Ultrastructure of spore development in <Emphasis Type="Italic">Scutellospora heterogama</Emphasis>

The ultrastructural detail of spore development in Scutellospora heterogama is described. Although the main ontogenetic events are similar to those described from light microscopy, the complexity of wall layering is greater when examined at an ultrastructural level. The basic concept of a rigid spore wall enclosing two inner, flexible walls still holds true, but there are additional zones within these three walls distinguishable using electron microscopy, including an inner layer that is involved in the formation of the germination shield. The spore wall has three layers rather than the two reported previously. An outer, thin ornamented layer and an inner, thicker layer are both derived from the hyphal wall and present at all stages of development. These layers differentiate into the outer spore layer visible at the light microscope level. A third inner layer unique to the spore develops during spore swelling and rapidly expands before contracting back to form the second wall layer visible by light microscopy. The two inner flexible walls also are more complex than light microscopy suggests. The close association with the inner flexible walls with germination shield formation consolidates the preferred use of the term ‘germinal walls’ for these structures. A thin electron-dense layer separates the two germinal walls and is the region in which the germination shield forms. The inner germinal wall develops at least two sub-layers, one of which has an appearance similar to that of the expanding layer of the outer spore wall. An electron-dense layer is formed on the inner surface of the inner germinal wall as the germination shield develops, and this forms the wall surrounding the germination shield as well as the germination tube. At maturity, the outer germinal wall develops a thin, striate layer within its substructure.     

ULTRASTRUCTURAL STUDY ON SPORE WALL MORPHOGENESIS IN LYCOPODIUM CLAVATUM (LYCOPODIACEAE)

Spore wall morphogenesis of Lycopodium clavatum was observed by transmission electron microscopy. The spore plasma membrane indicates the reticulate spore sculpture shortly after meiosis. The mature spore wall of this species consists of two layers, inner endospore and outer exospore. There is no perispore in the sporoderm of this species. The exospore formation begins during the tetrad stage; and this layer is divided into two distinct sublayers, an outer lamellar layer and an inner granular layer. The lamellar layer is formed on the sculptured spore plasma membrane. Additional lamellae attach to this layer in a centripetal direction. For that reason, this layer may be derived from spore cytoplasm. The granular layer is formed only in the proximal region following lamellar layer formation, and it also may be derived from spore cytoplasm. The endospore is formed lastly and seems to be derived from spore cytoplasm as well. Accordingly, the spore sculpture of this species may be under the genetic control of the spore nucleus.     

ULTRASTRUCTURE OF DISPERSED AND IN SITU SPECIMENS OF THE DEVONIAN SPORE RHABDOSPORITES LANGII: EVIDENCE FOR THE EVOLUTIONARY RELATIONSHIPS OF PROGYMNOSPERMS

Abstract: The spore Rhabdosporites (Triletes) langii (Eisenack) Richardson, 1960 is abundant and well preserved in Middle Devonian (Eifelian) ‘Middle Old Red Sandstone’ deposits from the Orcadian Basin, Scotland. Here it occurs as dispersed individual spores and in situ in isolated sporangia. This paper reports on a detailed light microscope (LM), scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis of both dispersed and in situ spores. The dispersed spores are pseudosaccate with a thick walled inner body enclosed within an outer layer that was originally attached only over the proximal face. The inner body has lamellate/laminate ultrastructure consisting of fine lamellae that are continuous around the spore and parallel stacked. Towards the outer part of the inner body these group to form thicker laminate structures that are also continuous and parallel stacked. The outer layer has spongy ultrastructure. In situ spores preserved in the isolated sporangia are identical to the dispersed forms in terms of morphology, gross structure and wall ultrastructure. The sporangium wall is two‐layered. A thick coalified outer layer is cellular and represents the main sporangium wall. This layer is readily lost if oxidation is applied during processing. A thin inner layer is interpreted as a peritapetal membrane. This layer survives oxidation as a tightly adherent membranous covering of the spore mass. Ultrastructurally it consists of three layers, with the innermost layer composed of material similar to that comprising the outer layer of the spores. Based on the new LM, SEM and TEM information, consideration is given to spore wall formation. The inner body of the spores is interpreted as developing by centripetal accumulation of lamellae at the plasma membrane. The outer layer is interpreted as forming by accretion of sporopollenin units derived from a tapetum. The inner layer of the sporangium wall is considered to represent a peritapetal membrane formed from the remnants of this tapetum. The spore R. langii derives from aneurophytalean progymnosperms. In light of the new evidence on spore/sporangium characters, and hypotheses of spore wall development based on interpretation of these, the evolutionary relationships of the progymnosperms are considered in terms of their origins and relationship to the seed plants. It is concluded that there is a smooth evolutionary transition between Apiculiretusispora‐type spores of certain basal euphyllophytes, Rhabdosporites‐type spores of aneurophytalean progymnosperms and Geminospora‐/Contagisporites‐type spores of heterosporous archaeopteridalean progymnosperms. Prepollen of basal seed plants (hydrasperman, medullosan and callistophytalean pteridosperms) are easily derived from the spores of either homosporous or heterosporous progymnosperms. The proposed evolutionary transition was sequential with increasing complexity of the spore/pollen wall probably reflecting increasing sophistication of reproductive strategy. The pollen wall of crown group seed plants appears to incorporate a completely new developmental mechanism: tectum and infratectum initiation within a glycocalyx‐like Microspore Surface Coat. It is unclear when this feature evolved, but it appears likely that it was not present in the most basal stem group seed plants.     

Formation and organization of the spore coat ofDictyostelium discoideum

Macromolecular components of the spore coat ofDictyostelium discoideum have been localized by gold-labeled affinity cytochemistry. The outer electron-dense layer is the residence of three prominent glycoproteins that express a fucose-dependent epitope, whereas the inner electron-dense layer includes SP85 and the galactose/N-acetylgalactosamine-containing polysaccharide (GPS). The cellulosic layers are interposed between them. The outer-layer glycoproteins and the GPS also can be found in the interspore fluid, which is usually lost during collection of the spores. Assembly of the spore coat, examined over time, showed that all components, except for the cellulose, are found in an internal secretory vesicle population. All components are found in each vesicle but are not uniformly intermixed within them. Cellulose does not appear until after the outer electron-dense layer of the spore coat has been organized following secretion. The GPS is excluded from the outer dense layer and largely from the cellulosic layer, being more concentrated in the inner layer. SP85 remains localized in the inner dense layer near the cell surface with a circumferentially focal distribution. The distinct distributions of these macromolecular species in the mature spore coat are foreshadowed by their mosaic distribution in the prespore vesicles from which they originate.     

AN ULTRASTRUCTURAL STUDY OF SPORE WALL MORPHOGENESIS IN EQUISETUM ARVENSE

Spore wall morphogenesis of Equisetum arvense was observed by transmission electron microscopy. The spore wall of E. arvense consists of four layers: intine, exine, middle layer, and elater. The exine is formed after meiosis and consists of two distinct layers. The inner portion of the exine is formed in advance of the outer layer of the exine. The middle layer is deposited after the exine. The elater can be subdivided into two distinct layers. The inner layer comprises longitudinal microfibrils that surround the spore in spiral fashion. The elater appears as thin beltlike structures at the beginning of development. Numerous microtubules were observed on the inner surface of the plasmodial plasma membrane opposite the inner layer of the elater, suggesting that these microtubules are involved with the synthesis of inner elater microfibrils. The matrix of the outer elater is formed by discharge of granules from the plasmodial cytoplasm. The intine is the last component of the sporoderm to be formed.     

Physiology and fine structure of sporangiospore germination in Piptocephalis unispora prior to infection

Germination of the sporangiospore of Piptocephalis unispora Benjamin, observed by means of light and electron microscopy, involved the formation of a new inner wall which became continous with the inner layer of the wall of the germ tube. The outer wall layer of the germ tube was continous with the original inner wall layer of the dormant spore. Preliminary details of appressorium structure were noted. Nutritional experiments indicated that sporangiospores required external sources of utilisable nitrogen and carbon compounds for maximal swelling and germ tube production. Limited development occurred when either nutrient was supplied singly. Comparison of germination of the asexual spore with that in other Mucorales, especially the Kickxellaceae, has been made, and the merosporangial status in P. unispora discussed.Non-Standard Abbreviations CH casein hydrolysate - Q spore quotient     

Honey-coloured,sessile Endogone spores

Summary Wall structure is described in the parent and resting spores of an Endogone sp. with honey-coloured, sessile spores. Wall thickness increases in the parent spore and subtending hypha by passage of material through the plasmalemma, or by formation of an apparently separate inner wall and degeneration of the trapped cytoplasm. Structure and development of the multi-layered wall of the mature resting spore are described. Unusual features are: 1. the incorporation of many pigment granules into the coloured outer wall, 2. the presence between the outer coloured and inner transparent walls of a tripartite membrane and adjacent layer with a regular periodicity and 3. a sectored layer with a crystalline component. The structure of the wall is discussed with reference to that of other mucoraceous fungi, to spore germination and to the mechanism of wall formation.     

THE ULTRASTRUCTURE OF CARPOSPOROGENESIS IN CALOGLOSSA LEPRIEURII (DELESSERIACEAE,CERAMIALES)

Carposporogenesis in Caloglossa leprieurii is divided into three cytological stages. At stage I, the young spores have few plastids and little starch. Abundant dictyosomes secrete a gelatinous wall layer in scale-like units. At stage II, dictyosomes produce a second fibrillar wall component in addition to the gelatinous constituent. Large fibrillar vesicles accumulate in the cytoplasm. Production of gelatinous material decreases in this stage. By stage III, starch grains and fully developed plastids are abundant. Rough endoplasmic reticulum occupies much of the peripheral cytoplasm. A dense, granular proteinaceous component appears in the wall in association with the fibrillar layer. Arrays of randomly oriented tubules are scattered in the cytoplasm. The mature carpospore is surrounded by an outer gelatinous wall layer and an inner fibrillar layer. Few dictyosomes persist in the mature spore. Carposporogenesis in Caloglossa is compared with that in other red algae.     

The tapetum inSchizaea pectinata (Schizaeaceae) and a comparison with the tapetum inPsilotum nudum (Psilotaceae)

A combination tapetum consisting of a cellular, parietal component and a plasmodial component occurs inSchizaea pectinata. A single, tapetal initial layer divides to form an outer parietal layer which maintains its cellular integrity until late in spore wall development. The inner tapetal layer differentiates into a plasmodium which disappears after the outer exospore has developed. In the final stages of spore wall development, granular material occurs in large masses and is dispersed as small granules throughout the sporangial loculus. No tapetal membrane develops. Comparisons are drawn with the combination tapetum found inPsilotum nudum.     

Electron microscopy of germinating ascospores ofSaccharomyces cerevisiae

The wall of mature ascospores ofSaccharomyces cerevisiae showed in sections under the electron microscope a dark outer layer and a lighter inner layer. The latter was composed of a greyish inner part and a light outer part. During germination, the spore grew out at one side and the dark outer layer was broken. Of the light inner layer, the inner greyish part became the wall of the vegetative cell, but the extented part of the cell had a new wall.     

Formation of the Dictyostelium spore coat

The spore coat forms as a rigid extracellular wall around each spore cell during culmination. Coats purified from germinated spores contain multiple protein species and an approximately equal mass of polysaccharide, consisting mostly of cellulose and a galactose/N-acetylgalactosamine polysaccharide (GPS). All but the cellulose are prepackaged during prespore cell differentiation in a regulated secretory compartment, the prespore vesicle. The morphology of this compartment resembles an anastomosing, tubular network rather than a spherical vesicle. The molecules of the prespore vesicles are not uniformly mixed but are segregated into partially overlapping domains. Although lysosomal enzymes have been found in the prespore vesicle, this compartment does not function as a lysosome because it is not acidic, and a common antigen associated with acid hydrolases is found in another, acidic vesicle population. All the prespore vesicle profiles disappear at the time of appearance of their contents outside of the cell; this constitutes an early stage in spore coat formation, which can be detected both by microscopy and flow cytometry. As an electron-dense layer, the future outer layer of the coat, condenses, cellulose can be found and is located immediately beneath this outer layer. Certain proteins and the GPS become associated with either the outer or inner layers surrounding this middle cellulose layer. Assembly of the inner and outer layers occurs in part from a pool of glycoproteins that is shared between spores, and unincorporated molecules loosely reside in the interspore matrix, a location from which they can be easily washed away. When the glycosylation of several major protein species is disrupted by mutation, the coat is assembled, but differences are found in its porosity and the extractibility of certain proteins. In addition, the retention or loss of proteolytic fragments in the mutants indicates regions of spore coat proteins that are required for association with the coat. Comparative examination of the macrocyst demonstrates that patterns of molecular distributions are not conserved between the macrocyst and spore coats. Thus spore coat assembly is characterized by highly specific intermolecular interactions, leading to saturable associations of individual glycoproteins with specific layers and the exclusion of excess copies to the interspore space.     

Ultrastructure of hyphae and ascospores in the genus Eremascus eidam

Hyphae and ascospores of Eremascus fertilis and E. albus were studied in ultrathin sections. The lateral wall of the hyphae had a thick electron-light inner layer and a thin dark outer layer. The septa had a simple central pore with or without a plug, and there were Woronin bodies in the vicinity. The wall of the ascospores of E. fertilis showed a thick light inner layer and a thin dark outer layer. In the wall of the spores of E. albus a dark fibrillar layer was present between the light inner layer and the dark outer layer. The spores of this species germinated with a tube the wall of which was continuous with a newly formed layer inside the spore wall.This investigation was supported by the Netherlands Organization for the Advancement of Pure Research (Z. W. O.)     

扁绒泡菌孢子形成过程超微结构

诱导扁绒泡菌显型原质团形成子实体并观察在形成过程中孢囊的超微结构,结果表明,全部原质团参入形成孢子及孢丝;孢子形成初期原质团聚缩使原生质密度加大,脂滴密度也增加;液泡联合形成液泡网体分割原质团,孢子及孢丝一同形成;相邻孢子初始形成的孢子壁可见吻合的突起和凹陷,这是孢子成熟后的表面纹饰部分;孢子壁随孢囊发育逐渐达到适宜位置,孢子壁由透明内层及电子密度较大的外层组成;随后可见外有疣突,内含脂滴的圆形孢子。     

Ultrastructural study of spore wall morphogenesis inOphioglossum thermale kom. var.nipponicum (Miyabe et Kudo) nishida

Spore wall morphogenesis ofOphioglossum thermale var.nipponicum was examined by transmission electron microscopy. The spore wall of this species consists of three layers: endospore, exospore, and perispore. The spore wall development begins at the tetrad stage. At first, the outer undulating lamellar layer of the exospore (Lo) is formed on the spore plasma membrane in advance of the inner accumulating lamellar layer (Li) of the exospore. Next, the homogeneous layer of the exospore (H) is deposited on the outer lamellar layer. Both lamellar layers may be derived from spore cytoplasm; and the homogeneous layer, from the tapetum. Then the endospore (EN) is formed. It may be derived from spore cytoplasm. The membranous perispore (PE), derived from the tapetum, covers the exospore surface as the final layer. Though the ornamentation of this species differs distinctly from that ofO. vulgatum, the results mentioned above are fundamentally in accordance with the data obtained fromO. vulgatum (Lugardon, 1971). Therefore, the pattern of spore wall morphogenesis appears to be very stable in the genusOphioglossum.     

Microscopic observations of germination and septum formation in pycnidiospores of Botryodiplodia theobromae

A population of aseptate pycnidiospores of the fungus Botryodiplodia theobromae can be induced to germinate or to form septa delimiting two cells; this developmental process is dependent upon nutritional and environmental factors. Transmission electron microscope investigations indicate that during germination of the aseptate spore, a new inner wall layer is synthesized de novo at the site of germ tube emergence. Formation of the septum also involves the de novo synthesis of an inner wall layer which comprises the majority of the septum and completely surrounds the spore. The wall of the germ tube emerging from the septate spore is a direct extension of this inner layer deposited during the formation of the septum. Although the early stages of spore germination may involve localized enzymatic degradation of the internal layers of the spore wall, transmission and scanning electron micrographs of germinating spores show that the outer wall layers are physically fractured by the emerging germ tube. It is suggested that spore germination and septum formation are initially similar processes regarding cell wall genesis but that some mechanism responsive to environmental and nutritional conditions determines the course of development.     

Electron microscopy of sporulation inSchwanniomyces alluvius

Sporulation inSchwanniomyces alluvius appeared to be preceded by fusion of a mother and a daughter cell. Meiosis probably occurred in the mother cell and one or two spores were formed in the latter. A study of thin sections showed that the spore wall developed from a prospore wall. The mature spore wall consisted of a broad light inner layer and a thinner dark outer layer including warts. An equatorial ledge was present. During germination in the ascus, a new light inner layer was formed and the old layers of the spore wall partly broke up. Ascospores in a strain ofS. persoonii had a different wall structure in that the dark layer had changed into light areas separated by dark material which formed bulges at the surface.     

BASIDIOSPOROGENESIS IN BOLETUS RUBINELLUS. I. STERIGMAL INITIATION AND EARLY SPORE DEVELOPMENT

Sterigmal initiation in Boletus rubinellus resembled hyphal tip growth. Four stages in early basidiospore development have been delineated based on gross morphology, and changes in wall layers and cytoplasm. Changes in wall layers and cytoplasm during spore development were stage-specific. During Stage 1 the spore wall consisted of two layers identical to those of the sterigmal wall with occasional pellicle remnants on the outer surface. The onset of wall differentiation began in Stage 2, and during Stage 3 wall layers characteristic of the mature spore developed. At Stage 4 there was a pronounced gradient in wall thickness from the apex to the base of the spore. Small vesicles (30–60 nm diam) were uniformly distributed in the cytoplasm of spherically enlarging spores (Stage 2), but during spore elongation (Stages 3 and 4) numerous larger vesicles as well as small vesicles aggregated at the spore apex. A variety of cytoplasmic organelles entered the spore during Stage 3; however, migration of storage materials and the nucleus to the spore did not occur until late basidiospore development. The hilar appendix body developed in the earliest spore primordium and persisted until Stage 3. Development of wall layers and their differential thickening, distribution of vesicles, and probable function of the hilar appendix body are discussed with reference to the control of spore shape. Systematic implications of the data are considered.     

Fine Structure of Bacillus megaterium during Microcycle Sporogenesis

Ultrathin sections were prepared from cultures of Bacillus megaterium QM B1551 undergoing microcycle sporogenesis (initial spore to primary cell to second-stage spore without intervening cell division) on a chemically defined medium. The cytoplasmic core of the dormant spore was surrounded by plasma membrane, cell-wall primordium, cortex, outer cortical layer, and spore coats. Early in the cycle, the coat opened at the germinal groove, the cortex swelled, ribosomes and a chromatinic area associated with large mesosomes (which may later be incorporated into the expanding plasma membrane) appeared in the core, and the cell wall became defined at the site of the cell wall primordium. Poly-β-hydroxybutyrate granules began to appear in the primary cell at about 3 hr. By 7 hr, the forespore of the second-stage spore was delineated by typical double membranes. Between 7 and 12 hr, second-stage cell-wall primordium and cortex developed between the separating forespore membranes. The inner membrane became the plasma membrane of the second-stage spore, and the outer membrane eventually disintegrated within the second-stage spore cortex. A densely staining double layer (spore-coat primordium) developed external to the outer forespore membrane. The inner spore coat and the outer cortical layer of the second-stage spore developed from this primordium. The outer part of the spore coat, probably of sporangial origin, was laid down on the external surface of the inner spore coat. By 12 hr, second-stage spores were almost mature. By 20 hr, the mature endospores, with a thickened outer coat, were often still enclosed by degenerate primary cell wall and by the outer cortical layer and spore coat of the initial spore.     

Ultrastructure of the Wall of the Germinating Sporangiospore of Linderina pennispora (Mucorales)

Carbon replicas of germinating sporangiospores of Linderinapennispora show the outer wall complex to break open basally,during the phase of swelling, and the surface of the germ tubeto be smooth. Chemical treatment reveal the microfibrillar wallof the germ tube to be continuous with the microfibrillar innerwall complex of the spore. Microfibrils of the germ tube arerandornly arranged and appear to be finer than those of thespore wall. Ultra-thin sections reveal the wall of the germtube to consist of an outer electron-dense layer and an innermicrofibrillar electron-transparent layer and both layers originatein the basal region of the spore between the plasmalemma andthe inner wall complex of the spore.     

SPORE WALL DEVELOPMENT IN ANDREAEA (MUSCI: ANDREAEOPSIDA)

The spore wall of Andreaea rothii (Andreaeopsida) is unique among mosses studied by transmission electron microscopy. The exine of other mosses is typically initiated on trilaminar structures of near unit membrane dimensions just outside the plasma membrane. The exine of Andreaea is initiated in the absence of such structures as discrete globules within the coarsely fibrillar network of the sporocyte wall. The sequence of wall layer development, nevertheless, is essentially like that of other mosses. The intine is deposited within the exine and the perine accumulates on the surface of the exine during the latter stages of spore maturation. The mature spore is weakly trilete and inaperturate. The wall consists of three layers, the inner intine, the spongy exine consisting of loosely compacted irregular globules of sporopollenin, and an outer layer of perine. The perine differs ultrastructurally from the exine only in its greater degree of electron opacity. This ultrastructural evidence of departure from the fundamental pattern of exine development in mosses supports the taxonomic isolation of Andreaea from mosses of the Sphagnopsida and Bryopsida.