Pattern morphogenesis in cell walls of diatoms and pollen grains: a comparison

Summary Mechanisms acting in pattern morphogenesis in the cell walls of two distant groups of plants, pollen of spermatophytes and diatoms, are compared in order to discriminate common principles from plant group- and wall material-specific features. The exinous wall in pollen is sequentially deposited on the exocellular side of the plasmalemma, while the siliceous wall in diatoms is formed intracellularly within an expanding silica deposition vesicle (SDV) which is attached to the internal face of the plasmalemma. Two levels of patterning occur in diatom and pollen walls: the overall pattern stabilises the wall mechanically and is apparently initiated in both groups by the parent cell, and a microtubule-dependent aperture and portula pattern created by the new mitotic (diatoms) or meiotic (pollen) cells. The parent wall in diatoms, and also the callosic wall in microspores, functions as anchor surfaces for transient, species-specific patterned adhesions of the plasmalemma to these walls, involved in pattern and shape creation. Patterned adhesion and exocytosis is blocked in pollen walls where the plasmalemma is shielded by the endoplasmic reticulum at the sites of the future apertures. In diatoms, wall patterning is uncoupled from the formation of a siliceous wall per se when the SDV and its wall is formed without contact to the the plasmalemma. Conversely, a blue-print pattern laid out in advance along the plasmalemma can be found in several diatoms. This highlights the key function of the plasmalemma and its associated membrane skeleton (fibrous lamina), and its orchestrated co-operation with elements of the radial filamentous cytoskeleton (actin?) in pattern formation. The role of microtubules during generation of the overall pattern may be primarily a transport and stabilizing function. Auxiliary organelles (spacer vesicles, endoplasmic reticulum, mitochondria) involved in diatoms for shaping the SDV, and a mechanism adhering and disconnecting this SDV together with spacer organelles in a species-specifically controlled sequence to and from the plasmalemma, are unnecessary for pollen wall patterning. The precise positioning of the portula pattern in diatom walls is discussed with respect to their role as permanent anchors of the cytoplasm to its wall, and in providing spatial information for nucelar migration and the next cell division, whereas apertures in pollen are for single use only.Abbreviations AF actin filaments - C/Ca callose - CF cleavage furrow - cPL cleavage plasmalemma - DV dense vesicles - ER endoplasmic reticulum - ET epitheca - HT hypotheca - mPL folded plasmalemma - MT microtubules - MTOC microtubule organising centre - PEV primexine (matrix) vesicles - PL plasmalemma - SDV silica deposition vesicle - Si silica - SL SDV-membrane - SPV spacer vesicles Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement     

PIT CONNECTION FORMATION IN THE RED ALGA PSEUDOGLOIOPHLOEA

Pit connection formation in the marine red alga Pseudogloiophloea confusa was studied with the electron microscope. The process of formation occurs in 2 stages. First, a septum forms as an annular ingrowth from the lateral walls. Lomasomes are associated with the centripetal accretion of wall material. The completed septum contains a large rimmed aperture, bounded by the continuous plasmalemma, and through which the cytoplasm is continuous from cell to cell. In the second stage, a highly structured plug is formed which completely blocks the aperture. The plug is condensed on flattened vesicles which lie parallel to one another and which traverse the aperture. The mature plug is composed of granules 50–100 A in diameter and surrounded by several dense layers which appear to enclose an 80 A limiting membrane. Once the pit connection is formed, no material is seen to traverse it.     

Discharge papilla development in the phycomycete Allomyces

The process of discharge papilla (DP) formation in Allomyces macrogynus was studied by light and electron microscopy. The plug of the DP was first deposited between the plasmalemma and the wall of the zoosporangium (ZS). The wall above the plug subsequently was eroded away. Deposition of a new inner wall layer in the sporangium held the plug in place and thickening of the layer formed a collar around the plug. Further deposition of material after this stage resulted in the characteristic pulley-shape. The plug material appeared homogeneous in electron micrographs but there was evidence of an outer layer. Digestion of the plug at the time of spore release was from within.Abbreviations DP discharge papilla - ZS zoosporangium     

Cotton fibre differentiation: Occurrence and distribution of coated and smooth vesicles during primary and secondary wall formation

Summary Coated vesicles occur in differentiating cotton fibres during primary and secondary wall formation. The coated vesicles are often associated with the plasmalemma, or with membranes at the secreting face of dictyosomes, corresponding positionally to GERL. During secondary wall formation the number of dictyosome-associated coated vesicles seems to be smaller than during primary wall formation. When sections are stained for periodateoxidizable polysaccharides (Thiéry reaction) the membrane of plasmalemma-associated coated vesicles is intensely stained. The membrane of dictyosome-associated coated vesicles is only weakly stained. On the basis of the present evidence it is not possible to clearly decide, whether the staining in plasmalemma-associated coated vesicles is due to obliquely cut membrane or to vesicle contents. The vesicle coat material is not stained. Possible functions of coated vesicles in differentiating cotton fibres are discussed.Vesicles with contents positively stained with the Thiéry reaction are observed only during primary wall formation. The membrane of these vesicles is smooth and seems to bud from the same cisternae, probably GERL, as do the coated vesicles. During secondary wall formation no vesicles containing periodate-oxidizable polysaccharides could be detected, even under conditions that result in a strong, specific reaction in the cellulosic secondary wall. In some instances polysaccharidic material, resembling secondary wall material, has been seen to adhere to the outside of the plasmalemma. These results are consistent with the hypothesis that, in higher plants, at least part of primary wall material may already be synthesized in dictyosome vesicles, whereas cellulose biosynthesis occurs at the cell surface.     

银耳（Tremella fuciformis）原基分化前期双核菌丝细胞和分生孢子的边缘体与质膜体

本文报道银耳（Ｔｒｅｍｅｌｌａｆｕｃｉｆｏｒｍｉｓ）原基分化前期．在双核菌丝的幼细胞、成熟细胞和分生孢子中,与质膜相关联的两类膜结构──边缘体和质膜体的形成与功能。根据相似结构的存在．支持小泡或多泡体排出质膜之外附在细胞壁上成为边缘体和参于细胞壁合成的假定。银耳原基分化前期．双核菌丝迅速分裂的幼细胞．其质膜内陷产生泡状质膜体,内含数个小泡,或产生膜状质膜体;在成熟细胞中．质膜内陷通常形成回旋的膜结构──膜状质膜体．内含１—２个电子致密小泡．当这两类质膜体脱离质膜进入细胞质后,有的膜层和小泡局部被消化．因此,推断质膜体具有内吞和输送养料的作用。另外．在桶孔隔膜闭塞一侧电子致密度高的细胞质中．还观察到一种罕见的只有单个膜层的质膜体．其内充满３个电子致密小泡．估计它的形成与功能同膜状质膜体相似。作者认为．桶孔闭塞和质膜体的出现是与银耳原基细胞分化有关联的两个重要特征。最后,在成熟细胞中,尚可以观察到质膜体的膜层能够散开形成内质网．因此．内质网也可以来源于质膜体。     

Budding in the Dimorphic Fungus Phialophora dermatitidis

Ultrastructural comparisons of yeast and hyphal bud formation in Phialophora dermatitidis reveal that bud initiation is characterized by a blastic rupture of the outer portion of the yeast or hyphal wall and the emergence of a bud protuberance through the resulting opening. The wall of the emerging bud is continuous, with only an inner wall layer of the parental yeast or hypha. The outer, ruptured portion of the parental wall typically forms a collar around the constricted emergence region of the developing bud. The cytoplasm within the very young emerging bud invariably contains a small number of membrane-bound vesicles. The septum formed between the daughter bud and the parental yeast or hypha is a complete septum devoid of a septal pore, septal pore plug, or any associated Woronin bodies characteristic of simple septa of the moniliform or true hyphae. These observations suggest that yeast bud formation and lateral hyphal bud formation in the dimorphic fungus P. dermatitidis involve a growth process which occurs identically in both the yeast and mold phase of this human pathogenic organism.     

The relationship of echinate inclusions and coated vesicles on wound healing inNitella flexilis (Characeae)

Summary Wound healing in internodal cells of the freshwater algaNitella flexilis (Characeae) was studied in the light and electron microscope. Immediately after punctation of the cell wall a wound plug is formed which stops outflow of cytoplasm. The plug consists of echinate inclusions which are normally located in the central vacuole. A wound wall consisting of pectin and cellulose microfibrils is formed beneath the plug within one to several hours. During that time the wound shows intensive fluorescence when treated with chlorotetracycline indicating transmembrane Ca²⁺ fluxes. Numerous coated pits and vesicles are found at the plasmalemma. The glycosomes undergo pronounced structural changes. Neither plug nor wound wall formation depend on actin filaments or microtubules as shown by inhibitor experiments with cytochalasin and amiprophos-methyl. The function of the coated vesicles and their interrelationship with other cell organelles is discussed.     

Development of the discharge apparatus in the fungus Entophlyctis

Correlative light and electron microscopic observations were used to reconstruct the morphological events involved in the development of the discharge apparatus of Entophlyctis zoosporangia. A discharge plug formed as vesicles containing fibrillar material fused with the plasma membrane and deposited their matrices between the plasma membrane and zoosporangial wall. At the apex of the enlarging plug, the zoosporangial wall lost its microfibrillar appearance, became diffuse, and left an inoperculate discharge pore. The discharge plug exuded through this pore and then expanded into a sphere which rested at the tip of the discharge papilla or tube. After the release of the discharge plug, the number of fibrilla containing vesicles decreased and abundant endoplasmic reticulum appeared in the cytoplasm below the plug. Granular material then accumulated at the interface of the discharge plug and the plasma membrane. This was the endo-operculum. A single layer of endoplasmic reticulum subtended the area of plasma membrane which the endo-operculum covered. Later, dictyosomes appeared in the cytoplasm below the endo-operculum. Fusion of Golgi vesicles with the plasma membrane below the endo-operculum coincided with the initiation of cytoplasmic cleavage. This sequence of events indicates that, unlike the discharge plug, the endo-operculum does not originate by vesicular addition of preformed material.     

Formation of the Fertilization Pore during Oogenesis of the Fern Ceratopteris thalictroides

The development of the fertilization pore during oogenesis of the fern Ceratopteris thalictroides was followed using transmission electron microscopy. The newly formed egg is appressed closely to the adjacent cells. There are well-developed plasmodesmata between the egg and the ventral canal cell, but none between the egg and the jacket cells of the archegonium. During maturation, a separation cavity is formed around the egg. However, a pore region persistently connects the egg and the ventral canal cell. The extra egg membrane is formed by deposition of sheets of endoplasmic reticulum (ER), but no ER is deposited on the inner surface of the pore region. Thus, a fertilization pore, covered by a layer of plasmalemma, is formed. The ventral canal cell undoubtedly participates the formation of the fertilization pore, probably by absorbing the sheets of ER beneath the pore region. The functional significance of the ventral canal cell in formation of the fertilization pore is discussed. The features of the mature egg include that abundant concentric membranes and osmiophilic vesicles occur in the cytoplasm of the mature egg. The initial, round nucleus of the egg eventually becomes cup-shaped. This investigation gives some new insights about the cells participating oogenesis in ferns.     

CHLORTETRACYCLINE-INDUCED FORMATION OF WALL APPOSITIONS (CALLOSE PLUGS) IN INTERNODAL CELLS OF NITELLA FLEXILIS (CHARACEAE)1

The formation of chlortetracycline (CTC)-induced wall appositions or plugs in internodal cells of Nitella flexilis (L.) Ag. was studied with light, fluorescence and electron microscopy. These plugs contain callose and pectin. A few minutes after CTC addition plug formation starts by fusion of polysaccharide-containing vesicles (glycosomes) with the plasmalemma. Plug growth is continued by incorporation of glycosome-endoplasmic reticulum (ER) complexes. The cytoplasm near the plug appears dense because of the accumulation of glycosomes and the increased electron density of plasma matrix and organelles. About 1 h after CTC addition plug growth ceases, the cytoplasm recovers to its pretreatment appearance, and a few glycosomes fuse singly with the plug membrane. Crystalline inclusions which consist of hexagonally packed rods are found near the plug. Coated vesicles and coated pits are clearly seen only in very early and late stages of plug formation. Callose is also found in parts of wound plugs produced after mechanical injury. No callose is present in the underlying, highly ordered wound wall. The failure to produce a wound wall beneath CTC-induced plugs appears to be related to the lower number of coated vesicles during plug formation. The possible significance of the partially coated reticulum in plug and wound wall formation is discussed.     

Aspects of morphogenesis and function of diatom cell walls with implications for taxonomy

Summary Aspects of morphogenesis and morphology of diatom cell walls are reviewed to highlight functional correlations between wall structures and three-dimensional cytoplasmic activities during the cell cycle. Morphogenesis of the siliceous valve within the silica deposition vesicle is discussed in the light of the dependency on a precisely orchestrated moulding machinery, involving the cytoskeleton, mitochondria, endoplasmic reticulum, spacer vesicles produced by the Golgi apparatus, and the plasmalemma, in combination with adhesion of the cells to parts of the parental wall and localized plasmolyses. Sensitivity of morphogenetic events to fluctuations of external factors has implications for taxonomy.Abbreviations CF cleavage furrows - cPL cleavage plasmalemma - GB girdle bands - LP labiate process - LPA labiate process apparatus - MC microtubule center - mLP macro labiate process - MT microtubule - MTOC microtubules organizing center - PL plasmalemma - SDV silica deposition vesicle - SL SDV membrane - SpV spacer vesicles Dedicated to Professor Peter Sitte on the occasion of his 65th birthday     

The ultrastructure of thePuccinia sorghi aecial stage

Summary In the aecium ofPuccinia sorghi the substantial outer layer of cell walls in the base of the stroma, together with the thick wall of peridial cells where they adjoin the compressed cell debris, may play a role in the water balance of the fruiting structure. With few exceptions binucleate cells are found only in the aecium. Cytoplasm of the binucleate peridial cells is densely filled with organelles and fat globules. Large multinucleate cells with inclusions, that are apparently proteinaceous in nature, are found in the aecial base. The ontogeny of aeciospore initials is annellidic and aeciosporophores have a well developed collar consisting of annellations. Sporophores, aeciospore initials, intercalary cells and young aeciospores are enveloped by a matrix. During aeciospore ontogeny fat globules are formed in the immediate vicinity of enlarging vacuoles and vesicle-containing structures, suggesting that the formation of fat globules is associated with these cytoplasmic components. Spine initials are first observed as electron-lucent areas between the plasmalemma and the wall of immature aeciospores. During further development the inter- and supraspinal wall of the spore is degraded, a process in which small, well-defined granules appear to be implicated. The spines become separated from the plasmalemma by wall apposition. Apposition also seals off the channel leading to the septal pore in the aeciospore base. The spore wall contains several germ pores. The vacuolate and often degenerate nature of inter- and intracellular hyphae in thalli with young sporulating aecia points to accumulation of food reserves in young aecia.     

Ultrastructure of Aerial Hyphae in Linderina pennispora

Ultrathin sections of the aerial hyphae of Linderina pennispora,fixed in glutaraldehyde, show the wall to be composed of anouter electron-dense layer and an inner less-dense one. Thefully developed septum is plugged by electrondense material.A similar septum exists at the base of the pseudophialide. Cytoplasmiccontinuity on both sides of the septum is maintained by theplasmalemma which passes through the septal pore around theperiphery of the plug. Membrane-lined vesicles may occur inthe wall, between the wall and the plasmalemma and internalto the plasmalemma. Ribosome-like particles are numerous inthe cytoplasm and endoplasmic reticulum is virtually absent.     

On the ultrastructure of differentiating secondary xylem in willow

Summary Studies of differentiating xylem inSalix fragilis L. show the immediate cambial derivatives to be ultrastructurally similar. The Golgi apparatus is important at all stages of wall synthesis, possibly producing (amongst other substances) hemicellulose material which is carried to the wall in vesicles or multivesicular bodies. The endoplasmic reticulum also contributes one or more components to the developing wall: at some stages during differentiation the endoplasmic reticulum produces electron opaque bodies which appear to be guided towards the wall by microtubules. Compact structures formed from concentric membranes (myelin-like bodies) have been found joined to rough endoplasmic reticulum, but their presence is not explained.Two types of plasmalemma elaboration occur: invagination of the plasmalemma itself to form vesicles which may contain cytoplasmic material; and vesicles between the plasmalemma and cell wall which are the result of single vesicles or multivesicular bodies traversing the plasmalemma. Both systems provide a means for transporting cytoplasmic material across the plasmalemma.Microtubules have been seen associated with all vesicles derived from the cytoplasm which appear to be moving towards the wall. The presence of microtubules may generally be explained in terms of orientation of vesicles, even if they also happen coincidentally to parallel the supposed orientation of microfibrils in the wall itself. It is possible to resolve connections between the microtubules and the plasmalemma.     

AN ULTRASTRUCTURAL STUDY OF VEGETATIVE CELL DIVISION IN OEDOGONIUM BORISIANUM12

Vegetative cell division in Oedogonium borisianum is initiated by the formation of a 3-layered ring adjacent to the wall in the upper portion of the cell. This structure enlarges by the coalescence of vesicles. When the ring is fully developed, the parent wall splits adjacent to the ring, and the ring expands into a cylinder, which becomes the cuticle of the upper daughter cell. The lateral wall then forms between this cuticle and the plasmalemma of the cell. Concurrent with ring development and expansion, the nucleus migrates to a position in the center of the cell and karyokinesis occurs. Commencing with late telophase, evidence of transverse wall formation becomes apparent. The zone between the daughter nuclei contains a layer of microtubules in a plane parallel to the plane in which the transverse wall will develop. Subsequently a random coalescence of vesicles occurs along this plane. During the latter stages of this process, the ring expands and the plane of the transverse wall moves upward to the base of the ring cylinder. The completed transverse wall then fuses at is periphery with the newly formed lateral wall.     

Studies on metacercarial excystment in Himasthla leptosoma (Trematoda: Echinostomatidae) and newly emerged metacercariae

Metacercarial cysts of Himasthla leptosoma were subjected to a solution containing bile salts, trypsin and l cysteine at 41°C. The treatment induced intense metacercarial activity and after 20 min metacercariae burst through the cyst walls and emerged. Electron microscopy demonstrated that organisms burst through a small area of cyst wall which was devoid of a layer of lamellae present elsewhere towards the innermost surface. The appearance of ruptured cyst walls indicated that they had been softened by the excystment medium. Newly emerged metacercariae possessed a reniform collar of 29 cephalic spines and these were sometimes withdrawn into pits, presumably by action of a muscle complex in the head region. Sensory papillae were distributed in a bilaterally symmetrical arrangement around the anterior sucker and none were visible on the surface of the ventral sucker. Tegumental spines were found only from a point some distance behind the head collar to the region of the ventral sucker. The most anterior spines were simple and peg-like and they quickly merged posteriorly into more complex palmate forms.     

THE CELL WALL OF COSMARIUM BOTRYTIS12

The cell wall of Cosmarium botrytis was studied through the use of the freeze-etch technique. The cell wall consists of many thin layers. Fracturing along one layer reveals the positioning of the wall sculpturing, wall pores, and wall microfibrils. The individual microfibrils are grouped together in bands of parallel oriented fibrils. The different bands of parallel microfibrils were apparently arranged at random angles with regard to each other. Small particles may also be present in the cell walls. The cell wall pore unit of Cosmarium botrytis was studied through the use of scanning, freeze-etching, and thin sectioning techniques. The pore sheaths, on the outside of the cell wall, form a collar around the mouth of each pore. The pore sheath is composed of needle-like fibrils radiating outward from the pore. A pore channel traverses the cell wall and leads to a complex pore bulb region between the cell wall and the plasmalemma. The pore bulb contains many small fibrils which radiate toward the plasmalemma from a number of net-like fibril layers which in turn merge into a very electron dense region near the base of the pore.     

Ultrastructure of Cysts of Naegleria spp: A Comparative Study*

SYNOPSIS. Ultrastructure of cysts of Naegleria gruberi, Naegleria fowleri, and Naegleria jadini was compared by transmission electron microscopy. Pores in the cyst wall were observed in all 3 species. In N. gruberi they were surrounded by a collar and sealed with a relatively large mucoid plug; no such collar was seen around the pores in the other 2 species, in which the plug was smaller than that in N. gruberi. An electron-dense plaque serving as an additional pore closure was present in all 3 species. In N. gruberi, the cyst wall consisted of an inner thick and an outer thin layer; however, only the inner component was present in cysts of N. fowleri and N. jadini, which had a smooth appearance. At the ultrastructural level, excystment of N. fowleri involved digestion of the mucoid plug and emergence of the trophozoite through the pore. Some digestion of the cyst wall also appeared to take place during excystment.     

Localization of lead accumulated by corn plants

Light and electron microscopic studies of corn plants (Zea mays L.) exposed to Pb in hydroponic solution showed that the roots generally accumulated a surface Pb precipitate and slowly accumulated Pb crystals in the cell walls. The root surface precipitate formed without the apparent influence of any cell organelles. In contrast, Pb taken up by roots was concentrated in dictyosome vesicles. Dictyosome vesicles containing cell wall material fused with one another to encase the Pb deposit. This encased deposit which was surrounded by a membrane migrated toward the outside of the cell where the membrane surrounding the deposit fused with the plasmalemma. The material surrounding the deposit then fused with the cell wall. The result of this process was a concentration of Pb deposits in the cell wall outside the plasmalemma. Similar deposits were observed in stems and leaves suggesting that Pb was transported and deposited in a similar manner.     

A freeze-etch study of the ultrastructure of red algal pit plugs

Summary Freeze-etch studies indicate that the fully-formed pit plug ofPalmaria palmata consists of a homogeneous plug core, a pair of plug caps, and associated membranes. A single cap membrane separates the two layers of each plug cap and is the only membrane found between the cytoplasm and the plug core. Due to the apparent junction of the plasmalemma and cap membranes, the plug core, along with the adjacent plug cap layer on either face of the core, is entirely membrane-bounded and essentially extracellular. Fracture faces of the cap membrane bear large populations of particles which are inconspicuous due to their low profile. The plasmalemma lining the plug core from cell to cell has numerous, very obvious particles on its fracture faces, more even than does the cytoplasmic plasmalemma.The relationship of the rhodophycean pit plug to the fungal septal pore plug is discussed.This report represents a portion of a thesis submitted in partial fulfillment of the requirements of a Master of Science degree, Cornell University, Ithaca, N.Y.