THE GENUS ACTINOCYCLUS (BACILLARIOPHYCEAE): FRUSTULE MORPHOLOGY OF A. SAGITTULUS SP. NOV. AND TWO RELATED SPECIES1

Actinocyclus octonarius var. tenellus (Bréb.) Hendey, A. actinochilus (Ehr.) Simonsen and A. sagittulus sp. nov. were examined by light and scanning electron microscopy from field samples and/or culture material. Consistent cingulum patterns of a wide valvocopula and one narrow pleura were found. In A. actinochilus, a small third band was found filling the opening of the pleura and is probably present in the other two taxa. The observed variability of the pseudonodulus, especially in A. actinochilus but also in other members of the genus, was confirmed and our results support the literature definition of Actinocyclus as having: 1) a marginal ring of large labiate processes, laterally expanded internally, 2) no external tubes, 3) essentially radial areolation arranged in fascicles, 4) external cribrum, internal foramen, and 5) a pseudonodulus usually present, but may be absent or difficult to detect on individual valves. On the basis of the present study another characteristic should be added; 6) hyaline bands with a wide valvocopula and at least one pleura. This characteristic appears to extend throughout the family Hemidiscaceae, suggesting a close relationship to those Coscinodiscus species with a single marginal row of large labiate processes and zero or one central labiate processes. In addition, Actinocyclus has been noted to have a thin valve overhang extending outside the valvocopula for as much as one-third of its width.     

CELLULAR MOVEMENT IN THE CENTRIC DIATOM ODONTELLA SINENIS1

Cultured, actively growing cells of Odentella sinensis secrete mucilage, forming gelatinous masses; the mucilage can be visualised with Alcian Blue. When examined live with the light microscope, many cells exhibited continuous small shuffing and rocking movements that could last for long periods (30-40 min); the cells, however, were not translocated and remained relatively fixed in position with respect to their neighbours. Ultrastructural examination of these cells showed prominent aggregations of mucilage vesicles, derived from the Golgi bodies, at the base of the labiate processes, each of which is close to an elevation bearing an ocellus. In Ditylum brightwellii, similar aggreations of these vesides were also located at teh labiate processes; this diatom, too, secretes mucilage but does not have ocelli. We conclude that the movements observed in O. sinenisis are an indirect result of active muilage secretion through the labiate process. It has been suggested that the raphe may have evolved from the labiate process; our conclusion, therefore, has phylogenetic implications, suggesting a functional as well as a morphological relationship between the two valve structures.     

VALVE AND BAND MORPHOLOGY OF SOME FRESHWATER DIATOMS I. FRAGILARIA CAPUCINA VAR. MESOLEPTA1,2

Acid cleaned cells from clonal cultures of Fragilaria capucina var. mesolepta Rabh. were examined with light and scanning electron microscopy. Recently isolated cells are linear-lanceolate in shape with a median constriction. After several transfers over 25 mo, cells exhibit size diminution resulting in small elliptically shaped valves. Adjacent valves are united to one another by interlocking marginal spines. Every valve has an apical pore field at each apex. A single labiate process is present infrequtently, appearing underdeveloped most often in size-reduced cells. The girdle region consists of two cingula, each composed of a series of underlapping bands. Each pleura in the series is a discontinuous ring with a central ligula. A survey of past ultrastructure studies on the freshwater Fragilariaceae reveals that the occurrence of the apical pore field and labiate process are likely key characteristics for the family. The apical pore field of Diatoma, Asterionella and Tabellaria is positioned on the valve face, whereas the apical pore field of F. capucina var. mesolepta is located on the valve mantle, the girdle region of F. capucina var. mesolepta is basically similar to that of Gomphonema parvulum (Kütz.) Grun.     

Cell division and morphogenesis of the labiate process in the centric diatomDitylum brightwellii

Summary Cells of the centric diatomDitylum brightwellii were filmed undergoing cell division and valve secretion, and were fixed for transmission electron microscopy. Attention was directed particularly at the origin of the Labiate Process Apparatus (LPA).As reported previously (li andVolcani 1985 a), the nucleus, centrally situated during interphase, moves laterally to undergo mitosis against the girdle bands. We describe the spindle which splits up into numerous fibres of overlapped polar microtubules (MTs) by metaphase. The chromosomes are diffuse and the spindle elongates rapidly during anaphase. A complex of organelles is found at the poles and ill-defined, dense material extends to the nearby plasmalemma from prophase on. The two Silica Deposition Vesicles (SDVs) are initiated during anaphase close to the poles and by midcleavage, the dense LPA arises on each SDV close to dense polar material. After cleavage, the daughter protoplasts round up and the SDV, already containing a nascent valve, expands over the cleavage furrow. The labiate process, a long straight hollow tube of silica, is rapidly (ca. 25 minutes) secreted from directly under the LPA; a fibrous plug (polysaccharide?) always appears in the SDV immediately adjacent to the LPA during the initiation of this secretion. The ill-defined Microtubule-Organizing Center (MC) from the spindle pole remains close to the LPA and in it can be seen the tiny presumptive primordial spindle on the nuclear envelope.The raphe and the labiate process (LP), both highly differentiated apertures in the valve, probably function in a specialized form of the mucilage secretion involved in generation of movement in raphid diatoms, and in a simple form of movement in some centrics. Morphogenesis of the LP is associated with the LPA while differentiation of the raphe is almost associated with the MC; both MC and LPA have an intimate ontological relationship with the spindle pole and the postmitotic cytoskeletal system of MTs. This association also is seen in the formation of the LP in an araphid pennate,Diatoma (work in progress). Therefore, from functional, morphogenetic and ontogenetic observations, we support the proposal that the raphe of pennate diatoms arose from the LP of centric diatoms.     

NANOSTRUCTURE OF THE DIATOM FRUSTULE AS REVEALED BY ATOMIC FORCE AND SCANNING ELECTRON MICROSCOPY

The cell wall (frustule) of the freshwater diatom Pinnularia viridis (Nitzsch) Ehrenberg is composed of an assembly of highly silicified components and associated organic layers. We used atomic force microscopy (AFM) to investigate the nanostructure and relationship between the outermost surface organics and the siliceous frustule components of live diatoms under natural hydrated conditions. Contact mode AFM imaging revealed that the walls were coated in a thick mucilaginous material that was interrupted only in the vicinity of the raphe fissure. Analysis of this mucilage by force mode AFM demonstrated it to be a nonadhesive, soft, and compressible material. Application of greater force to the sample during repeated scanning enabled the mucilage to be swept from the hard underlying siliceous components and piled into columns on either side of the scan area by the scanning action of the tip. The mucilage columns remained intact for several hours without dissolving or settling back onto the cleaned valve surface, thereby revealing a cohesiveness that suggested a degree of cross-linking. The hard silicified surfaces of the diatom frustule appeared to be relatively smooth when living cells were imaged by AFM or when field-emission SEM was used to image chemically cleaned walls. AFM analysis of P. viridis frustules cleaved in cross-section revealed the nanostructure of the valve silica to be composed of a conglomerate of packed silica spheres that were 44.8 ± 0.7 nm in diameter. The silica spheres that comprised the girdle band biosilica were 40.3 ± 0.8 nm in diameter. Analysis of another heavily silicified diatom, Hantzschia amphioxys (Ehrenberg) Grunow, showed that the valve biosilica was composed of packed silica spheres that were 37.1 ± 1.4 nm and that silica particles from the girdle bands were 38.1 ± 0.5 nm. These results showed little variation in the size range of the silica particles within a particular frustule component (valve or girdle band), but there may be differences in particle size between these components within a diatom frustule and significant differences are found between species.     

ACTIVE GLIDING MOTILITY IN AN ARAPHID MARINE DIATOM,ARDISSONEA (FORMERLY SYNEDRA) CRYSTALLINA1

Active gliding movement over long distances was observed and filmed in the marine pennate diatom Ardissonea (Synedra) crystallina (Agardh) Kütz. Typical speeds measured ca. 1–2 μm-s^?1. Motion wax often smooth and steady; however, discontinuous jerky motions and rolling movements were common. Motion, was associated with secretion of twin or, less commonly, single straight trails of mucilage from one end of the cell. In a few instances, reversal in direction was related to cessation of mucilage secretion at one end and commencement at the other. Temporary cessation of movement due to an obstruction was accompanied by a build-up of mucilage at one end of the cell. Mucilage was apparently secreted at two specific sites at each end of the cell and was stained by alcian blue. Persistent trails were visible under scanning electron microscopy (SEM). SEM confirmed that cells had no raphes or labiate processes. The apparent site of secretion was a deep groove formed at the junction of the valve and valvocopula (first girdle band) at each end of the cell. Transmission electron microscopy confirmed the presence of mucilage vesicles in the cytoplasm, but these were not in any manner obviously related to secretion nor was any morphological structure associated with secretion. Cells often become epiphytic through secretion of a terminal stipe. Both stipe secretion and movement may involve the same structural differentiation of the frustule. These results demonstrate a previously unrecorded type of diatom motility. The mechanism, involves mucilage secretion and appears similar to that seen, for example, in some other algae such as the desmids (green algae).     

VALVE AND BAND MORPHOLOGY OF SOME FRESHWATER DIATOMS. III. PRE- AND POST-AUXOSPORE FRUSTULES AND THE INITIAL CELL OF MELOSIRA ROESEANA1

Frustules of a clonal culture of Melosira roeseana Rabenh. were examined with light and scanning electron microscopy. Vegetative valves in the post-auxospore (full size) stage exhibit a larger width/length ratio than those in the pre-auxospore (size-reduced) stage. Cells form chains by linking spines of adjacent valves which occur at the periphery of the valve face-mantle junction. Three or jour large pores occur at the center of the valve face, with the diameter of each pore tapering from the inner to the outer valve surface; these pores are often occluded by siliceous processes. Features of M. roeseana, not shown previously for Melosira, include a “stepped” mantle, on only one of the two valves resulting from the same cell division, flattened processes attached to short siliceous stalks on the valve face, disk-like processes on the mantle, and an open girdle band with up to eight antiligulae. Siliceous scales on the surface of the initial cell are remnants of the auxospore wall. The epivalve of the initial cell is larger in diameter than the hypovalve, and both valves lack linking spines and a step on the valve surface. The initial, cell epicingulum consists of only two bands; the hypocingulum has up to seven. Initial cells with four or more hypocingular bands divide to form new post-auxospore filaments. Melosira roeseana should not be included in the genus Melosira as it is presently defined by the type species, M. nurnmuloides C. Ag. Major differences include irregular linking spines, a closed pseudoloculate valve construction, and labiate processes on the valve face and mantle of M. nummuloides, compared with well-defined linking spines, a valve constructed of a basal siliceous layer perforated by poroid areolae, and labiate processes lacking on the valve of M. roeseana.     

PHYLOGENETIC POSITION OF TOXARIUM,A PENNATE‐LIKE LINEAGE WITHIN CENTRIC DIATOMS (BACILLARIOPHYCEAE)1

The diatom genus Toxarium Bailey has been treated as a pennate because of its elongate shape and benthic lifestyle (it grows attached to solid substrata in the marine sublittoral). Yet its valve face lacks all structures that would ally it with the pennates, such as apical labiate processes, a midrib (sternum) subtending secondary ribs and rows of pores extending perpendicularly out from the midrib, or a raphe system. Instead, pores are scattered irregularly over the valve face and only form two distinct rows along the perimeter of the valve face. In our nuclear small subunit rDNA phylogenies, Toxarium groups with bi‐ and multipolar centrics, as sister to Lampriscus A. Schmidt. Thus, the genus acquired a pennate‐like shape and lifestyle independently from that of the true pennates. The two species known, T. hennedyanum Grunow and T. undulatum Bailey, differ only in a single feature: the valve perimeter of the former shows only a central expansion, whereas that of the latter possesses in addition a regular undulation. Yet both forms were observed in our monoclonal cultures, indicating that the two taxa represent extremes in a plasticity range. Toxarium resembles another elongate and supposedly araphid diatom, Ardissonea De Notaris, in being motile. Cells can move at speeds of up to 4 μm·s ^{? 1} 1 Received 7 June 2002. Accepted 4 October 2002. through secretion of mucilage from the cell poles or they remain stationary for longer periods, when they form short polysaccharide stalks. Division during longer periods of quiescence leads to the formation of small colonies of linked or radiating cells.     

Studies on the biochemistry and fine structure of silica shell formation in diatoms

Summary The sequential wall formation in the centric diatom,Ditylum brightwellii (West) Grunow, is described. The silica deposition vesicle is formed by the coalescence of small vesicles. Silicification of the new valve starts from the central labiate process area prior to the completion of cytokinesis, and the developing valve grows in a centrifugal direction. The initiation of the structures on the valve follows the sequence: labiate process, marginal ridge, and rota. A novel labiate process apparatus, which is situated in the cytoplasm close to the developing labiate process, appears prior to the initiation of the labiate process and disappears upon its maturation. Segments of the girdle bands are formed in individual silica deposition vesicles after the valve matures and is exocytosed. Three morphological forms of deposited silica have been determined: thin base layers, microfibrils, and hexagonal columns. The involvement of cytoplasmic structures in the patterning of the siliceous wall is discussed.     

CHARACTERIZATION OF MUCILAGE ON THE PROXIMAL CELLS OF YOUNG GAMETOPHYTES OF BOTRYCHIUM DISSECTUM (OPHIOGLOSSACEAE)

Young gametophytes of Botrychium dissectum produce a mucopolysaccharide coating on the external surface of the proximal cell. Scanning electron microscopy reveals deposition of the mucilage, which initially has a patchy distribution, in the shape of a thick triangular ring. Young gametophytes in the hydrated condition have a thin coating of mucilage over the other areas of the proximal cell wall. Histochemical staining indicates that the mucilage contains sugars with vicinal hydroxyl groups, carboxylated sugars, and small amounts of sulfated sugars. Protein and phenolic materials are also found in the mucilage. Lipids, β, 1-3 glucans and β,1-4 glucans are not present. Fluorescein labelled lectin binding shows the presence of terminal galactose and terminal fucose units. Germinating spores in the presence of the microtubule inhibitors CIPC (3-chloro-N-phenyl isporopyl carbamate) and griesiofulvin cause abnormal mucilage secretion. The inhibitor CIPC prevents mucilage secretion whereas griesiofulvin disrupts the pattern of deposition. Several functions of the mucilage are postulated.     

STUDIES ON MARINE PLANKTON DIATOMS. III. STRUCTURE AND CLASSIFICATION OF GOSSLERIELLA TROPICA1

The structure of the tropical oceanic diatom Gossleriella tropica Schütt was examined in the light and electron microscopes. The species is characterized by a primarily poroid valve, surface, it single central labiate process and a unique ring of spines attached to the cingulum. Placement of the genus in the family Rhizosoleniaceae is suggested. Although G. tropica is widely distributed in the world's oceans, it is usually confined to the lower photic zone, salinities of 34.5–35.5% and temperatures of 15–30 C.     

ASSOCIATION OF MUCILAGE WITH THE LIGULE OF SEVERAL SPECIES OF SELAGINELLA

Mucilage bodies of protein and some polysaccharide were closely associated with ligules of Selaginella kraussiana (Kunze) A. Braun grown in the greenhouse, and Selaginella wallacei Hieron. and Selaginella oregana D.C. Eat. from natural habitats. With S. wallacei and S. oregana, several mucilage bodies were associated with some ligules. No mucilaginous material was associated with the ligules of Selaginella douglasii Hook, et Grev., Selaginella densa Rydb. or Selaginella apoda (L.) Fernald from natural habitats. When stem tips of S. kraussiana and S. douglasii were placed in sterile culture on a medium with sucrose, massive amounts of mucilage appeared around the ligules in the tight interstices of the stem tip leaves and extended out on the surface of the leaves. The mucilage from a cultured stem tip of S. kraussiana contained 0.034 μgm of protein, 14.6 μgm of apparent glucose equivalents as polysaccharide and no significant amounts of phenolics. In stem tips with tightly appressed leaves, either ligules or young leaves could have produced the mucilage; however, in more mature regions of the stem where internodal elongation had separated the leaves, mucilage coated only the apical portions of the ligules and was not associated with leaves. No other possible source of mucilage (bacteria or fungi) was detected among the leaves. When stem tips of S. wallacei, S. densa and S. apoda were placed in culture, mucilage was not produced. Since mucilage is closely associated with the ligule of some species of Selaginella growing in nature, the greenhouse, or tissue culture, the ligule may be glandular.     

A LIGHT- AND ELECTRON-MICROSCOPIC STUDY OF PRIMARY HETEROGENEITY IN THE EGGS OF TWO BROWN ALGAE

The mature eggs of Zonaria farlowii and Dictyota binghamiae exhibit a distinctively symmetrical arrangement of organelles. In mature oogonia plastids occur in two groups in the central region of the cell and vacuoles of several types are arranged in peripheral layers. Before the mature egg is released, an amorphous “mucilage layer” developes inside the wall of the oogonium. After fertilization the plastids become uniformly distributed. When the eggs of Zonaria develop parthenogenetically the primary heterogeneity is retained. The symmetrical arrangement of membranes seen in the oocytes of these plants may represent an organizing system that influences cytodifferentiation and perhaps subsequent embryology.     

STUDIES IN PENNATE DIATOMS: VALVE MORPHOLOGIES OF LICMOPHORA AND CAMPYLOSTYLUS1

Using scanning electron and light, microscopy, several hundred specimens of Campylostylus normanianus (Grev.) Gerloff, Licmophora abbreviata Agardh, L. gracilis var. anglica (Kütz) Per. et Per., and L. flabellata (Carm.) Agardh were examined to elucidate their valve morphology. These species were found to be heterovalvar with respect to the presence of the labiate process in the basal apices of the cell, although one was always present at the head pole of each valve. This form of heterovalvar may be one basis for partitioning the large and variable family Diatomaceae. Because of the similar valve morphologies exhibited by C. normanianus and Licmophora species examined, it is recommended that Campylostylus normanianus be placed in the older genus Licmophora, as L. normaniana (Grev.) Wahrer.     

Studies on the biochemistry and fine structure of silica shell formation in diatoms

Summary The morphogenesis of the different types of labiate processes is compared among three species of centric diatoms [Stephanopyxis turns (Greville) Ralfs,Odontella sinensis (Greville) Grunow, andOdontella aurita (Lyngbye) Agardh]. In all species, a cytoplasmic structure,i.e., the labiate process apparatus, situated close to the developing labiate process, appears before the labiate process begins to form and disappears upon its maturation. The possibility that the labiate process apparatus is implicated in the phylogeny of the labiate process is 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 MARINE,PLANKTONIC DIATOMS THALASSIOSIRA OCEANICA SP. NOV. AND T. PARTHENEIA1

The diatom clone 13–1 isolated from the Sargasso Sea by Dr. R. R. L. Guillard as Cyclotella nana Hust. or Thalassiosira pseudonana Hasle and Heimdal is described as a separate species, Thalassiosira oceanica sp. nov. An amplified diagnosis of Thalassiosira partheneia Schrader is given, and a comparison of the two species is made based on light and electron microscopy. Similarities are present in the apparent distribution pattern, cell size, and girdle structure. Differences are present in the shape of the areolae of the valvocopula and the copula, in the structure of the vela of these bands, in the texture of the external valve surface, in the morphology of the strutted processes, in the location of the labiate process, in the distance between the marginal strutted processes, and by the presence of a marginal ridge in T. oceanica. Fifteen nanoplanktonic (maximum diameter = 20 UmUm) Thalassiosira species are listed, among them T. oceanica and T. partheneia.     

Resting spore formation in the antarctic diatoms Coscinodiscus furcatus Karsten and Thalassiosira australis Peragallo

Summary The resting spore morphology of the antarctic diatoms Coscinodiscus furcatus Karsten and Thalassiosira australis Peragallo are described. Both species form endogenous resting spores. The spore valve of C. furcatus differ from those of the vegetative cells primarily by (i) a greater convexity and (ii) a coarser and more distinctly fasciculated areolation. This resting spore is identical to the diatom traditionally identified as C. stellaris var. symbolophorus (Grunow) Jørgensen in the Antarctic. The resting spore of T. australis differs from the vegetative cells by (i) a lack of clusters of strutted processes in a modified ring on valve face, (ii) a coarser areolation and tangential rows of areolae and (iii) a narrower and more simply structured girdle. The resting spore valve of T. australis has been described as belonging to a separate species, Actinocyclus excentricus Peragallo.     

DEVELOPMENT OF MUCILAGINOUS SURFACES IN EUGLENOIDS. II. FLAGELLATED,CREEPING AND PALMELLOID CELLS OF EUGLENA1

Critical-point dried (CPD) cells from clonal cultures of Euglena gracilis Klebs (Z strain), E. deses Ehrb., E. tripteris (Duj.) Klebs and E. myxocylindracea Bold & MacEntee were examined by scanning electron microscopy. Flagellated motile cells of E. gracilis are naked except for a few strands of mucilage on the posterior tip. Flagellated cells of E. tripteris have a permanent mucilage coating often of uneven distribution and usually not as well developed as that of nonflagellated creeping cells which have a distinctive mucilage. In E. deses the coating appears rough due to the aggregation of isolated groups of strands above the cell surface. In E. tripteris the coating appears smooth except for breaks near the articulation of the pellicular strips where the mucilage may rise above the surface to form waves. At high magnification this mucilage consists of a network of strands generally lying parallel to the cell surface; the strands become obscure in some specimens. In E. myxocylindracea elongated, mucilage-coated cells contract to form spheres which undergo further mucilage deposition producing the mucilage covering of palmellae. As palmellae mature, the mucilage surface becomes less porous and the individuality of most mucilage strands is lost.     

Actinocyclus exiguus sp. nov. from the southern parts of the Indian and Atlantic Oceans

A new species, Actinocyclus exiguus, a small centric diatom, is described from the southern Indian and Atlantic Oceans. The pseudonodulus, a diagnostic feature of the genus and family, has not been seen in the light microscope and is more easily seen on the inside of eroded valves with the aid of the scanning electron microscope. A. exiguus is compared to closely related species.