Auffällige ER-Konfigurationen in Protein-Polysaccharidschleim-sezernierenden pflanzlichen Drüsen

In the ovary of Aptenia cordifolia and Platythyra haeckeliana placentary papillae produce a slime containing polysaccharides and proteins. These papillae show two types of conspicuous vacuoles enclosed by rough ER cisternae and complexes of concentrically arranged rough ER. The enclosed vacuoles probably play an important role in the accumulation of the polysaccharide-protein slime. In the case of storage vesicles (first vacuole type) derivates of the Golgi apparatus are enclosed by ER. In other instances (second vacuole type) ER cisternae which have lost their membrane-bound ribosomes seem to delimit protoplasmic regions free of organelles.

     

Genetic diversity of Nostoc microsymbionts from Gunnera tinctoria revealed by PCR-STRR fingerprinting

The cyanobacteria belonging to the genus Nostoc fix atmospheric nitrogen, both as free-living organisms and in symbiotic associations with a wide range of hosts, including bryophytes, gymnosperms (cycads), the small water fern Azolla (Pteridophyte), the angiosperm genus Gunnera, and fungi (lichens). The Gunnera–Nostoc symbiosis is the only one that involves a flowering plant. In Chile, 12 species of Gunnera have been described with a broad distribution in the temperate region. We examined the genetic diversity of Nostoc symbionts from three populations of Gunnera tinctoria from Abtao, Chiloé Island, southern Chile, and microsymbionts from other two species of Gunnera from southern Chile, using PCR amplification of STRR (short tandemly repeated repetitive) sequences of the Nostoc infected tissue. To our knowledge, this is the first report of PCR fingerprinting obtained directly from symbiotic tissue of Gunnera. Genetic analyses revealed that Nostoc symbionts exhibit important genetic diversity among host plants, both within and between Gunnera populations. It was also found that only one Nostoc strain, or closely related strains, established symbiosis with an individual plant host.     

Surface ultrastructure of the tegument of Clonorchis sinensis newly excysted juveniles and adult worms.

The tegumental structures of newly excysted juveniles and adult worms of Clonorchis sinensis were studied using scanning and transmission electron microscopy. After excystation the juvenile's tegumental surface is characterized by knoblike protuberances and is armed almost entirely with numerous rows of small spines encircling the body. These spines are double- or triple-pointed on the anterior portion of the body and become single-pointed posteriorly. Four types of presumed sensory structures were observed as follow: A) ciliated knoblike papillae and B) nonciliated platelike papillae, both of which are arranged in rougly a bilaterally symmetrical pattern dorsally, ventrally, and laterally; C) rounded swellings of nonciliated papillae on the lips of the ventral and oral suckers, which are characterized in the transmission electron microscope by a rounded dense body in the apical bulb; and D) a sensory receptor with a bulbous projection having the appearance of a modified cilium, which was not found with SEM likely owing to its being enclosed by an extension of the tegument. In full-grown adult worms, the tegumental surface is knobbed or lobulated in various forms without surface spines. The tegumental structures in the adults appear to be clearly differentiated from those in the juveniles. Upraised, buttonlike papillae, each topped by a short cilium, which are similar to the Type A papillae in the juveniles, are distributed thickly around the oral and ventral suckers, and are rather randomly scattered over the remainder of the body. Some nonciliated swollen papillae were found on the lip of the ventral sucker.     

Nitrogen deprivation stimulates symbiotic gland development in Gunnera manicata

Gunnera is the only genus of angiosperms known to host cyanobacteria and the only group of land plants that hosts cyanobacteria intracellularly. Motile filaments of cyanobacteria, known as hormogonia, colonize Gunnera plants through cells in the plant's specialized stem glands. It is commonly held that Gunnera plants always possess functional glands for symbiosis. We found, however, that stem gland development did not occur when Gunnera manicata plants were grown on nitrogen (N)-replete medium but, rather, was initiated at predetermined positions when plants were deprived of combined N. While N status was the main determinant for gland development, an exogenous carbon source (sucrose) accelerated the process. Furthermore, a high level of sucrose stimulated the formation of callus-like tissue in place of the gland under N-replete conditions. Treatment of plants with the auxin transport inhibitor 1-naphthylphthalamic acid prevented gland development on N-limited medium, most likely by preventing resource reallocation from leaves to the stem. Optimized conditions were found for in vitro establishment of the Nostoc-Gunnera symbiosis by inoculating mature glands with hormogonia from Nostoc punctiforme, a cyanobacterium strain for which the full genome sequence is available. In contrast to uninoculated plants, G. manicata plants colonized by N. punctiforme were able to continue their growth on N-limited medium. Understanding the nature of the Gunnera plant's unusual adaptation to an N-limited environment may shed light on the evolution of plant-cyanobacterium symbioses and may suggest a route to establish productive associations between N-fixing cyanobacteria and crop plants.     

The α-amylase inhibitor of bean seed: two-step proteolytic maturation in the protein storage vacuoles of the developing cotyledon

In the nitrogen fixing symbiosis between Nostoc and the angiosperm Gunnera , the cyanobiont is found in stem glands and is thought to have a heterotrophic mode of nutrition. To investigate whether the photosynthetic machinery in the cyanobiont is down-regulated in the symbiosis, the presence of the phycobiliproteins, phycoerythrin and phycocyanin, and ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco, EC 4.1.1.39) in cyanobionts of Gunnera magellanica Lam. and in a free-living (cultured) isolate of the cyanobacterium was studied by immunoelectron microscopy. Carboxysomes were numerous in all vegetative cells (ca 3.5 per cell section), and on an area basis they showed a high Rubisco label compared to the cytoplasm; but recalculation on a volume basis demonstrated that the carboxysomal fraction of Rubisco decreased in the cyanobiont along the plant stem. Along the whole Gunnera stem both types of phycobiliproteins were present in the symbiotic Nostoc and in amounts equivalent to or above those detected in the free-living isolate. As the symbiotic Nostoc is located intracellularly, out of reach of light in the plant stem, the findings indicate a lack of regulation of the photosynthetic protein synthesis in the symbiotic state.     

Observations on the innervated face of the electrocyte of the main organ of the electric eel (Electrophorus electricus L.)

Summary The innervated face of electrocytes in the main electric organ of Electrophorus electricus L. was examined by light microscopy, both conventional and with Nomarski contrast, and by transmission and scanning electron microscopy. Acetylcholinesterase cytochemistry was used in the demonstration of the greater density of synapses over the caudal papillae. The various techniques contributed to a better understanding of the distribution and form of papillae and synapses at the posterior face of the electrocyte. Caudal papillae are longer and thinner than those at the rostral face, but it was not possible to recognize a different type sometimes referred to in the literature as small papillae. The contact of nerve endings with the electrocyte seems to be made predominantly on the terminal half of caudal papillae, however a smaller number occur elsewhere on the posterior face. Synaptic terminals frequently appear as round profiles, but may be also elongated, with or without bulges, usually occupying a depression, and separated from the post-synaptic membrane by a space of 60–100 nm, where an expansion may be found.     

The first record of Dirofilaria immitis infection in a Humboldt penguin, Spheniscus humboldti

Dirofilaria immitis infection is an important parastic disease in many mammals, especially canids, but has not been reported in bird hosts. Filarial worms were isolated from the lumen of the right atrium of the heart and the connective tissue of the lung of a captive female Humboldt penguin, Spheniscus humboldti, that died at a zoo in Japan. One of these worms was observed morphologically and identified as D. immitis by features such as 4 pairs of cephalic papillae, 1 pair of cervical papillae, esophagus divided into 2 regions, 4 pairs of pre-anal papillae, 5 pairs of post-anal papillae, and unequal spicules. In addition, the partial DNA sequence (234 bp) of mitochondrial CO / gene of the filarial worm was identical to that of D. immitis. This is the first report of D. immitis infection in a bird.     

Studies on the tardigrades. I. Fine structure of the anterior foregut of Milnesium tardigradum Doyère

The fine structure of the anterior foregut of the tardigrade Milnesium tardigradum is presented. The oral region consists of a terminal mouth opening surrounded by six plate-like lips lying within a circlet of six prominent papillae. The buccal cavity is enclosed within a thick cuticular tube which possesses appendage structures, the stylet sheaths, stylet supports and paired protrusible stylets. Two large salivary glands envelop the buccal structures and contain voluminous amounts of secretory product. The arrangement, possible functions and phyletic significance of these structures are discussed.     

Characterization by genotypic methods of symbiotic Nostoc strains isolated from five species of Gunnera

The genetic diversity of ten symbiotic Nostoc strains isolated from different Gunnera species was investigated. The strains were analyzed using molecular methods with different taxonomic resolutions, including restriction fragment length polymorphisms (RFLP) of the PCR-amplified 16S ribosomal gene and the 16S-23S internal transcribed spacer (ITS) region combined with computer-assisted analyses. The functional gene hetR, assigned to heterocyst differentiation, was used for denaturing gradient gel electrophoresis. A high genetic diversity was observed among the isolates even in the conserved gene coding for the small ribosomal unit. No correlation was observed between clustering of cyanobacteria and the host species of Gunnera.     

Neuro-histochemical study of the tongue of Gallus domesticus (White Leghorn)

An investigation was undertaken to study the neuro-histochemistry of tonue of Gallus domesticus (white Leghorn) by cholinesterase technique under maintained pH of 5.2, temperature 40 degrees C and incubation period 20 h, and Ungewitter's silver impregnation method. Fungiform (8--16 in number), filiform (5--16 in number), circumvallate and foliate (numerous) were recorded profusely innervated in the various regions of the tongue. Neuro-muscular spindles related either with the non-myelinated, myelinated nerve fibres or with the ganglia via pre- or postganglionic nerve fibre showed cholinesterase (AChE) positive activity in the form of dark brown patches. Neural network over the glands, formed the basket envelope likestructures. Glands showed positive cholinesterase (AChE) reaction on the periphery. Dot-like nerve endings were connected with the neural terminal network in the foliate papillae region and motor end plate like endings on blood vessel, were occasionally recorded. Parasympathetic ganglia of various shape and sizes, arranged in chain-like fashion were observed in the connective tissue space between the fungiform and circumvallate papillae and foliate papillae region. The pre- and postganglionic fibres of these ganglia were related with the anteriorly and posteriorly located papillae via the neural terminal network. Cholinesterase (AChE) activity was marked either in the centre, of the fungiform or on the periphery of the filiform, circumvallate and foliate papillae.     

On the homology of the scales in Gunnera (Gunneraceae)

In the genus Gunnera , the species of subgenus Panke are unique in having large, triangular scales between the leaves on the rhizomes. The morphological significance of these scales has been extensively debated in the past. They have been interpreted as stipules, ligules, or cataphylls, with attempts made to identify homologous structures in representatives of the other subgenera. In the stoloniferous subgenus Misandra, the shoot apex is covered by a hoodlike structure, usually termed ochrea, generally considered homologous to the scales in Panke . Efforts to find similar structures in the remaining subgenera were unsuccessful. Due to the lack of a phylogenetic hypothesis, the homology of the scales has remained unresolved. In this study, the shoot apices of species from all subgenera were examined by light and scanning electron microscopy and the results interpreted in a phylogenetic context. The tip of the stolons in subgenera Pseudogunnera and Milligania carries two opposite cataphylls at the base of the new shoot. These are homologous to the ochrea in Misandra which is in turn homologous to the scales in Panke ; the latter are thus neither stipules nor ligules but cataphylls. The stems of subgenera Ostenigunnera and Gunnera have no similar structures. © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003, 142 , 301−308.     

Ultrastructural studies on Diphyllobothrium ditremum and D. dendriticum (Cestoda, Pseudophyllidea), with emphasis on the scolex tegument and the tegument in the area around the genital atrium.

The scolices of D. ditremum and D. dendriticum are similar in regard to general morphology and ultrastructure of the tegument. At the scolex apex and along the upper bothrial edges sensory endings are numerous. The inner bothrium surface of D. ditremum appears to be plain or smooth, the distal cytoplasm is only about 0.6 mum thick and covered with long and slender microtriches. When D. ditremum is attached to the host gut an adhesive layer of secreted substance exists between the inner bothrium surface and the host intestinal villi. In D. dendriticum, however, no secreted layer exists between the inner bothrium surface and the host intestinal villi and the inner bothrium surface of this species appears to be lobed or lappet formed. The lobes are protrusions of the distal cytoplasm and are covered with long and slender microtriches. Large conglomerations of secretion globules appearing to be enclosed by a double membrane, are observed in the scolices of D. dendriticum and D. ditremum, although more frequent in the latter. Differences existing between the distal cytoplasm of the scolex and the gravid proglottid are described. The papillae around the genital atrium in mature and gravid segments were studied. Each papilla appears as a thickening of the distal cytoplasm, which in this region is 10-12 mum thick, when studied with transmission electron microscope. From the area between the papillae sensory endings appearing in sections to be either single, double or triple are described.     

葡萄糖和氢对Gunnera／Nostoc固氮共生体固氮活力的影响

Ｇｕｎｎｅｒａ／Ｎｏｓｔｏｃ固氮共生体固氮相对效率（ＲＥ）可在０．２６～０．８０之间变动,而不是一个常数。外加１．５％葡萄糖液可使其固氮活力提高约１００％,同时也使组织的呼吸速率提高了近１６０％。加外源Ｈ２可使其固氮活力提高近１００％,但却使组织的呼吸速率降低了近５０％。正常生长条件下的组织净放Ｈ２量较低．而外源２％葡萄糖液可使组织净放Ｈ２量提高近２倍。外加５ｍｍｏｌ／Ｌ的ＮＨ１Ｃｌ溶液可使其固氮活力下降约７０％。故认为Ｇｕｎｎｅｒａ／Ｎｏｓｔｏｃ共生体固氮活力受碳水化合物供应状况及比代谢两者构成的“还原力库”或“电子库”的调节,在此“还原力库”中,Ｈ２代谢起到了一个“中间调节者”的作用。     

Skin structure in the snout of the Australian lungfish,Neoceratodus forsteri (Osteichthyes: Dipnoi)

Many fossil lungfish have a system of mineralised tubules in the dermis of the snout, branching extensively and radiating towards the epidermis. The tubules anastomose in the superficial layer of the dermis, forming a plexus consisting of two layers of vessels, with branches that expand into pore canals and flask organs, flanked by cosmine nodules where these are present. Traces of this system are found in the Australian lungfish, Neoceratodus forsteri, consisting of branching tubules in the dermis, a double plexus below the epidermis and dermal papillae entering the epidermis without reaching the surface. In N. forsteri, the tubules, the plexus and the dermal papillae consist of thick, unmineralised connective tissue, enclosing fine blood vessels packed with lymphocytes. Tissues in the epidermis and the dermis of N. forsteri are not associated with deposits of calcium, which is below detectable limits in the skin of the snout at all stages of the life cycle. Canals of the sensory line system, with mechanoreceptors, are separate from the tubules, the plexus and the dermal papillae, as are the electroreceptors in the epidermis. The system of tubules, plexus, dermal papillae and lymphatic capillaries may function to protect the tissues of the snout from infection.     

Wheat Streak Mosaic Virus-induced Cytoplasmic Expansion and Membrane Proliferation

The cytoplasmic volume of wheat streak mosaic virus-infected cells was significantly greater than that of cells in healthy control tissue. Ultrastructural examination revealed that mainly cellular membranes and ribosomes filled the expanded cytoplasm. Imperfectly spherical inclusions, containing continuous endoplasmic reticulum membranes at the periphery and a mixture of membranes and ribosomes in the centre, were observed near nuclei at early infection stages. The inclusions became larger as infection progressed. Membranes and ribosomes proliferated also throughout the cell, forming a matrix in which organelles and various cytopathic structures were enclosed. Numerous vesicles were observed in the cytoplasm of other WSMV-infected cells. Multi-layered membrane bodies were found at later infection stages. Virus particles were present in the central space of these myelin-like structures. The presence of apparently intermediate stages in myelin-like structure development in chloroplasts suggest that at least some of the myelin-like structures originated from the chloroplasts.     

The Nostoc-Gunnera symbiosis: carbon fixation and translocation

The in vitro specific activity of ribulose-1,5-bisphosphate carboxylase (Rubisco; EC 4. 1. 1. 39) and the dark and light in vivo CO₂ fixation activities were determined in the cyanobiont of Gunnera . Compared to the free-living isolate Nostoc PCC 9231, the in vitro Rubisco activity was high, while the in vivo CO₂ fixation was very low. Light did not significantly influence CO₂ fixation if the cyanobiont was left in the sliced Gunnera tissues, while a small light stimulation was found for CO₂ fixation of the freshly-isolated cyanobiont. The adjacent non-infected Gunnera tissue showed a very low CO₂ fixation. A rapid translocation of fixed ¹⁴CO₂ from leaves towards apical parts of the plant was apparent, in particular to the symbiotic tissue. The ¹⁴C label appeared mainly in soluble form in this tissue and was rapidly catabolised as shown by ¹⁴C chase experiments. Also, short-term experiments revealed that maximum ¹⁴C accumulation occurred in the symbiotic tissue showing the highest rates of nitrogen fixation (Söderbäck et al. 1990), about 10–15 mm from the plant apex. The data were taken to indicate that there is a modification in the photosynthetic light reaction of the cyanobiont and that the cyanobiont lives heterotrophically in the dark on photo-synthate rapidly delivered from nearby leaves of the host plant.     

Life cycle and description of a new species of brachylaimid (Trematoda: Digenea) in Spain

The life cycle of Brachylaima llobregatensis n. sp. (Trematoda: Brachylaimidae) is elucidated. Embryonated, operculated, asymmetric eggs (30.9 x 18.2 microm) are eliminated with feces of natural hosts wood mouse Mus spretus; white-toothed shrew, Crocidura russula; and an experimental host, domestic mouse, Mus musculus var. domesticus. The eggs are ingested by the helicid gastropod Helix (Cornu) aspersa, the only natural and experimental first intermediate host. The miracidium hatches from the egg, infects the snails, and develops into a branched sporocyst in the digestive gland. Microcaudate cercariae emerge from this snail and develop into unencysted metacercariae in the kidney of second intermediate host snails H. (C.) aspersa and Otala punctata (natural hosts) and Theba pisana (experimental host). Ingestion of infected snails leads to the infection of definitive hosts, with the adults inhabiting the middle part of the small intestine. There is a chaetotaxic pattern specific on the acetabular (S(II) 5-6 papillae) and body (papillae absent on P(II)) levels. Three types of cercaria papillae were observed by scanning electron microscopy: argentophilic papillae with fingerlike processes (cephalic, body, and acetabular levels); argentophilic papillae with opening (2 papillae in the M body level); and nonargentophilic dome-shaped papillae (on the cephalic C(II) level, alternating with argentophilic S(II) papillae on the ventral sucker). Suckers are subequal, with the acetabulum located in the posterior part of the anterior third of body. Vitellaria extend from anterior margin of acetabulum to between middle level and anterior margin of anterior testis.     

Development of mechanical papillae of the tongue in the domestic goose (<Emphasis Type="Italic">Anser anser f. domestica</Emphasis>) during the embryonic period

Three types of mechanical papillae, i.e., conical, filiform, and hair-like papillae, are present on the tongue in the domestic goose. Within conical papillae, we distinguish three categories: large and small conical papillae on the body and conical papillae on the lingual prominence. The arrangement of mechanical papillae on the tongue in Anseriformes is connected functionally with different feeding mechanisms such as grazing and filter-feeding. The present work aims to determine whether morphology of three types of mechanical papillae in goose at the time of hatching is the same as in an adult bird and if the tongue is prepared to fulfill feeding function. Our results revealed that the primordia of the large conical papillae start to develop during the differentiation stage. The primordia of the small conical papillae and conical papillae of the lingual papillae start to develop during the growth stage. At the end of the growth stage, only large conical papillae, three pairs of small conical papillae, and conical papillae of the lingual prominence have similar arrangement as in an adult bird. The shape and arrangement of the remaining small conical papillae probably will be changed after hatching. During embryonic period, the filiform papillae and hair-like papillae are not formed. The embryonic epithelium that covered the mechanical papillae undergoes transformation leading to the formation of multilayered epithelium. During prehatching stage, epithelium becomes orthokeratinized epithelium. In conclusion, the tongue of the domestic goose after hatching is well prepared only for grazing. The filtration of food from water is limited due to the lack of filiform papillae.     

Morphology, and muscle- and papilla-volume ratios, of the tongue of Laudakia stellio (Agamidae, Squamata): a histological and stereological study

We examined the histological structure of the tongue of Laudakia stellio, the starred agama lizard (Agamidae, Squamata), under light microscopy. We also investigated the muscle and papilla volume ratios, with volumes of each aspect of interest estimated according to the Cavalieri method. The macroscopically short, thick and muscle-rich front tip of the tongue of L. stellio does not show any bifurcation, and under light microscopy, the oval-shaped papilla-free front tip was seen to be covered by keratinized stratified epithelium. The dorsal and ventral parts were different, with the former partially covered by keratinized stratified epithelium and rich in secretory glands and secretory cells. The ventral part, which contained keratinized stratified cells, had a flat surface with no papillae. The dorsal surface of the anterior and posterior parts contained fungiform papillae, with the apical parts of these papillae containing minimal keratin; the interpapillar space was covered by keratin-free squamous stratified epithelium. The middle section of the tongue contained cylindrical-type papillae, with serous and mucous secretory glands and ducts at their base. Finally, the frontal and middle parts of the ventral and dorsal surfaces did not contain any taste buds, although there were some in the hind part of the dorsal surface. As morphometric estimates of volumes of the muscles and papillae, the mean volume ratios (relative to total tongue volume)+/-standard deviation were 0.66+/-0.03 and 0.33+/-0.03, with mean coefficients of error of 0.02 and 0.03, respectively.