THE DEVELOPMENT OF THE SPOROCARP OF MARSILEA VESTITA

The tissues of the sporocarp of Marsilea vestita undergo profound changes during development. Early in development, the cells of the peripheral tissues, epidermis, hypodermis and layers of the transitional zone between the hypodermis and more internal tissues contain prominent vacuolar bodies. As development proceeds, these vacuolar bodies disappear. Prominent amyloplasts are found only in the guard cells and in the cells of the transitional zone. Later in development the cells of the hypodermis divide periclinally forming two layers which differentiate as macrosclereids. The cells of the outermost layer of the transitional zone differentiate as osteosclereids. Internally, the cells of the sorophore accumulate large amounts of mucilage in the central vacuoles. The peripheral cytoplasm ultimately degenerates leaving just hygroscopic mucilage. The mucilage carbohydrate contains the sugars, rhamnose and arabinose. In the young sorus, only the spore mother cells and the cells of the indusium contain amyloplasts. By the time of meiosis, there is a massive accumulation of starch in the receptacle, stalk and jacket but not in the tapetum of the sporangia. Late in development, the starch disappears and the mega- and microspores become coated with carbohydrate.     

Aspects of spore production in the red algaCeramium

Summary Tetraspore development from the post-meiotic to the mature stage has been studied using light and electron microscopy and histochemistry. The structure of the mature carpospore is identical to that of the tetraspore suggesting a similar developmental sequence.The tetrasporangial wall consists of 3 main fibrillar layers, the origin of the inner of which appears to be the wall-plasmalemma interface. The development of furrows cleaving the protoplast into 4 results in the formation of new plasmalemma and subsequently new wall fibrils. The Golgi apparatus is important in the formation of two well-defined substances. The first is fibrillar and is secretedvia vacuole-like structures into the sporangial wall. After spore release, this functions as a protective mucilaginous layer. The second has a distinctive fine structural morphology and probably functions as an adhesive.Observations on spore releasein vivo reveals a similar process for both types of spore. Each spore is surrounded by mucilage which may assist in initial attachment prior to the secretion of the adhesive.     

Life Cycles of Myxogastria Stemonitopsis typhina and Stemonitis fusca on Agar Culture

Myxogastria is a group of protozoa characterized by cellular uninucleate amoeboflagellates (myxamoebae and flagellated swarm cell), acellular multinucleate plasmodia, and stationary spore‐bearing sporocarps. The Stemonitales is a large order in the Myxogastria and contains approximately 230 species, but only 13 species have their completed life cycles observed so far. Here, we described the life cycles of two species in Stemonitales, Stemonitopsis typhina and Stemonitis fusca by culturing in water agar medium and observing the morphogenesis of their spore germination, plasmodium, and sporocarp development. The spore‐to‐spore life cycles of Ste. typhina and S. fusca were completed in approximately 67 and 12 d, respectively. Both species possessed an aphanoplasmodium. However, the spores of Ste. typhina and S. fusca germinated by the V‐shape split and pore methods, respectively. Unlike S. fusca with an evanescent peridium, Ste. typhina produced a shiny persistent peridium which was continuous with the membrane surrounding its stalk. The information will contribute to a better understanding of their taxonomy and phylogeny.     

MORPHOGENESIS OF THE SPORANGIUM OF COMATRICHA

Goodwin , Donna C. (State U. Iowa, Iowa City.) Morphogenesis of the sporangium of Comatricha. Amer. Jour. Bot. 48(2): 148–154. IIlus. 1961.—Three species of the myxomycete genus, Comatricha, were studied: Comatricha nigra, C. fimbriata, and C. elegans. The sporangia developed on living bark of Ulmus americana in moist chamber. The hypothallus is formed under the homogeneous protoplasmic mass of the sporangial initial. The fibrous threads of the hypothallus bend upward, lengthening at the apices to become the fibers of the stalk and columella. The undifferentiated protoplasm is carried upward as the stalk elongates. When the columella has attained its mature height, threads bend out from the columella and grow toward the periphery of the sporangium. These threads form the capillitium. Simultaneous with the appearance of the capillitial initials, the peridium, a delicate membrane, forms. After the capillitium is mature, the protoplast cleaves into many cells, the future spores. The peridium evanesces early in the stage of spore maturation. Cellulose is present in the stalk, capillitium, and spore walls but is not found in the peridium or hypothallus. The capillitium of these species follows a developmental pattern designated as the “Comatricha-type” by Ross (1957) from a study of Comatricha typhoides. The taxonomic implications of the sporangial developmental pattern are discussed.     

A new species of Lamproderma (Myxomycetes) from Costa Rica

Lamproderma magniretisporum, a new species of myxomycete from Costa Rica, is described and illustrated. This new species is characterized by its lignicolous habitat, long stalk, dark radial capillitium, large spores, and reticulate spore ornamentation. The stability of the taxonomic characters of L. magniretisporum is supported by two well-developed collections. The morphology of the sporocarp was subjected to detailed study with both the light microscope and the scanning electron microscope. Micrographs of all relevant features are presented. Taxonomic novelty Lamproderma magniretisporum G. Moreno, C. Rojas, S.L. Stephenson & H. Singer     

The ultrastructure of the zygospore in Endogone flammicorona Trappe & Gerdemann

The ultrastructural organization of the spores of the sporocarp of Endogone flammicorona was studied. Two types of organization are described. Initially the spore possessed a vacuolate protoplasm and was bound by two cell wall layers. The spore was surrounded by a hyphal mantle formed of a sheet of vacuolized hyphae with uniformly thin walls. Secondly, although the ultrastructural features of the spore appeared the same, it was now surrounded by a hyphal mantle with unevenly thickened walls (i. e., the so-called flaming crown) due to the gradual and irregular deposition of granules and lamellae. This crown gives the spore its most commonly observed morphological feature and is the preminent character employed taxonomically to speciate Endogone flammicorona Trappe & Gerdemann.     

Spores of microorganisms

Spores ofBacillus cereus (strain NCIB 8122) were germinated in a synthetic germination limited medium (GL-medium), which permitted germination but did not make the termination of post-germinative development possible. Incorporation of¹⁴C-diaminopimelic acid into the newly formed cell wall was followed in this culture. Morphological changes were studied by optical and electron microscopy. Germination was associated with the usual germination changes,i.e. depolymerization of the “bulky” cortex, differentiation of nuclear structure and mesosomes and ribosomes in the cytoplasm. At this stage the spore protoplast is surrounded by several layers: exosporium, laminated coat with four layers, residual spore wall and the protoplast membrane. During incubation in this limited medium the residual wall layer thickens and the nuclear structure, mesosomes and ribosomes were not more detectable. After enrichment of the GL medium (shift up) the thick-walled cells can form additional cell wall material, elongate and an atypical septum formation can occur. The cell wall material forms local thickenings. On long-term cultivation in the GL medium some of the cells in the GL medium lyze. If, in the course of 3–6 h the cells are transferred from the GL-medium to a solid complex medium (Difco Nutrient Agar) the thickwalled cells are transformed into dividing cells. When the cells are transferred later, their colony-forming ability rapidly decreases. The decrease of viability of the thick-walled cells derived directly from spores after their germination in the limited medium indicates that these cellular forms probably do not represent more stable cellular types that would be of considerable importance for survival of the populat ion of bacilli.     

NUCLEAR MIGRATION AND ASYMMETRIC CELL DIVISION IN ONOCLEA SENSIBILIS SPORES: AN ULTRASTRUCTURAL AND CYTOCHEMICAL STUDY

A method of preparation for electron microscopy of fern spores in early stages of germination is presented. The cytochemistry and fine structure of Onoclea spores during the early stages of germination are described. The cytoplasm of the hydrated spore is filled with lipid droplets, protein granules and chloroplasts. During the early stages of development ribosomes and mitochondria increase in the area surrounding the central nucleus, and a new peripheral wall forms around the protoplast. Microtubules and large, branching mitochondria are associated with the nucleus during migration from its original central position in the spore to the proximal face and then to one end of the spore. There is no morphological polarization of cytoplasmic organelles of the spore before migration of the nucleus.     

The induction of spore cells in vitro by membranes of Dictyostelium discoideum

A mutant of Dictyostelium discoideum, HM18, will differentiate into both stalk and spore cells when plated at high cell density (10⁵ cells/cm²) as a monolayer on non-nutrient agar containing 5 mM cAMP [6]. At low cell density (10³ cells/cm²) neither stalk nor spore cells are produced, but the addition of a cytosol fraction leads to stalk cell formation, and the addition of a membrane fraction leads to spore cell formation. The spore cell-inducing activity of the cell membranes is developmentally regulated; it is first detectable during late aggregation and increases to a maximum level in the pseudoplasmodial stage of development. The activity is sensitive to proteolysis and insensitive to periodate treatment. It is partially inactivated by incubation at 100 °C for 5 min. Variable amounts of the activity can be removed from the membrane by washing, suggesting that at least part of the activity is loosely membrane-bound. Activity is enriched in plasma membrane fractions, suggesting that the inducing factor is located at the cell surface. It is possible that the membranes are replacing a cell-cell contact requirement for spore formation.     

GIBBERELLIC ACID-INDUCED GERMINATION OF SPORES OF ANEMIA PHYLLITIDIS: NUCLEIC ACID AND PROTEIN SYNTHESIS DURING GERMINATION

The distribution and synthesis of nucleic acids and proteins during gibberellic acid-induced germination of spores of Anemia phyllitidis were studied in order to relate biochemical activity with morphogenetic aspects of germination. Germination is accompanied by the hydrolysis of storage protein granules and the localized appearance of cytoplasmic RNA, protein, and insoluble carbohydrates in a small area adjoining the spore wall and surrounding the nucleus. The protoplast of the spore enlarges in this region, the spore wall breaks and a protonemal cell is formed which contains many chloroplasts. A second division in the spore at right angles to the first yields a rhizoid cell. Autoradiography of ³H-thymidine incorporation has shown that DNA is synthesized both in the nucleus and in the immediately surrounding cytoplasm of the germinating spore until some time after the first division, although a strictly nuclear DNA synthesis is observed later. Synthesis of RNA and proteins is limited to the presumptive regions of the germinating spore which become the protonema and rhizoid, shifting to specific sites in these cells as germination proceeds. The nucleus of the spore continues to be biosynthetically active long after it ceases to divide.     

A new ballistosporous species of Protostelium

During surveys of the protostelids of the Hawaiian Islands and the South Island of New Zealand, an undescribed species of Protostelium was discovered fruiting on collections of substrates found in several sites on the southern and western parts of the island of Hawaii and from one site near Port Elizabeth, New Zealand. The new species, P. okumukumu, has a sporocarp with a bipartite stalk that supports a single, spherical spore. The basal portion of the stalk is straight and rigid. The upper part of the stalk is a nearly spherical apophysis. The junction between the stalk base and the apophysis is flexible such that the spore and apophysis swing back and forth as a unit. Spores are forcibly discharged from the stalk, and only the straight base of the stalk is left behind. Amoebae typical of the taxon Protostelium germinate from the spores, and when an amoeba differentiates into a prespore cell, it becomes lozenge shaped (nearly ellipsoid) in top view, as is typical for species of Protostelium. This represents the seventh species of protostelids described to have forcible spore discharge, and the possibility of forcible discharge needs to be examined in several other species.     

Electron Microscope Observations on Spore Formation in the True Slime Mold Didymium nigripes

SYNOPSIS. Developing and mature sporangia of the true slime mold Didymium nigripes were studied with the electron microscope to follow the course of spore formation. The sporangium forms from the plasmodium as a protoplasmic bleb which differentiates into a stalk and an apical sphere containing a mass of protoplasm. Nuclei within this protoplasmic mass undergo synchronous division (presumably meiosis). The division spindle forms within the nuclear membrane which is retained intact throughout the division; centrioles have not been observed at the spindle poles. At the same time the nuclei are dividing, the protoplasm cleaves to give ultimately uninucleate spheres—the incipient spores. Capillitial threads come to lie in the furrows created by the cleaving protoplasm. A wall consisting of an inner thick component and an outer thin component forms about each sphere. Cyto-chemical tests suggest that the inner wall of the spore is cellulose-containing and that the outer component might contain chitin.     

SPORE DEVELOPMENT IN THE LIVERWORT RICCARDIA PINGUIS

The ontogeny of spores of the liverwort Riccardia pinguis was studied at the light and electron microscope levels. Three stages of development were arbitrarily defined: spore mother cell (SMC); early tetrad with nonpigmented and unsculptured walls; and mature tetrad with pigmented and sculptured spore walls. The SMC is quadrilobed with a two-layered SMC wall, containing a central nucleus, many chloroplasts, spherosomes, and other organelles. During and following meiosis cell plates form from coalescing Golgi vesicles. These plates by continued coalescence eventually form a septum, completing the tetrad. This septum comprises middle lamella and primexine; within the latter the exine forms. By continued addition of vesicle contents to the septum and dorsal surfaces of the tetrad, the exine (sexine and nexine) and intine layers of the spore wall are laid down. The contents of the vesicles change successively during wall formation, corresponding to the different wall layers being formed. It is concluded that wall formation is under the exclusive control of the spore protoplast, and that the pattern of the mature exine is determined by the primexine. Rearrangement of organelles and other cellular components during sporogenesis is described.     

Euopsis and Harpidium,Genera of the Lichinaceae (Lichenes) with Rostrate Asci*

Based on corresponding ascocarp ontogeny and thallus structure, the genera Euopsis and Harpidium are included in the family Lichinaceae. In the two species of Euopsis, E. granatina and E. pulvinata, the apothecia develop from ascogonia in generative tissue, while in H. rutilans they are pycnoascocarps. In thallus anatomy, the species of Euopsis resemble Pyrenopsis haematopsis and allied species, while H. rutilans corresponds in structure and development of the thallus and apothecia to Pyrenopsis haemaleella (syn. P. sphinctotricha). H. rutilans is the first member of Lichinaceae known to have only a green algal symbiont. In E. granatina, two phycobionts are always present, a species of Gloeocapsa and a chlorococcalean alga. In Euopsis and Harpidium, the ascus wall is composed of an outer, non-expansible and an inner, expansible layer; the latter surrounds the protoplast as an amyloid collar, which expands during spore release into a long, tapering rostrum. In Euopsis, the outer wall layer is strongly amyloid and the upper part separated from the expanded amyloid rostrum by a non-amyloid zone, appearing like a slit in LM studies. The ultrastructure and function of the ascus in E. granatina has been studied in TEM and is interpreted as functionally unitunicate-rostrate. Unitunicate asci with short rostrum are described for P. haemaleella and P. haematopsis.     

Electron microscopy of spore germination and cell wall formation in Mucor rouxii

Summary The fine structure of ungerminated and aerobically germinated sporangiospores of Mucor rouxii was compared. The germination process may be divided into two stages: I, spherical growth; II, emergence of a germ tube. In both stages, germination is growth in its strictest sense with overall increases in cell organelles; e.g., the increase in mitochondria is commensurate with the overall increase in protoplasmic mass. Noticeable changes occurring during germination are the disappearance of electron-dense lipoid bodies, formation of a large central vacuole and, most strikingly, formation of a new cell wall. Unlike many other fungi, M. rouxii does not germinate by converting the spore wall into a vegetative wall. Instead, as in other Mucorales, a vegetative wall is formed de novo under the spore wall during germination stage I. This new wall grows out, rupturing the spore wall, to become the germ tube wall. Associated with the apical wall of the germ tube is an apical corpuscle previously described. The vegetative wall exhibits a nonlayered, uniformly microfibrillar appearance in marked distinction to the spore wall which is triple-layered, with two thin electron dense outer layers, and a thick transparent inner stratum. The lack of continuity between the spore and vegetative walls is correlated with marked differences in wall chemistry previously reported. A separate new wall is also formed under the spore wall during anaerobic germination leading to yeast cell formation. On the other hand, in the development of one vegetative cell from another, such as in the formation of hyphae from yeast cells, the cell wall is structurally continuous. This continuity is correlated with a similarity in chemical composition of the cell wall reported earlier.     

MORPHOLOGY AND LABORATORY CULTIVATION OF ECHINOSTELIUM MINUTUM

Alexopoulos , Constantine J. (State U. Iowa, Iowa City.) Morphology and laboratory cultivation of Echinostelium minutum. Amer. Jour. Bot. 47(1): 37—43. Illus. 1960.—The morphology of the sporangium, spores, swarm cells and Plasmodium of the white form of Echinostelium minutum is described. A peridium is present in the early stages of sporangial formation. It eventually disappears leaving only a small collar at the base of the columella. The structure of the spore wall is unique in this genus. The spore case may be described as consisting of a thin wall with several thickened portions distributed over its surface. These are particularly evident in germinated spores. Spore germination and swarm cells are described for the first time. Swarm cells are biflagellate with two long anterior flagella of nearly equal length. The Plasmodium remains microscopic until fruiting time, when it gives rise to but a single sporangium. The plasmodial protoplast never becomes differentiated into veins but remains more or less homogeneous. It exhibits almost imperceptibly slow, irregular streaming instead of the reversible, rapid, rhythmic motion characteristic of plasmodia of most other Myxomycetes which have been studied. It typifies, therefore, a third type of Plasmodium which may be placed alongside that of the Physarales, and that of Stemonitis flavogenita. The laboratory cultivation of E. minutum from spore to spore on agar media is reported here for the first time.     

The anterior-like cells in Dictyostelium are required for the elevation of the spores during culmination

 Shortly after initiation of Dictyostelium fruiting body formation, prespore cells begin to differentiate into non-motile spores. Although these cells lose their ability to move, they are, nevertheless, elevated to the tip of the stalk. Removal of the amoeboid anterior-like cells, located above the differentiating spores in the developing fruiting body, prevents further spore elevation although the stalk continues to elongate. Furthermore, replacement of the anterior-like cells with anterior-like cells from another fruiting body largely restores the ability to lift the spores to the top of the stalk. However, if amoeboid prestalk cells are used to replace the anterior-like cells, there is no restoration of spore elevation. Finally, when a droplet of mineral oil replaces differentiating spores, it is treated as are the spores: the mineral oil is elevated in the presence of anterior-like cells and becomes arrested on the stalk in the absence of anterior-like cells. Because a similar droplet of mineral oil is totally ignored by slug tissue, it appears that there is a dramatic transformation in the treatment of non-motile matter at this point in Dictyostelium development. Received: 26 January 1998 / Accepted: 27 May 1998     

适用于双向电泳的米曲霉孢子破壁方法研究

为探究米曲霉(Aspergillus oryzae)孢子适用于双向电泳的最佳破壁方法,采用5种不同的破壁方法对米曲霉孢子进行破壁,用血球计数板进行破壁率计算,Bradford方法测定释出的可溶性蛋白含量,并进行双向电泳可行性验证。结果表明,在普通光学显微镜下,破壁后的米曲霉孢子多为碎片,极少数为孢壁内空圆球。5种破壁方法中石英砂研磨+超声、液氮研磨、MP·Fast-prep均质器法在孢子浓度较低(107个/m L)时破壁效果较佳,但是随着孢子浓度的不断提升(109个/m L),只有均质器法能保证较高的破壁率,破壁率高达90%,且适用于双向电泳的蛋白质提取。     

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.     

A DEVELOPMENTAL STUDY OF GUTTULINOPSIS VULGARIS (ACRASIALES)

The cellular slime mold, Guttulinopsis vulgaris E. W. Olive, is a common and widely distributed species. Its myxamoebae have lobose pseudopodia and aggregate, not by streaming, but by movement singly or in small groups into the aggregation center. The resulting shield-shaped pseudoplasmodium gives rise to one (or possibly more on occasion) whitish, simple or compound sorocarp. Sorocarps are 95–465 μ in height, typically with relatively short, stout stalks and with mucilaginous spore heads that measure 75–350 μ broad. The spores are mostly irregular, with flattened or concave sides, and are surrounded by a thin spore wall. They measure 3.8–9.2 μ diam., or 2.6–7.8 × 3.9–9.2 μ. During sorocarp development the myxamoebae become grouped within membranous compartments both in the sorocarp and pseudoplasmodium. In the upper part of the sorocarp they develop almost entirely into spores, while in the lower stalk region of the sorocarp and in the pseudoplasmodium, some myxamoebae develop into spores while others degenerate. The phylogenetic position of Guttulinopsis is unclear, but the genus is probably not closely related to the Dictyosteliaceae. This investigation has been supported by National Science Foundation Grants G–44263 and GB-501. The writer is grateful to Miss Carmen Stoianovitch for her assistance.