Localization of the beet mild yellowing virus inSinapis alba L.

Virus particles of isometric shape with a diameter of 26 nm were found in the sieve tubes and accompanying phloem cells in ultrathin sections prepared from the nerves of white mustard (Sinapis alba L.) leaves and roots infected with the beet mild yellowing virus (BMYV). BMYV particles were much more frequent in the roots ofSinapis alba plants. Isometric particles were not found in the leaves and roots of healthy mustard plants.     

Virus-like particles in phloem tissue of chervil (Anthriscus cerefolium) infected with anthriscus yellows virus

Clusters of isometric virus-like particles c. 22 nm in diameter, embedded in amorphous electron-dense material, were found in the phloem tissue of Anthriscus cerefolium (chervil) plants infected with the semi-persistent aphid-borne virus, anthriscus yellows (AYV). The particles resembled those seen previously in thin sections of AYV-transmitting aphids (Cavariella aegopodii). The particles were found only in the central vascular bundle of the petiole and its continuation in the leaf midrib. They were also found in extracts made by grinding petiole and midrib tissue in 10 % sucrose using Carborundum. These results confirm earlier studies which suggest that AYV is confined to deeper-lying tissues.     

Properties of cocksfoot streak and cocksfoot cryptic, two viruses infecting cocksfoot (Dactylis glomerate) in Scotland

A Scottish isolate of cocksfoot streak virus (CSV-S) was found to have flexuous filamentous particles which, in sap of infected cocksfoot plants, had a modal length of 712 nm. It was transmitted from infected to healthy cocksfoot plants in a non-persistent manner by Myzus persicae and by mechanical inoculation of infective sap extracts containing an anti-oxidant. Apart from cocksfoot, mechanical inoculation of infective sap succeeded in infecting only four of 22 plant species tested. The infectivity of sap extracts containing 0.2% thioglycerol was lost after heating for 10 min at 55^oC but not 50^oC, storage at room temperature for 48 but not 24 hours, and after diluting 10-2 to 10-3. Highly purified preparations of CSV-S particles sedimented as a single component with a sedimentation coefficient of 139S and had a buoyant density in rubidium bromide of 1.31 g/cm3. Virus particles were composed of one protein and one ssRNA species with estimated Mr of 31 000 and 3.2 times 106 respectively. In ELISA, an antiserum prepared to CSV-S detected the virus in all aerial parts of infected cocksfoot plants and, when present in the ratio of 1 infected leaf: 1000 healthy leaves. Both CSV-S-infected and -uninfected cocksfoot also contained a previously undescribed virus with isometric particles c. 30 nm in diameter. This virus, named cocksfoot cryptic virus (CCV), was seed-borne in two cvs of cocksfoot tested and its particles contained two dsRNA species of estimated Mt of 1.14 times 106 and 1.27 times 106. Despite the fact that particles of CSV-S were largely free from CCV particles following exclusion chromatography on agarose beads prior to immunisation, immunoelectron microscopy (IEM) showed that the antiserum prepared to CSV-S also contained some antibodies to CCV. Evidence from IEM suggested a possible distant serological relationship of CCV to ryegrass and beet (BCV 1 or BCV 2, or both) cryptoviruses, all members of sub-group A of cryptoviruses.     

Electron Microscopy of an Isometric Caulimo-Like Virus from Sweet Potato (Ipomoea batatas)

A newly recognised virus with isometric particles 50 nm in diameter was detected in Ipomoea batatas (sweet potato) by graft-transmission to I. setosa. Virus particles and intracellular inclusions, although not seen in infected I. batatus, were found in the cytoplasm of most cell types of I. setosa. The caulimo-like particles were most abundant with vesicles immediately adjacent to inclusions. The ovoid or spherical inclusions, which differ markedly from those of caulimovi, ruses, have a large central lacuna and usually several smaller peripheral lacunae. Infected vascular parenchyma cells sometimes protrude into, and occasionally completely occlude, adjacent xylem vessels, observations possibly explaining the sudden wilting and premature senescence of infected leaves.     

Western Flower Thrips (Frankliniella occidentalis) Transmits Maize Chlorotic Mottle Virus

Maize chlorotic mottle virus (MCMV) is one of the co‐infection pathogens that cause corn (maize) lethal necrosis, but the transmission mechanism of MCMV is not yet clear. In order to determine the ability of western flower thrips (Frankliniella occidentalis; WFT) to transmit MCMV, imported maize seeds from Thailand were germinated in an insect‐free greenhouse and the seedlings were tested for the transmission by WFT of chlorotic mottle virus disease. The thrips (WFT), starved for 48 h then allowed to feed for 30 min on maize plants infected with MCMV or asymptomatic maize plants, were transferred to healthy seedlings. After 35 days, the seedlings with WFT from diseased maize plants showed chlorotic mottle symptoms, whereas seedlings with WFT from asymptomatic maize plants remained healthy. A single band of 711 bp was amplified by RT‐PCR using primers MCMV‐F/MCMV‐R from the MCMV‐infected plants and WFT collected from the diseased plants. Sequencing of the amplified product and further sequence comparison indicated that the two viruses from both sources showed 99% similarity of nucleotides and they should be regarded as identical. In addition, isometric particles c. 30 nm in diameter, characteristic of MCMV, were found in the WFT samples from diseased maize plants. Thus, it is concluded that WFT transmits MCMV. Our findings suggest that corn lethal necrosis disease can be controlled or minimized by the eradication of WFT from the field or greenhouses.     

Properties of cocksfoot streak and cocksfoot cryptic, two viruses infecting cocksfoot (Dactylis glomerata) in Scotland

A Scottish isolate of cocksfoot streak virus (CSV-S) was found to have flexuous filamentous particles which, in sap of infected cocksfoot plants, had a modal length of 712 nm. It was transmitted from infected to healthy cocksfoot plants in a non-persistent manner by Myzus persicae and by mechanical inoculation of infective sap extracts containing an anti-oxidant. Apart from cocksfoot, mechanical inoculation of infective sap succeeded in infecting only four of 22 plant species tested. The infectivity of sap extracts containing 0.2% thioglycerol was lost after heating for 10 min at 55^oC but not 50^oC, storage at room temperature for 48 but not 24 hours, and after diluting 10^-2to 10^-3. Highly purified preparations of CSV-S particles sedimented as a single component with a sedimentation coefficient of 139S and had a buoyant density in rubidium bromide of 1.31 g/cm³. Virus particles were composed of one protein and one ssRNA species with estimated M_r of 31 000 and 3.2 times 10⁶respectively. In ELISA, an antiserum prepared to CSV-S detected the virus in all aerial parts of infected cocksfoot plants and, when present in the ratio of 1 infected leaf: 1000 healthy leaves. Both CSV-S-infected and -uninfected cocksfoot also contained a previously undescribed virus with isometric particles c. 30 nm in diameter. This virus, named cocksfoot cryptic virus (CCV), was seed-borne in two cvs of cocksfoot tested and its particles contained two dsRNA species of estimated M_r of 1.14 times 10⁶and 1.27 times 10⁶. Despite the fact that particles of CSV-S were largely free from CCV particles following exclusion chromatography on agarose beads prior to immunisation, immunoelectron microscopy (IEM) showed that the antiserum prepared to CSV-S also contained some antibodies to CCV. Evidence from IEM suggested a possible distant serological relationship of CCV to ryegrass and beet (BCV 1 or BCV 2, or both) cryptoviruses, all members of sub-group A of crypto viruses.     

Identification and Characterization of Pothos latent virus Causing Necrotic Ringspots and Line Patterns on Lisianthus (Eustoma grandiflorum) in Taiwan

Lisianthus (Eustoma grandiflorum) grown in screenhouses in Taiwan showed ringspots and concentric line patterns on leaves. A virus having isometric particles approximately 30–32 nm in diameter was isolated from affected lisianthus. Combined results of biological, cytological, serological, molecular and phylogenetic analyses show that the virus can be identified as Pothos latent virus (PoLV), genus Aureusvirus, family Tombusviridae. Inoculating the virus on non‐infected lisianthus plants reproduced the symptoms previously observed in the field. So, this is the first report of PoLV causing disease in lisianthus and the first report of the virus in Taiwan.     

小麦黄花叶病毒的分离提纯及其血清学等特性

应用梯度离心和超速离心浓缩获得部分提纯的病毒制剂,产量约为7.45g/kg病叶提纯的病毒制剂的紫外吸收曲线呈典型的核蛋白吸收曲线,OD260/OD242和OD260/OD280的比值分别为1.24和1.38。病毒粒子呈线状,宽13—14nm,长度主要分布于250—300nm和550—700nm之间,1000nm以上的粒子也有检到。病毒外壳蛋白仅由一个分子量约为30Kd的亚基组成。在免疫电镜试验中、病毒粒子与日本WYMV抗血清发生强烈的血清学反应。新鲜病叶的超薄切片中可看到大量风轮体和膜状体。     

ULTRASTRUCTURAL CHARACTERIZATION OF THE LYTIC CYCLE OF AN INTRANUCLEAR VIRUS INFECTING THE DIATOM CHAETOCEROS CF. WIGHAMII(BACILLARIOPHYCEAE) FROM CHESAPEAKE BAY,USA1

Numerous microalgal species are infected by viruses that have the potential to control phytoplankton dynamics by reducing host populations, preventing bloom formation, or causing the collapse of blooms. Here we describe a virus infecting the diatom Chaetoceros cf. wighamii Brightw. from the Chesapeake Bay. To characterize the morphology and lytic cycle of this virus, we conducted a time‐course experiment, sampling every 4 h over 72 h following viral inoculation. In vivo fluorescence began to decline 16 h after inoculation and was reduced to <19% of control cultures by the end of experiment. TEM confirmed infection within the first 8 h of inoculation, as indicated by the presence of virus‐like particles (VLP) in the nuclei. VLP were present in two different arrangements: rod‐like structures that appeared in cross‐section as paracrystalline arrays of hexagonal‐shaped profiles measuring 12 ± 2 nm in diameter and uniformly electron‐dense hexagonal‐shaped particles measuring ～ 22–28 nm in diameter. Nuclei containing paracrystalline arrays were most prevalent early in the infection cycle, while cells containing VLP increased and then declined toward the end of the cycle. The proportion of nuclei containing both paracrystalline arrays and VLP remained relatively constant. This pattern suggests that rod‐like paracrystalline arrays fragmented to produce icosahedral VLP. C. cf. wighamii nuclear inclusion virus (CwNIV) is characterized by a high burst size (averaged 26,400 viruses per infected cell) and fast generation time that could have ecological implications on C. cf. wighamii population control.     

Geminate structures of African cassava mosaic virus

Two types of geminate structures were purified from African cassava mosaic geminivirus (ACMV)-infected Nicotiana benthamiana plants and analyzed by electron cryomicroscopy and image reconstruction. After cesium sulfate density gradient centrifugation, they were separated into lighter top (T) and heavier bottom (B) components. T particles comigrated with host proteins, whereas B particles were concentrated in a cesium density typical for complete virions. Both particles were composed of two incomplete icosahedra of 11 capsomers each, but T particles were slightly larger (diameter, 22.5 nm) and less dense in the interior than B particles (diameter, 21.5 nm). T particles were frequently associated with small globules of approximately 14 nm diameter of unknown origin. The overall structure of ACMV, a begomovirus transmitted by whiteflies, was similar to that of Maize streak virus (MSV), a mastrevirus transmitted by leafhoppers, although the vertices of the icosahedra were less pronounced. Models of ACMV coat proteins based on Satellite tobacco necrosis virus support the exposure of parts of the molecule essential for transmission specificity by whiteflies and provide possible structural explanations for the smaller protrusion of the ACMV capsid relative to MSV. The differences of ACMV and MSV virion shapes are discussed with reference to their different animal vectors.     

A Virus Infection in the Marine Brown Alga Ectocarpus siliculosus (Phaeophyceae)

Laboratory cultures of Ectocarpus siliculosus originating from New Zealand showed a defect in gametangium formation. Nuclear divisions in gametangium initials are not followed by cell wall formation. In the resulting multinucleate cells nuclear DNA increases dramatically, and nuclear membranes disintegrate. Eventually, the entire structure is filled with hexagonal particles of approximately 130 nm diameter. These were isolated and shown by EM to consist of a dense core surrounded by a 3-layered shell. They are released into the culture medium when the host cells burst. Ectocarpus gametes from healthy cultures could be infected by these particles. The resulting partheno-sporophytes developed pathological symptoms, suggesting that the particles are viruses. The expression of the defect is temperature dependent. At 10°C all gametangia are abnormal, while between 15 and 20 °C defective and normal gametangia and gametes are formed on the same plant. Partheno-sporophytes developing from such gametes carry the viral particles expressed in deformed unilocular and plurilocular sporangia.     

Electron microscopic study of teleost (Pimelodus maculatus) pancreatic islet cells

The endocrine pancreas of the freshwater teleost Pimelodus maculatus was studied by electron microscopy. Based on the granule morphology 2 cell types were described: Secretory granules of type I cells are rounded, nearly completely filling the limiting membranous sac which measures from 120 to 150 nm in diameter; the type II granules are also rounded and measure from 220 to 270 nm in diameter; they consist of an eccentrical electron dense core separated from the limiting membrane by a wide electron lucent halo. These characteristics are correlated with those found in other teleosts.     

A new virus disease in the housefly, Musca domestica (Diptera)

Reovirus particles were isolated from adults in laboratory colonies of the housefly, Musca domestica. These particles were spherical in outline, 57–76 nm in diameter, and were found only in hemocyte cytoplasm, where virions have been disclosed by a new technique. Virions were present in large numbers, and viral inclusion bodies were identified. The virus particles had pentagonal and hexagonal shapes resembling a simple icosahedral structure. The virus was shown to be infectious and pathogenic to adult flies through injection or by feeding them suspensions from flies that had died of the virus. Electron micrographs of midgut sections from infected flies showed that the midgut cells were packed with dark undulating threads which were not present in uninfected flies. However, no virus particles or inclusion bodies could be seen in these cells. On the basis of their association with infected flies, and the similarity to results from other studies on reoviruses and insect viruses, it is suggested that these threads are an alternative replicative form of the reovirus. When the virus suspensions from heavily infected flies were dialyzed against weak alkaline solutions, the threads showed an inner component of coiled material, 12 nm in diameter, inside an envelope with a diameter of 50–83 nm, mean 60.3 ± 7.5, composed of subunits 7–8 nm long and 7–8 nm across.     

A virus infection inMyriotrichia clavaeformis (Dictyosiphonales,Phaeophyceae) from Argentina

Summary Virus-infected plants ofMyriotrichia clavaeformis from the coast of Argentina are sterile and produce viras particles in lateral vesicles homologous to plurilocular sporangia of normal plants. Virions are released into the surrounding sea water and can infect swimming spores of healthyMyriotrichia plants. Virus particles are hexagonal in cross section with a diameter of 170–180 nm and have an envelope with an electron dense core. Similar viruses are known for several genera in the order Ectocarpales. This is the first record of an infectious agent in the order Dictyosiphonales, which contains thalli with more complex tissues and life histories.Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement     

Auswirkungen des Anbaus einer Wirts- und einer Nichtwirtspflanze unter wechselnden ökologischen Bedingungen auf die Populationsdynamik von Rhizoctonia solani Kühn und Fusarium solani f. pisi (Jones) Snyd. et Hans

An isometric virus was isolated from Helianthus annuus L. plants showing a yellow leaf spot mosaic on affected leaves. Infected plants were found in different ecological regions of Ukraine. A procedure of virus purification is described. The diametres of the virus particles were nonuniform and ranged from 50 to 120 nm. The sedimentation coefficient of the virus was 518–540 S and the floating density in the CsCl gradient was 1.22 g/cm³. The MW of proteins separated by electrophoresis amounted to 78±0.9, 58±0.8, 52±0.2, and 27±0.8 kDa, respectively. The virus was assigned to the tospoviruses for which sunflower is a new previously undescribed natural host plant.     

Ultrastructural Evidence for Neuromuscular Systems in Coelenterates

Cellular interrelationships and synaptic connections in tentaclesof several species of coelenterates were examined by means ofelectron microscopy to determine if neuromuscular pathways werepresent. The presence of sensory cells, ganglion cells, epitheliomuscularcells, interneuronal synapses, and neuromuscular junctions suggeststhat neuromuscular pathways are present in coelenterates. Nakedaxons without sheath cells form several synapses en passantwith the same and with different epitheliomuscular cells aswell as with nematocytes and other neurons. Interneuronal synapsesand neuromuscular and neuronematocyte junctions have clear ordense-cored vesicles (700–1500 Å in diameter) associatedwith a dense cytoplasmic coat on the presynaptic membrane, acleft (100–300 Å in width) with intracleft filaments,and a subsynaptic membrane with a dense cytoplasmic coat. Atscyphozoan neuromuscular junctions there is a subsurface cisternaof endoplasmic reticulum, which is separated from the epitheliomuscularcell membrane by a narrow cytoplasmic gap (100–300 Åin width) . Neuromuscular junctions in coelenterates resembleen passant axonal junctions with smooth muscle in higher animals. Morphological evidence is presented for a simple reflex involvinga two-cell (sensory or ganglion-epitheliomuscular cell) or three-cell(sensory-ganglion-epitheliomuscular cell) pathway that may resultin the coordinated contraction of the longitudinal muscle intentacles of coelenterates.     

Purification and some properties of strawberry mottle virus

Strawberry mottle virus (SMoV) (three isolates: HJ, 3E and N) were transmitted to Chenopodium quinoa plants by sap inoculation. All three isolates induced very similar symptoms consisting of chlorotic spots and ringspots in inoculated leaves, and vein chlorosis, mottling, and dwarfing of the upper leaves. SMoV isolate HJ was purified from infected C. quinoa by homogenisation with 10 mM phosphate buffer, pH 7.2 containing 5% Triton X-100, followed by differential, sucrose density-gradient and CsCl equilibrium density-gradient centrifugations. A fraction with a buoyant density of 1.42g^- cm^-3 after CsCl density-gradient centrifugation was highly infectious to C. quinoa and contained many isometric virus-like particles c. 37 nm in diameter. These virus-like particles were never found in fractions from uninfected preparations. Electrophoretic analysis of a fraction containing virus-like particles revealed that these particles might have a single coat protein subunit with the apparent molecular mass of 26 K daltons and one nucleic acid of 6.6 kilobases. Double-stranded RNA analysis of isolate HJ-infected or uninfected C. quinoa and Fragaria vesca var. semperflorens seedling line ‘Alpine’ plants showed that both infected plants had two infection-specific dsRNA bands of mol. wts 4.5 and 3.9 × 10⁶.     

Effects of Temperature on BYDV Concentration in Oat Plants and Virus Purification Efficiency, and Detection of a Putative Associated Satellite Virus

RPV and MAV-like serotypes of barley yellow dwarf virus (BYDV), designated R-568 and F, were found during sucrose density gradient centrifugation to suspend at 10 °C and 4 °C but to totally sediment at 15 °C and 12 °C, respectively. These properties were used to purify these serotypes, and antisera were then prepared.
Partially purified IgG from antiserum was used in immunosorbent electron microscopy (ISEM) and in enzyme-labelled immunosorbent (ELISA) tests to detect BYDV RPV-like serotypes. Using anti-BYDV R-568 polyclonal antiserum and the BYDV R-568 serotype in ISEM tests, isometric virus particles of two sizes were trapped: the 28 nm particles of BYDV R-568, and others 17 nm in diameter which may be those of a satellite virus.
The effects of temperatures on virus concentrations in oat plants infected with BYDV serotypes F and R-568 were investigated. BYDV F and R-568 concentrations in the roots and shoots were sensitive to changes in temperature between 10 °C and 25 °C. The concentrations of both viruses in the roots and shoots of infected plants could be manipulated by varying the temperature at which plants were grown. The ELISA absorbance values related to detection of F MAV-like serotypes were higher in roots and shoots of oats grown at 10 °C than for oats grown at 25 °C. Conversely, cool temperatures reduced the absorbance values for R-568 RPV-like serotype in the roots, but less significantly in the shoots.     

Viruslike particles in Gyratrix hermaphroditus (Turbellaria: Rhabdocoela)

During an ultrastructural examination, viruslike particles were observed in the turbellarian Gyratrix hermaphroditus. This is the first time viruslike particles have been found in a noncultivated platyhelminth species. The particles are 70 nm in diameter and have a capsidlike outer layer and an inner core measuring 40–50 nm in diameter. They occur in a crystalline arrangement in the nucleus as well as in the cytoplasm. Numerous cytoplasmic abnormalities were seen in connection with the particles. The occurrence of the particles in different tissues and their significance for the host are discussed.     

Topography and functional information of plasma membrane

By using atomic force microscope (AFM), the topography and function of the plasmalemma surface of the isolated protoplasts from winter wheat mesophyll cells were observed, and compared with dead protoplasts induced by dehydrating stress. The observational results revealed that the plasma membrane of living protoplasts was in a state of polarization. Lipid layers of different cells and membrane areas exhibited distinct active states. The surfaces of plasma membranes were unequal, and were characterized of regionalisation. In addition, lattice structures were visualized in some regions of the membrane surface. These typical structures were assumed to be lipid molecular complexes, which were measured to be 15.8±0.09 nm in diameter and 1.9±0.3 nm in height. Both two-dimensional and three-dimensional imaging showed that the plasmalemma surfaces of winter wheat protoplasts were covered with numerous protruding particles. In order to determine the chemical nature of the protruding particles, living protoplasts were treated by proteolytic enzyme. Under the effect of enzyme, large particles became relatively looser, resulting that their width was increased and their height decreased. The results demonstrated that these particles were likely to be of protein nature. These protein particles at plasmalemma surface were different in size and unequal in distribution. The diameter of large protein particles ranged from 200 to 440 nm, with a central micropore, and the apparent height of them was found to vary from 12 to 40 nm. The diameter of mid-sized protein particles was between 40―60 nm, and a range of 1.8―5 nm was given for the apparent height of them. As for small protein particles, obtained values were 12―40 nm for their diameter and 0.7―2.2 nm for height. Some invaginated pits were also observed at the plasma membrane. They were formed by the endocytosis of protoplast. Distribution density of them at plasmalemma was about 16 pits per 15 μm2. According to their size, we classified the invaginated pits into two types―larger pits measuring 139 nm in diameter and 7.2 nm in depth, and smaller pits measuring 96 nm in diameter and 2.3 nm in depth. On dehydration-induced dead pro-toplasts, the degree of polarization of plasma membranes decreased. Lipid molecular layers appeared relatively smooth, and the quantity of integral proteins reduced a lot. Invaginated pits were still de-tectable at the membrane surface, but due to dehydration-induced protoplast contraction, the orifice diameter of pits reduced, and their depth increased. Larger pits averagely measuring 47.4 nm in di-ameter and 31.9 nm in depth, and smaller pits measuring 26.5 nm in diameter and 43 nm in depth at average. The measured thickness of plasma membranes of mesophyll cells from winter wheat examined by AFM was 6.6―9.8 nm, thicker in regions covered with proteins.