THE LABORATORY CULTIVATION OF STEMONITIS

Alexopoulos , Constantine J. (State U. Iowa, Iowa City.) The laboratory cultivation of Stemonitis. Amer. Jour. Bot. 46(2): 140-142. Illus. 1959.—The cultivation of a species of Stemonitis, probably S. flavogenita, in laboratory culture is reported here for the first time. The organism completed its entire life cycle on artificial media from spore to spore, in the presence of contaminating bacteria, in 36 days. The plasmodium, characteristically different from those of the Physarales, is described and illustrated.     

LABORATORY CULTIVATION OF CLASTODERMA DEBARYANUM

Mc Manus , Sister Mary Annunciata , R.S.M. (Mount Mercy Coll., Cedar Rapids, Iowa.) Laboratory cultivation of Clastoderma debaryanum. Amer. Jour. Bot. 48(10): 884–888. Illus. 1961.—Clastoderma debaryanum has been cultivated on laboratory media for the first time and its life cycle studied. Spores germinate to produce a single protoplast which may remain a myxamoeba or may become a swarm cell. The swarm cells fuse in pairs to form a zygote, which grows into a microscopic, spherical plasmodium. The plasmodium never develops a network of veins and shows only irregular streaming. At maturity each plasmodium gives rise to 1 sporangium. There is always a sphere of gelatinous material about 4/5 the distance up the stalk, which appears late in the development of the sporangium, and which dries to become a wrinkled enlargement. The peridium dehisces early, leaving only a few plates attached to some of the peripheral tips of the capillitial twigs, and a peridial collar. The solid capillitium branches from the tip of a very short columella.     

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.     

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.     

SPORE ULTRASTRUCTURE OF THE MYXOMYCETE PHYSARUM POLYCEPHALUM

Spores of the true slime mold Physarum polycephalum were examined at several stages of their development by means of scanning and transmission electron microscopy. The spores were globose, spine-covered structures produced within a sporangium enclosed in a tough, noncellular peridium. Cytologically, the spore represented a typical eukaryotic cell, having discrete organelles similar to spores of other myxomycetes. The presence of dictyosomes, helical filaments, and microbodies in these cells, as well as the further elucidation of the cell wall and the “polysaccharide-containing” areas, represent new contributions to the ultrastructure of the myxomycete spore. Of special interest were observations of metaphase nuclei just prior to spore cleavage, interphase nuclei in young spores, and nuclei in mature spores containing synaptonemal complexes. These observations indicate that in Physarum polycephalum mitosis occurs just prior to spore cleavage, and meiosis takes place after spore cleavage.     

ULTRASTRUCTURAL STUDIES OF IN SITU DEVONIAN SPORES: BARINOPHYTON CITRULLIFORME

Each sporangium in the Upper Devonian taxon Barinophyton citrulliforme contains both microspores and megaspores. Microspores range up to 50 μm in diam and possess a homogeneous sporoderm characterized by an outer separable layer. The sporoderm of the megaspores (up to 900 μm) is constructed of sporopollenin units that are loosely arranged in the outer portion of the wall, and that give the megaspore wall a spongy organization. Ultrastructural evidence suggests that the small spores were not abortive megaspores, but that both spore types were functional. The spores of this plant, as well as other Devonian spores that show less dramatic size differences, are suggested as demonstrating a phase in the evolution of heterospory where sex determination was established in spores within the same sporangium prior to the evolution of micro- and megasporangia.     

狗脊孢壁发育超微结构的研究

鳞毛蕨型孢子类型众多,初步研究表明形态相似的孢子类型其孢壁发育特征存在差异,因此有必要对各代表类群的孢壁发育进行深入地研究。该文利用透射电镜对乌毛蕨科(Blechnaceae)狗脊(Woodwardia japonica)孢壁结构和发育的超微结构进行研究。结果表明:(1)狗脊孢子囊的结构由外向内分别为孢子囊壁细胞、两层绒毡层细胞和孢子母细胞;(2)狗脊孢子具乌毛蕨型(Blechnoid type)外壁,表面光滑,由两层构成,裂缝区域具辐射状的槽;(3)周壁属于空心型(cavity type),由四层构成,从内向外分别为P1、P2、P3和P4层,前三层叠合在一起,层间有不同程度的空隙,P4层与前三层之间具有明显而连续的空腔,并隆起形成片状褶皱纹饰;(4)有小球体和小杆共同参与孢子周壁的形成,周壁部分或全部来源于孢子囊壁细胞。综上所述,狗脊孢子与同属于鳞毛蕨型的贯众(Cyrtomium fortunei)和朝鲜介蕨(Dryoathyrium coreanum)孢壁的发育在周壁结构、周壁各层的发育顺序、周壁来源和参与成壁的特征物质等方面存在差异。该研究有利于进一步理解蕨类植物孢壁所蕴含的分类和演化上的科学意义和价值。     

A study on the amoebo-flagellate transformation in the slime moldEchinostelium minutum de Bary

Summary The structure and behavior of the non-flagellate and flagellate phases of the slime moldEchinostelium minutum de Bary are here described from living cultures examined with phasecontrast microscopy. The ultrastructure of the myxamoebal and swarm cell phases was studied in sectioned material fixed sequentially with glutaraldehyde-acrolein and OsO₄.The nearly spherical myxamoeba has two pairs of juxtanuclear centrioles with associated microtubular arrays. During the amoebo-flagellate transformation each pair of centrioles assumes an anterior position in the cell and becomes arranged at right angles to one another within a cone of microtubules. This microtubular cytoskeleton extending under the plasmalemma establishes the twisted, narrowly ovoid form of the swarm cell. Each centriole functions potentially as a basal body. When transformed in phosphate buffer pH 6.8 at 12 °C and a cell concentration of 5×10⁵/ml, the myxamoebae develop 1 to 8 flagella. The average number of flagella per swarm cell is 2.7. After approximately 2 hours the swarm cells begin to revert to myxamoebae by resorption of their flagella. The phylogenetic implications of these light and ultrastructural observations are discussed with regard to possible evolutionary relationships between theProtostelia andMyxomycetes.     

Sporulation of protoplasts

Vegetative cells ofBacillus megaterium formed protoplasts in a sucrose-stabilized medium under the influence of lysozyme. The protoplasts sporulated during subsequent incubation. The morphological properties, germination, resistance to u. v. irradiation and thermo-resistance of protoplast spores were the same as with normal cells. It thus appears that in the later sporulation stages the spore formation occurs, without participation of the sporangium cell wall.     

Spore Dormancy Mutants of Phycomyces

Rivero, F., and Cerdá-Olmedo, E. 1994. Spore dormancy mutants of Phycomyces. Experimental Mycology 18, 221-229. The spores of the Zygomycete Phycomyces blakesleeanus are called dormant because few of them germinate when placed in a medium that sustains mycelial growth and development. Nearly all the spores germinate after activation, that is, exposure to heat or certain chemicals. We have looked for mutants whose spores would not need activation. Nine mutants formed authentic, but transient spores, which germinated spontaneously in the sporangium. Mutant mycelia had lower alcohol and aldehyde dehydrogenase activities and less glycogen than wild-type mycelia. The spontaneous germination and the metabolic alterations are attributed to the same recessive mutations. No differences were found between mutants and wild type in the cyclic AMP and fructose 2,6-bisphosphate concentrations in immature sporangia and the trehalase activity in the mycelia. In another mutant the spore primordia did not form spores, but remained viable for some time in the sporangium. The mutants were difficult to keep in the laboratory (except as lyophils); this stresses the importance of preventing spore germination in the sporangium.     

紫萁孢子囊发育中多糖和脂滴的细胞化学观察

采用光镜、透射电镜和细胞化学技术,对紫萁孢子囊发育过程中孢壁的超微结构和孢子囊内多糖和脂滴的分布及其动态变化进行研究,以探讨紫萁孢子囊发育过程中多糖和脂滴的代谢特征,为蕨类孢子发生的研究提供基础资料。结果表明:(1)紫萁孢子囊由1层囊壁细胞、2层绒毡层和产孢组织构成。(2)紫萁孢子壁由发达而分2层的外壁(外壁内层和外壁外层)和薄的不连续的周壁构成,由外壁形成棒状纹饰的轮廓;孢子外壁内层由多糖类物质构成,外壁外层和周壁均含有脂类物质。(3)在紫萁孢原细胞中观察到少量脂滴;随着紫萁孢壁的形成,囊壁细胞中淀粉粒的大小逐渐变小、数目先增加后减少,它们转运到内层绒毡层原生质团并转化为孢粉素前体物质,再穿过原生质团内膜表面进入囊腔,成为孢粉素团块或以小球形式填加到孢子表面形成孢壁。(4)紫萁孢子囊将多糖类营养物质转化为脂类,以脂滴的形式储藏在孢子中。     

Two cryptospore-bearing land plants from the Lower Devonian (Lochkovian) of the Welsh Borderland

Two examples of fragmentary, coalified plant fossils with cellular preservation and in situ spores are described using scanning and transmission electron microscopy (SEM and TEM), from a Lower Devonian (Lochkovian) locality in the Welsh Borderland. A sporophyte which produced dyads of ?Cymbohilates of. horridus is unique in that stomata are numerous on the supporting axis, and are comparable with those described from contemporary vascular plant remains. The in situ dyads possess a bilayered exospore wall, with an outer exosporal envelope present over the distal faces. A fusiform sporangium, with externally smooth epidermis, contains specimens of the tetrad Velatitetras sp. Each tetrad is encompassed by a laevigate, folded, exosporal envelope of uniform thickness, which contains a layer of regular voids. Spores within the tetrads are ultrastructurally bilayered, with a complex, digitate outer margin presumably representing spore wall ornamentation. Neither the tetrads nor dyads reported in this paper are lamellate in ultrastructure. The combination of stomata, branching and dyads in the same sporophyte holds significance for the understanding of cryptospore affinities.     

THE LABORATORY CULTIVATION OF PHYSARUM FLAVICOMUM

Gray , William D. (Ohio State U., Columbus.) The laboratory cultivation of Physarum flavicomum. Amer. Jour. Bot. 48(3): 242–243. 1961.—The laboratory cultivation of P. flavicomum on artificial medium is reported here for the first time. The plasmodium of this species can be easily cultivated on dry rolled oats in a moist chamber in the same manner as plasmodia of P. polycephalum, or on 3% rolled-oats agar. Plasmodia form sclerotia from which new cultures can later be started or they can be obtained by sowing spores on rolled oats agar. Like P. polycephalum, the plasmodia of P. flavicomum must be illuminated with visible light in order to induce the onset of the sporulating stage. Sporangia which develop in culture vary most noticeably with respect to the type of peridium. The nature of the peridium is largely determined by whether the sporangium develops under moist or dry conditions; dry conditions favor the formation of a lime-encrusted peridium, while humid conditions favor the development of a thin, limeless peridium which has a metallic luster.     

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。     

Protostelids from German Beech forests

A survey for protostelids from old-growth beech forests of northeastern Germany resulted in 14 species found in both ground and aerial litter, constituting the first survey of these organisms from Central Europe. Additionally the myxomycete Echinostelium bisporum was recorded as new for Germany. A detailed investigation of randomly sampled dead Fagus leaves (16 leaves on ground, 16 aerial leaves, three replicate cultures per leaf) resulted in 7 species; a hyperbolic regression of the respective species accumulation curve gives the theoretical value of 8.3 species. Protostelids occurred especially at aerial leaves with a high frequency (0.94). A program simulating random spore hits was written to calculate the minimum spore fallout necessary to explain this frequency. The resulting average value of about 3 spores per leaf can well be matched by the potential spore productivity estimated to be about 1000 spores per leaf for the most common species, Protostelium mycophaga.     

A light microscopical investigation of amoebal and plasmodial mitosis in the MyxomyceteEchinostelium minutum

Summary Light microscopical observations on mitosis in living material of the amoebal and plasmodial phases of the MyxomyceteEchinostelium minutum de Bary (orderEchinosteliales) are reported for the first time. The uninucleate amoebal cell undergoes centric, open spindle mitosis whereas the multinucleate plasmodium exhibits acentric, closed spindle mitosis.     

Development of the Mycetozoan Echinosteliopsis oligospora*

SYNOPSIS. The mycetozoan genus Echinosteliopsis, resembling the myxomycete Echinostelium in some of its features, is described. The single species, E. oligospora Reinhardt & Olive, forms small sporocarps which consist of a basal disk, stalk and a sporangium with only 1–8 spores. Spores form progressively, not simultaneously, by segmentation. The spores germinate to release non-flagellate amebae which, in liquid, assume a characteristic broad, fan shape. Each ameba has one or more nuclei. The nucleus is distinctive because of refractile, globular to elongate peripheral bodies which cytochemical tests indicate to be primarily RNA. At the time of nuclear division the characteristic RNA bodies disappear and, as observed with the phase microscope and in stained preparations, optically dense material accumulates in the middle area of the nucleus. Threads, either a spindle or actual chromatin, can be seen attached to the nuclear membrane. The threads separate to opposite poles as the nucleus elongates. During this division process the nuclear membrane apparently remains intact. Synchronous binucleate divisions, as well as a tripolar nuclear division, have been observed. Uninucleate and synchronous binucleate divisions may or may not be followed by cytokinesis. The absence of cell division after nuclear division leads to the production of cells with varying numbers of nuclei. Nuclear divisions in early sporangial stages and in spores have not been observed. The spores are uni- to multinucleate. In 8-spored sporangia and in most 4-spored sporangia there is a characteristic small “stalk spore” at the apex of the stalk. The stalk spore germinates slowly, if at all, but the larger spores germinate readily. No evidence of a sexual process has been found.     

SPORE MORPHOLOGY IN THE CYATHEACEAE. I. THE PERINE AND SPORANGIAL CAPACITY: GENERAL CONSIDERATIONS

The literature on cyatheaceous spore morphology relative to the presence of a perine layer is reviewed, and evidence based on a sodium-hydroxide assay is presented indicating that the outer scultpine layer in certain cyatheaceous spores is perine. Perine so defined characterizes Metaxya, paleotropical and certain neotropical species of Sphaeropteris, nearly all species of Alsophila, all species of Nephelea, and certain species of Trichipteris and Cyathea. It is lacking in Lophosoria, many species of Trichipteris and Cyathea, and all species of Cnemidaria. Two major patterns of spore number per sporangium in the family are reported. Lophosoria, Sphaeropteris, Trichipteris, Cyathea, Cnemidaria, and probably Metaxya are characterized by 64-spored sporangia, whereas most species of Alsophila and all species of Nephelea are characterized by 16-spored sporangia. The congruence of this generic distribution of sporangial-capacity types with Tryon's phyletic arrangement of cyatheaceous genera supports the naturalness of his system. The intrasporangial germination of spores retained in dehisced and dispersed sporangia supports the suggestion that decreased spore number per sporangium in Alsophila and Nephelea may relate to the role of the sporangia as dispersal units. The decreased number of spores per sporangium is associated with a trend toward increase in the number of sporangia per sores, with the highest known count approaching 1000 sporangia per sorus. The Alsophila-Nephelea evolutionary line has probably not been ancestral in the phylogeny of the more advanced groups of ferns.     

中华水韭孢子囊发育研究

采用半薄切片法,连续观察了极度濒危级(CR)植物中华水韭大小孢子囊的发育过程,以期从无性生殖的角度,为探讨其濒危原因提供直观可靠的理论根据。结果显示:(1)中华水韭的大小孢子叶相间排列,无混生孢子囊。(2)隔丝为孢子供给营养,其体积直接影响孢子的大小、产量和育性。(3)大小孢子囊都近半数败育,小孢子囊为整齐发育,大孢子囊为不整齐发育。(4)大小孢子囊均无柄,且都不存在开裂结构,只有孢子囊壁腐烂后才能散播孢子。研究认为,中华水韭的濒危与孢子囊的发育特征密切相关,孢子囊的高频率败育、没有开裂结构以及对环境的依赖,是造成中华水韭濒危的重要因素之一;通过与近缘类群孢子囊的比较,发现仅水韭孢子的散播借助外力,对生境要求较高,即验证了水韭古老的系统学地位,同时说明水韭更具有监测生境地区环境指标的能力。     

Sporangium Exposure and Spore Release in the Peruvian Maidenhair Fern (Adiantum peruvianum,Pteridaceae)

We investigated the different processes involved in spore liberation in the polypod fern Adiantum peruvianum (Pteridaceae). Sporangia are being produced on the undersides of so-called false indusia, which are situated at the abaxial surface of the pinnule margins, and become exposed by a desiccation-induced movement of these pinnule flaps. The complex folding kinematics and functional morphology of false indusia are being described, and we discuss scenarios of movement initiation and passive hydraulic actuation of these structures. High-speed cinematography allowed for analyses of fast sporangium motion and for tracking ejected spores. Separation and liberation of spores from the sporangia are induced by relaxation of the annulus (the ‘throwing arm’ of the sporangium catapult) and conservation of momentum generated during this process, which leads to sporangium bouncing. The ultra-lightweight spores travel through air with a maximum velocity of ~5 m s^-1, and a launch acceleration of ~6300g is measured. In some cases, the whole sporangium, or parts of it, together with contained spores break away from the false indusium and are shed as a whole. Also, spores can stick together and form spore clumps. Both findings are discussed in the context of wind dispersal.