototrophy, i.e. the use of chloroplasts from ingested prey, can be found among some species of dinoflagellates and ciliates. The best studied examples of this phenomenon in these groups are within the ciliate genus Mesodinium and the dinoflagellate genus Dinophysis, both ecologically important genera with a worldwide distribution. Mesodinium species differ considerably in their carbon metabolism. Some species rely almost exclusively on food uptake, while other species rely mostly on photosynthesis. In Mesodinium with acquired phototrophy, a number of prey organelles in addition to chloroplasts may be retained, and the host ciliate has considerable control over the acquired chloroplasts; Mesodinium rubrum is capable of dividing its acquired chloroplasts and can also photoacclimate. In Dinophysis spp., the contents of ciliate prey are sucked out, but only the chloroplasts are retained from the ingested prey. Some chloroplast house-keeping genes have been found in the nucleus of Dinophysis and some preliminary evidence suggests that Dinophysis may be capable for photoacclimation. Both genera have been claimed to take up inorganic nutrients, including NO₃⁻, indicating that processes beyond photosynthesis have been acquired. M. rubrum seems to depend upon prey species within the Teleaulax/Plagioselmis/Geminigera clade of marine cryptophytes. Up until now, Dinophysis species have only been maintained cultured on M. rubrum as food, but other ciliates may also be ingested. Dinophysis spp. and M. rubrum are obligate mixotrophs, depending upon both prey and light for sustained growth. However, while M. rubrum only needs to ingest 1–2% of its carbon demand per day to attain maximum growth, Dinophysis spp. need to obtain about half of their carbon demand from ingestion for maximum growth. Both Mesodinium and Dinophysis spp. can survive for months in the light without food. The potential role for modeling in exploring the complex balance of phototrophy and phago-heterotrophy, and its ecological implications for the mixotroph and their prey, is discussed.     

Ultrastructure of the Oral Apparatus of Mesodinium rubrum from Korea

Mesodinium rubrum Lohmann is a photosynthetic marine ciliate that has functional chloroplasts of cryptophyte origin. Little is known about the oral ultrastructure of M. rubrum compared with several reports on the sequestration of nuclei and plastids from prey organisms, such as Geminigera cryophila and Teleaulax species. Here, we describe the fine structure of the oral apparatus of a M. rubrum strain from Gomso Bay, Korea. The cytopharynx was cone‐shaped and supported by 20–22 ribbons of triplet microtubules. At the anterior end of the cytopharynx, an annulus anchored small cylinders composed of 11 microtubules. The small cylinders were spaced at regular intervals, each reinforced by one set of the triplet microtubules. At the opening of the cytostome, larger 14‐membered microtubular cylinders were set adjacent to the small, 11‐membered microtubular cylinders, each pair surrounded by separate membranes, however, only the large cylinders extended into the oral tentacles. There were 20–22 oral tentacles each having one to five extrusomes at its tip. At the anterior end of the oral apparatus, microtubular bands supporting the cytostome curved posteriad, extending beneath the cell cortex to the kinetosomes of the somatic cirri. The microtubular bands were connected by striated fibers and originated from kinetosomes anchored by fibers. Each cirrus consisted of eight cilia associated with 16 kinetosomes. The ultrastructure of M. rubrum from Korea provides information useful for taxonomic characterization of the genus Mesodinium and relevant to developing a better understanding of the acquisition of foreign organelles through phagocytosis by M. rubrum.     

Sequestered plastids in Mesodinium rubrum are functionally active up to 80 days of phototrophic growth without cryptomonad prey

The red tide ciliate Mesodinium rubrum is an obligate mixotroph which requires feeding on cryptomonad prey mainly to retain its photosynthetic apparatus. Functionality of the sequestered plastids has been known to be lowered within a few weeks. The upper limit of the functionally active duration for the newly retained plastid, however, has been rarely estimated or determined. In parallel with genetic analysis, we investigated dynamics of population density, orange fluorescence of the plastids, and DCMU ((3-(3,4-dichlorophenyl)-1,1-dimethylurea) photosynthetic capacity of phototrophically growing M. rubrum (strain MR-MAL01) for 100 days. M. rubrum populations continued their phototrophic growth for the first 6 weeks, with gradually decreasing growth rates. Rapid decline of population density began from the 8th week. The photosynthetic capacity remained quite stable, ranging from 0.7 during the 1st week down to 0.5 during the 11th week. On day 87, the photosynthetic capacity steeply decreased to 0.05. The orange fluorescence of the retained plastids became very weak during the 4th week, to be almost undetectable on day 98. Only plastid 16S rRNA gene kept strong band intensity of PCR products throughout the whole period of 100 day experiment. Interestingly, the band intensities from psaA and psbA genes all become dramatically weakened after day 77. After new prey cryptomonads (strain CR-MAL03) were offered to M. rubrum starved for 80 days, ‘CR-MAL03 type’ 1192-bp PCR product of plastid 16S rRNA gene was detected in most experimental single M. rubrum cells. Here, we demonstrate that M. rubrum can grow for ∼6 weeks in the absence of cryptomonad prey, and photosynthetic capacity of M. rubrum can be maintained active for ∼11 weeks without prey. Additionally, M. rubrum starved for 80 days was shown to be physiologically healthy enough to ingest cryptomonad preys and retain new plastids.     

Physiological Responses of Mesodinium major to Irradiance,Prey Concentration and Prey Starvation

Ciliates within the Mesodinium rubrum/Mesodinium major species complex harbor chloroplasts and other cell organelles from specific cryptophyte species. Mesodinium major was recently described, and new studies indicate that blooms of M. major are just as common as blooms of M. rubrum. Despite this, the physiology of M. major has never been studied and compared to M. rubrum. In this study, growth, food uptake, chlorophyll a and photosynthesis were measured at six different irradiances, when fed the cryptophyte, Teleaulax amphioxeia. The results show that the light compensation point for growth of M. major was significantly higher than for M. rubrum. Inorganic carbon uptake via photosynthesis contributed by far most of total carbon uptake at most irradiances, similar to M. rubrum. Mesodinium major cells contain ~four times as many chloroplast as M. rubrum leading to up to ~four times higher rates of photosynthesis. The responses of M. major to prey starvation and refeeding were also studied. Mesodinium major was well adapted to prey starvation, and 51 d without prey did not lead to mortality. Mesodinium major quickly recovered from prey starvation when refed, due to high ingestion rates of > 150 prey/predator/d.     

A Dinoflagellate Amylax triacantha with Plastids of the Cryptophyte Origin: Phylogeny,Feeding Mechanism,and Growth and Grazing Responses

The gonyaulacalean dinoflagellates Amylax spp. were recently found to contain plastids of the cryptophyte origin, more specifically of Teleaulax amphioxeia. However, not only how the dinoflagellates get the plastids of the cryptophyte origin is unknown but also their ecophysiology, including growth and feeding responses as functions of both light and prey concentration, remain unknown. Here, we report the establishment of Amylax triacantha in culture, its feeding mechanism, and its growth rate using the ciliate prey Mesodinium rubrum (= Myrionecta rubra) in light and dark, and growth and grazing responses to prey concentration and light intensity. The strain established in culture in this study was assigned to A. triacantha, based on morphological characteristics (particularly, a prominent apical horn and three antapical spines) and nuclear SSU and LSU rDNA sequences. Amylax triacantha grew well in laboratory culture when supplied with the marine mixotrophic ciliate M. rubrum as prey, reaching densities of over 7.5 × 10³ cells/ml. Amylax triacantha captured its prey using a tow filament, and then ingested the whole prey by direct engulfment through the sulcus. The dinoflagellate was able to grow heterotrophically in the dark, but the growth rate was approximately two times lower than in the light. Although mixotrophic growth rates of A. triacantha increased sharply with mean prey concentrations, with maximum growth rate being 0.68/d, phototrophic growth (i.e. growth in the absence of prey) was ?0.08/d. The maximum ingestion rate was 2.54 ng C/Amylax/d (5.9 cells/Amylax/d). Growth rate also increased with increasing light intensity, but the effect was evident only when prey was supplied. Increased growth with increasing light intensity was accompanied by a corresponding increase in ingestion. In mixed cultures of two predators, A. triacantha and Dinophysis acuminata, with M. rubrum as prey, A. triacantha outgrew D. acuminata due to its approximately three times higher growth rate, suggesting that it can outcompete D. acuminata. Our results would help better understand the ecophysiology of dinoflagellates retaining foreign plastids.     

Mass entrapment and lysis of Mesodinium rubrum cells in mucus threads observed in cultures with Dinophysis

The entrapment and death of the ciliate Mesodinium rubrum in the mucus threads in cultures with Dinophysis is described and quantified. Feeding experiments with different concentrations and predator–prey ratios of Dinophysis acuta, Dinophysis acuminata and M. rubrum to study the motility loss and aggregate formation of the ciliates and the feeding behaviour of Dinophysis were carried out. In cultures of either Dinophysis species, the ciliates became entrapped in the mucus, which led to the formation of immobile aggregates of M. rubrum and subsequent cell lysis. The proportion of entrapped ciliates was influenced by the concentration of Dinophysis and the ratio of predator and prey in the cultures. At high cell concentrations of prey (136 cells mL⁻¹) and predator (100 cells mL⁻¹), a maximum of 17% of M. rubrum cells became immobile and went through cell lysis. Ciliates were observed trapped in the mucus even when a single D. acuminata cell was present in a 3.4 mL growth medium. Both Dinophysis species were able to detect immobile or partly immobile ciliates at a distance and circled around the prey prior to the capture with a stretched out peduncle. Relatively high entrapment and lysis of M. rubrum cells in the mucus threads indicates that under certain conditions Dinophysis might have a considerable impact on the population of M. rubrum.     

SEQUESTRATION,PERFORMANCE, AND FUNCTIONAL CONTROL OF CRYPTOPHYTE PLASTIDS IN THE CILIATE MYRIONECTA RUBRA (CILIOPHORA)1

Myrionecta rubra (Lohmann 1908, Jankowski 1976 ) is a photosynthetic ciliate with a global distribution in neritic and estuarine habitats and has long been recognized to possess organelles of cryptophycean origin. Here we show, using nucleomorph (Nm) small subunit rRNA gene sequence data, quantitative PCR, and pigment absorption scans, that an M. rubra culture has plastids identical to those of its cryptophyte prey, Geminigera cf. cryophila (Taylor and Lee 1971, Hill 1991). Using quantitative PCR, we demonstrate that G. cf. cryophila plastids undergo division in growing M. rubra and are regulated by the ciliate. M. rubra maintained chl per cell and maximum cellular photosynthetic rates (P_max^cell) that were 6–8 times that of G. cf. cryophila. While maximum chl‐specific photosynthetic rates (P_max^chl) are identical between the two, M. rubra is less efficient at light harvesting in low light (LL) and has lower overall quantum efficiency. The photosynthetic saturation parameter (E_k) was not different between taxa in high light and was significantly higher in M. rubra in LL. Lower Chl:carbon ratios (θ), and hence P_max^C rates, in M. rubra resulted in lower growth rates compared with G. cf. cryophila. G. cf. cryophila possessed a greater capacity for synthesizing protein from photosynthate, while M. rubra used 3.2 times more fixed C for synthesizing lipids. Although cryptophyte plastids in M. rubra may not be permanently genetically integrated, they undergo replication and are regulated by M. rubra, allowing the ciliate to function as a phototroph.     

Growth and Chloroplast Replacement of the Benthic Mixotrophic Ciliate Mesodinium coatsi

While the ecophysiology of planktonic Mesodinium rubrum species complex has been relatively well studied, very little is known about that of benthic Mesodinium species. In this study, we examined the growth response of the benthic ciliate Mesodinium coatsi to different cryptophyte prey using an established culture of this species. M. coatsi was able to ingest all of the offered cryptophyte prey types, but not all cryptophytes supported its positive, sustained growth. While M. coatsi achieved sustained growth on all of the phycocyanin‐containing Chroomonas spp. it was offered, it showed different growth responses to the phycoerythrin‐containing cryptophytes Rhodomonas spp., Storeatula sp., and Teleaulax amphioxeia. M. coatsi was able to easily replace previously ingested prey chloroplasts with newly ingested ones within 4 d, irrespective of prey type, if cryptophyte prey were available. Once retained, the ingested prey chloroplasts seemed to be photosynthetically active. When fed, M. coatsi was capable of heterotrophic growth in darkness, but its growth was enhanced significantly in the light (14:10 h light:dark cycle), suggesting that photosynthesis by ingested prey chloroplast leads to a significant increase in the growth of M. coatsi. Our results expand the knowledge of autecology and ecophysiology of the benthic M. coatsi.     

The use of a mucus trap by Dinophysis acuta for the capture of Mesodinium rubrum prey under culture conditions

A capture mechanism observed in a culture of the dinoflagellate Dinophysis acuta when preying on the ciliate Mesodinium rubrum (also sometimes referred to as Myrionecta rubra) is described. The dinoflagellate released cohesive clumps of mucilage into the culture media. When M. rubrum cells came into contact with this mucilage, they were immediately immobilized, but remained alive for a short period of time. Observations of D. acuta cells ‘visiting and probing’ trapped M. rubrum cells were made and at a critical point D. acuta cells removed individual M. rubrum cells from the mucus to swim away with them. The removal of M. rubrum from the mucus coincided with the cells losing all their cilia and becoming swollen, presumably signifying the death of the cell. These changes may enable the D. acuta peduncle to penetrate the ciliate cell cortex. It is hypothesized that toxins produced by D. acuta play a role in the immobilization process within the mucilage trap.     

Acquired phototrophy in ciliates: a review of cellular interactions and structural adaptations

Many ciliates acquire the capacity for photosynthesis through stealing plastids or harboring intact endosymbiotic algae. Both phenomena are a form of mixotrophy and are widespread among ciliates. Mixotrophic ciliates may be abundant in freshwater and marine ecosystems, sometimes making substantial contributions toward community primary productivity. While mixotrophic ciliates utilize phagotrophy to capture algal cells, their endomembrane system has evolved to partially bypass typical heterotrophic digestion pathways, enabling metabolic interaction with foreign cells or organelles. Unique adaptations may also be found in certain algal endosymbionts, facilitating establishment of symbiosis and nutritional interactions, while reducing their fitness for survival as free-living cells. Plastid retaining oligotrich ciliates possess little selectivity from which algae they sequester plastids, resulting in unstable kleptoplastids that require frequent ingestion of algal cells to replace them. Mesodinium rubrum (=Myrionecta rubra) possesses cryptophyte organelles that resemble a reduced endosymbont, and is the only ciliate capable of functional phototrophy and plastid division. Certain strains of M. rubrum may have a stable association with their cryptophyte organelles, while others need to acquire a cryptophyte nucleus through feeding. This process of stealing a nucleus, termed karyoklepty, was first described in M. rubrum and may be an evolutionary precursor to a stable, reduced endosymbiont, and perhaps eventually a tertiary plastid. The newly described Mesodinium"chamaeleon," however, is less selective of which cryptophyte species it will retain organelles, and appears less capable of sustained phototrophy. Ciliates likely stem from a phototrophic ancestry, which may explain their propensity to practice acquired phototrophy.     

Maintenance and regeneration of cytoplasmic organelles in hybrid amebae formed by nuclear transplantation

The fine structure of cytoplasmic organelles was studied in hybrid amebae that were formed by transplanting the nucleus from one kind of ameba into the cytoplasm of a different type of ameba. Four different species or strains of amebae were utilized; Amoeba proteus, A. discoides, A. dubia, and A. amazonas. The fine structure of A. discoides was indistinguishable from that of A. proteus, but A. dubia and A. amazonas had distinctive features. To determine if the fine structure of cytoplasmic organelles in hybrid cells is like that of the nuclear parent or the cytoplasmic parent, nuclei were transferred between A. proteus and A. amazonas, which differ in their cytoplasmic ultrastructure, especially in the morphology of mitochondria. The cytoplasm of these hybrids resembled most closely that of the cytoplasmic parent. The degree to which different heterologous nuclei are capable of maintaining normal cytoplasmic organization was then studied by replacing an A. proteus nucleus with a nucleus from one of the other kinds of amebae. An A. discoides nucleus maintained A. proteus cytoplasm for several weeks. A nucleus from A. dubia or A. amazonas was able to maintain A. proteus cytoplasm for a few days, but then the cytoplasm began to show ultrastructural changes characteristic of enucleated amebae. The ability of a heterologous nucleus to promote the regeneration of Golgi bodies and endoplasmic reticulum after their decline in the absence of the nucleus was tested by transplanting a heterologous nucleus into A. proteus cytoplasm that had been enucleate for 5 days. A. discoides was the most effective heterologous nucleus donor but was less successful than normal A. proteus. Insertion of a nucleus from either A. amazonas or A. dubia resulted in “reactivation” of the cytoplasm with extension of pseudopods and resumption of motility but little or no regeneration of cell organelles. Thus, the ability of a heterologous nucleus both to maintain and to regenerate normal features of cytoplasmic organelles varied with the type of heterologous nucleus and was related to the degree of morphological resemblance between the donor and recipient cells. The role of the nucleus in motility and its role in regeneration of cytoplasmic organelles were dissociated from one another in the case of some hybrid renucleates.     

The bloom-forming ciliate Mesodinium rubrum harbours a single permanent endosymbiont

Whether the red tide Mesodinium rubrum contains a permanent cryptophyte symbiont or whether it only sequesters chloroplasts from cryptophyte prey was addressed using electron microscopy and the dynamics of photosynthesis, chloroplasts and nuclei. Mesodinium rubrum contains a branched cryptophyte symbiont consisting of many chloroplasts, mitochondria, nucleomorphs, an endoplasmic reticulum and one nucleus. The volume of the symbiont constitutes 36% of the consortium and it is separated from its host by a single-cell membrane. The chloroplasts of Mesodium are larger and morphologically different from two Teleaulax species that served as prey. The symbiont nucleus is also much larger than Teleaulax nuclei. Although M. rubrum is functionally a phototroph, sustained growth beyond two to four generations requires ingestion of prey, but less than one prey cell per generation suffices for maximum growth. This suggests that either the ciliate or its symbiont needs an essential growth factor for continuous growth.