Virus-like Particles in the Calyx of Phanerotoma flavitestacea (Hymenoptera: Braconidae) and Their Transfer Into Host Tissues

Virus-like particles occur in the calyx fluid and cells of the braconid parasitoid, Phanerotoma flavitestacea Fischer. These particles occur in the cytoplasm, nucleus, and cytoplasmic vacuoles of the calyx wall cells as well as in the lumen of the calyx. They were subsequently transferred to the host, Paramyelois transitella (Walker), during oviposition of the parasitoid. Within 2 hours after the wasps had oviposited, similar virus-like particles were found in the host in the cytoplasm of developing muscle tissue, fat body, and nerve tissue. Such particles were absent in unparasitized hosts. The significance of these findings as related to the avoidance of a host response is discussed.     

THE DEVELOPMENT AND POTENTIAL ROLES OF CELL‐WALL TRABECULAE IN CAULERPA MEXICANA (CHLOROPHYTA)1

Siphonous plants represent an alternate scheme to the way most macroscopic plants are constructed. They are single, often large (1–2 m), sometimes morphologically complex, multinucleate (coenocytic) cells where the whole of the cytoplasm is a continuum. Caulerpa mexicana Sond. ex Kütz. is a siphonous tropical marine green alga characterized by four morphologically distinct regions and, as with other members of the genus, by the presence of a dense network of anastomosing cylindrical cell wall in growths called trabeculae. Based on the results of this study, we propose several roles for trabeculae: (i) They are structural components, which likely add some small amount of support in compression but add considerable strength in tension. (ii) As extensions of the cell wall and plasma membrane, they act as diffusion channels from the cell exterior to the interior cytoplasm. It is possible that trabeculae also play a role in determining cell shape through developmental positioning and placement patterns, thus facilitating the diverse shapes found in the morphologically distinct regions of Caulerpa sp.     

Rhabdovirus-like and closterovirus-like particles in ultrathin sections of Ribes species with symptoms of blackcurrant reversion and gooseberry veinbanding diseases

In attempts to determine the causal agents of blackcurrant reversion (BCRD) and gooseberry veinbanding (GVBD) diseases of Ribes species, details of the ultrastructure of different kinds of tissue from plants affected with these different diseases were studied. In three of 12 blackcurrant plants affected with BCRD, leaves and flowers of plants showing symptoms typical of the severe (R) form of the disease, contained rhabdovirus-like particles c. 65–80 nm × 215–485 nm. They were seen most often in the nucleus of cells as single particles but were also found in clusters or rafts. In leaves, these virus-like particles (VLPs) were present only in cells associated with the xylem parenchyma where they occurred as membrane-bound clusters within the nucleus. In flowers, they were also found in phloem parenchyma cells in the peripheral cytoplasm and very occasionally in the cytoplasm of epidermal cells. All non-nuclear VLPs were membrane-bound, either singly or in groups and the membrane seemed to be part of the endoplasmic reticulum. The proportion of vascular cells containing these VLPs was very low (< 1%). In a few cells, smaller bacilliform particles, c. 40–50 nm × 200–250 nm, were found in the nucleus together with the larger particles. Double-membrane bodies, detected in fig leaves affected with fig mosaic (the agent of which is also mite-transmitted), were not detected in any BCRD-affected plants. In leaf tissue of one of three gooseberry and one of two blackcurrant plants affected with GVBD, two kinds of VLPs were found. Rhabdovirus-like particles, similar to those in BCRD-affected material, were present in the nuclei, perinuclear space and cytoplasm of xylem parenchyma cells. They were c. 60–72 nm × 155–230 nm but there was no evidence of the smaller rhabdovirus-like particles detected in a few cells of BCRD-affected tissues. The second kind of VLP was found in noncrystalline masses, with a mean centre-centre spacing of c. 10 nm, in the cytoplasm of phloem cells. These particles, together with other ultrastructural changes, were typical of those reported for aphid-transmitted closteroviruses. No badnavirus-like particles, reported previously from GVBD-affected plants, were observed in any of the plants studied. The significance of these findings in relation to these two important diseases of commercial Ribes species is discussed.     

DEVELOPMENT OF PERIPHERAL VACUOLES IN PLANT CELLS

The vacuolar apparatus of various plant cells consists of two distinct features: the large central vacuole and peripheral vacuoles which are derived from invaginations of the plasma membrane. Peripheral vacuoles are conspicuous structures in both living and fixed hair or filament cells of Tradescantia virginiana. They occur as spherical structures along the inner boundary of the peripheral cytoplasm and can be recognized as projections into the central vacuole. These structures are variable in size and number within a cell and can represent a significant proportion of the volume of the vacuole. Peripheral vacuoles most frequently are observed in motion with the streaming cytoplasm although their velocity is usually somewhat slower that that of the cytoplasmic organelles. Ultrastructural studies show two closely approximated membranes, one for each vacuole, in areas where a peripheral vacuole projects into the central vacuole. These are separated by an intermembrane zone continuous with the peripheral cytoplasm. The movement of organelles over the perimeter of the peripheral vacuole is presumed to occur along this intermembrane zone. The internal area of the peripheral vacuoles may appear empty although some contain a vesicular content of unknown origin and function.     

Occurrence of unusual heterogeneous lipid-containing granule-storing cells in the main excretory duct epithelium of the male mouse submandibular gland

Summary The fine structure of the main excretory duct epithelium of the male mouse submandibular glands was investigated by scanning and transmission electron microscopy. Three principal cell-types were observed: type I and II, and basal cells. This epithelium was characterized by the presence of intercellular canaliculi. Type-I cells were the most numerous. They had an abundance of mitochondria, well-developed Golgi apparatus, a few electron-lucent lipid-containing granules and poorly developed basal infoldings. These cells were also characterized by many glycogen granules throughout the cytoplasm and abundant smooth endoplasmic reticulum in the apical cytoplasm. Type-II cells were the second most numerous. Their most characteristic feature was the presence of abundant heterogeneous lipid-containing granules having acid phosphatase activity at the periphery. They were concentrated in the infra- and supranuclear cytoplasm. The granules may be derived from mitochondrial transformation and seem to be a special kind of secondary autolysosome. Type-II cells also contained abundant mitochondria throughout the cytoplasm, much smooth endoplasmic reticulum in the apical cytoplasm, a well developed Golgi apparatus adjacent to the heterogeneous lipid-containing granules and no basal infoldings. Basal cells were situated adjacent to the basal lamina. They had a large nucleus and the cytoplasm was filled with glycogen granules.     

Using fluorescence lifetime microscopy to study the subcellular localization of anthocyanins

Anthocyanins are flavonoid pigments that accumulate in most seed plants. They are synthesized in the cytoplasm but accumulate inside the vacuoles. Anthocyanins are pigmented at the lower vacuolar pH, but in the cytoplasm they can be visualized based on their fluorescence properties. Thus, anthocyanins provide an ideal system for the development of new methods to investigate cytoplasmic pools and association with other molecular components. We have analyzed the fluorescence decay of anthocyanins by fluorescence lifetime imaging microscopy (FLIM), in both in vitro and in vivo conditions, using wild‐type and mutant Arabidopsis thaliana seedlings. Within plant cells, the amplitude‐weighted mean fluorescence lifetime (τ_m) correlated with distinct subcellular localizations of anthocyanins. The vacuolar pool of anthocyanins exhibited shorter τ_m than the cytoplasmic pool. Consistently, lowering the pH of anthocyanins in solution shortened their fluorescence decay. We propose that FLIM is a useful tool for understanding the trafficking of anthocyanins and, potentially, for estimating vacuolar pH inside intact plant cells.     

Ultrastructural investigations of mature embryo sacs of Daaucus carota,D. aureus and D. muricatus — possible cytological explanations of paternal plastid inheritance

Summary The embryo sacs of Daucus carota, D. Aureus and D. muricatus are of the Polygonum-type. They contain one egg cell, two synergids, a giant central cell and three antipodal cells that are to a great extent degenerated. The species of Daucus investigated have an egg cell with a vacuole that is large in comparison to the amount of cytoplasm. The most extreme case of reduced cytoplasm with respect to the volume of the vacuole occurs in D. muricatus. The egg cell of this species contains only very few intact plastids and other cytoplasmic organelles. Paternal plastid inheritance in the cross D. muricatus × D. carota is discussed in connection with the small number of cytoplasmic organelles in the female gamete of D. muricatus.     

STIGMA,STYLE, AND OBTURATOR OF ORNITHOGALUM CAUDATUM (LILIACEAE) AND THEIR FUNCTION IN THE REPRODUCTIVE PROCESS

Transmitting tissue in Ornithogalum is divided into three regions corresponding to classical divisions of the gynoecium: stigma, style, and ovary. The stigma differentiates from epidermal cells of the stylar apex. These cells form the stigmal papillae and have dense cytoplasm with abundant ER and lipid bodies. Papillae have walls with small transfer-ingrowths. At floral receptivity, papillae secrete a small amount of surface exudate. Epidermal cells of the style contain numerous spherosomes and have thin filaments of cytoplasm traversing the central vacuole. The stylar cortex is composed of 3-6 layers of parenchyma cells which contain numerous spherosomes and often have secondary vacuoles. Vascular tissue in the style consists of one collateral bundle in each lobe. Cells of the epidermal layer lining the stylar canal are secretory. They are initially vacuolate but fill progressively with dense cytoplasm as their secretory activity increases. Secretory activity occurs in three phases, each characterized by a particular organelle population and secretory product. At anthesis, the canal is filled with an exudate consisting of carbohydrate, protein, and lipid. In the ovary, the obturator differentiates from cells at the base of the funiculus and the tip of the carpel margins. It forms a pad of tissue which covers most of the former placenta. The obturator is secretory and produces a surface exudate. We believe our observations on Ornithogalum support the hypothesis that all transmitting tissue is of the same morphological origin and that it provides nutritive and chemotropic factors for pollen tube growth.     

Association of different kinds of bacilliform particle with vein chlorosis and mosaic diseases of raspberry (Rubus idaeus)

Thin sections of diseased raspberry (Rubus idaeus) were examined by electron microscopy. Plants of the cv. Baumforth's B and of an aphid (Amphorophora rubi)-resistant breeding selection (6820/54), both infected with raspberry vein chlorosis virus (RVCV) but not with other detectable viruses, contained large bacilliform particles c. 430 × 65 nm. Particles occurred in the cytoplasm and perinuclear space of a small proportion of xylem parenchyma cells. They had an inner core c. 25–30 nm in diameter with cross-banding of periodicity 4·5 nm, and were bounded by an outer membrane. They are probably the particles of RVCV. Plants of cv. Mailing Jewel and of a selection (M14) both showing symptoms of raspberry mosaic (veinbanding) disease contained smaller bacilliform particles c. 125 × 30 nm, which occurred singly or in clusters in the cytoplasm of a small proportion of vascular parenchyma cells. It is not known which, if any, of the viruses associated with raspberry mosaic are represented by the particles.     

Functional morphology of the neck organ in Artemia salina nauplii

Fine structure of the ion transporting epithelium of the neck organ in the brine shrimp (Artemia salina) nauplius is described. The neck organ is a dome-like gland situated atop the cephalothorax of the larva and is composed of 50 to 60 cuboidal epithelial cells. These cells possess many of the characteristics of salt-secretory cells from other tissues. They contain many mitochondria and exhibit a high degree of plasma membrane elaboration. This membrane amplification takes two forms; the apical plasmalemma is infolded into irregular loops, while the basal and lateral membranes penetrate the cytoplasm in the form of branching sinusoids. The labyrinth of tubular reticulum thus formed fills most of the cell volume. Mitochondria in the labyrinth are often in intimate contact with these tubular membranes and regular arrays of parallel mitochondria with constricted intervening sinusoids are often observed. Other organelles including Golgi complexes, multivesicular bodies, and rough endoplasmic reticulum are also numerous, particularly in the narrow rim of cytoplasm which lies between the apical infolds and the labyrinth. Yolk platelets and glycogen fields are conspicuous in the basal perinuclear regions of the cells.     

Confocal pH Topography in Plant Cells — Acidic Layers in the Peripheral Cytoplasm and the Apoplast

The distribution of intracellular pH was studied in cultured cells of Gossypium hirsutum by con-focal pH topography using the fluorescent probe car-boxy SNARF1 and a ratio imaging procedure. The resulting pH maps can visualize pH differences with an accuracy of 0.1 unit in the investigated range between 7.5 and 5.6. They reveal the following characteristic features of the Gossypium cells: – the pH of the cytoplasmic core regions ranges from near 7.4 in younger to near 6.0 in older cells; – vacuoles show the expected acidity with pH < 5.6; – the cell wall/apoplastic region is acidic with a pH near 5.6 or below, especially in young, growing cells; – interestingly, acidic areas appear also at the periphery of the cytoplasm, i.e. beneath the plasma membrane. They remain stable in the presence of 5/μmol/I of the protonophore CCCP. Acidic layers of peripheral cytoplasm were also detected in protoplasts of Penicillium cyclopium, i.e. eukaryotic cells of simpler structure, which served as a reference object. This ronfirms earlier findings obtained with classical fluorescence microscopy and another fluoroprobe (fluorescein diacetate). Though additional experimental support is needed, low pH regions at the cytoplasm/plasma membrane interface should be considered a real contribution to the pH control of plant and fungal cells, facilitating e.g. the maintenance of cytosolic pH in acidic environments.     

An ultrastructural investigation of the hypocerebral organ of the adult Euperipatoides kanangrensis (Onychophora, Peripatopsidae)

The hypocerebral organs of Euperipatoides kanangrensis are a pair of spherical vesicles located ventral to the cerebral ganglia. They develop in the embryo from the most anterior pair of ventral organs, in the antennal segment. The wall of each hypocerebral organ is a dense epithelium of elongate cells with peripheral nuclei. The cytoplasm of the cells includes numerous mitochondria, Golgi bodies and microtubules. The small lumen, located eccentrically within the organ, contains concentrically layered electron-dense material resembling cuticle.Each hypocerebral organ is enclosed by a layer of extracellular matrix continuous with that surrounding the adjacent cerebral ganglion. There are no nerve connections between ganglion and organ, but cellular connections traverse the intervening matrix and could serve as a communication pathway. The ultrastructure of the hypocerebral organs indicates that they are glands.     

Endothelial cells of the meningeal capillaries in the rainbow trout: A freeze-fracture study

Meningeal capillaries are unfenestrated. They are made up of endothelial cells that have a pinocytotic caveolae density of 41 ± 11/μm² and 89 ± 21/μm² on the abluminal and luminal sides respectively. The total density of intramembrane particles is not significantly different between the luminal and the abluminal membranes; however, the coefficients of partition are significantly different (P< 0.001). One or two strands of tight junctions occur between adjacent cells but no gap junctions nor desmosomes exist. The density of nuclear pores is less than 3.2/μm². An abundance of intermediate filaments and free vesicles, some of which are seriated, characterize the cytoplasm. The functional significance of these findings is discussed.     

Immunohistochemical studies with antibodies to myosins from the cytoplasm and membrane fraction of human blood platelets

Summary Antibodies were raised to myosins extracted from the cytoplasm and solubilized membranes of human blood platelets. Both antibodies had similar titers as shown by enzyme-immunoassay and bound to the same sites as shown by immunohistochemistry. They were specific for cytoplasmic myosins (e.g., in human white blood cells, platelets and fibroblasts and rat endothelial cells). They showed no crossreaction with human or rat smooth muscle.Abbreviations ATPase adenosine triphosphatase EC 3.6.1.3 - FITC fluorescein isothiocyanate     

Kristalloide Einschlüsse im Zytoplasma pflanzlicher Zellen

Summary In healthy as well as dahlia mosaic sick plants ofVerbesina encelioides, Sanvitalia procumbens, Zinnia elegans, Calendula spec. andDahlia hybrids, leaf cell vacuoles are found in the marginal cytoplasm which contain protein crystals. They are single membrane-limited products of the endoplasmatic reticulum. They may be found mainly in the older leaves and especially in those of virus infected plants. The crystalline structures consist of tetragonally arranged tubules of 105 Å in diameter, separated by an interspace about 35 Å wide. There are similar structures in virus infected plants ofFragaria vesca, but not inChenopodium quinoa, where the vacuoles contain no bodies. This cell organelle is compared with crystalline inclusions already described. Its significance and relations to the virus disease are discussed.     

The involvement of nurse cells in oogenesis and embryonic development in the marine sponge,Haliclona ecbasis

Oogenesis and embryonic development in the marine sponge, Haliclona ecbasis, were studied using standard histological procedures. When the oocytes reach a diameter of about 30 μ, nurse cells begin to aggregate around them. Then when the oocytes are about 36 μ in diameter, they begin to engulf the associated nurse cells. Whole nurse cells are engulfed; and although the nucleus of the nurse cells disappears either as or soon after the cells are engulfed, the cytoplasm remains essentially unchanged. The accumulation of these cells within the oocytes most of the cytoplasm is nurse cell cytoplasm. During cleavage of the egg, the engulfed nurse cells are gradually fragmented, but otherwise appear unchanged. At the same time the cytoplasm of the nurse cells is progressively incorporated into that of the blastomeres by what appears to be fusion process. When the latter process is complete, the embryo develops into a typical parenchymula larva.     

An ultrastructural investigation of the early stages of oocyte differentiation in Actinia fragacea (Cnidaria; Anthozoa)

Individuals from a population of the intertidal sea anemone Actinia fragacea (Tugwell) were collected at approximately monthly intervals over an 18 month period. Samples of gonad were removed from each anemone and examined by light and electron microscopy. During late spring and early summer, large numbers of small cells were seen in the endoderm of the female gonads, lying close to the mesoglea. For convenience, these cells were classified into three types. Type I cells are 6–9 μm in diameter, with relatively very large nuclei, which may contain synaptinemal complexes, and scant cytoplasm containing few organelles. Type II cells are larger, reaching 15 μ m in diameter, with more abundant cytoplasm containing more organelles and inclusions. The nucleus is more dense, but may also contain synaptinemal complexes. Type III cells are less common. They are similar in size to Type II cells, but their nuclei contain irregular dense chromatin masses, and the nuclear envelope is incomplete or absent. The possible significance of the various cell types is discussed. It is suggested that Type I cells are oocytes at a very early stage of differentiation and that Type II cells are rather later oocytes. The status of the Type III cells is uncertain.     

The Structure and Origin of the Plasmodium of Rhopalura ophiocomae(Phylum Orthonectida)

Abstract Adult orthonectids develop from germinal cells within a cytoplasmic matrix called a plasmodium. This is generally assumed to be formed by the parasite. In the case of Rhopalura ophiocomae, which lives in the brittle star Amphipholis squamata, the plasmodia occupying the perivisceral coelom are closely associated with the walls of the genital bursae or the gut, and they are covered by peritoneum. They have been reported to contain scattered small nuclei distinct from those within germinal cells, embryos, and adults, but the results of the present study indicate that such nuclei probably do not exist. Furthermore, electron micrographs show that some plasmodia are in continuity with the cytoplasm of contractile cells that lie beneath the peritoneum of a genital bursa or the gut of the host. The matrix of a plasmodium of R. ophiocomaeappears, therefore, to consist of cytoplasm of a contractile cell. It is proposed that after a contractile cell has been entered by an infective cell of the parasite, it hypertrophies, bulging progressively farther into the perivisceral coelom and lifting up the peritoneum, which remains in intimate contact with it.     

The Subchordal Cells of Oikopleura dioica and O. albicans (Appendicularia,Chordata)

Abstract Oikopleura dioica has two large subchordal cells which were studied in vivo and with light and electron microscopy. They have fixed positions within the haemocoel of the tail but change their morphology continually by protruding and withdrawing processes in an amoeboid manner. Also the fine structure varies considerably from one animal to the next. The cell surface sometimes indicates a strong pinocytotic activity with many coated pits and vesicles. In other cases there are many small vesicles which are interpreted as exocytotic. They are found both in the cytoplasm close to the plasma membrane and in the haemocoelic fluid. Subchordal cells with no pronounced surface activity have a large amount of rough endoplasmic reticulum, which suggests that they synthesize proteins. The presence of subchordal cells in the tail is correlated with the presence of the ontogenetically related bioluminescent oral gland cells in the pharynx; either both types exist simultaneously or both are lacking. It is speculated that the two cell types are also functionally co-ordinated. Oikopleura albicans has a multitude of tiny subchordal cells which have essentially the same fine structure as the two large Oikopleura dioica cells.