A comparative study of periplast structure inCryptomonas cryophila andC. ovata (Cryptophyceae)

Summary Freeze-fracture followed by deep-etch was used with transmission electron microscopy to characterize and compare the periplasts of two cryptomonads,Cryptomonas ovata andC. cryophila. The periplast ofC. ovata consists of a dense surface mat of granular/fibrillar material overlying a series of polygonal plates attached to the undersurface of the plasma membrane (PM) at their upturned edges. Fracture faces of the PM reveal a highly stable substructure with distinct patterns of intra-membrane particles (IMPs) associated with the underlying plates; a role for the PM in plate development is indicated. The surface periplast component ofC. cryophila exhibits a cover of morphologically complex, overlapping heptagonal scales (termed rosette scales) in addition to elongate fibrils. The arrangement of IMPs within the PM is predominantly random and the inner periplast component consists of a sheet with regular pores where ejectisomes are located. The sheet does not appear closely associated with the PM. The combination of features exhibited by the periplast ofC. cryophila warrants its inclusion as a new type within theCryptophyceae.     

Cytokinesis inCryptomonas ovata (Cryptophyceae)

Summary We describe the pattern of cytokinesis inCryptomonas ovata. Cell division begins at the posterior of an enlarged cell and proceeds through the gullet and between four active flagella. This pattern of cytokinesis inCryptomonas ovata differs from that previously described for other cryptomonads and might be of importance in establishing taxonomic affinities for the group.     

Observations with the Electron Microscope on a Species of Saprolegnia

Electron micrographs are presented to show the morphology ofthe flagella in each of the two motile phases of Saprolegniaferax, and one visual light photograph of a stained cell ofthe second stage is added. The front flagellum of both phasesis a Flimmergeissel with hairs between 2µ and 3µlong arranged in two rows and each ending in a thin hair-point.The axis is covered by a wide transparent sheath made of somematerial which appears to liquefy after death. The hind flagellum,which is devoid of Flimmer, is covered by a similar sheath,though there are signs that this is flattened into a fin inthe hind flagellum of the second stage. Some very fine shorthairs 0?5µ long are interpreted as a possible internalframework to this fin. The axis of both flagella is fibrillar,but accurate numerical details have not yet been obtained. Theskins discarded from cysts have also been examined and shownto be covered with characteristic double-headed hooks, whichare strong enough to attach a cyst to passing objects.     

EFFECTS OF TEMPERATURE AND IRRADIANCE ON THE GROWTH OF TWO FRESHWATER PHOTOSYNTHET1C CRYPTOPHYTES1

Effects of light and temperature on growth of two freshwater photosynthetic cryptophytes of different cell size were studied in batch cultures. For the smaller Cryptomonas 979/67, Steele's model and equation of Platt et al. described the relationship between growth rate and photon flux density (PFD), whereas a hyperbolic tangent function gave a better fit for the larger Cryptomonas 979/62. Maximum growth rates given by the three models were consistent with each other, but the hyperbolic tangent function gave slightly lower estimates. Maximum growth rates in relation to temperature were well described for both species by the model of Logan et al. The optimum temperature for growth for Cryptomonas 979/67 was ca. 24.5° C and 19.0° C for Cryptomonas 979/62. The lethal temperatures were 30.4° C and 23.1° C for 979/67 and 979/62, respectively. The estimated maximum growth rates were 1.38 div.·day^?1 for Cryptomonas 979/67 and 0.87 div.·day ^?1 for Cryptomonas 979/62. There were interspecific differences in photoadaptation strategies, as Cryptomonas 979/67 required relatively high PFDs to show net growth, whereas Cryptomonas 979/62 grew at lower irradiances. Cryptomonas 979/67 showed photoinhibition soon after the saturation point, but Cryptomonas 979/62 tolerated a much wider range of irradiance. From their growth responses to light, Cryptomonas 979/ 67 appears to be a stenotopic and Cryptomonas 979/ 62 a eurytopic strain.     

STRUCTURE OF THE PERIPLAST OF CRYPTOMONAS OVATA VAR. PALUSTRIS1,2

The periplast of Cryptomonas ovata var. palustris is composed of polygonal plates which are delineated by shallow ridges. A small ejectosome is located at each corner of the plate area. The plate areas vary in size; they are smallest at the anterior and posterior ends and are largest in the middle of the cell with an average length of 0.5 μ and of width 0.4 μ. In cross section a plate area is composed of 2 distinct layers, an outer plasma membrane layer with a fine particulate, appearance, and an inner layer consisting of two sheets. The sheets of the inner layer have a striated lattice pattern with a periodicity of about 20 nm. In negatively stained preparations one lattice appears to underlie another at certain angles. Protease digestion removed polygonal shaped inner layer.     

Lectin binding in Cryptomonas and Chroomonas (Cryptophyceae)

The cell envelopes of Cryptomonas and Chroomonas exhibited significant fluorescence using FITC-labelled concanavalin A and wheat germ agglutinin when the cells were fixed prior to lectin binding. The periplast became intensely labelled in Chroomonas whereas Cryptomonas showed fluorescing granula in its gullet/furrow region and on the cell surface. Lectin labelling followed by fixation showed only label of periplast remnants of lysed cells and of the flagella of Chroomonas. Isolated periplasts of Cryptomonas and Chroomonas were intensively labelled with both concanavalin A and wheat germ agglutinin. Glycostaining of gels, onto which total cell protein extracts were loaded, showed a glycoprotein of high molecular weight for Cryptomonas and Chroomonas and an additional glycoprotein for Cryptomonas species.     

Further Observations with the Electron Microscope on Spermatozoids in the Brown Algae

Observations are recorded for Ascophyllum, Pelvetia, Himanthalia,and Dictyota in addition to Fucus which has already been describedin a previous paper. Fibrillar disintegration of cilia has beenobtained in all except Pelvetia, in each case to give nine peripheralstrands and a central pair. This corrects a previous error forthe hind flagellum of Fucus. Some facts are given regardingthe internal organs associated with the parts of the cilia insidethe body in Himanthalia and Dictyota. A proboscis similar tothat previously described for Fucus has been demohstrated inAscophyllum and Pelvetia, but is absent from Himanthalia andDictyota. Himanthalia differs from the other Fucoids in therelative lengths of front and hind flagella. Dictyota has onlya single flagellum. In all, the front flagellum is a Flimmergeisselwith two rows of hairs, which, in certain cases, notably Himanthalia;have very long hair points. In Himanthalia there is a largespine near the distal end of the front flagellum borne on onefibril of the peripheral series. In Dictyota there is a rowof smaller spines at the front end of the flagellum borne ina line between the two lateral rows of hairs. These spines inDictyot can be used as evidence regarding the internal symmetryof the whole cilium which is summarized in a diagram.     

Localization of calmodulin in flagella of zoospores ofPhytophthora cinnamomi

Summary Calmodulin distribution in the tinsel and whiplash flagella of zoospores ofPhytophthora cinnamomi has been studied by immunofluorescence microscopy and immunogold labelling. In whole zoospores labelled with a monoclonal antibody raised against pea calmodulin, followed by a second antibody-FITC, both flagella appear to be weakly stained except for a region at the base of the tinsel flagellum which was stained intensely. A similar staining pattern was also detected in isolated flagella labelled with anti-calmodulin. To identify the calmodulin rich region of the tinsel flagellum, we labelled sections of zoospores embedded in Lowicryl K4M with anti-calmodulin followed by a second antibody gold probe. In the tinsel flagellum, the gold labelling was restricted to a paraxonemal swelling close to the base. Very little gold labelling was detected elsewhere. The swelling extends for 1.5–2.0 n from the base of the tinsel flagellum and is hook shaped in cross section. Immunoblot analysis confirmed that the staining was specific for calmodulin.     

CRYPTOGLENA PIGRA: A EUGLENOID WITH ONE CHLOROPLAST12

The taxonomic status of Cryptoglena pigra Ehrb., interpreted from observations based on bright-field microscopy, has been uncertain. Examination with the electron microscope of a clone of C pigra isolated by E. G. Pringsheim reveals certain features which, collectively, are distinctly euglenoid: periplast associated with muciferous bodies and subpellicular microtubules; canal and reservoir with microtubules; one flagellum with a swelling and emergent through a canal, and a second flagellum without a swelling and nonremergent; stigma (eyetpot) closely apprrssed to but not part of the chloroplast; nucleus with permanently condensed chromosomes attached to the inner nuclrar membrane; mitochondria with disc-shaped cristae constricted at the base; chloroplast with thylakoids often in triplets; and paramylon grains in the cytoplasm. Unlike most euglenoids, C. pigra possesses a single chloroplast that in transverse thin sections is U-shaped.     

Occurrence of bacterivory in Cryptomonas,a common freshwater phytoplankter

Summary Bacterivory was detected by incorporation of 0.57 m diameter, fluorescent polystyrene beads and fluorescently labeled bacteria (FLB) in two cultured species of Cryptomonas (C. ovata and C. erosa), and a population of Cryptomonas sp in a humic, mesotrophic lake. Rates of ingestion and clearance were very low, and similar for the cultures and the in situ population. The in situ population incorporated 0.7–1.7 bacteria cell^-1 h^-1, thereby ingesting 0.3%–2.0% of the total bacterial numbers present in the water per day, and receiving less than 2% of its carbon content per day through bacterivory. Thus, bacterivory by Cryptomonas was quantitatively important neither as a sink for bacterial biomass, nor as a carbon source for the algal cells. Possibly, it served in the uptake of essential nutrients.     

Periplast development in Cryptophyceae II. Development of the inner periplast component inRhinomonas pauca,Proteomonas sulcata [haplomorph],Rhodomonas baltica,andCryptomonas ovata

Summary The inner periplast component (IPC) of numerous cryptomonads is composed of discrete inner plates, situated beneath (and intimately associated with) the plasma membrane (PM). Freeze-fracture images reveal that the PM is organized into a series of ordered structural domains, which directly correspond in size and shape to the underlying inner plates. Freeze-fracture images are used here to compare IPC arrangement inRhinomonas pauca, Proteomonas sulcata [haplomorph],Rhodomonas baltica, andCryptomonas ovata, and to examine development of inner plates in these cryptomonads. In all genera examined, the IPC is highly ordered across most of the cell periphery but appears to be modified adjacent to the vestibulum and mid-ventral line, which represent the anamorphic zones. Variations in the size and shape of PM domains in these regions suggest that development of the IPC occurs within anamorphic zones, by the de novo formation and enlargement of inner plates throughout the cell cycle.     

ULTRASTRUCTURAL VARIATIONS IN CRYPTOMONAD FLAGELLA1

The arrangement of flagellar appendages in 19 cryptomonad species was examined and four new flagellar types are described. The first new type has a single row of mastigonemes on both flagella and hairs on the side opposite the mastigonemes. The second type, which is common, has unilateral rows of mastigonemes on both flagella, but no hairs. A third type has an acronematic short flagellum and a single row of mastigonemes on the long flagellum. A fourth type lacks mastigonemes but has a unilateral row of curved “spikes” on the short flagellum and hairs on both flagella. These additional flagellar variations may contribute to a more natural system of classification for cryptomonads.     

Transformation and development of the flagellar apparatus ofCryptomonas ovata (Cryptophyceae) during cell division

Summary InCryptomonas ovata, long, dorsal flagella are produced which transform during the following cell division into short, ventral flagella. At division there is a reorientation in cell polarity, and the parental basal apparatus, which comprises the basal bodies and associated roots, is distributed to the daughter cells via a complex sequence of events. Flagellar apparatus development includes the transformation of a four-stranded microtubular root into a mature root of different structure and function. Each newly formed basal body nucleates new microtubular roots, but receives a striated fibrous root from a parental basal body. The striated roots are originally produced on the transforming basal body and are transferred to the new basal bodies at each successive division. The development of the asymmetric flagellar apparatus throughout the cell cycle is described.     

Flagella of a chrysophycean alga play an active role in prey capture and selection

Summary The flagella of the pigmented algaEpipyxis pulchra (Chrysophyceae) were observed with image enhanced video microscopy to play an active role in gathering, physically seizing and selecting prey prior to phagocytosis. Vegetative unicells of this sessile, freshwater species possess two structurally and functionally distinct flagella, both active in feeding. During prey gathering the long flagellum, which is adorned with stiff hairs, beats rapidly to direct a strong water current towards the cell while the short, smooth flagellum moves very little. When a potential food particle is drawn by the current to contact the flagellar surfaces, the long flagellum stops beating and positions itself, in concert with the short flagellum, to seize the prey between them. Both flagella then briefly rotate the prey before selecting or rejecting it. If rejected, the particle is discarded by the coordinated activity of both flagella. If selected as food, the prey is held in place until a complex collecting cup emanates out from a position near the basal bodies and engulfs it. The cup plus enclosed food particle, now a food vacuole, is then retracted back to the cell proper.     

FLAGELLAR HAIRS IN PRASINOPHYTES (CHLOROPHYTA): ULTRASTRUCTURE AND DISTRIBUTION ON THE FLAGELLAR SURFACE1

The flagellar hair ultrastructure of 16 strains of species of the prasinophycean genera Mantoniella, Mamiella, Pseudoscourfieldia, Nephroselmis, Tetraselmis, Scherffelia, Pterosperma, and Pyraminonas was examined in detail by whole-mount electron microscopy. The flagellar hairs of all genera displayed a high degree of ultrastructural complexity that was completely conserved within each strain. In all strains, flagellar hairs occurred on the sides of the flagella (lateral hairs); in several strains, special flagellar hairs also were found on the flagellar tips (tip hairs; absent in the Chlorodendrales and in Nephroselmis). Two groups of lateral hairs were distinguished: 1) T-hairs (“Tetraselmis-type” flagellar hairs), characterized by a smooth, tubular shaft of ca. 15 nm diameter and an overall length of 0.5–1.3 μm, and 2) P_t-hairs (“Pterosperma-type lateral flagellar hairs”), which were considerably longer (ca. 1.5–5.4 μm), characterized by a thick shaft of ca. 30 nm diameter, which was covered with a layer of regularly spaced small particles of ca. 10 nm diameter. In both groups of flagellar hairs, a strain-specific number of subunits (1–101) in linear arrangement was attached to the distal end of the shaft. Tip hairs were either structurally related to T-hairs (Mamiellales, Pseudoscourfieldia) or represented a separate group, P_t-hairs (“Pterosperma-type flagellar tip hairs”; Pterosperma, Pyramimonas). In four genera (Mantoniella, Mamiella, Pseudoscourfieldia, Nephroselmis), both groups of lateral hairs occurred together on the same cell. Interestingly in these taxa the P_t-hairs were exclusively attached to the shorter immature flagella (no. 2), but, in contrast, in Mantoniella and Pseudoscourfieldia the tip hairs were restricted to the longer mature flagellum (no. 1). Thus, flagella of different developmental status differ in their hair-scale complement. The occurrence, distribution, and ultrastructure of flagellar hairs can be used to identify and classify prasinophytes at all taxonomic levels.     

Über einen intrazellulären Parasiten beiCryptomonas (Cryptophyceae), I

An intracellular parasite occurring inCryptomonas rostratiformis and less numerously also inC. erosa andC. phaseolus. The parasite is described. It grows in the dorsal side of the host near the nucleus from which it is optically indistinguishable in young stages. When mature the parasite fills 1/3 to 1/2 the volume of theCryptomonas cell. It is seen as a colourless blister, which pushes back the plastid of the host. Reproduction occurs by separation of the protoplast into a hundred or moreBodo-like swarmers which perhaps represent the infectious phase. Under certain conditions, however, such as during decline of theCryptomonas population, the parasite transforms into thick-walled spindle-shaped cysts. Like the swarmers these cysts are released by rupture of the cryptomonad cells. The fate of the cysts is not known. TheCryptomonas population is destroyed by the infection in the course of a few days. Literature studies have shown that the parasite has been known for a long time, but considered incorrectly by different authors as part of theCryptomonas, or as a result of phagotrophic uptake of theCryptomonas. The parasite is compared with a somewhat similar parasite inMallomonas, and with certain parasitic dinoflagellates. The similarity with the latter is superficial only as it posesses a eucaryotic nucleus. The parasite shows some similarity with the genusParadinium (Mycetozoa) as well as with certainSporozoa.

     

Changes in pigmentation of phytoplankton species during growth and stationary phase — consequences for reliability of pigment-based methods of biomass determination

In applied water ecology several methods for estimating the biomass or activity of phytoplankton depend on the proportion of accessory pigments (xanthophylls) to chlorophyll a. Therefore, changes in pigmentation during growth and stationary phase were investigated in four different species (Amphidinium klebsii, Euglena gracilis, Prymnesium parvum, Cryptomonas ovata) typical representatives of the major algal groups. The ratios of the different xanthophylls to chlorophyll a depended not only on the growth phase, but also on the species. InAmphidinium andEuglena, the ratio of xanthophylls to chlorophyll rises continuously during the growth phase and declined during the stationary phase. InPrymnesium, quantitative pigmentation was found to be nearly independent of the growth phase. InCryptomonas, however, this ratio was relatively constant during growth, but increased in the stationary phase. In contrast to higher plants, in which the breakdown of chlorophylls occurs before that of the xanthophylls, in three of the species both pigment classes were reduced in parallel when the cultures were in the stationary phase. AgingCryptomonas, however, exhibited a pigment breakdown pattern similar to higher plants. The use of these findings for the widely applied biomass determination by chlorophyll fluorescence and for other pigment-based methods is discussed.     

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.     

Die Zweiteiligkeit der Chromatophoren bei Cryptomonaden

In theChrysophyceae as well as in different species ofCryptomonas bilobed chromatophores are present. These chromatophores consist of two large parietal lobes closed to the lateral sides of the cell and joined by a narrow bridge on its dorsal part. A survey of all species with a single bilobed chromatophore is given. Besides, also species with two separate chromatophores have been found. The presence of several chromatophores inCryptomonas cells is doubtful. The morphology of the chromatophores has to be taken into consideration in the taxonomy ofCryptomonas.