KATABLEPHARIS OVALIS,A COLORLESS FLAGELLATE WITH INTERESTING CYTOLOGICAL CHARACTERISTICS1

Katablepharis ovalis Skuja, isolated from an impoundment in Colorado, has a cell covering composed of two layers over the cell body and flagella. The outer component of the cell covering contains 25-nm-diameter hexagonal scales arranged in rows. The inner component of the cell covering is composed of a layer of interwoven microfibrils. The inner component of the cell covering is joined to the plasma membrane by one or more attachment strips that always occur outside, and along, one of the microtubular groups of the outer array. The attachment strips resemble hemidesmosomes and are composed of rows of electron-dense material, 12 nm apart, that protrude through the plasma membrane into the extracellular space, to attach to the inner wall. The two flagella are inserted subapically into a raised area of the cell. The flagella do not have any fibrillar or tubular hairs and are covered only by the two-layered cell covering. The cell has an inner and outer array of microtubules, both of which are spindle-shaped, arising at the anterior end of the cell and continuing into the posterior end of the cell. A single large Golgi apparatus occurs in the anterior cytoplasm. The nucleus is in the center of the cell. Two rows of large ejectisomes occur posterior to the area of flagellar attachment. Smaller ejectisomes occur under the plasma membrane in the posterior and medial areas of the cell. Each ejectisome is composed of a single body containing a spirally wound, tapered ribbon. On discharge, the ejectisome ribbon rolls inward, creating a tubular structure. The possible relationship between Katablepharis, the green algae, and the cryptophytes is discussed.     

THE FLAGELLAR APPARATUS OF CHILOMONAS PARAMECIUM (CRYPTOPHYCEAE) AND ITS COMPARISON WITH CERTAIN ZOOFLAGELLATES1

The major components of the internal flagellar apparatus of Chilomonas paramecium Ehr. are two large microtubular roots and a striated root paralleled by three microtubules. The two microtubular roots overlap at the basal bodies. One microtubular root follows a curved path in the anterior of the cell, and the other extends straight to the posterior passing through a groove in the nucleus. The striated root extends laterally from the basal bodies. Except that it is smaller, the posteriorly directed root bears a strong resemblance to the axostyle of oxymonads. The overall arrangement and structure of the flagellar roots is similar to the pelta, axostyle and costa of trichomonads and the pelta and axostyle of oxymonads, groups of mitochondrion-less, largely parasitic or symbiotic protozoans. An affinity between cryptomonads and oxymonads or trichomonads would have many phylogenetic implications, some of which are discussed.     

ULTRASTRUCTURE OF THE FEEDING APPARATUS AND MYONEMAL SYSTEM OF THE HETEROTROPHIC DINOFLAGELLATE PROTOPERIDINIUM SPINULOSUM1

The feeding veil or pallium of the thecate heterotrophic dinoflagellate Protoperidinium spinulosum Schiller is a highly vesiculate membranous sac containing several arched, sometimes bifurcated microtubular ribbons. It originates from an internal microtubular basket, passes through a sphincter-like osmiophilic ring located inside the posterior flagellar pore, and emerges from the cell at that pore. The osmiophilic ring is part of an interconnected myonemal system (composed of two striated collars and several striated connectives) that is anchored to the pore plate and to two inward protrusions composed of minute sulcal plates. A related species, Protoperidinium punctulatum (Paulsen) Balech, also possesses a microtubular basket/osmiophilic ring complex. Elongate electron-dense bodies within the basket resemble digestive secretory granules found in other protists. Granular, electron-lucent microbodies clustered at the anterior end of the basket may also have a role in prey digestion. Dense membranous whorls observed within a P. spinulosum cell presented as it was preparing to initiate feeding indicate a condensed storage site for pallium membranes. A narrow microtubule-strengthened pseudopodal appendage found in two non-feeding cells constitutes the tow filament that serves as the initial linkage between the dinoflagellate and its food. The structures that constitute the pallium and pallium precursors, described here for the first time, are unlike those of other known protists, although some similarities with the dinoflagellate peduncle are evident. The existence of this unique system of organelles may have important ramifications in the search for evolutionary relationships among protists.     

THE FLAGELLAR APPARATUS OF OXYRRHIS MARINA (PYRROPHYTA)1

The three-dimensional structure of the flagellar apparatus in the dinoflagellate Oxyrrhis marina has been reinvestigated and found to consist of several previously unknown components and component combinations that appear strikingly similar to those of some gymnodinoid taxa. The flagellar apparatus of this dinoflagellate is asymmetric and extremely complex consisting of a longitudinal and a transverse basal body that gives rise to eight structurally different components. The only posteriorly directed component is the large microtubular root that consists of 45–50 microtubules at its origin and is attached proximally to a perpendicularly oriented striated fibrous component. Arising from each basal body, two striated fibrous roots with different periodicities extend to the cell's left. A single stranded microtubular root with associated electron dense material emanates from the transverse basal body and also extends to the cell's left. A striated fibrous connective arises from the longitudinal basal body and extends toward the cell's right ventral surface and terminates near the sub-thecal microtubular system. A compound root consisting of microtubules and electron dense material also originates from the longitudinal basal body and extends ventrally into the anterior region of the tentacle. Structural similarities between the parallel striated fibrous roots of Oxyrrhis and Polykrikos are discussed as are flagellar apparatus similarities among other gymnodinoid dinoflagellates. A diagrammatic reconstruction of the Oxyrrhis flagellar apparatus is also presented.     

EFFECTS OF LIGHT AND GLYCEROL ON THE ORGANIZATION OF THE PHOTOSYNTHETIC APPARATUS IN THE FACULTATIVE HETEROTROPH PYRENOMONAS SALINA (CRYPTOPHYCEAE)1

The marine cryptophyte Pyrenomonas salina Santore is capable of autotrophic and heterotrophic nutrition. We studied the physiological and ultrastructural changes that accompany the shift between these nutritional modes. The addition of glycerol to batch cultures of P. salina, grown at an irradiance limiting for photoautotrophic growth, increased its growth rate and induced specific biochemical and structural changes in its photosynthetic system. Results from extracted pigment analyses, thin-section electron microscopy, and freeze-fracture electron microscopy indicated that glycerol addition reduced the cell phycoerythrin content, phycoerythrin to chlorophyll a ratio, degree of thylakoid packing, number of thylakoids · cell^?1, and PSII particle size. These properties were reduced to a similar extent in cells grown photoautotrophically under an irradiance saturating for growth. These results are consistent with the hypothesis that enhancement of heterotrophic potential occurs at the expense of light-harvesting ability in glycerol-grown P. salina.     

STRUCTURE OF EXTRACELLULAR POLYSACCHARIDES PRODUCED BY A SOIL CRYPTOMONAS SP. (CRYPTOPHYCEAE)1

The Hindak strain of a Cryptomonas species (Cryptophyceae) produces extracellular polysaccharides. Because there is no information on the structure of these compounds in the Cryptophyceae we conducted structural studies. Gas–liquid chromatographic analyses showed that the polysaccharide is composed of fucose, rhamnose, xylose, mannose, glucose, galactose, galacturonic acid, glucuronic acid, and traces of 3-O-methyl galactose. The polysaccharide was separated into two subtractions by ion-exchange chromatography. Fraction A consisted mainly of 1,3-linked galactose units and 1,4-linked galacturonic acid. Unlike fraction B, fraction A did not have xylose, 3-O-methyl galactose, or glucuronic acid. Also, its degree of branching was low compared to that of fraction B. Only traces of sulfate were present infraction A, but fraction B was 10–15% sulfated. Protein was approximately 1% in both fractions. These polysaccharides appear to be a novel type of polymer in algae.     

INGESTION AND RETENTION OF CHROOMONAS SPP. (CRYPTOPHYCEAE) BY GYMNODINIUM ACIDOTUM (DINOPHYCEAE)1

Gymnodinium acidotum Nygaard, a blue-green dinoflagellate previously shown to contain cryptophycean chloroplasts and other organelles, was observed from water and soil samples and in culture. The dinoflagellate excysts from soil samples as a mononucleated colorless cell that is positively phototactic. Colorless cells in unialgal culture remain colorless and can only be maintained less than one week whereas pigmented cells cultured unialgally grow for 10 days but then decline rapidly. Colorless cells cultured with Chroomonas spp. regain chloroplasts and have been maintained in mixed cultures for nine months. Fifty-seven percent of the dinoflagellates from mixed cultures are bi-nucleated, and three cells have been observed possibly ingesting cryptophytes. We suggest that cryptophycean chloroplasts are retained and possibly utilized by G. acidotum for at least ten days and then digested.     

STRUCTURE AND SIGNIFICANCE OF THE CRYPTOMONAD FLAGELLAR APPARATUS. I. CRYPTOMONAS OVATA (CRYPTOPHYTA)1

The absolute configuration of the flagellar apparatus in Cryptomonas ovata has been elucidated and found to be similar to that reported for Chilomonas paramecium. Variations apparent in the flagellar apparatus of Cryptomonas ovata include the presence of striations in the mitochondrion associated lamella, a rhizostyle which does not bear wing-like extensions from the microtubules and does not lie close to the nucleus, and a striated fibrous anchoring structure associated with one basal body which has not hitherto been described. The flagellar apparatus also includes a four stranded microtubular root which traverses into the anterior dorsal lobe of the cell, a striated fibrous root which is associated with a five stranded microtubular root, and a two stranded Cr root. The homologous nature of these roots to those in the larger cryptomonads is discussed in relation to the apparent reduction in flagellar apparatus size and complexity among the smaller cryptomonads. A diagrammatic reconstruction of the flagellar apparatus of Cryptomonas ovata is also presented.     

ULTRASTRUCTURE OF THE BASAL APPARATUS AND PUTATIVE VESTIGIAL FEEDING APPARATUSES IN A QUADRIFLAGELLATE EUGLENOID (EUGLENOPHYTA)1

The flagellar apparatus and presumptive vestigial feeding apparatuses of a cold-water, photosynthetic, quadriflagellate euglenoid is described. The organism possesses two similar sets of flagella each consisting of one short and one long flagellum. Each pair of flagella is associated with three microtubular roots for a total of six roots in the basal apparatus. At the level of the ventral basal bodies, each intermediate root is nine-membered, while the ventral roots are composed of eight to nine microtubules. Only one of the ventral roots lines the single microtubule reinforced pocket. A four-membered dorsal root attaches to each dorsal basal body, and at the level of the reservoir each gives rise to a dorsal band. An additional bundle of microtubules, not arising from the microtubular roots of the basal apparatus, begins posterior to the basal apparatus as a small group of a few microtubules and extends anteriorly on the right ventral side of the reservoir ending at the canal. At the level of the stigma, the microtubules are organized into a multi-layered bundle that continues to increase in size and eventually splits to form two bundles at the level of the canal. We postulate that these bundles may represent the remnants of a rod-and-vane-type feeding apparatus like that found in many phagotrophic euglenoids.     

RECONSTRUCTION OF THE FEEDING APPARATUS IN PLOEOTIA COSTATA (EUGLENOPHYTA) AND ITS RELATIONSHIP TO OTHER EUGLENOID FEEDING APPARATUSES

The ultrastructure of the feeding apparatus in Ploeotia costata Farmer and Triemer was determined and compared to other euglenoid feeding apparatuses. The feeding apparatus opened subapically onto the ventral surface and extended nearly the entire length of the cell. It consisted of four parts at the anterior surface: a comb, cytostome/pocket, vanes, and supporting rods. The comb was a multilayered structure of three horizontal microtubular rows encased in cement and formed the dorsal lip of the apparatus. The cytostome/pocket was located between the comb and the supporting rods, tapered into the cell as the cytopharynx and was surrounded by five vanes. The electron-opaque vanes extended the entire length of the feeding apparatus and were lined with microtubules for most of their length. Finally, two cement supporting rods that were joined by a crosspiece at the anterior end formed the ventral lip. The rods separated briefly before merging with the vanes. As the merged rods and vanes descended into the cell, they gradually narrowed and terminated. Comparisons of the feeding apparatus with Ploeotia vitrea, Diplonema ambulator, Lentomonas applanatum, and other euglenoids have led to the conclusion that the Type II feeding apparatus is found only in Ploeotia species.     

NEW MARINE MEMBERS OF THE GENUS HEMISELMIS (CRYPTOMONADALES,CRYPTOPHYCEAE)1

Cryptomonads are a ubiquitous and diverse assemblage of aquatic flagellates. The relatively obscure genus Hemiselmis includes some of the smallest of these cells. This genus contained only two species until 1967, when Butcher described seven new marine species mainly on the basis of observations with the light microscope. However, from these seven taxa, only H. amylifera and H. oculata were validly published. Additionally, the features Butcher used to distinguish species have since been questioned, and the taxonomy within Hemiselmis has remained clouded due to the difficulty in unambiguously applying his classification and validating many of his species. As a result, marine strains are often placed into one of three species—H. rufescens Parke, H. virescens Droop, or the invalid H. brunnescens Butcher—based on cell color alone. Here we applied microscopic and molecular tools to 13 publicly available Hemiselmis strains in an effort to clarify species boundaries. SEM failed to provide sufficient morphological variation to distinguish species of Hemiselmis, and results from LM did not correlate with clades found using both molecular phylogenetic and nucleomorph genome karyotype analysis, indicating a high degree of morphological plasticity within species. On the basis of molecular characters and collection geography we recognize four new marine species of Hemiselmis—H. cryptochromatica sp. nov., H. andersenii sp. nov., H. pacifica sp. nov., and H. tepida sp. nov.—from the waters around North America.     

ULTRASTRUCTURE OF THE FLAGELLAR APPARATUS OF PYROBOTRYS (CHLOROPHYCEAE)1

The flagellar apparatus of Pyrobotrys has a number of features that are typical of the Chlorophyceae, but others that are unusual for this class. The two flagella are inserted at the apex, but they extend to the side of the cell toward the outside of the colony, here designated as the ventral side. Four basal bodies are present, two of which extend into flagella. Four microtubular rootlets alternate between the functional and accessory basal bodies. In each cell, the two ventral rootlets are nearly parallel, but the dorsal rootlets are more widely divergent. The rootlets alternate between two and four microtubules each. A striated distal fiber connects the two functional basal bodies in the plane of the flagella. Two additional, apparently nonstriated, fibers connect the basal bodies proximal to the distal fiber. Another striated fiber is associated with each four-membered rootlet near its insertion into the flagellar apparatus. A fine periodic component is associated with each two-membered rootlet. A rhizoplast-like structure extends into the cell from each of the functional basal bodies. The arrangement of these components does not reflect the 180° rotational symmetry that is usually present in the Chlorophyceae, but appears to be derived from a more symmetrical ancestor. It is suggested that the form of the flagellar apparatus is associated with the unusual colony structure of Pyrobotrys.     

THE FLAGELLAR APPARATUS OF CHRYSOLEPIDOMONAS DENDROLEPIDOTA (CHRYSOPHYCEAE), A SINGLE-CELLED MONAD COVED WITH ORGANIC SCALES1

The flagellar apparatus of Chrysolepidomonas dedrolepidota Peters et Andersen is similar to that of other members of the Ochromonadales, Chrysophyceae. there are four microtubular roots (R_1-4) and a system II fiber (= rhizoplast). the R₁ root consists of three microtubules that nucleate many cytoplasmic microtubules. One compressed band of 10 or more cytoplasmic microtubules is directed black along the R1 root in an anti-parallel direction. The R₂ root consists of one to two microtubules, and it extends toward the distal end of the R₁ root. The R₃ root consists of six (?seven) microtubules near its proximal end. The “a” and “f” microtubules of the R₃ root are under the short flagellum, and the “f” microtubule loops back and under the basal body, extending down to the nucleus. The R₄ root consists of one to two microtubules extending along the left side of the shot flagellum and curving under the short flagellum where it terminates near the “a” microtubule of R₃ Both flagella have a transitional plate and a transitional helix with five gyres. There is a thin, second plate in the basal body at the level of the distal end of the “c” tubules of the basal body triplets. The tripartite flagellar hairs have long lateral filaments but lack short lateral filaments. We compare the flagellar apparatus with that of other members of the Ochromonadales and members of the Hydrurales and Hibberdiales.     

ULTRASTRUCTURE OF GLOEODINIUM MONTANUM (DINOPHYCEAE)1

The freshwater dinoflagellate Gloeodinium montanum Klebs (1912) was examined with transmission and scanning electron microscopy. Micrographs of ultrathin sections revealed a series of membrane layers rather than the usual dinoflagellate theca in vegetative cysts and in legates. Swarmers had distinct pellicles but appeared to be devoid of thecal plates and vesicles. The organization of cysts and swarmers appeared remarkably similar. All cell types had typical dinoflagellate nuclei with condensed chromosomes. Chloraplasts had girdle lamellae. One pyrenoid per cell was also present in chloroplasts of vegetative cysts. Starch grains and oil globules were distributed throughout the cytoplasm. Large accumulation bodies and polyvesicular vacuoles were found in aging cysts. Trichocysts and flagellar hairs were absent. Two types of intra-cellular prokaryotic organisms were discovered.     

IDENTITY OF THE ENDOSYMBIONT OF PERIDINIUM FOLIACEUM (PYRROPHYTA): ANALYSIS OF THE rbcLS OPERON1

Extant chromophytic algae have been suggested to have originated via the engulfment of a photo synthetic alga by a colorless protist. The dinoflagellate Peridinium foliaceum (Stein) Biecheler contains a reduced chlorophyll c–containing endosymbiont and, thus, represents an evolutionary intermediate stage in the establishment of chloroplasts. Although the exact phylogenetic relationship of the symbiont to extant algal species is unknown, it had been suggested that the P. foliaceum symbiont was either a diatom or a chrysophyte. Identification of the closest living relative of the P. foliaceum symbiont would provide a free-living model system with which the photosynthetic symbiont could be compared. Nucleotide sequence analysis of rbcL and rbcS (encoding the large and small subunits ofribulose-1,5-bisphosphate carboxylase/oxygenase) by the P. foliaceum symbiont was performed to provide insights into its identity. Cloned restriction fragments from a chloroplast DNA library were screened, and clones encoding the rbcLS operon were sequenced. Parsimony phylogenetic analysis was performed for each gene. Our data strongly suggest that the symbiont originated from a photosynthetic diatom.     

PHOTOINHIBITION OF MECHANICALLY STIMULABLE BIOLUMINESCENCE IN THE AUTOTROPHIC DINOFLAGELLATE CERATIUM FUSUS (PYRROPHYTA)1

Ceratium fusus (Ehrenb.) Dujardin was exposed to light of different wavelengths and photon flux densities (PFDs) to examine their effects on mechanically stimulable bioluminescence (MSL). Photoinhibition of MSL was proportional to the logarithm of PFD. Exposure to I μmol photons·m^?2s^?1 of broadband blue light (ca. 400–500 nm) produced near-complete photoinhibition (≥90% reduction in MSL) with a threshold at ca. 0.01 μmol photons·m^?2·s^?1. The threshold of photoinhibition was ca. an order of magnitude greater for both broadband green (ca. 500–580 nm) and red light (ca. 660–700 nm). Exposure to narrow spectral bands (ca. 10 nm half bandwidth) from 400 and 700 nm at a PFD of 0.1 μmol photons·m^?2·s^?1 produced a maximal response of photoinhibition in the blue wavelengths (peak ca. 490 nm). A photoinhibition response (≥ 10%) in the green (ca. 500–540 nm) and red wavelengths (ca. 680 nm) occurred only at higher PFDs (1 and 10 μmol photons·m^?2·s^?1). The spectral response is similar to that reported for Gonyaulax polyedra Stein and Pyrocystis lunula Schütt and unlike that of Alexandrium tamarense (Lebour) Balech et Tangen. The dinoflagellate's own bioluminescence is two orders of magnitude too low to result in self-photoinhibition. The quantitative relationships developed in the laboratory predict photoinhibition of bioluminescence in populations of C. fusus in the North Atlantic Ocean.     

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.     

THE FLAGELLAR APPARATUS OF THE ZOOSPORE OF UROSPORA PENICILLIFORMIS(CHLOROPHYTA)1

The flagellar apparatus of Urospora penicilliformis (Roth) Aresch. is unique, or at least very unusual among green algae. The flagellar axonemes are rigid, and contain wing-like projections. There are no central microtubules in the most proximal part of the axoneme. The transition region contains a series of electron dense transverse lamellae rather than a single septum, and lacks a stellate pattern. There is no cartwheel pattern in the proximal part of the basal bodies. The latter are associated with four different types of fibrous elements: ascending striated fibers that attach to an electron dense plate in the papillar center, lateral striated fibers that parallel microtubular roots, fibrous elements that link adjacent basal bodies, and finally two massive striated fibers that descend into the cell, passing closely along the nucleus (system II fibers, or rhizoplasts). Each of the four microtubular flagellar roots is sandwiched between two system I striated structures. The roots are probably equal; they contain proximally four, and distally up to eight microtubules. Based on the zoospore flagellar apparatus, it is concluded that the multinucleate U. penicilliformis is related to the Ulvaphyceae. Finally, a possible explanation in functional terms is given for the peculiar external morphology and behavior of the zoospore.     

ULTRASTRUCTURE OF THE BASAL BODY COMPLEX AND PUTATIVE VESTIGIAL FEEDING APPARATUS IN PHACUS PLEURONECTES (EUGLENOPHYCEAE)

Phacus pleuronectes (O. F. Müller) Dujardin is a phototrophic euglenoid with small discoid chloroplasts, a flat rigid body, and longitudinally arranged pellicular strips. The flagellar apparatus consisted of two basal bodies and three flagellar roots typical of many phototrophic euglenoids but also had a large striated fiber that connected the two basal bodies and associated with the ventral root. The three roots, in combination with the dorsal microtubular band, extended anteriorly and formed the major cytoskeletal elements supporting the reservoir membrane and ultimately the pellicle. A cytoplasmic pocket arose in the reservoir/canal transition region. It was supported by the ventral root and a C-shaped band of electron-opaque material that lined the cytoplasmic side of the pocket. A large striated fiber extended from this C-shaped band toward the reservoir membrane. The striated fibers in the basal apparatus and associated with the microtubule-reinforced pocket in P. pleuronecte s appear to be similar to those of the phagotrophic euglenoids.