DEVELOPMENT OF MUCILAGINOUS SURFACES IN EUGLENOIDS. I. STALK MORPHOLOGY OF COLACIUM MUCRONATUM1

The envelope and stalk of Colacium mucronatum Bourr. & Chad, were examined in living cells with light microscopy and in fixed preparations with scanning electron microscopy using critically point dried (CPD) and freeze dried (FD) preparations. The envelope of palmelloid cells is formed over the entire cell surface by many individual strands attached at right angles to areas of articulation of the pellicular strips. Strands were observed to anastomose on the posterior tip of otherwise naked cells. Stalks of living cells in India ink preparations had an optically dark inner core with a lighter outer sheath. In FD stalks a definite inner core was not evident, whereas CPD stalks had an outer surface composed of thick strands which may be the collapsed and aggregated strands of the FD stalks. In both there was also an amorphous matrix. The stalk forms from the aggregation of many strands from the anterior cell tip back to a point encompassing the cell surface anterior to a cross section of the tip 9 μm diam. The outer surface of the stalk comes from the pellicular surface joining that area and the core from the cell tip in the area of the canal opening. Any possible participation of the inner canal surface in stalk formation could not be determined because of the great density of the mucilage at the cell-tip/stalk junction.     

The Fine Structure of the Stalk of the Pentacrinoid Larva of a Feather Star,Comanthus japonica (Echinodermata: Crinoidea)

The stalk of the pentacrinoid larva of a feather star (Comanthus japonica) is described for the first time by transmission electron microscopy. One end of the stalk bears the calyx and the other end is cemented to the substrate by attachment cement consisting of a meshwork of 5 nm filaments. The stalk is supported by a scries of skeletal ossicles pierced by a central canal: short intercolumnal ligaments connect adjacent skeletal ossicles and central through-going ligaments run the length of the central canal. At the end of the stalk nearest the calyx, the chambered organ and the closely associated axial organ are histologically similar to those of adult crinoids. Presumed neurosecretory neurons are associated with the intercolumnal ligaments, and the following kinds of nerves run down the central canal: (1) a large stalk nerve in each of the five interradii; (2) smaller coelomic nerves in each of the five radii in association with the epithelium of tubular aboral extensions of the chambered organ; (3) a very small nerve associated with the aboral extension of the axial organ in the stalk axis. This axial organ extension is surrounded by a haemal channel. Because of the small size of the stalk, none of the nerves or the haemal channel were described in previous light microscopic studies. The discussion gives special attention to the controversial motility of the pentacrinoid stalk.     

THE MORPHOLOGY AND DEVELOPMENT OF SOME PROMINENTLY STALKED SOUTHERN AUSTRALIAN HALYMENIACEAE (CRYPTONEMIALES,RHODOPHYTA). I. CRYPTONEMIA KALLYMENIOIDES (HARVEY) KRAFT COMB. NOV. AND C. UNDULATA SONDER1

Several southern Australian red algae of the family Halymeniaceae (Cryptonemiales) are differentiated into hard, massive stalks and considerably softer laminar blades or phyllodes. The taxonomy, morphology and pit-connection ultrastructure of one such species, Cryptonemia kallymenioides (Harvey) Kraft comb. nov., are compared to C. undulata Sonder, which lacks massive stalks. In both species there is extensive periodic secondary cortication of the stalks, resulting in the formation of distinct “growth rings.” The blades of C. kallymenioides appear to be seasonal and its stalks perennial, while plants of C. undulata are apparently perennial but shorter lived than C. kallymenioides. As a result, stalks in the latter can reach 2–3 cm in diameter with up to 18 growth rings, compared to the 1–2 mm diameters and up to 6 rings within the stalks of C. undulata. Heavy secondary thickening of cortical cell walls occurs in both species and confers a “woody” texture to the stalks of C. kallymeniodes. Regardless of the large differences in average stalk diameters between the two species, the pit-connection ultrastructure from cortex to medulla shows much the same sequence of morphological modification. Pit-connections are standard red algal structures in the outer cortex, but become increasingly convoluted on the membrane-bound surfaces abutting cytoplasm and develop wider apertures and less dense cores with increasing distance from the stalk surface. In occasional medullary cells of C. kallymenioides, the cytoplasm disintegrates, leaving cell walls and pit-connections to play an apparently structural role which has not been reported in other red algae. It is suggested that the increase in aperture size and surface areas of pit-connections is compatible with their playing a role in the intercellular transport of solutes towards the inner cell layers which may, in C. kallymenioides, lie many millimeters distant.     

Secretory organelle biogenesis: Trichocyst formation in a ciliated protozoan

Summary— By classical electron microscopy and immunoelectron microscopy, the biogenesis of trichocyst secretory granules has been followed in the ciliated protozoan Pseudomicrothorax dubius. The very early pre-trichocysts form by fusion of bristle-coated, electron-dense vesicles (dense vesicles) with electron-translucent vesicles (clear vesicles), both of which originate in a well-developed trans-Golgi network (TGN). The pre-trichocyst grows by further fusion with dense and clear vesicles as well as with other pre-trichocysts until it reaches its maximum diameter of about 2 μm. Dense and clear vesicle formation from the TGN has been followed, and the fusion sequence of dense vesicles with the pre-trichocyst has been documented. The contents of the dense vesicles are the precursors of the trichocyst tip, which is composed of four arm-like rods, whereas the shaft precursors are supplied by the clear vesicles. The first evidence of trichocyst shaft formation is the appearance of a paracrystalline, dense core condensation center in the pre-trichocyst. Following shaft formation, the trichocyst tip forms by fusion and condensation of the dense arm precursors along each of the four sides of the shaft. Docking of the fully formed trichocyst in the cell cortex is described. Pre-trichocyst biogenesis in cells grown with and without Se is compared.     

Differential treatment ofAcetabularia with cytochalasin B and N-Ethylmaleimide with special reference to their effects on cytoplasmic streaming

Summary Cytoplasmic streaming in the stalk ofAcetabularia, ryukyuensis at the vegetative stage was reversibly inhibited by cytochalasin B (cB) of 50 g/ml and irreversibly by N-Ethylmaleimide (NEM) above concentrations of 0.25 mM.After the endoplasm and the chloroplasts were pushed forward one end of the stalk by gentle centrifugation at about 500 × g for 3 minutes, numerous ectoplasmic striations remainedin situ in the stalk cortex. The striations ran in parallel with the longitudinal axis of the stalk at unequal intervals. The endoplasm streamed back only along these striations.By combining centrifugation and a double chamber technique, the endoplasm and the cortex of the stalk were treated separately with CB or NEM. CB treatment of the cortex arrested streaming; when treatment was restricted to the endoplasm, streaming continued at an normal rate. NEM treatment restricted to the cortex permitted normal streaming rates. Treatment restricted to the moving endoplasm inhibited streaming.These results suggest that microfilaments and a moiety, possibly myosin, play an active role in the streaming. Microfilaments must reside in the cortex, especially in the ectoplasmic striations, while the putative myosin must reside in the moving endoplasm.     

DEVELOPMENT AND DISTRIBUTION OF MUCILAGE CANALS IN LYCOPODIUM

Two distinct types of mucilage canals are found in Lycopodium. One type, the veinal canal, is found in both sporophylls and in vegetative leaves, and is always in close proximity to the leaf trace. The other, the basal canal, is restricted to the strobilus where it forms a complex and extensive mucilaginous cylinder in the outer cortex and extends into the base of the sporophylls. Protruding secretory cells are formed in both types during a lysigenous developmental process. The occurrence of these two types of canals correlates with the three subgenera. Urostachys which lacks canals, Lepidotis which possesses both veinal and basal canals, and Lycopodium which possesses only basal canals.     

THE LEAF OF CALYCADENIA AND ITS GLANDULAR APPENDAGES

Carlquist , Sherwin . (Rancho Santa Ana Botanic Garden, Claremont, Calif.) The leaf of Calycadenia and its glandular appendages. Amer. Jour. Bot. 46(2) : 70-80. Illus. 1959.—Large tack-shaped glands are characteristic of the leaves of Calycadenia which are associated with the inflorescence. These glands may be divided into those which are terminal on leaves and those which occur laterally on the surface of the leaf. Lateral glands show stages early in their development which are identical with those of simpler trichomes of Madinae. Terminal glands, which possess more vascularization of the stalk, show a more modified form of development. Vascularization is not derived from protoderm, but from more deeply-seated cells. These cells are included in a zone of elongation which forms the stalk. Vascular bundles may extend to the base of glands which lack vascularization in their stalks. Tack-shaped glands are considered an advanced form of trichome in which internal tissues of the leaf are involved. Within the genus Calycadenia, ontogenetic and comparative studies suggest that the following characters are advanced: reduction to a single terminal gland, “inrolling” of margins to form a cylinder of bundles, concomitant with a central core of fibers or a pectic channel. Systematic distribution of gland occurrence and of types of foliar structure are given.     

ULTRASTRUCTURE OF TETRASPOROGENESIS IN PALMARIA PALMATA (RHODOPHYTA)1

The tetrasporangial initial in Palmaria palmata (L.) O. Kuntze (formerly Rhodymenia palmata (L.) Greville) arises from a cortex cell which enlarges and deposits a protein-rich wall layer. This cell undergoes mitosis to form a tetrasporocyte and a stalk cell. Synaptonemal complexes are formed in the sporocyte nucleus while in the cytoplasm floridean starch is deposited in association with ER or with particles presumed to be ribosomes. Microbody-like structures become numerous between the nuclear envelope and perinuclear ER, and clusters of non-membranous, spherical structures also are associated with the nucleus. Chromatin condensation is reversed following pachytene and a prolonged diffuse stage ensues, when dictyosomes and ER produce vesicles which deposit mucilage rich in sulfated and acidic polysaccharides around the tetrasporocyte. A conspicuous lenticular thickening of the mucilage sheath develops at the apical end of the sporangium. Dictyosomes are frequently associated with mitochondria which may be associated with chloroplasts. Following nuclear divisions the tetrasporocyte is cleaved into four spores by sequentially initiated, but simultaneously completed periclinal and anticlinal furrows. When mucilage deposition ceases, the dictyosomes begin to produce vesicles with glycoprotein-rich contents. These vesicles are abundant in released tetraspores, and they probably contain adhesive material aiding in the attachment of the liberated spores.     

SECRETORY CELLS OF LILY PISTILS. I. FINE STRUCTURE AND FUNCTION

The fine structure of the cells which line the canal of Lilium longiflorum pistils confirms the secretory function which has been ascribed to them. The cells differ in structure from the secretory cells which cover the stigma surface and are therefore referred to as canal cells rather than stigmatoid cells. Their most striking feature is an elaborate wall, 8–14 μ in maximum depth, on the side facing the canal, which with associated structures, we term the secretion zone. Pollination, which triggers chemotropic activity in the style and secretory activity in the canal cells, is not correlated with marked changes in the fine structure of the canal cells. The canal cells appear to fit well into that category which Gunning and Pate have termed “transfer cells.”     

Austropeltum glareosum gen. et sp. nov., a New Lichen from Mountain Plateaux in Tasmania and New Zealand*

Austropeltum gen. nov. is described as a monotypic genus of the family Stereocaulaceae. It differs from other genera of the family by a sqamulose habit. The single species, Austropeltum glareosum sp. nov. is characterized by an heteromerous thallus with a thick, heavily gelatinized upper cortex, and stalked, lecideine, glomerate apothecia. The apothecial stalk is a pseudopodetium, which is delimited from the subhymenium by a pigmented boundary tissue. Asci are eight-spored and possess an amyloid tube-structure; the simple ascospores are fusiform and colourless. Filiform conidia are produced laterally and terminally in marginal pycnidia. A. glareosum grows over gritty soil on exposed mountain summits and plateaux in New Zealand and Tasmania.     

Ultrastructure of the larval compound setae of the polychaete Nereis vexillosa grube

Larval compound (jointed) setae of the polychaete Nereis vexillosa Grube were examined by scanning and transmission electron microscopy and by polarization microscopy. Long-bladed spinigers and short-bladed falcigers are described. The proximal shaft of each of these types of setae flares distally into a serrated collar and encloses the proximal end of a toothed blade. The collar projects on one side as a boss. The blade and the cortex of the shaft have longitudinal channels. A large excentric cavity in the shaft (the medullary channel) contains a loose meshwork of trabeculae. In the distal part of the shaft these trabeculae are aggregated into diaphragms. The seta is invested with an electron dense layer of enamel. Juvenile setae contain both chitin and protein. With respect to the long axis of the seta, the blade and the cortex of the shaft are positively birefringent and the medullary diaphragms are negatively birefringent. KOH extraction renders the setae negative to a test for protein and reverses the sign of birefringence of the cortical material of the shaft.     

Bravohollisia Bychowsky & Nagibina, 1970 and Caballeria Bychowsky & Nagibina, 1970 (Monogenea: Ancyrocephalidae) from Pomadasys hasta (Bloch) (Pomadasyidae), with the description of a new attachment mechanism

Nine species of monogeneans were collected from the fish Pomadasys hasta (Bloch): Bravohollisia magna Bychowsky & Nagibina, 1970; B. rosetta n. sp.; B. reticulata n. sp.; B. gussevi n. sp.; B. kritskyi n. sp.; Caballeria pedunculata Bychowsky & Nagibina, 1970; C. robusta Bychowsky & Nagibina, 1970; C. liewi n. sp.; and C. intermedius n. sp. The anchors of these species contain a canal beginning in the shaft and ending near the point; ducts of haptoral glands enter the canal in the shaft and are associated with net-like structures near the anchor tip. The net-like structures of Bravohollisia spp. are extensive, while those of Caballeria spp. are smaller and more compact. These nets probably assist the monogeneans in attaching to the gill filaments. Caballeria spp. possess four pairs of extensible haptoral digits. Species of Bravohollisia and Caballeria posses two seminal vesicles.     

BRIEF INCUBATION OF GAMETANGIA-BEARING CAPS IN ANTIBIOTICS ELIMINATES BRANCHING IN PROGENY OF ACETABULARIA ACETABULUM (CHLOROPHYTA)1

Branching of the stalk of Acetabularia acetabulum L. (Silva) was investigated by inbreeding and by a brief treatment of gametangia with a variety of antibiotics. The position of the branch along the stalk varied, implying that branching was not restricted to any one time in development (base is oldest and apex is youngest). The branching phenotype was not inherited in Mendelian fashion. Although three microscopic structures (“bubbles,”“pustules,” and “scars”) occurred on the stalks of cells that had branched, these structures were not statistically correlated with branching in the population (n=699 cells). However, brief treatment of gametangia with a new antibiotic mixture did eliminate all macro- and microscopic structures associated with branching of the stalk in the subsequent generation. We could not fulfill Koch's postulates or provide clear evidence for the pathogenic nature of cell branching. Our brief antibiotic treatment of gametangaa of Acetabularia acetabulum was rapid, had no adverse effects, and virtually eliminated branching (and any potential pathogens) from laboratory cultures in the subsequent generations. Our method allows biochemical and molecular analyses to proceed uncomplicated by the possible presence of other organisms and provides a clean baseline for the future selection of mutations that may induce heritable branching.     

Surface porous fibre-reinforced composite bulk bone substitute

The aim of this study was to describe and evaluate the significance of a porous surface with bioactive glass granules (S53P4) covering an artificial bulk material based on polymethylmetacrylate (PMMA) and fibre-reinforced composite (FRC) technology. Effort was focused particularly on characters of the porous surface and biomechanical properties of the material in vitro, and test in vivo the implant in reconstruction in an experimental long bone segment defect model. The defect, 10 mm in length, created in the shaft of rabbit tibia, was reconstructed by the implant and fixed by intramedullary K-wires. The implant was incorporated within 4 weeks by new bone growth from the host bone covering particularly its posterior surface and cortex/implant junctions with bridging trabecular bone. Later, at 8 weeks, new bone was found also at the cortex/implant interface and in the medullary canal of the implant. Histometric measurements revealed direct bone/implant surface contact in 34% at the interface. Bioactive glass granules in the porous surface evoked the most direct contact with bone. The implants manufactured from PMMA only served as a control group, and showed significantly lower osteoconductive properties. Biomechanical measurements in vitro of fibre-reinforced PMMA specimens revealed values for bending strength and the flexural modulus to match them to human bone. This artificial bulk bone material based on PMMA/FRC technology seems to have proposing properties to be used as a bone substitute on load-bearing conditions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Correlative 3D‐imaging of Pipistrellus penis micromorphology: Validating quantitative microCT images with undecalcified serial ground section histomorphology

Detailed knowledge of histomorphology is a prerequisite for the understanding of function, variation, and development. In bats, as in other mammals, penis and baculum morphology are important in species discrimination and phylogenetic studies. In this study, nondestructive 3D‐microtomographic (microCT, µCT) images of bacula and iodine‐stained penes of Pipistrellus pipistrellus were correlated with light microscopic images from undecalcified surface‐stained ground sections of three of these penes of P. pipistrellus (1 juvenile). The results were then compared with µCT‐images of bacula of P. pygmaeus, P. hanaki, and P. nathusii. The Y‐shaped baculum in all studied Pipistrellus species has a proximal base with two club‐shaped branches, a long slender shaft, and a forked distal tip. The branches contain a medullary cavity of variable size, which tapers into a central canal of variable length in the proximal baculum shaft. Both are surrounded by a lamellar and a woven bone layer and contain fatty marrow and blood vessels. The distal shaft consists of woven bone only, without a vascular canal. The proximal ends of the branches are connected with the tunica albuginea of the corpora cavernosa via entheses. In the penis shaft, the corpus spongiosum‐surrounded urethra lies in a ventral grove of the corpora cavernosa, and continues in the glans under the baculum. The glans penis predominantly comprises an enlarged corpus spongiosum, which surrounds urethra and baculum. In the 12 studied juvenile and subadult P. pipistrellus specimens the proximal branches of the baculum were shorter and without marrow cavity, while shaft and distal tip appeared already fully developed. The present combination with light microscopic images from one species enabled a more reliable interpretation of histomorphological structures in the µCT‐images from all four Pipistrellus species. J. Morphol. 276:695–706, 2015. © 2015 Wiley Periodicals, Inc.     

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.     

The cytoskeleton in the unique cell reproduction by conidiogenesis of the long-neck yeast Fellomyces (Sterigmatomyces) fuzhouensis

Summary. The morphology of conidiogenesis and associated changes in microtubules, actin distribution and ultrastructure were studied in the basidiomycetous yeast Fellomyces fuzhouensis by phase-contrast, fluorescence, and electron microscopy. The interphase cell showed a central nucleus with randomly distributed bundles of microtubules and actin, and actin patches in the cortex. The conidiogenous mother cell developed a slender projection, or stalk, that contained cytoplasmic microtubules and actin cables stretched parallel to the longitudinal axis and actin patches accumulated in the tip. The conidium was produced on this stalk. It contained dispersed cytoplasmic microtubules, actin cables, and patches concentrated in the cortex. Before mitosis, the nucleus migrated through the stalk into the conidium and cytoplasmic microtubules were replaced by a spindle. Mitosis started in the conidium, and one daughter nucleus then returned to the mother via an eccentrically elongated spindle. The cytoplasmic microtubules reappeared after mitosis. A strong fluorescence indicating accumulated actin appeared at the base of the conidium, where the cytoplasm cleaved eccentrically. Actin patches then moved from the stalk together with the retracting cytoplasm to the mother and conidium. No septum was detected in the long neck by electron microscopy, only a small amount of fine “wall material” between the conidium and mother cell. Both cells developed a new wall layer, separating them from the empty neck. The mature conidium disconnected from the empty neck at the end-break, which remained on the mother as a tubular outgrowth. Asexual reproduction by conidiogenesis in the long-neck yeast F. fuzhouensis has unique features distinguishing it from known asexual forms of reproduction in the budding and fission yeasts. Fellomyces fuzhouensis develops a unique long and narrow neck during conidiogenesis, through which the nucleus must migrate into the conidium for eccentric mitosis. This is followed by eccentric cytokinesis. We found neither an actin cytokinetic ring nor a septum in the long neck, from which cytoplasm retracted back to mother cell after cytokinesis. Both the conidium and mother were separated from the empty neck by the development of a new lateral wall (initiated as a wall plug). The cytoskeleton is clearly involved in all these processes. Correspondence and reprints: Department of Biology, Faculty of Medicine, Masaryk University, Tomešova 12, 602 00 Brno, Czech Republic.     

The spermatogenous body cell of the coniferPicea abies (Norway spruce) contains actin microfilaments

Summary In conifer pollen, the generative cell divides into a sterile stalk cell and a body cell, which subsequently divides to produce two sperm. InPicea abies (Norway spruce, Pinaceae) this spermatogenous body cell contains actin microfilaments. Microfilament bundles follow the spherical contour of the body cell within the cell cortex, and also traverse the cytoplasm and enmesh amyloplasts and other organelles. In addition, microfilaments are associated with the surface of the body cell nucleus. The sterile stalk cell also contains microfilament bundles in the cytoplasm, around organelles, and along the nuclear surface. Within the pollen grain, microfilament bundles traverse the vegetative-cell cytoplasm and are enriched in a webbed cage which surrounds the body cell. Microfilaments were identified with rhodamine-phalloidin and with indirect immunofluo-rescence using a monoclonal antibody to actin. The majority of evidence in literature suggests that the spermatogenous generative cell in angiosperms does not contain actin microfilaments, so the presence of microfilaments within the spermatogenous body cell inP. abies appears to be a fundamental difference in sexual reproduction between conifers and angiosperms.     

THE FINE STRUCTURE OF STALKED BACTERIA BELONGING TO THE FAMILY CAULOBACTERACEAE

The fine structure of a series of stalked bacteria belonging to the genera Caulobacter and Asticcacaulis has been examined in thin sections. The cell wall has the multilayered structure typical of many Gram-negative bacteria, and continues without interruption throughout the length of the stalk. The core of the stalk, continuous with the cytoplasmic region of the cell, is enclosed in an extension of the cell membrane, and contains a system of internal membranes: it is devoid of ribosomes and nucleoplasm. A membranous organelle occupies the juncture of stalk and cell, separating the ribosomal region from the core of the stalk. Typical mesosomes also occur in the cell, being particularly frequent at the plane of division. The secreted holdfast is located at the tip of the stalk in Caulobacter, and at the pole of the cell adjacent to the stalk in Asticcacaulis.     

Myoanatomy of <Emphasis Type="Italic">Barentsia discreta</Emphasis> (Busk, 1886) (Kamptozoa: Coloniales)

The anatomy of the muscular system of Barentsia discreta (Kamptozoa) was studied by confocal laser scanning and transmission electron microscopy. The calyx musculature, muscles associated with the digestive tract, atrial ring muscles, and tentacle muscles are described. The structure of the muscular bulbus located in the upper part of the stalk and the muscle base of the stalk were examined. The middle part of the stalk and the stolon lack musculature. The structure of the star-cell complex lying at the boundary of the stalk and calyx was examined in detail. Emschermann’s (1969) opinion was confirmed that the star-cell complex performs the function of a heart, providing the transport of substances from the calyx to the stalk and stolon. The general plan of the muscle arrangement is similar in all Kamptozoa; it consists of central muscles of the calyx, atrial ring muscles, tentacle muscles, and muscles associated with the digestive tract. Oral, lateral, and aboral muscles extending from the stalk into the calyx, which were described for solitary forms, are lacking in the calyx of colonial B. discreta. The calyx of B. discreta is separated from the stalk by a septum, through which muscles do not penetrate from the stalk.