Ultrastructure and taxonomy of a marine photosynthetic stramenopile Phaeomonas parva gen. et sp. nov. (Pinguiophyceae) with emphasis on the flagellar apparatus architecture

Phaeomonas parva gen. et sp. nov., a marine photosynthetic stramenopile from oceanic water near the Caroline Islands, is described. Cells are naked and spherical to ovoid. The alga is motile with two laterally inserted flagella during the light period, whereas during the dark period, it absorbs the flagella and rounds up. The anterior (immature, No. 2) long flagellum possesses tubular tripartite mastigonemes. The posterior (mature, No. 1) short flagellum is smooth and has autofluorescence at the base. The cupshaped, yellowish‐brown chloroplast occupies the posterior half of the cell, and a pyrenoid occurs in the inner cavity of the cup‐shaped chloroplast. The flagellar apparatus has several unusual features. Two basal plates and a two‐gyred proximal helix in the flagellar transitional region may suggest that P. parva is related to the Pelagophyceae, Dictyochophyceae and Sulcochrysis biplastida, a photosynthetic stramenopile of uncertain taxonomic position. The R3 and R4 roots form a loop that resembles phagotrophic chrysophytes. However, this resemblance is superficial because Phaeomonas is not phagotrophic, its R3 root has a different number of microtubules and its R3 root does not split to form a food‐uptake mouth. Phaeomonas has a ‘bypassing root’, which is found only with the Phaeophyceae, Giraudyopsis stellifera (Chrysomerophyceae), and Ankylochrysis lutea (probably a member of the Pelagophyceae). The taxonomic position of P. parva could not be determined solely from ultrastructural features. However, molecular phylogeny and biochemical analyses (published separately) strongly supported a relationship between P. parva and four other monotypic strameno‐piles, Glossomastix, Pinguiochrysis, Pinguiococcus and Polypodochrysis. Although these algae are morphologically distinct, they have unusually high percentages of polyunsaturated fatty acids, especially eicosapentoic acid. This unusual assemblage of stramenopiles is classified in a new class, the Pinguiophyceae (published separately), and P. parva is its only biflagellate member.     

Symbiomonas scintillans gen. et sp. nov. and Picophagus flagellatus gen. et sp. nov. (Heterokonta): two new heterotrophic flagellates of picoplanktonic size

Two new oceanic free-living heterotrophic Heterokonta species with picoplanktonic size (< 2 microm) are described. Symbiomonas scintillans Guillou et Chrétiennot-Dinet gen. et sp. nov. was isolated from samples collected both in the equatorial Pacific Ocean and the Mediterranean Sea. This new species possesses ultrastructural features of the bicosoecids, such as the absence of a helix in the flagellar transitional region (found in Cafeteria roenbergensis and in a few bicosoecids), and a flagellar root system very similar to that of C. roenbergensis, Acronema sippewissettensis, and Bicosoeca maris. This new species is characterized by a single flagellum with mastigonemes, the presence of endosymbiotic bacteria located close to the nucleus, the absence of a lorica and a R3 root composed of a 6+3+x microtubular structure. Phylogenetical analyses of nuclear-encoded SSU rDNA gene sequences indicate that this species is close to the bicosoecids C. roenbergensis and Siluania monomastiga. Picophagus flagellatus Guillou et Chrétiennot-Dinet gen. et sp. nov. was collected in the equatorial Pacific Ocean. Cells are naked and possess two flagella. This species is characterized by the lack of a transitional helix and lateral filaments on the flagellar tubular hairs, the absence of siliceous scales, two unequal flagella, R1 + R3 roots, and the absence of a rhizoplast. SSU rDNA analyses place this strain at the base of the Chrysophyceae/Synurophyceae lineages.     

A "Total Evidence" Analysis of the Phylogenetic Relationships among the Photosynthetic Stramenopiles

Phylogenetic relationships among the nine major autotrophic stramenopile taxa were inferred in a combined analysis of the rbcL, SSU rDNA, partial LSU rRNA, carotenoid, and ultrastructural data sets. The structure of the shortest combined tree is: (Outgroup, ((((Bacillariophyceae, (Pelagophyceae, Dictyochophyceae)),((Phaeophyceae, Xanthophyceae), Raphidophyceae)), Eustigmatophyceae),(Chrysophyceae, Synurophyceae))). The Synurophyceae/Chrysophyceae is the best supported group followed by the Phaeophyceae/Xanthophyceae and the Pelagophyceae/Dictyochophyceae clades. The monophyletic groups composed of Bacillariophyceae/Pelagophyceae/Dictyochophyceae and Phaeophyceae/Xanthophyceae/Raphidophyceae received the lowest Bremer support values. The optimal combined tree suggests that the diatom frustule is derived from the siliceous "skeleton" in Dictyochophyceae, that the reduced flagellar apparatus arose once in the Bacillariophyceae/Dictyochophyceae/Pelagophyceae clade, and that the specific photoreceptor-eyespot apparatus in Chrysophyceae and the Phaeophyceae/Xantophyceae clade originated independently within the autotrophic stramenopiles. Despite conflicts in tree structure between the most-parsimonious combined phylogeny and the optimal tree(s) of each data partition, it cannot be concluded that extensive incongruence exists between the data sets.     

PHYLOGENETIC AFFINITIES OF THE SARCINOCHRYSIDALES AND CHRYSOMERIDALES (HETEROKONTA) BASED ON ANALYSES OF MOLECULAR AND COMBINED DATA1

Small-subunit ribosomal RNA nucleotide sequences were inferred for Giraudyopsis stellifera Dangeard (Chrysomeridales), as well as for Pulvinaria sp. and Sarcinochrysis marina Geitler (Sarcinochrysidales,). Phylogenetic analyses of the molecular data indicate that the former is weakly related to the Phaeophyceae/Xanthophyceae clade, whereas the latter two have affinities to the Pelagophyceae, and the Sarcinochrysidales sensu stricto is transferred to this class. A recent study proposed that the Pelagophyceae belongs to a larger assemblage of chromophytic species characterized by reduced flagellar apparatuses. Although the flagellar apparatus characterizing the Sarcinochrysidales is reduced relative to the Chysomeridaels and some other chromophytes, it is the most complicated to be associated with “the reduced flagellar apparatus” lineage. Cladistic analyses of a traditional data set (largely ultrastructural features of the flagellar apparatus) and a combined traditional/molecular data set were used to assess the evolutionary trends of reduction in the flagellar apparatus within the heterokont chromophytes.     

Proposal of Pseudochattonella verruculosa gen. nov., comb. nov. (Dictyochophyceae) for a formar raphidophycean alga Chattonella verruculosa, based on 18S rDNA phylogeny and ultrastructural characteristics

Chattonella verruculosa Y. Hara et Chihara was re‐examined by molecular methods and microscopic examination. The 18S rDNA phylogenetic analysis clearly indicated that C. verruculosa is a member of the Dictyochophyceae, with a specific affinity to Florenciella parvula. The morphological features in C. verruculosa– namely the proximal helix with two gyres and many scattered DNA‐containing areas in the chloroplasts – display the evolutionary link to the Dictyochophyceae, instead of the Raphidophyceae. Similarly, unique pyrenoid morphologies are shared between C. verruculosa and the dictyochophycean algae. Combining the molecular data and morphological characteristics, C. verruculosa is transferred to Pseudochattonella gen. nov. of the class Dictyochophyceae as Pseudochattonella verruculosa (Y. Hara et Chihara) Hosoi‐Tanabe, Honda, Fukaya, Inagaki et Sako comb. nov.     

The flagellar root apparatus,the microtubular system and associated organelles in the chrysophycean flagellate,Poterioochromonas malhamensis Peterfi (syn.Poteriochromonas stipitata Scherffel andOchromonas malhamensis pringsheim)

Summary The two flagella ofPoterioochromonas are inserted in an apical platform which is shaped by six long flagellar root fibres. The arrangement and structure of these root fibres are described in detail. One of these fibres is the single nucleating site for cytoplasmic interphase microtubules which extend peripherally down to the cytoplasmic tail. Another fibre proceeds toward the centre of the cell and passes the nucleus but is different in structure, position and function from the striated rhizoplast found in many chrysophycean flagellates which is observed but vestigial inPoterioochromonas.A specific kinetosomal mitochondrion has a threefold attachment to the flagellar root apparatus. The chloroplast is also bound to the root system. It has no stigma, but a special continuation of the periplastidial cisterna is developed instead. Another cisterna extends from the nuclear envelope-dictyosome interspace to the kinetosome of the long flagellum. The functional and taxonomic meanings of these structures and of their mutual arrangement are discussed. It is concluded that the present strain (no. 933-1 a of the Collection of Algal Cultures at the Institute of Plant Physiology, Göttingen) has to be excluded from the genusOchromonas.     

Homologs of the sexually induced gene 1 (sig1) product constitute the stramenopile mastigonemes

The tripartite tubular mastigoneme on the anterior flagellum is a morphological feature that characterizes the stramenopiles. Mastigonemes are significant and potentially informative structures not only from the viewpoint of systematics, but also of cell biology. Nevertheless, few biochemical studies have been reported on stramenopile mastigonemes. The flagella of Scytosiphon lomentaria (Phaeophyceae) were successfully isolated and analyzed using SDS-PAGE followed by protein sequencing. The partial amino acid sequence of one flagellar protein (115kDa) showed high similarity with the sexually induced gene 1 (sig1) product of centric diatoms. A polyclonal antibody against the 115-kDa protein reacted not only to the shaft of mastigonemes in Scytosiphon lomentaria, but also another distinctly different stramenopile flagellate, Sulcochrysis biplastida (Dictyochophyceae). Therefore, we propose that the 115-kDa protein (i.e. Sig1 homologs) is a constituent of the tubular shaft of the mastigoneme.     

Ultrastructure of the Harmful Unarmored Dinoflagellate Cochlodinium polykrikoides (Dinophyceae) with Reference to the Apical Groove and Flagellar Apparatus

ABSTRACT. The external and internal ultrastructure of the harmful unarmored dinoflagellate Cochlodinium polykrikoides Margalef has been examined with special reference to the apical groove and three‐dimensional structure of the flagellar apparatus. The apical groove is U‐shaped and connected to the anterior sulcal extension on the dorsal side of the epicone. The eyespot is located dorsally and composed of two layers of globules situated within the chloroplast. A narrow invagination of the plasma membrane is associated with the eyespot. The nuclear envelope has normal nuclear pores similar to other eukaryotes but different from the Gymnodinium group with diagnostic nuclear chambers. The longitudinal and transverse basal bodies are separated by approximately 0.5–1.0 μm and interconnected directly by a striated basal body connective and indirectly by microtubular and fibrous structures. Characteristic features of the flagellar apparatus are as follows: (1) a nuclear extension projects to the R1 (longitudinal microtubular root) and is connected to the root by thin fibrous material; (2) fibrillar structures are associated with the longitudinal and transverse flagellar canal; and (3) a striated ventral connective extends toward the posterior end of the cell along the longitudinal flagellar canal. We conclude, based on both morphological and molecular evidence, that Cochlodinium is only distantly related to Gymnodinium.     

THE FLAGELLAR APPARATUS OF GYMNODINIUM SP. (DINOPHYCEAE)1

The detailed structure of the flagellar apparatus has been determined in a small dinoflagellate of the genus Gymnodinium. Although diminutive, this dinoflagellate possesses a complex flagellar apparatus consisting of a posteriorly directed microtubular root, a transverse striated fibrous root, several striated fibrous connectives that attach the basal bodies to one another as well as to the different roots, and a conspicuous non-striated fibrous connective that directly links the posteriorly directded microtubular root with the extended lobe of the nucleus. This represents the second discovery of a nuclear connective linked to the flagellar apparatus in the Dinophyceae but is the first report to elucidate the spatial relationships of the connective with the flagellar apparatus and the cell. A detailed diagrammatic reconstruction is provided and the similarities between these flagellar apparatus features are compared with those known for other dinoflagellates. Additionally, the structure and displacement of the nuclear connective are compared with nuclear connectives described in other protists.     

The Morphology,Ultrastructure and SSU rRNA Gene Sequence of a New Freshwater Flagellate,Neobodo borokensis n. sp. (Kinetoplastea,Excavata)

A small free‐living freshwater bacteriotrophic flagellate Neobodo borokensis n. sp. was investigated by electron microscopy and analysis of its SSU ribosomal RNA gene. This protist has paraxonemal rods of typical bodonid structure in the flagella, mastigonemes on the proximal part of the posterior flagellum, two nearly parallel basal bodies, a compact kinetoplast, and discoid mitochondrial cristae. The flagellar pocket is supported by three microtubular roots (R1, R2 and R3) originating from the kinetosome. The cytopharynx is supported by the root R2, a microtubular prism, cytopharynx associated additional microtubules (CMT) and cytostome associated microtubules (FAS) bands. Symbiotic bacteria and small glycosomes were found in the cytoplasm. Cysts have not been found. The flagellate prefers freshwater habitats, but tolerates salinity up to 3–4‰. The overall morphological and ultrastructural features confirm that N. borokensis represents a new species of the genus Neobodo. Phylogenetic analysis of SSU rRNA genes is congruent with the ultrastructure and strongly supports the close relationship of N. borokensis to Neobodo saliens, N. designis, Actuariola, and a misidentified sequence of “Bodo curvifilus” within the class Kinetoplastea.     

Effects of the mixotrophic flagellate Ochromonas sp. on colony formation in Microcystis aeruginosa

The aim of this study was to investigate the interactions between the cyanobacteria (Microcystis aeruginosa) and the potential grazer (Ochromonas sp.) with regard to colony formation. Two kinds of treatments were carried out: (i) In the dialyse experiment Microcystis aeruginosa and Ochromonas sp. were physically separated by a dialyse tubing. (ii) In the contact experiment interactions between Microcystis aeruginosa, heterotrophic bacteria and Ochromonas sp. in different concentrations were investigated. In one treatment where the predator Ochromonas sp. came in direct contact with Microcystis, aggregates were formed.In the contact experiment, there were some interactions between the predator Ochromonas sp. and the two groups of prey, Microcystis aeruginosa and heterotrophic bacteria. When exposed to a low initial Ochromonas sp. concentration, Microcystis aeruginosa decreased and then remained stable in concentration. Ochromonas sp. switched to feed on heterotrophic bacteria and increased. At a high initial Ochromonas sp. concentration Microcystis was grazed down.     

THE FLAGELLAR APPARATUS OF CHRYSOCHROMULINA SP. (PRYMNESIOPHYCEAE)1

The flagellar apparatus of an undescribed species of Chrysochromulina Lackey that bears “eyelash” scales is reconstructed. The transitional region consists of two transitional plates each with an axosome, with no stellate pattern between them. Fine osmiophilic rings lie between the flagellar membrane and the outer doublets in the transitional region. The two jagella and the haptonema are inserted in a subapical depression that is lined ventrally by a spine-like projection formed by one of the parietal chloroplasts. The angles of insertion are similar to those of some other Chrysochromulina species in that both the haptonema and the right basal body lie at an extreme angle to the left basal body. The connectives of the apparatus consist of a striated distal band with a dorsal extension to the R1 and a ventral extension overlying the R2, a striated distal accessory band, an auxiliary connective from the right basal body to the adjacent ventral chloroplast, a well-developed intermediate band, two striated proximal bands, and a striated proximal accessory band. Of the microtubular roots in this Chrysochromulina species, three are associated with the left side of the cell (an R1 of 8+3; a small crystalline compound root, the R1C, associated with the R1; an R2 of three micro-tubules), and two are associated with the right basal body (an R3 of 2/2 microtubules with which the single-stranded R4 converges to form a 2/2+1 and then a 2/3 tiered arrangement). Comparisons are drawn with other species in the genus and related genera, particularly Prymne-sium.     

Dictyochophyceae Plastid Genomes Reveal Unusual Variability in Their Organization

Dictyochophyceae (silicoflagellates) are unicellular freshwater and marine algae (Heterokontophyta, stramenopiles). Despite their abundance in global oceans and potential ecological significance, discovered in recent years, neither nuclear nor organellar genomes of representatives of this group were sequenced until now. Here, we present the first complete plastid genome sequences of Dictyochophyceae, obtained from four species: Dictyocha speculum, Rhizochromulina marina, Florenciella parvula and Pseudopedinella elastica. Despite their comparable size and genetic content, these four plastid genomes exhibit variability in their organization: plastid genomes of F. parvula and P. elastica possess conventional quadripartite structure with a pair of inverted repeats, R. marina instead possesses two direct repeats with the same orientation and D. speculum possesses no repeats at all. We also observed a number of unusual traits in the plastid genome of D. speculum, including expansion of the intergenic regions, presence of an intron in the otherwise non‐intron‐bearing psaA gene, and an additional copy of the large subunit of RuBisCO gene (rbcL), the last of which has never been observed in any plastid genome. We conclude that despite noticeable gene content similarities between the plastid genomes of Dictyochophyceae and their relatives (pelagophytes, diatoms), the number of distinctive features observed in this lineage strongly suggests that additional taxa require further investigation.     

38 The sysytematics of ochromonas (chrysophyceae)

Ochromonas sensu lato is the largest genus in the Chrysophyceae, containing over 100 names. Ochromonas species are biflagellate, naked, plastid‐bearing single cells, distinguished from loricate, scaled, colonial and colorless genera. Most, if not all, species of Ochromonas are mixotrophic, i.e., they photosynthesize but they also engulf bacteria and other small prey. Preliminary evidence from SSU rRNA sequences show that Ochromonas is a polyphyletic genus. Ochromonas tuberculata is the most distinct from all other Ochromonas species. The other Ochromonas species (examined thus far) are scattered in three clades. For example, O. danica and O. sphaerocystis are sister to Poterioochromonas stipitata and P. malhamensis. Four additional species (identified by light microscopy as O. elegans Doflein, O. globosa Skuja, O. ovalis Dolfein, O. sociabilis Pringheim) have SSU rRNA sequences identical to P. malhamensis. Of these, only O. sociabilis has been transferred to Poterioochromonas. Thus, at least some species may be synonymous with others. Two clades of marine species are also known, one containing coastal species and the other containing open ocean species. A number of genera (some also polyphyletic) are interspersed amongst the Ochromonas species (e.g., Chrysolepidomonas, Chrysonephele, Chrysoxys, Cyclonexis, Dinobryon, Epipyxis, Uroglena, Uroglenopsis). The goal of this research (just beginning) is to establish a monophyletic Ochromonas, probably by assigning some species to other genera (existing or new). One major problem is that the type species, O. triangulata Vysotskii, hasn't been observed in over 100 years, and it is unclear which of several clades of Ochromonas contains the type. Results will be discussed.     

Mixotrophy and the toxicity of Ochromonas in pelagic food webs

1. Toxic compounds produced by many phytoplankton taxa are known to have negative effects on competitors (allelopathy), anti‐predatory effects on grazers (mortality or impaired reproduction) or both. Although mixotrophs of the genus Ochromonas are known to be toxic to zooplankton, it has often been assumed in studies of plankton community processes that all flagellates in the size range of this taxon are edible to typical zooplankton grazers (i.e. cells ≤30 μm for Daphnia, ≤6 μm for rotifers). 2. We explored the toxicity of a species of Ochromonas to other planktonic taxa, including its competitors (two species of phytoplankton and protists) and consumers (two species of zooplankton). To test if mode of nutrition by this mixotroph influences its toxicity to other taxa, we exposed each test species to Ochromonas cultured in chemostats under four different nutritional regimes: osmotrophy (labile dissolved organic carbon) and phagotrophy (bacterial prey) in both light and dark conditions (i.e. with or without photosynthesis). 3. Filtrate from osmotrophically fed Ochromonas had a significant negative effect on the population growth rate of two obligate phototrophic phytoplankton, Cryptomonasozolini and Chlamydomonas reinhardtii. The protists Tetrahymena tetrahymena and Paramecium aurelia were also negatively affected by Ochromonas filtrate. Ochromonas cells were toxic to both the rotifer Brachionus calicyflorus and the cladoceran Daphnia pulicaria, with the toxic effects significantly more severe when fed at high cell densities (75 000 cells mL⁻¹) than at low densities (7500 cells mL⁻¹). Ochromonas cultured osmotrophically in the light was more toxic to the Daphnia than cells cultured under other conditions. In contrast, Ochromonas from all nutritional conditions was equally highly toxic to Brachionus. 4. Our findings support the view that Ochromonas can be toxic to other components of the food web with which it interacts. It is especially toxic to zooplankton that directly consume it, although the effect depends upon Ochromonas cell density and whether or not a good food source is simultaneously present. Our results call into question the common practice of pooling flagellates into a single ‘functional group’ included in an ‘edible phytoplankton’ category of cells <30 μm in diameter.     

THE FLAGELLAR APPARATUS OF HYMENOMONAS CORONATA (PRYMNESIOPHYTA)1

The ultrastructure of Hymenomonas coronata Mills was reinvestigated to determine the microarchitecture of the flagellar apparatus. Cell morphology and flagellar apparatus structure are very similar to those of Pleurochrysis. Some important variations occur. First, a crystalline root (= compound root) is absent on microtubular root 1. Second, a two-stranded microtubular root emanates at a right angle from microtubular root 2. Third, a fibrous root emanates from the dorsal region between the basal bodies and extends to the cell's right, paralleling microtubular root 3. These similarities and variations in flagellar apparatus characters are discussed in reference to known variations in the Prymnesiophyta.     

COMPARATIVE ANALYSES OF THE DINOFLAGELLATE FLAGELLAR APPARATUS. II. CERATIUM HIRUNDINELLA1

The three-dimensional structure of the flagellar apparatus in the gonyaulacoid dinoflagellate. Ceratium hirundinella var. furcoïdes (Schröder) Hub.-Pest. was determined using serial section electron microscopy. The flagellar apparatus is quite large and consists of several components. The two basal bodies nearly abut at their proximal ends and are separated by an angle of approximately 120° The broad longitudinal microtubular root extends from the cell's left edge of the longitudinal basal body and bends around the sulcal/cingular depression into the cell's left antapical horn. A transverse striated fibrous root is associated with the transverse basal body and a narrow electron dense extension is present along the anterior edge of the transverse basal body. This study revealed severa1 hitherto unreported fibrous components of the flagellar apparatus that link the various microtubular and fibrous components to themselves and to the two striated collars. A large striated fibrous connective links the two striated collars to one another. This fibrous connective is linked to another striated fibrous connective that originates from the longitudinal basal body and lies perpendicular to the longitudinal microtubular root. The readily identifiable and numerous components of the Ceratium flagellar apparatus are comparable to those of other dinoflagellates. The combined presence of well dpveloped striated collars, a striated collar connective, and a basal body angle of approximately 120° indicates that this flagellar apparatus is most like that described for Peridinioid dinoflagellates. Important similarities are also noticeable between this flagellar apparatus and that of Oxyrrhis marina.     

Ultrastructure of Euglena anabaena var. minor (Euglenophyceae)

The freshwater green euglenoid Euglena anabaena var. minor has a pellicle with groove‐ridge articulation, a chloroplast with pyrenoids doubly sheathed by two paramylon caps, and a nucleus with permanently condensed chromosomes and nucleolus. The flagellar apparatus basically resembles that of Euglena. The dorsal root (DR) originates at the dorsal basal body of the emergent flagellum, while both the intermediate root (IR) and ventral root (VR) originate at the ventral basal body of the non‐emergent flagellum. The cytoplasmic pocket is associated with the ventral root/ reinforcing microtubular band. However, ultrastructural characterization of E. anabaena var. minor shows the pocket to consist of five to seven microtubules, and flagellar roots with microtubule configuration of 3–4–6 in the DR‐IR‐VR. The dorsal band microtubules pair at the reservoir‐canal transition level. The doublet microtubules are formed into triplets and doublets at the lower canal level and then make pellicular microtubules at the upper canal level.     

An ultrastructural study of Marsupiomonas pelliculata gen. et sp. nov., a new member of the Pedinophyceae

A new species, Marsupiomonas pelliculata gen. et sp. nov. (Pedinophyceae), is described. A single flagellum emerges from a deep pit with a distinctive thickened margin. The flagellum has rigid fibrillar hairs which are probably formed in the perinuclear space. A short second flagellar basal body lies within the cell close to the basal body of the emergent flagellum and the flagellar root system consists of striated and microtubular roots. There is a distinctive theca covering all but the anterior end of the cell and also a single large bright green chloroplast with an immersed pyrenoid surrounded by a starch shell. The wide salinity tolerance of the species is discussed in relation to its distribution in estuarine and salt marsh habitats. The salient features of the new species—the insertion of the emergent flagellum into a deep pit and the possession of a theca—are also seen in Pedinomonas tenuis, and it is suggested that P. tenuis could be transferred to the new genus Marsupiomonas. The class Pedinophyceae now includes three genera (Pedinomonas, Resultor and Marsupiomonas) and the distinguishing features are discussed.     

Isolation of the Flagellar Swelling and Identification of Retinal in the Phototactic Flagellate, Ochromonas danica (Chrysophyceae)

Ochromonas danica, a freshwater, planktonic chrysophyte, is capable of sensing the light conditions of its environment. This biflagellate alga has a swelling near the base of the short flagellum and a chloroplastidic stigma in close association with it. A procedure is described for the isolation of this three dimensional flagellar swelling, the presumed photoreceptor. In contrast to an earlier method developed for the isolation of the paraflagellar swelling from Euglena gracilis, the protocol reported here for Ochromonas results in higher yields that should facilitate future biochemical investigations and could open avenues of investigation for the isolation and purification of the presumptive receptor protein. To verify the hypothesis that a rhodopsin-like protein might be present in this alga, we applied a standard extraction procedure successfully used in the identification of retinal. We here report the purification and identification of all-trans retinal in Ochromonas cells by column chromatography, HPLC and GC-MS. Since retinal is the chromophore of rhodopsin-like proteins, this finding may suggest that in these unicellular algae, too, a rhodopsin-like protein could be the photoreceptor pigment.