Effect of restrictive conditions on the growth and morphology of a temperature-sensitive mannose-requiring mutant of Aspergillus nidulans

When incubated at 45 degrees C in the absence of added mannose, pregrown hyphae of a temperature-sensitive, mannose-relief mutant (mnrA455) of Aspergillus nidulans grew normally for a short time (4-5 h) before exhibiting an abnormal morphology consisting of the production by hyphae of discrete spherical swellings called balloons. These swellings could be up to 10 microns in diameter and were produced either at or behind the hyphal apex. Often only one swelling was produced in association with each hyphal tip, but in a significant minority of cases (approximately 19.6%) a second balloon was produced in close association with the first. Hyphal tip extension slowed before and during balloon formation, but growth at individual tips did not usually stop when a balloon began to be formed in the same hypha. All tip extension ceased after approximately 8 h in cultures maintained at 45 degrees C. However, normal growth resumed 45-60 min after transfer of such a culture to the permissive temperature of 37 degrees C even after 48 h at 45 degrees C. Electron microscopic examination indicated that balloons consistently had thicker walls than the surrounding hyphae but that no accumulation of cytoplasmic vesicles was apparent within them. This indicates that a modification of wall structure, probably including deposition of new wall material, was caused by a mannose deficiency, but that this altered wall synthesis and attendant hyphal swelling was not due to diversion of the normal vesicle-mediated tip-extension system to the side walls of hyphae.     

Observations on the ultrastructure ofFrankia sp. in root nodules ofDatisca cannabina L.

Summary The fine structures of the microsymbiont inside the root nodules ofDatisca cannabina have been studied by light, by transmission- and by scanning-electron microscopy. The endophyte is prokaryotic and actinomycetal in nature. The hyphae are septate and branched, diameter 0.3–0.5 m. The tips of hyphae are swollen to form electron-dense, clubshaped to filamentous vesicles, ranging in diameter: 0.4–1.4 m. The endophyte penetrates through walls of the cortial cells. The infected zone is kidney shaped and confined to one side of the acentric stele. The orientation of infection is reversed from other actinorhizae exceptCoriaria. The hyphae are near the host cell wall and vesicles are directed towards the central vacuole. Vesicles are aseptate and no collapsing of the vesicle cell wall (void area) has been observed. Vesicle clusters structures are globular with an opening at one side of the cluster. The host cell is multinucleate or contains a lobed nucleus. Groups of mitochondria are located in between the hyphae, suggesting a strong association between the host and the endophyte for energy supply and amino acid production. The consequences of the inability to separate the mitochondria from the vesicle clusters in nodule homogenates in physiological studies have been discussed.Isolated vesicles clusters showed dehydrogenase activity, indicated by the presence of formazan crystals, after incubation with NADH and NBT. Strongest reducing activity was found within the vesicles. The possible role of filamentous vesicles in nitrogen fixation has been discussed.     

Ultrastructure of freeze-substituted hyphae of the basidiomyceteLaetisaria arvalis

Summary The ultrastructure of freeze-substituted (FS) hyphae ofLaetisaria arvalis is described and compared to that of similar hyphae preserved by conventional chemical fixation (CF). The outline of membrane-bound organelles as well as the plasma membrane was smooth in FS cells. In contrast, hyphae preserved by CF exhibited membrane profiles that were extremely irregular. Centers of presumed Golgi activity were best preserved by FS. Microvesicles, 27–45 nm diameter and hexagonal in transverse section, were observed most readily in FS cells. Filasomes (= microvesicles within a filamentous matrix) were only observed in FS cells. Apical vesicles, 70–120 nm diameter, associated with the centers of Golgi activity and within the Spitzenkörper region exhibited finely granular matrices in FS hyphae, whereas in CF hyphae the contents were coarsely fibrous and less electron-dense. Microvesicles were present at hyphal apices and regions of septa formation. Filasomes were also found at regions of septa formation as well as along lateral hyphal tip cell walls. Microvesicles, but not filasomes, were observed in membrane-bound vesicles (= multivesicular bodies) and in larger vacuoles. Filaments, 5.2–5.4 nm wide, were juxtaposed with centripetally developing septa. Cytoplasmic inclusions, 20–40 m in length, composed of bundles of 6.7–8.0 nm wide filaments were observed in both FS and CF hyphae.     

Anterior adhesive areas and adjacent secretions in the parasitic flatworms Decacotyle lymmae and D. tetrakordyle (Monogenea: Monocotylidae) from the gills of stingrays

Abstract. The monogeneans Decacotyle lymmae and D. tetrakordyle (Monocotylidae: Decacotylinae), from gills of the dasyatid stingrays Taeniura lymma and Pastinachus sephen , respectively, have a single aperture for adhesive secretion on each side of the anterior ventrolateral region. Rod-shaped bodies (S1) and electron-dense spherical secretion (S2) exit through specialised ducts opening adjacent to one another within these apertures. The S1 bodies are 230 ± 11 nm wide and ≥4 μm long in D. lymmae and 240 ± 9 nm wide and ≥3.3 μm long in D. tetrakordyle . The S2 bodies have a diameter of 88 ± 7 nm in D. lymmae and 65 ± 6 nm in D. tetrakordyle . The apertures are unusual in being extremely small (internal diameter, 3–5 μm). Each aperture has a slit-like surface opening as small as 160 nm wide, surrounded by muscle fibres indicating that they may be opened and closed. The aperture is also surrounded and underlain by muscle fibres that may aid in secretion from, or even eversion of, the tissue within the aperture. Sensilla/cilia are also found within the apertures. Additional secretions from anteromedian and anterolateral glands (body glands), each containing granular secretions, occur in profusion and exit anteriorly and posteriorly to the position of the apertures, through duct openings in the general body tegument. These granular secretions do not appear to be associated with anterior adhesion. Both species show similarities in aperture, underlying tissue, sense organ, and secretion detail, in accordance with findings from other monogenean genera, and which supports the importance of such data for phylogenetic studies.     

Ultrastructure of Wall Swellings of Germinating Sporangia of Phytophthora palmivora (Butl.) Butl.

Germ tubes of directly germinating sporangia of P. palmivoraincubated in yeast extract solution at 30 ?C usually developedinto prominent swellings from which hyphae later emerged. Thegerm tubes arose as an extension of a new germination wall formedinternal to the sporangial wall prior to germination. The germtube swellings contained typical hyphal organelles. The germtube swelling possessed a thicker wall than both hyphae growingout of it and germ tubes that did not form swellings.     

Deciphering interfungal relationships in the 410-million-yr-old Rhynie chert: Morphology and development of vesicle-colonizing microfungi

The intraradical portion of arbuscular mycorrhizal (AM) fungi comprises mycelium, vesicles, and special physiological interfaces termed arbuscules; sometimes mycorrhizal fungi also produce spores within their hosts. Arbuscules are ephemeral structures that collapse after a few days, while the hyphae and vesicles appear to remain intact for some time after arbuscule senescence (post-arbuscule stage). However, little is known about the fate of mycorrhizal fungi in the post-arbuscule state. The Lower Devonian Rhynie chert yields the oldest fossil evidence of arbuscular mycorrhizas, including multiple specimens of mycorrhizal axes in the post-arbuscule stage. These fossils indicate that many older vesicles of Rhynie chert mycorrhizal fungi are colonized by other microfungi. Three types of fungal remains in vesicles are distinguished based on morphology and development: (1) spheroidal propagules up to 55 μm in diameter extending from short, distal branches of a hypha; (2) spheroidal propagules up to 23 μm in diameter produced within a tenuous mycelium; and (3) thin-walled propagules up to 10 μm in diameter within a hyphal inflation. The abundance of microfungal propagules in vesicles of Rhynie chert mycorrhizal fungi suggests that the mycorrhizal vesicles in some way positively affected the development and reproduction of the intrusive microfungi. Although the systematic affinities of the intrusive microfungi remain unresolved, this discovery is important because it broadens our understanding of the levels of organismal interactions that existed in early non-marine ecosystems.     

Ripening-related changes in the cell walls of Spanish pear (Pyrus communis)

Unripe Spanish pears ( Pyras commanis L. ev. Blanquilla ) were ripened at 18°C for 5 and 10 days. Softening of the cortical tissues was associated with swelling of parenchyma cell walls from 1 to more than 5 μm in 10 day ripe pears, by which time the pears were over ripe. However, there was little indication of cell separation and the middle lamella could be detected between most cell walls. Furthermore, cell separation was constrained by regions rich in plasmodesmata where wall swelling was prevented. Parenchyma cells in the 500 μm of tissue underlying the epidermis did not undergo ripening-related changes to the same extent as those of the cortex. These cells, in combination with a sub-epidermal layer of lignified sclereid clusters, constituted a relatively tough and protective skin. Ripening of the cortical tissues was associated with a depletion of alcohol-insoluble pectic polysaccharides, as indicated by the decrease in arabinose and uronic acid. Analysis of alcohol-insoluble cell wall preparations enriched in either parenchyma or sclereid cell walls indicated that this change was predominantly associated with the parenchyma walls. Such changes were less prominent in the peel. The decrease in pectic polysaccharides was accompanied by an increase in their solubility. During ripening, the sclereid clusters of the cortex continued in develop, as indicated by an increase in their size and yield of cell wall xylose and glucose. Cortical parenchyma cells radiating from the sclereids were firmly attached to the lignified cells. This was due to lignification extending from the sclereids into the primary walls of the parenchyma cells. We conclude that dissolution of pectic polysaccharides is one of several factors which determine softening during ripening of Spanish pears.     

Reproduction and fecundity of tucunaré, Cichla kelberi (Perciformes: Cichlidae), an exotic species in Três Marias Reservoir, Southeastern Brazil

To evaluate the reproductive cycle and fecundity of tucunaré ( Cichla kelberi Kullander & Ferreira, 2006), 697 specimens were captured in Três Marias Reservoir, São Francisco River, Brazil during 1994–1995 and 2005–2006. Reproductive activity was recorded throughout the sampling periods, with females exhibiting asynchronous oocyte development and multiple spawnings with a peak in September–October. Vitellogenic oocytes of the tucunaré were elliptical in shape with the longest diameter approximately 1230 μm and the shortest 700 μm, yolk globules with ellipsoid inclusions, lipid vesicles, small cortical alveoli and thin zona radiata (9.3 ± 2.0 μm thickness). Follicular cells were prisma-shaped (49.0 ± 16.4 μm) in the vegetative pole, progressively becoming cubic cells in the animal pole toward the micropyle. Histochemical analyses indicated the presence of mucosubstances in the outer zona radiata and follicular cells that could be contributors to egg adhesiveness. Batch fecundity ranged from 4450 to 13 900 oocytes for females 31.5–43.5 cm total length, respectively, and correlated to gonadal weight ( r ² = 0.80) and body weight ( r ² = 0.70). Mean relative fecundity was 10.6 vitellogenic oocytes per gram body weight. As tucunaré is an exotic piscivorous species well-adapted to the Três Marias Reservoir, the present work may be considered a contribution toward future strategies for population control.     

The relationship between food particle size and larval size in Simulium noelleri Friederichs

SUMMARY. 1. In a laboratory experiment, larvae of Simulium noelleri were fed on polystyrene latex microspheres of a range of diameters from 5 to 100 μm.
2. Examination of the particle size distribution in the water used in the experiment showed those <13μm to be the most numerous (87% of all particles present). Particles of this size made up 57±2% (mean ±SE)of the total of those in the gut of larvae.
3. Comparison of particle composition in the gut contents and the water of the experiment, using Jacob's index of electivity, showed that larvae of all sizes filtered proportionally fewer of the particles <13 μm and more of those >13μm.
4. As larvae increased in size they became better able to filter the largest particles present (>52μm in diameter) and less well able to filter the smallest particles (<13μm in diameter).
5. Gut retention time was longer in larger larvae.
6. The biology of S. noelleri , which inhabits lake outlets in high population densities, is considered in the light of these findings.     

Otolith microstructure and daily age of Anguilla japonica, Temminck & Schlegel elvers from the estuaries of Taiwan with reference to unit stock and larval migration

To test the hypothesis that the Japanese eel, Anguillu japonica , elvers in the eastern and western coasts of Taiwan are recruited from two different spawning grounds and to increase the knowledge of the early life history of the eel, the otolith microstructure and daily age of elvers collected from five estuaries in the coast of Taiwan during December 1989 through February 1990 were examined by scanning electron microscopy. The total length of elvers at arrival in the estuaries was similar among estuaries, averaging c . 56.0 mm and showed a seasonal decrease. The maximum radius of the sagittal otolith of elvers ranged from 124.46 to 181.82μm with a mean of 143.15 ± 12.72 μm. The otolith from centre to edge included an organic-rich primordium (9.20 ± 2.02 μm in diameter), a diffusively calcified core (20.94 ± 1.99 μm), and the daily growth increments; these three layers were probably deposited during the embryonic, yolk sac and feeding period respectively. The growth rate of the otolith was higher at the beginning of early life (0.5−1.0 μm day ⁻¹), lowest at approximately 100-days old (<0.5 μm day⁻¹), and highest 1 month before arrival at the estuary (> 1.0 pm day⁻¹). The mean age for elvers arriving in the estuaries of the coasts of Taiwan was approximately 170.4 ± 21.02 days. Neither the growth pattern of the otolith nor the age of elvers arriving in the estuary were significantly different among estuaries, indicating that the elvers in both eastern and western Taiwan were probably recruited from the same spawning ground. The growth pattern of the otolith in relation to larval migration was analysed.     

The ultrastructure of the stroma of the brown rot fungi

Summary Most of the hyphae forming the medulla of the stroma of the brown rot fungi are 4–7 in diameter and contain food reserves in large vacuoles and lipid bodies. Some stromatal hyphae have very thick walls and perform a protective function. Smaller hyphae (1–2 in diameter) form a network through the medulla and their structure suggests that they initiate the growth of vegetative hyphae and spores after the stroma has passed through a period of rest.     

Cell surface appendages of lactobacilli

Abstract Seventeen vaginal isolates of lactobacillus were negatively stained using 1% phosphotungstic acid and examined by transmission electron microscopy. One or two flagella were observed in a polar or, less frequently, sub-polar position on 14 of the 17 isolates. The structures, measuring 12 nm in diameter and up to 12 μm long, were seen on cells cultured in both solid and liquid MRS medium. Fimbriae, measuring 3.5 nm in diameter and 0.1 μm to 1.0 μm in length, extended from the cell surface of 15 of the 17 lactobacillus isolates. Production of fimbriae was maximal when the lactobacilli were cultured on solid medium in an anaerobic atmosphere. Fimbriation significantly increased adhesion of lactobacilli to vaginal epithelial cells in vitro.     

Pollen morphology of Croton sect. Lamprocroton (Müll. Arg.) Pax (Euphorbiaceae) and its taxonomic implications

A palynological analysis of 22 taxa of Croton sect. Lamprocroton (Müll. Arg.) Pax was accomplished in order to obtain evidence for a good interspecific delimitation and phylogenetic characterization of this group. Samples of 22 taxa (from which approximately 90% of the species belonged to this section) were obtained from herbarium specimens; moreover, pollen grains were analyzed with light and scanning electron microscopy. Characters observed from pollen grain diameter and from exine ornamentation were registered and compared. Pollen grains were found to be spherical, apolar, inaperturate and with a diameter ranging from 49.32±0.94 μm in C. ehrenbergii Schltdl. to 86.42±1.33 μm in C. muellerianus L. R. Lima . Pollen morphology of three recently described species of Croton was analyzed for the first time herein. In addition the results of this study confirm the distinctness of these species but do not support a recently proposed new variety of C. pallidulus Baill.     

A simple method for the preparation of small unilamellar vesicles

A simple and quick method for the preparation of small unilamellar vesicles (SUV) was developed. SUV are spontaneously formed by swelling of the specially prepared phospholipid film in water/buffer. Normally, large multilamellar vesicles (MLV) are formed when a phospholipid film is dissolved in water. To prevent the formation of multilamellar structures we used the slightly charged phospholipids which exhibit infinite swelling while the formation of large structures was prevented by the deposition of the phospholipid film on the support with small surfaces. These two requirements were met by mixing a small amount of ionic detergent into phospholipid which was deposited on microcrystals. The size and size distribution of the produced vesicles depend on the size and homogeneity of the microcrystals. When 1.5 wt% of cetyltetramethylammonium bromide (CTAB) in egg yolk phosphatidylcholine was deposited on zeolite X microcrystals with crystallite sizes of approx. 0.4 μm a homogeneous population of vesicles with average diameter 21.5 nm was obtained.     

Three New Limax Amoebae Isolated from Marine Surface Sediments: Vahlkampfia caledonica N. Sp., Saccamoeba marina N. Sp., and Hartmannella vacuolata N. Sp.

ABSTRACT. Three new limax amoebae, isolated from marine, surface sediment samples are described using light microscopic and fine structural features. One species, characterized by eruptive locomotion typical of the family Vahlkampfiidae, is assigned the name Vahlkampfia caledonica (47.4 ± 16.0 μm × 12.1 ± 3.2 μm). The other two monopodial species move with steady locomotion characteristic of the family Hartmannellidae. One is a Saccamoeba with a distinct posterior bulbous uriod, vaculoes containing prominent crystals, glycocalyx with cup-like components, and spherical nucleus with central nucleolus. It is assigned the name Saccamoeba marina (72.5 ± 14.9 ±μm × 20. 7 ± 4. 5 μM). The other hartmannellid limax amoeba moves by steady locomotion and has a rather constant monopodial from, lacks a uroid, but has occasional trailing masses of cytoplasm, contains cup-like structures in the glycocalyx, and is characterized μm). Few limax amoebae have been described from marine environment and these data provide additional evidence that limax amoebae may be more abundant in marine sediments that realized previously.     

木耳菌丝老化过程的微形态学研究

分别从显微和超微结构观察木耳菌种老化过程中菌丝细胞的形态变化。结果显示：接种后30d时,光镜下观察到菌丝结构均匀紧凑,细胞壁光滑;电镜下观察到细胞结构完整,内含物丰富,各种细胞器形态规整,没有老化现象。接种后60d时,光镜下菌丝部分肿胀,色泽加深;电镜下细胞壁疏松,线粒体和液泡肿大,细胞核不规则肿胀,核仁消失,脂肪滴和囊泡增多,并有少量电子致密度高的嗜锇性黑色颗粒状物质出现,表明菌丝开始老化。90d时,光镜下部分菌丝严重肿胀,且色泽更深;电镜下线粒体和液泡肿胀明显,部分细胞核破裂,脂肪滴、囊泡和电子致密度高的嗜锇性黑色颗粒状物质显著增多,细胞壁更加疏松。120d时,光镜下许多菌丝开始断裂,色泽进一步加深;电镜下细胞壁塌陷,膜系统也随之解体,线粒体等细胞器部分溶解消失。150d时,光镜下大部分菌丝完全断裂,并失去菌丝形态;电镜下细胞膜及其内含物已基本消失,只剩部分严重塌陷的细胞壁残骸。由此表明,木耳菌丝的老化是一个由个别向整体逐渐过渡的不可逆的过程。     

A Cytological Study on the Nutrient Uptake Mechanism of a Saprophytic Orchid Gastrodia elata

There are three type cells infected by the mycorrhizal fungus, ArmiUaria mellea (Vahl ex Fr. ) Karst in the tube cortex of Gastrodia elata BI., namely the passage cells, host cells and digesting cells. Ultrastructural study demonstrated that the passage cells were distroyed by the hyphae, the host cells kept symbiotic with the hyphae, but the digesting cells could become inversely parasitic on the hyphae from which nutrient were being uptaken. The detail process of the digesting cells obtaining nutrient'from the fungus is described as follows: Firstly the digesting cells began to attack the invading hyphae by releasing numerious electron-transparent vesicles of lysosomal property, secondly they took up the soluble organic material leaked out from the digested hyphae by forming many electron-dense endocytic tubes and vesicles, and finally they endocytosed and hydrolysed the insoluble hyphal walls by forming large digesting vacuole in which a piece of hyphal wall was completely enveloped.     

Three-dimensional architecture of the tubular endocytic apparatus and paramembranous networks of the endoplasmic reticulum in the rat visceral yolk-sac endoderm

The three-dimensional architecture of the tubular endocytic apparatus and the endoplasmic reticulum in the rat yolk-sac endoderm was investigated after loading with horseradish peroxidase-conjugated concanavalin A by intrauterine administration. After 30 min, small vesicles (50–150 nm in diameter), small tubules (80–100 nm in diameter) and large vacuoles (0.2–1.0 m in diameter) in the apical cytoplasm were labeled with the tracer, but lysosomes (1.0–3.5 m in diameter) in the supranuclear cytoplasm were not labeled until 60 min after loading. Stereo-viewing of the labeled small tubules in thick sections revealed that they were not isolated structures but formed three-dimensional anastomosing networks, which were also confirmed by scanning electron microscopy after maceration with diluted osmium tetroxide. Their earlier labeling with the endocytic tracer, localization in the apical cytoplasm and three-dimensional network formation indicated that the labeled small tubules represented tubular endosomes (tubular endocytic apparatus). These well-developed membranous networks provided by the tubular endosomes are suggested to facilitate the receptor-mediated endocytosis and transcytosis of the maternal immunoglobulin in the rat yolk-sac endoderm. Scanning electron microscopy further revealed lace-like networks of the smooth endoplasmic reticulum near the lateral plasma membrane. Their possible involvement in transport of small molecules or electrolytes is discussed.     

Eimeria tarentolae n. sp. from the Moorish Gecko, Tarentola mauritanica

ABSTRACT. A new species of Eimeria from the Moorish gecko, Tarentola mauritanica , of Minorca, Spain, is described. Oocysts of Eimeria tarentolae are ellipsoidal and measure 17.8 (17.6–18.7) μm by 13.5 (12.9–14.0) μm. Micropyle, oocyst residuum, and polar granule are absent. Each oocyst contains four spherical to slightly subspherical sporocysts 6.8 (6.4–7.0) μm in diameter. A sporocyst residuum is present, while a Stieda body is lacking. Sporulation is completed within 24–32 h at 21 ± 2°C.