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.     

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.     

A combined morphological and molecular approach to Nitzschia varelae sp. nov., with discussion of symmetry in Bacillariaceae

A previously unknown member of the Bacillariaceae was discovered almost simultaneously in four different brackish coastal wetlands on the Atlantic and Mediterranean coasts of the Iberian Peninsula. It appears to tolerate a wide range of salinities but was never common in samples where it occurred. The frustules were consistently hantzschioid (i.e. with the raphe systems always on the same side of the frustule) and the valve outline was asymmetrical about the apical plane, two features that have until recently been considered characteristic of Hantzschia. Molecular phylogenies based on rbcL and LSU rDNA indicated, however, that the new species does not belong in Hantzschia but among the several disparate lineages that comprise the paraphyletic genus Nitzschia. This finding, coupled with the recent discovery of other diatoms with constant hantzschioid symmetry but with a morphology very similar to the type species of Nitzschia, is discussed in relation to the status and characterization of Hantzschia as an independent genus. It is concluded that, while a core of hantzschioid species may exist that can be classified together, corresponding to the traditional understanding of the genus Hantzschia, there is no single morphological feature common to all of them that can be used to diagnose the group and differentiate it from the various hantzschioid lineages that are separate from true Hantzschia and currently placed in e.g. Nitzschia or Cymbellonitzschia. Testing whether a hantzschioid species does or does not belong to Hantzschia will in many cases require molecular evidence. Although the new coastal species does not belong to the same lineage as the type species of Nitzschia, N. sigmoidea, it is described for the moment as N. varelae Carballeira, D.G. Mann & Trobajo, sp. nov., until there is a better understanding of generic limits in the Bacillariaceae following a wider molecular and morphological survey of that family.     

The effect of drugs on diatom valve morphogenesis

Summary The effects of various drugs on cell wall (valve) morphogenesis was investigated in three species of diatoms (Pinnularia spp., Surirella robusta, andHantzschia amphioxys) using light microscopy (LM) and scanning electron microscopy (SEM). Treatment ofSurirella with the microtubule (MT) disrupting agent colchicine during early valve formation results in a characteristic malformation of the valve, whereby part of the normally circumferential raphe canal forms as an abnormal protruding lip on the valve surface, located up to 20 m from the edge of the valve. The position of this malformed lip coincides with the location of a microtubule center (MC) at the time of colchicine addition, suggesting that the MC may play a direct role in positioning the tip of the raphe canal during valve formation. The migration of this MC to the tip of the cell during early valve morphogenesis is reversibly inhibited by the metabolic inhibitor 2-4-dinitrophenol (DNP). The effect of colchicine onPinnularia valve formation is less severe, causing occasional malformation of the raphe, but little if any lateral displacement. InHantzschia, colchicine has no effect on the positioning of the raphe, but prolonged exposure causes fusion of the raphe canal with the valve face. Cochicine treatment also results in the absence of the normal curvature at the central interruption in the raphe, as well as abnormal pore formation in this central area. Addition of cytochalasin D during early valve formation inHantzschia causes the raphe canal to form in the center of the valve face, suggesting that the normal translocation of the raphe canal to the valve edge is actindependent. Comparison of valves from control and cytochalasintreatmentHantzschia suggest that the pore spacing within the valve is determined by the position relative to the raphe, and does not depend on whether to pores form on the side (mantle) or the face of the mature valve.Abbreviations DM diatom medium - DNP dinitrophenol - MT microtubule - MC microtubule center - PSS primary silicification site - SDV silica deposition vesicle     

THE “PARADOX” DIATOM BACILLARIA PAXILLIFER (BACILLARIOPHYTA) REVISITED1

Defined by its unique colonial locomotion, Bacillaria paxillifer (O. F. Müll.) Hendey was recognized as a single, pandemic species by many phycologists. However, reinvestigation of colonies from different habitats revealed three distinct groups: (A) brackish/freshwater, (B) marine littoral, and (C) marine planktonic taxa. Groups differed in colony and cell form, raphe flanges (RFs), shape and position of transapical ribs (Tr's), and morphogenesis. Linear‐shaped species were restricted to group A: Tr's thickened principally to the interior. Lanceolate forms were confined to groups B and C: valve formation proceeded from an internal base layer to the exterior. The planktonic species differed in the shape of its raphe slit, and the transformation of girdle bands (GBs) into “winglets.” Taxa also differed in chloroplast shape and number. All species formed motile colonies. Siblings adhered via elastic fibrils secreted through their raphe. Raphe ribs were held in position by siliceous clamps (fibulae), anchored in an extra pair of axial ribs (fibular ribs) parallel to the raphe ribs. This raphe system resembled that of Cylindrotheca rather than the “canal raphe” of Nitzschia. Many valves were asymmetric along the apical axis due to protruding RFs shuttling in a 1:1 ratio within a colony, but raphe slits were mirror images, as were the growth direction of fibulae and position of plastids, with pyrenoids tilted in the same direction. Species possessed four open GBs per epitheca; the third band invariably bore an internal, organic ridge to aid in adhesion of the plasmalemma during cleavage. The results suggested that these taxa are a natural phylogenetic group, requiring precise determination of their taxonomic position.     

VALVE MORPHOGENESIS IN SURIRELLA (BACILLARIOPHYCEAE)1

Valve morphogenesis in two Surirellae (S. ovalis Brebisson and S. robusta Ehrenberg) is described. Mitosis takes place at the broad end of the cell. After cleavage, a new Microtubule Center (MC) arises near each spindle pole and moves to the adjacent plasmalemma. Soon, a specific group of microtubules (MTs) extends from very near the MC around the periphery of the cell. Concurrently, the new tubular Silica Deposition Vesicle (SDV) grows around the periphery of the cell close to these MTs. A double rib of silica is rapidly formed inside the SDV; the space between the ribs becomes the raphe. Mitochondria line up along the MTs, and the SDV may be molded around these to create the canal raphe. Soon, the SDV expands in two directions to create the face and the mantle of the new valve. Meanwhile, each daughter nucleus, accompanied by the MC, moves to its interphase position at the center of the cell; this movement is colchicine-sensitive. As in several other pennate diatoms, an interruption in the raphe of the mature valve coincides with the initial position of the MC. The canal raphe thickens rapidly around the mitochondria; a rudimentary raphe fiber may be associated with the creation of a tiny curvature at the inner raphe fissure. As the SDV expands in the large S. robusta, the daughter cell protoplasts slowly shrink by plasmolysis, thereby creating the complex curved surface of the new valve surmounted by the arching canal raphes which are now quite rigid. In S. ovalis, the daughter cell protoplasts remain appressed and therefore the new valve surface is basically flat. The symmetry of Surirella is quite different from that of other pennate diatoms. However, the cytoplasmic events accompanying valve morphogenesis are similar in all important respects to those described in other raphid pennate diatoms, and clearly supports a naviculoid origin for this genus.     

OBSERVATIONS ON THE STRUCTURE OF SOME FORMS OF GOMPHONEMA PARVULUM KÜTZ. III. FRUSTULE FORMATION1

Gomphonema parvulum Kütz. was investigated by electron microscopy for details of frustule formation. An expansion of the cell along the pervalvar plane occurs prior to cell division. After nuclear division the organelles are, separated into 2 entities, either by division or by dispersion. The cell divides into 2 halves by the invagination of the plasmalemma which is derived from Golgi vesicular activity. When cytoplasmic cleavage, is complete, the Golgi actively produces electronlucent vesicles which collect and coalesce beneath the. plasmalemma to form the silicalemma around the silicon deposition vesicle. The endoplasmic reticulum is also closely associated with this vesicular activity. The vesicle gradually expands and becomes extremely electron dense as silica is deposited within it—first in the region, followed by the mantle edge. When the valve is mature, Golgi vesicles collect and fuse to form the silicalemma of the first girdle band. The first girdle band becomes aligned against the mantle edge on completion, by the “sloughing off” of the external silicalemma and plasmalemma. The second and third bands are formed, individually in a similar manner. Separation of the 2 daughter cells commences at the apical pole and progresses to the basal pole. The plasmalemma and external silicalemma are “sloughed off” so that the 2 cells can separate. The inner segment of the silicalemma becomes the new plasmalemma of the daughter cell.     

PSEUDO-NITZSCHIA AS A GENUS DISTINCT FROM NITZSCHIA (BACILLARIOPHYCEAE)1

Morphologic, diagnostic characters of the subgenus Nitzschia, genus Nitzschia Hassall 1845, and the marine planktonic genus Pseudo-nitzschia H. Peragallo in H. & M. Peragallo 1900 were compared. Colony formation by overlap of cell ends; weakly silicified, shallow, and flattened valves; an extremely eccentric raphe, not elevated above the general level of the valve; lack of con-opea; and striated girdle bands characterize Pseudo-nitzschia as a natural group and a genus separate from Nitzschia.     

MICROMORPHOGENESIS DURING DIATOM WALL FORMATION PRODUCES SILICEOUS NANOSTRUCTURES WITH DIFFERENT PROPERTIES1

Micromorphogenesis within the silica deposition vesicle (SDV) of the diatom Pinnularia viridis (Nitzsh) Ehrenb. resulted in distinct silica nanostructures and layers within forming valves and girdle bands. These siliceous components were similarly disclosed following alkaline etching of mature valves/girdle bands, where their different susceptibilities to dissolution over time resulted from apparent differences in silica density and/or chemistry. The bulk of silica appeared to be deposited at the interface of the forming valve or girdle band with the silicalemma and occurred by the outward expansion of microfibrils of silica that aligned perpendicularly to the silicalemma. Microfibrils originated from both sides of the “silica lamella,” the first nanostructure formed within the SDV, and several silica species of distinct nanostructure and density resulted, including distinctive inner and outermost silica “coverings” of mature valves/girdle bands and the central and terminal nodules. Not all silica deposition and micromorphogenesis occurred in contact with the expanding silicalemma, but was somehow directed within the SDV cavity, and resulted in the distinct silica layers that lined the raphe fissures and poroids. Following alkaline etching, the inner surfaces of valves/girdle bands, as well as the silica layers lining the raphes, poroids, and slits, were determined to be significantly more resistant to alkaline etching than the exterior surfaces, while the outer silica coating and the nodules were quickly dissolved. The processes of micromorphogenesis must have exerted precise control over the chemical nature of the silica formed at different positions within the SDV and affected the overall structure and function of the diatom wall.     

管壳缝类硅藻中国新记录属种——非洲南氏藻

利用光学和电子显微镜对采自黄海水域的1个管壳缝类硅藻——非洲南氏藻进行了形态学研究,并对其地理分布进行了讨论.结果表明:(1)该种壳体带面呈矩形,壳面窄椭圆形,具有钝圆的末端.(2)壳缝居中,由两条等长的分支组成.(3)管壳缝由复杂、接合的肋突支撑,但无龙骨.(4)每条线纹仅有1个孔纹,壳套上最多有1列孔纹.(5)目前...     

Studies on the biochemistry and fine structure of silicia shell formation in diatoms VII. Sequential cell wall development in the pennateNavicula pelliculosa

Summary The development of the wall of synchronized culture ofN. pelliculosa is described. The first step, modification of the 3-2 configuration of the girdle bands of the wall during interphase, occurs immediately before mitotic division by the addition of a third girdl band to the hypotheca. Following cytokenesis, the new valve is initiated when a primary central band is formed within a silica deposition vesicle. This band extends the length of the cell and contains a central nodule. Secondary arms extend from the central nodule, join with extensions of the primary central band, and constitute the raphe rib. Mounds or knolls are formed on the central nodule and disappear as the valve matures. Transapical ribs appear on both the primary central band and secondary arms, and cross extensions join to form the sieve plate areas. The wall appears to be released by a joining of the inner silicalemma and the plasmalemma. An organic coat covers the newly released wall. Two girdle bands are formed and released sequentially.     

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.     

Frustule morphogenesis of raphid pennate diatom <Emphasis Type="Italic">Encyonema ventricosum</Emphasis> (Agardh) Grunow

Diatoms stand out among other microalgae due to the high diversity of species-specific silica frustules whose components (valves and girdle bands) are formed within the cell in special organelles called silica deposition vesicles (SDVs). Research on cell structure and morphogenesis of frustule elements in diatoms of different taxonomic groups has been carried out since the 1950s but is still relevant today. Here, cytological features and valve morphogenesis in the freshwater raphid pennate diatom Encyonema ventricosum (Agardh) Grunow have been studied using light and transmission electron microscopy of cleaned frustules and ultrathin sections of cells, and scanning electron and atomic force microscopy of the frustule surface. Data have been obtained on chloroplast structure: the pyrenoid is spherical, penetrated by a lamella (a stack of two thylakoids); the girdle lamella consists of several short lamellae. The basic stages of frustule morphogenesis characteristic of raphid pennate diatoms have been traced, with the presence of cytoskeletal elements near SDVs being observed throughout this process. Degradation of the plasmalemma and silicalemma is shown to take place when the newly formed valve is released into the space between sister cells. The role of vesicular transport and exocytosis in the gliding of pennate diatoms is discussed.     

A polyphasic approach to the study of the genus Nitzschia (Bacillariophyta): three new planktonic species from the Adriatic Sea

The paraphyletic diatom genus Nitzschia comprises over 1000 morphologically distinct pennate taxa, known from the benthos and plankton of freshwater, brackish, and marine environments. The principal diagnostic characters for delimitation of Nitzschia species include valve shape, the position and structure of the raphe, presence/absence and shape of the proximal raphe endings and terminal raphe fissures, areola structure, and specific morphometric features such as cell size, and stria and fibula density. In this study, we isolated 12 diatom strains into culture from samples collected at the surface or greater depths of the southeastern Adriatic Sea. Morphological analyses included LM, SEM, and TEM observations, which, along with specific morphometric features, allowed us to distinguish three new Nitzschia species. These findings were congruent with the results of phylogenetic analyses performed on nuclear‐encoded SSU (18S) rDNA and chloroplast‐encoded rbcL and psbC genes. One of the new species (Nitzschia dalmatica sp. nov.) formed a lineage within a clade of Bacillariaceae containing members of the Nitzschia sect. Dubiae, which was sister to Psammodictyon. A second lineage was part of a novel clade that is significantly distinct from other Nitzschia species sequenced so far and includes Nitzschia adhaerens sp. nov. and N. cf. adhaerens. A further new species was found, Nitzschia inordinata sp. nov., which appeared as the sister group to the N. adhaerens clade and the conopeoid Nitzschia species in our phylogenetic trees. Our findings contribute to the overall diversity of genus Nitzschia, especially in identifying some deep branches within the Bacillariaceae, and highlight under‐scoring of this genus in marine plankton.     

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.     

SCALY INCUNABULA,AUXOSPORE DEVELOPMENT,AND GIRDLE POLYMORPHISM IN SELLAPHORA MARVANII SP. NOV. (BACILLARIOPHYCEAE)1

Uniparental auxosporulation was observed in a monoclonal culture of a Sellaphora clone isolated from the epipelon of a fishpond in the Czech Republic. The cox1 sequence for the clone confirmed that it belonged to the Sellaphora pupula–bacillum species complex but showed significant differences from all previously characterized Sellaphora species, and it is therefore described as S. marvanii sp. nov. Protoplast, valve, and girdle structure resembled those of other Sellaphora species, but a novel finding for all diatoms was a change in girdle structure during the life cycle: the most advalvar girdle band (valvocopula) bore a single line of pores in enlarged postauxospore cells but was entirely plain in small cells and gametangia. The young auxospores were covered by incunabula containing large, delicate, ± circular scales, resembling those of centric diatom auxospores; similar scales have been reported in a few other raphid diatoms (Pseudo‐nitzschia multiseries, Diploneis sp.) but contrast with the strip incunabula of some Nitzschia and Pinnularia and the helmet‐like caps of Neidium. The scales persisted during auxospore expansion, mostly as two caps over the auxospore poles. The transverse perizonium comprised a very wide, closed primary band, flanked by numerous secondary bands whose open ends were strongly incurved toward the center. Initial valves were differentiated from their immediate descendants by the very strong external demarcation of the raphe sternum, irregular shape, and curved transapical profile.     

Die Lage der Raphen in den Zellen von Nitzschia-Arten

Bei fünf Arten von Nitzschia liegen die beiden Raphen einer Zelle nicht immer diagonal einander gegenüber, sondern manchmal wie bei Hantzschia an derselben Seite. Die beiden Lagen treten im allgemeinen in natürlichen Populationen im Verhältnis von 1:1 auf. Die Tochterraphen werden immer in bezug auf die Mutterzelle gegeneinander versetzt, also nie in gleicher Höhe gebildet. Dies hat zur Folge, daß H-Zellen immer eine H- und eine N-Tochterzelle hervorbringen, während N-Zellen immer entweder zwei N-Zellen oder zwei H-Zellen entstehen lassen, und dies offenbar in gleicher Häufigkeit. Eine sechste untersuchte Nitzschia-Art, N. fonticola, bildet ausnahmslos Nitzschia-Nachkommen.     

DIATOMS LIVING INSIDE THE THALLUS OF THE GREEN ALGAL LICHEN COENOGONIUM LINKII IN NEOTROPICAL LOWLAND RAIN FORESTS1

Coenogonium linkii Ehrenb. is a very common filamentous lichen, growing on stems, hanging roots, and lianas in the understory of neotropical lowland rain forests. We investigated several thalli of this species from locations in Panama and French Guiana. All thalli were inhabited by various species of terrestrial diatoms, which were found between the thallus filaments on extracellular material of the mycobiont. We identified 18 species of diatoms belonging to nine genera: Diadesmis, Eunotia, Hantzschia, Luticola, Melosira, Nitzschia, Orthoseira, Pinnularia, and Stauroneis. The potential benefits both diatoms and lichens could derive from symbiosis in relation to water, irradiance, and nitrogen availability are discussed.     

STRUCTURE,LIFE HISTORY AND SYSTEMATICS OF RHOICOSPHENIA (BACILLARIOPHYTA). I. THE VEGETATIVE CELL OF RH. CURVATA1

Rhoicosphenia Grun. has been placed by some authors in the monoraphid group with Achnanthes Bory and Cocconeis Ehrenb., and by others near Gomphonema Ehrenb. In order to clarify the systematic position of the genus, the morphology and anatomy of the vegetative cells of Rh. curvata (Kütz.) Grun. were investigated using light and electron microscopy. The structure and formation of the two types of valve are described, and the heterovalvy shown to be of a different type from that of the monoraphids; on the basis of raphe, valve and girdle structure a close relationship between these and Rhoicosphenia is unlikely. Rhoicosphenia shows many resemblances to Gomphonema but the types of pore occlusion present, coupled with apparently slight differences in the mucilage-secreting structures and the girdle, suggest that classification in the same family is unwise. The cryptic asymmetry of the valves, and in particular of the raphe system, is noted and explained with reference to their formation; with respect to this asymmetry two configurations of the valves can occur (named cis and trans types) and the distribution of these in raphid genera is discussed briefly. In view of the lack of evidence in raphid diatoms supporting a classification of bands into copulae and pleurae, it is recommended that this practice be suspended.