VALVE MORPHOGENESIS IN THE CENTRIC DIATOM PROBOSCIA ALATA SUNDSTROM1

Valve morphogenesis in Proboscia alata Sundstrom was followed in living cells and during treatment with antiactin and antimicrotubule drugs. Once cleaved, sibling cells rounded up and retracted. Soon, a granular organizing center (OC) appeared adjacent to the stub of the initiated valve. Silicification started within a silicon deposition vesicle (SDV) adjacent to the OC. The elongating valve was initially tubular and sealed at one end, creating the proboscis of the conical valve. The edge of the SDV and thinnest region of the forming valve was lined by a sleeve of bundled microtubules (MTs) that terminated short of the older more rigid part of the valve. The growing proboscis of living cells treated with the anti‐MT drug oryzalin became grossly distorted. EM revealed dense material lining the growing edge of the SDV; immunofluorescence microscopy showed a ring of actin here. Applied to living cells, the antiactin drug cytochalasin D caused the very young proboscis to collapse; in older valves, the base of the proboscis expanded. Thus, valve morphogenesis appeared controlled by the MT cytoskeleton, keeping the proboscis straight while actin molded its conical outline. At the tip of the proboscis was a slit resembling a labiate process. Its morphogenesis involved striated fibers and two MTs, reminiscent of the fibers and MTs associated with raphe morphogenesis. In contrast to spine‐like processes that elongate by tip growth, the tip of the proboscis was formed first, and the consequent “antitip growth” suggests the tip was originally the center of the valve face.     

VALVE MORPHOGENESIS AND THE MICROTUBULE CENTER IN THREE SPECIES OF THE DIATOM NITZSCHIA1

The relationship of cell organelles to valve morphogenesis was investigated in three species of Nitzschia. One, N. sigmoidea (Nitzsch) W. Sm., showed consistent ability to generate both nitzschioid and hantzschioid symmetry in daughter cells following cytokinesis; the other two maintained nitzschioid symmetry stably. From previous work with Hantzschia, a certain sequence of events could be anticipated in the cytoplasm. In two significant areas–the behavior of the Microtubule Center (MC) and its microtubule (MT) system, and the central origin of the silicalemma–not only were the results unexpected, but the three species showed fundamental differences among themselves. In N. sigmoidea, the silicalemma (and the future raphe region) arises centrally on the cleavage furrow, and after some lateral expansion, the silicalemmas and their associated organelles move in opposite directions in daughter cells, so that the raphe and the raphe canals end up along the girdle side of the cell as expected. However, the MCs never become associated with their silicalemma, remaining throughout near the girdle bands. In N. sigma (Kütz) W. Sm., the silicalemmas arise centrally and after lateral growth, move in opposite directions to generate nitzschioid symmetry. In this case, the MCs move to the vicinity of but never close to the silicalemmas, and follow them distantly during their lateral movement. In N. tryblionella Hantzsch, the new silicalemmas arise opposite one another, on one side of the daughter cells; each MC soon moves very close to its silicalemma, and remains thus through most of valve morphogenesis. Later, only one silicalemma/MC complex moves laterally, establishing the nitzschioid symmetry in both daughter cells. In all three species, as in Hantzschia, linear arrays of mitochondria aligned along MTs occupy the forming raphe canal, and microfilaments line the outer edge of the expanding silicalemma. The fibulae (the wall struts arching across the raphe canal) in Hantzschia always grow from the valve surface to the girdle surface of the forming valves. In these three Nitzschiae, this invariably happens in only one daughter cell of any pair; in the other, all the fibulae grow from the girdle surface to the valve surface. An explanation of these variations is proposed: that the morphogenetic machinery of Nitzschia and Hantzschia have a common origin, with present Nitzschiae having undergone considerable diversification at the intracellular level, causing the unstable cell symmetry exhibited by several modern species. Perhaps a taxonomic distinction between Hantzschia and Nitzschia lies in whether the morphogenetic machinery associated with valve morphogenesis moves laterally in the same or in opposite directions.     

VARIATION IN PATTERNS OF VALVE MORPHOGENESIS BETWEEN REPRESENTATIVES OF SIX BIRAPHID DIATOM GENERA (BACILLARIOPHYCEAE)

Scanning electron microscopic studies of silica valve formation in naviculoid diatoms representing six different genera revealed that the precise sequence of depositional events varied among genera. Valve deposition begins with the formation of the raphe sternum, from which virgae (lateral outgrowths) extend. Areolae (pores) are formed between the virgae by the fusion of cross-extensions (vimines). In most of the species studied ( Craticula ambigua (Kützing) D. G. Mann, Frustulia vulgaris (Thwaites) De Toni, Craspedostauros australis E. J. Cox, and Gomphonema truncatum Ehrenberg), areola (pore) formation began near the raphe sternum before completion of the valve margin, but in Pinnularia gibba Ehrenberg the valve margin fused before the areolae were formed. Silica deposition in all these taxa was mainly distal to proximal (with respect to the cytoplasm), but in Haslea sp. it was mainly proximal to distal. Haslea also differed in that areolae were defined as the valve margin was completed. These data have also contributed to the interpretation of taxonomically important features, such as raphe endings. In P. gibba the internal central raphe fissures were laterally deflected but subsequently obscured by additional silicification of the valve, whereas in G. truncatum they were initially straight, becoming laterally deflected as valves mature. External raphe fissures in Frustulia became Y-shaped only just before maturity; in immature valves they were dotlike, as in Amphipleura Kützing. The comparison of developmental pathways in diatoms is a useful adjunct to morphological and other approaches in diatom systematics and warrants renewed attention.     

FUNCTIONAL VALVE MORPHOLOGY IN SOME SURIRELLA SPECIES (BACILLARIOPHYCEAE) AND A COMPARISON WITH THE NAVICULACEAE1

On the basis of results of stratigraphic and comparative morphological studies on the diatom frustule, the Surirellaceae is generally assumed to be the endpoint of the evolution of the Pennales. The present study shows that a line of development, based on frustule construction and which parallels the search for optimum design of comparable elements in engineering, can be traced from the Naviculaceae to the Surirellaceae. In both cases lightweight construction is achieved through economy of material and energy expenditure. This leads to structural stability and in the case of the diatom valve, a larger area for metabolic exchange. From the functional-morphological point of view, three construction principles can be distinguished in the genus Surirella: 1. valves with pennate costal framework, raphe keels and fibulae (Surirella gemma group); 2. frustules where all supporting elements are in the form of corrugations, with raphe keel and fibulae (Pinnatae, Fastuosae, Surirella striatula group); and 3. as in 2, but with true alae with alar canals (Robustae) instead of keels with fibulae.     

VALVE MORPHOGENESIS IN THE PENNATE DIATOM ACHNANTHES COARCTATA

Mitosis and valve morphogenesis in the pennate diatom Achnanthes coarctata (Bréb. in W. Sm.) Grun. are described. After cytokinesis, both daughter nuclei and their microtubule centers (MCs) are found near one side of the cell. Each new tubular silica deposition vesicle (SDV) arises centrally, forming a single rib running the length of the cell. Each MC then migrates around its nucleus and positions itself directly adjacent to the new SDV. The enlarging silicalemmas with their associated MCs, nuclei, microtubules (MTs) and microfilaments (MFs) appear in mirror image in the daughter cells. Both SDVs soon generate a second longitudinal rib alongside the first; the gap between the ribs ultimately becomes the future raphe fissure. The MC, MTs and nucleus are associated with each fissure. However, the subsequent behavior of the valve secreting machinery now becomes quite different in the daughter cells. In the cell that will form a raphid valve, the silicalemma, flanked by MFs, expands laterally in both directions over the cleavage furrow. Within the expanding SDV, silica secretion continues, eventually generating the structure of the mature valve, and during this phase the raphe fissure becomes delineated as in other raphid diatoms. In the other daughter cell, however, the MC and its MTs withdraw from the silicalemma, and the SDV moves laterally across the cleavage furrow until the double rib is at the corner of the cell. As silica is secreted into this expanding SDV, the raphe fissure completely fills in. This valve, therefore, lacks a raphe when mature and has a symmetry quite different from that of the valve formed in the other daughter cell. These events are compared with the course of morphogenesis described for other raphid diatoms.     

OVERLOOKED HETEROPOLARITY IN SURIRELLA CF. FASTUOSA (BACILLARIOPHYTA) AND RELATIONSHIPS BETWEEN VALVE MORPHOGENESIS AND AUXOSPORE DEVELOPMENT

Surirella cf. fastuosa is an apparently isopolar elliptic marine raphid diatom. We observed cells before and after sexual reproduction in monoclonal cultures using light and scanning electron microscopy (LM and SEM). After sexual reproduction cells were approximately twice as large as before, in valve length and width. The stria and infundibula densities were stable during the life cycle. Subtle morphological differences were detectable between the two poles of the frustule. One pole (pole A) was characterized by endings of the external raphe fissure that turned toward the valve face, continuity of the domed wall of the raphe canal externally, an elliptic chamber visible internally, a shallow nick in the interior of the valvocopula. The other pole (pole B) was with the following: straight endings of the external raphe fissures, a dent in the domed wall of the raphe canal externally, a double chamber internally, presence of the open ends of the valvocopula nearby, a deep nick in the valvocopula. Furthermore, at pole A virgae developed at an early stage in morphogenesis, whereas at pole B they were not formed. In the auxospores, pole A was situated beneath the primary transverse perizonial band. Pole A is suggested to be homologous with the head pole in heteropolar Surirella and is the “protopole” likely equivalent to the central nodule in naviculoid taxa. Pole B is homologous with the foot pole in heteropolar Surirella and is the “synaptic pole” formed by fusion of components equivalent to both poles of naviculoid taxa.     

VALVE MORPHOGENESIS IN THE PENNATE DIATOM NAVICULA CUSPIDATA1

The deposition of siliceous valves during asexual reproduction of the pennate diatom, Navicula cuspidata Kütz., is described with emphasis on the cytoplasmic components involved. The events accompanying valve secretion are similar to those already known from other pennate species. After mitosis, the microtubule centre (MC) moves to the center of the cleavage furrow where silica deposition is initiated inside a tubular silicalemma, and it remains associated with the prospective central nodule during valve growth. Microtubules (MTs), emanating from the MC, run parallel to the prospective raphe and together with the raphe fibres, appear to be involved with raphe development. Multiple raphe fibres occupy the maturing raphe fissure, in contrast to the single fibre of Pinnularia viridis, P. maior and Hantzschia amphioxys. The fibers exhibit a periodic substructure and are often opposed to the silicalemma where they may inhibit silica deposition and control the shaping of the raphe fissure. In contrast with the above species, in N. cuspidata MTs are clustered strictly opposite the raphe and lose their association with the MC which degenerates before the valves are mature. The primary role of MTs may be the stabilization of the cytoplasmic region where initial silicification occurs. Mitochondria and endoplasmic reticulum are not involved in molding valve growth in this species. Evidence for vesicle involvement in silica transport and deposition was limited. The possible contributions provided by comparative studies on the ultrastructure of valve morphogenesis towards elucidating the control of valve formation and the taxonomy of diatoms are discussed briefly.     

MONITORING RAPID VALVE FORMATION IN THE PENNATE DIATOM NAVICULA SALINARUM (BACILLARIOPHYCEAE)1

After each division of a diatom cell, a new siliceous hypovalve is formed inside the silica deposition vesicle (SDV). We present the sequence of this early formation of the new valve in the pennate marine diatom Navicula salinarum (Grunow) Hustedt, visualized by using the fluorescent probe 2‐(4‐pyridyl)‐5‐((4‐(2‐dimethylaminoethylamino‐carbamoyl)methoxy)phenyl)oxazole (PDMPO). Our observations confirm that two‐dimensional expansion of the growing valve is a rapid process of no more than 15 min; three‐dimensional completion of the valve appears to be slower, lasting most of the time valve formation takes. The results are relevant to studies of the timing of molecular processes involved in valve formation (i.e. the bio‐ and morphogenesis of the SDV) in relation to uptake and transport of silicic acid. Use of this probe helps us to identify specific developmental stages for further detail analysis of diatom basilica formation, which eventually could lead to obtaining enriched SDV fractions.     

PERIZONIUM AND INITIAL VALVE FORMATION IN THE DIATOM NAVICULA CUSPIDATA (BACILLARIOPHYCEAE)1

This paper describes the perizonium and initial valve formation in Navicula cuspidata Kütz., based on light microscope (LM) and scanning electron microscope (SEM) observations. The perizonium consists of concentric over-lapping bands, laid down sequentially at the tips of the expanding biconical auxospore during its elongation. The central perizonial band has fimbriate edges and is considerably more rigid than the more distal bands. During auxospore elongation and the band secretion, the chloroplasts continuously oscillate between the two ends of the cell; this oscillation ceases once the elongation is complete. The initial valves, formed within the perizonium, are molded into the basically biconical shape of the perizonium except for a central flattening of each valve face. In contrast to the raphes in gametangial and vegetative valves which are surrounded by a smooth axial area, the raphes in initial valves lie within a raised ridge running along the apical axis of the valve. The regular pattern of apically oriented ridges on the outer surface of vegetative valves is also lacking on initial valves. Comparison of pore–pore spacing within striae of gametangial valves, initial values and post-initial valves (first division and vegetative cells) reveals that the pore–pore distance within striae is conserved at all sexual stages. However, the distance between striae is considerably larger in initial valves than in gametangial and post-initial valves. Vegetative interstriae spacing as well as the planar morphology of the valve face is regained at the first division of the initial cell. This suggests that the spacing between striae is dependent on the sexual stage of the cell during valve formation (i.e. not directly dependent on the cell size) and can be altered independently of the pore–pore spacing.     

VALVE MORPHOLOGY OF SYNEDRA GOULARDI (BACILLARIOPHYCEAE)1

Specimens of Synedra goulardi Bréb. from samples taken in Costa Rica were acid cleaned and examined by light and scanning electron microscopy. This linear-lanceolate diatom displays an unusual pattern of markings in its central area which have been mistaken as striae but are actually internal costae. As is typical for members of its genus and that of many other genera in the Fragilariaceae, S. goulardi possesses non-stalked labiate processes, apical pore fields and a lack of marginal spines. There is a great deal of variability in the striae of the genus, ranging from single rows of puncta to areolate striae. Synedra goulardi has double punctate striae. The presence of fossil forms (Neogene) with single punctate striae suggest this may be a primitive condition in the genus.     

CELL DIVISION AND MORPHOGENESIS OF THE CENTRIC DIATOM CHAETOCEROS DECIPIENS (BACILLARIOPHYCEAE) I. LIVING CELLS

Like other diatoms, living cells of Chaetoceros decipiens Cleve expand lengthwise before they divide. During prophase, the nucleolus disappears in about 30 s. The spindle is very small but anaphase chromosome separation can be seen. Following rapid cleavage, the protoplasts contract, plasmolyzing slightly and transforming the cleavage furrow into a lens-shaped opening between daughter cells. During valve initiation, the surface of the furrow is molded slightly into the shape of the mature valve face. Then daughter cells expand further, becoming fully turgid as they open the slots in the girdle bands through which the setae will grow. Soon, delicate protrusions push through the girdle bands and develop into the setae, which are very sensitive: any disturbance will immediately stop their steady growth. Healthy setae display soft, mobile tips and tiny organelles (mitochondria) actively move along the lumen. Their curvature and uniform diameter is controlled during growth with exquisite precision, and in optimal conditions, they can become very long. At their initiation, cells appear fully turgid; however, many cells soon become slightly plasmolyzed during seta growth. This observation strongly suggests that turgor pressure cannot be responsible for driving extension; the possible mechanism is discussed in the following paper.     

MORPHOGENESIS OF THE LABIATE PROCESS IN THE ARAPHID PENNATE DIATOM DIATOM A VULGARE1

Each valve of the araphid pennate diatom Diatoma has a labiate process (LP) at one end; in a frustule, the LPs are at diagonally opposite ends. After mitosis is over, an elongated dense body detaches from the spindle pole and migrates to one end of the daughter cell, always diagonally opposite the LP of the parental valve. This dense body trails a cone-shaped array of microtubules (MTs). Meanwhile, the new valve has begun to form within the Silica Deposition Vesicle (SDV). Having reached the end of the cell, this dense body moves back slightly and then settles onto the SDV, developing a layered substructure as it does so. Immediately beneath it, the LP of the new daughter valve differentiates. This dense object is clearly the homologue of the fibrous Labiate Process Apparatus (LPA) involved in the differentiation of the LP in several centric diatoms. In a few cases, these LPAs also hair been shown to originate from some component of the spindle pole. Thus, the homologue of the LPA of centric diatoms has now been found in an araphid pennate diatom; in each case, the LPA apparently comes from the pole of the spindle and presumably uses a cytoskeleton of MTs to locate the LP in its correct position. These observations support the possibility that the raphe evolved from the LP.     

ULTRASTRUCTURE OF CYST FORMATION IN OCHROMONAS TUBERCULATA (CHRYSOPHYCEAE)1

The cysts (statospores) of Ochromonas tuberculata Hibberd are produced within a cytoplasmic silica deposition vesicle (SDV) whose membrane (silicalemma) appears to be formed by the coalescence of golgi vesicles. Silica is first deposited as small nodules and the collar and spines develop by centrifugal growth only after a complete but still thin wall has been laid down. Small vesicles appear to be attached to the SDV only in the region overlying the developing collar; a cap of radially arranged, moderately electron-dense material occurs at the tip of the growing spines. The cyst pore is formed at the anterior end of the flagellate cell, by lack of silica deposition over a small region of the SDV and rupture of the SDV and other membranes crossing this region. When the cyst wall is complete, an extracystic plug is formed in the pore, resulting in the loss of some extracystic cytoplasm and the plasmalemma, and the inner SDV membrane becomes the functional plasmalemma. The plug develops first by coalescence with the cell membrane of golgi-derived vesicles containing dense but apparently nonsiliceous spicules surrounded by amorphous material. During later stages of plug formation only fibrous material is deposited, some of which may be extruded through the pore forcing out some of the spiculate component. Scanning electron micrographs of the mature wall show it is smooth except for the concentrically wrinkled inner face of the flared collar and that the real pore diameter is only ca. half that of the collar. At germination the plug completely disappears in an unknown way and a single cell, similar to a normal vegetative cell emerges through the pore. Chrysophycean cyst formation generally resembles cell wall formation in diatoms, but differs in some details.     

CELL DIVISION AND MORPHOGENESIS OF THE CENTRIC DIATOM CHAETOCEROS DECIPIENS (BACILLARIOPHYCEAE) II. ELECTRON MICROSCOPY AND A NEW PARADIGM FOR TIP GROWTH

Valve morphogenesis starts when the silica deposition vesicle (SDV) expands across a cleavage furrow covered by an unidentified layer, which may aid in its shaping. A labiate process (LP) is present only in the outer valve of terminal cells in the filament. Before these particular cells form setae, a layered "labiate process apparatus" (LPA) appears on the SDV in the exact center of the forming valve, near the microtubule center arising after cleavage. The LPA thereafter surmounts the lips of the LP as it forms. After the girdle bands separate slightly, two lateral protrusions develop in the corners of the cell. These nascent setae are lined internally by a cylindrical, fibrous band (sleeve), which assembles immediately ahead of the expanding edge of the SDV, very close to the plasmalemma. Then these protrusions, lined by the fibrous band, the SDV, and the forming silica wall, grow through two gaps in the girdle bands. The cytoplasm at the tip of the growing seta is naked. Immediately behind the tip, this fibrous band is adpressed to the plasmalemma and thereby apparently defines the diameter of the seta; it extends to internally ensheath the tipmost edge of the SDV for a short distance, like a tight-fitting inner sleeve. This structure is considered the major organelle involved in seta morphogenesis. Microtubules (MTs), while present, are variable in extent and disposition within the seta. Turgor pressure is considered irrelevant in driving seta growth. Instead, a new paradigm proposed for tip-growing cells generally, may apply to seta morphogenesis, as follows. If, as is suspected, the fibrous band contains actin, cycling of this actin (as in animal cells undergoing ruffling or filopodial extension) could drive seta extension via attachment of the band to the just-formed silica wall. The band is visualized as a molecular treadmill whose support base, the new wall, is being continually extended; extension is controlled and generated strictly at the tip.     

EFFECT OF SILICATE LIMITATION ON VALVE MORPHOLOGY IN THALASSIOSIRA AND COSCINODISCUS (BACILLARIOPHYCEAE)1

Coscinodiscus radiatus Ehrenb. and Thalassiosira eccentrica (Ehrenb.) Cleve were grown in a silicate-limited chemostat at silicate concentrations below 1 μg-atoms · l^?1. The resulting abnormal valves of C. radiatus lacked a thickened ring around the foramina; their pore membranes were thinner and their loculi shallower than those in normal cells. Abnormal valves of T. eccentrica had a fasciculate areolae pattern; they lacked a silica covering over the foramina and some tangential areolae walls. Neither abnormal valve could be termed a new species.     

BACTERIA THAT INDUCE MORPHOGENESIS IN ULVA PERTUSA (CHLOROPHYTA) GROWN UNDER AXENIC CONDITIONS1

Marine foliaceous green macroalgae such as Ulva lose their typical morphology when cultured aseptically in defined synthetic media. However, after reinfection by certain marine bacteria (isolated from unialgal cultures of Ulva pertusa Kjellman), the organisms regain their typical foliaceous or tubular morphology. To investigate the morphogenesis (MG) induced in U. pertusa by bacteria, we isolated and identified bacteria with MG activity on U. pertusa and studied the distribution of such bacteria in seawater and on various marine macroalgae. We isolated 1555 bacterial strains from 18 species of marine macroalgae (six Chlorophyta, five Phaeophyta, and seven Rhodophyta), from seawater and from sediment collected at the beach at Omaezaki, Shizuoka Prefecture; Japan. Of these, 676 bacterial strains (43.5%) showed MG activity. They were classified into six bacterial groups, Flavobacterium, Vibrio, Pseudomonas, Deleya, Escherichia, and gram-positive cocci. These bacteria were ubiquitous among the samples and were not specific to U. pertusa. Several plant growth regulators had no MG activity. Filter-sterilized supernatants of culture media of MG-active bacteria strains did not induce MG. Cocultivation of Ulva with active bacterial strains is so far the only way to induce the MG effect, which suggests that for MG direct contact between Ulva and the bacterial strain is necessary.     

凤尾竹硅酸体形态及其发生初探

利用光学显微镜、扫描电子显微镜对自然生长凤尾竹(Bambusa multiplex vat．nana(Roxb．)Keng f．)营养体各部分及生长发育各时期叶片的硅酸体进行观察研究。凤尾竹营养体各部分硅酸体形态分别为：叶片竹节形;叶鞘鞍形、钝方形;箨叶哑铃形;箨叶鞘鞍形;杆的表皮部位有少量椭圃形硅酸体,而内部来见成形的硅酸体;地下茎及根皆来见成形硅酸体。成熟硅酸体均沿脉横向排列。硅酸体在叶片、叶鞘、箨叶和箨叶鞘中基本上是按生长发育顺序从不成形到成形,由小到大变化的,分布由不规则到沿脉横列。同一叶从叶鞘到叶片呈现出由鞍形→钝方形(椭圆形)→竹节形渐变的趋势。光镜下,硅酸体内一般有折光率不同于其它部位的l到多个细小颗粒聚在一起形成的核状结构。凤尾竹不同生长发育期各部分硅酸体的晶核存在状况、硅酸体的形态和大小的演变趋势体现了硅质在植物器官表皮细胞上的连续沉积过程。     

COLONY FORMATION AND SEXUAL MORPHOGENESIS IN THE COCCOLITHOPHORE PLEUROCHRYSIS SP. (HAPTOPHYTA)1

Pleurochrysis sp. formed two types of symmetrical, diploid colonies on solid media: (i) single‐cell lineage (SCL) colonies and (ii) aggregation (AG) colonies. The first division of a single mother cell was asymmetric in ～54% of SCL colonies. These colonies developed at a slower rate than AG colonies. Diffusible molecules released from the cells acted like morphogens enhancing formation of AG colonies; their influence on chemotaxis of aggregating cells was dependent on concentration of the inoculum. Nitrogen depletion of diploid colonies induced sexual morphogenesis and colony patterning into inner and outer regions. The smaller innermost cells were surrounded by outer larger cells. Developmental mechanisms of colony formation were examined in relation to the heteromorphic, haplo‐diploid life cycle.     

THE RELATIONSHIP BETWEEN VALVE DIAMETER AND NUMBER OF CENTRAL FULTOPORTULAE IN THALASSIOSIRA WEISSFLOGII (BACILLARIOPHYCEAE)1

Cells well below the reported minimum size for Thalassiosira weisslogii (Grunow) Fryxell et Hasle were observed in two unialgal cultures. The relationship between diameter and number of central fultoportulae was found to be very regular. Regresion analysis of three different populations showed equivalent slopes and r² values above 80%. The y-intercepts were significantly different and support observations in the litrature that nonmarine valeve of T. Weissflogii have fewer central fultoportulae than marine forms.