Transferzellen im Xylem derValerianaceae

Stems, incl. rhizomes, and roots of 42 species ofValerianaceae were investigated in order to reveal the occurrence, structure and distribution of xylem transfer cells. Within nodes and internodes their frequency, distribution and gradients of development are similar to other families. — Within the secondary xylem of some species transfer cells can develop from cambial derivates, inValeriana tuberosa andPatrinia villosa even from pith cells. Within the turnip ofV. tuberosa transfer cells are very frequent and well developed. Here, after degradation of the cell-wall ingrowths they can be redifferentiated into storage cells which usually contain starch grains (Hüllenstärkekörner). In the transitional zone between stem and root of some predominantly herbaceous taxa transfer cells are often very frequent and form large protuberances before they degrade and lignify. SEM observations inValeriana decussata show that the cell-wall ingrowths are degradated at the beginning of lignification with the exception of brush-like protuberances remaining in the half-bordered pit-pairs. During the subsequent process of lignification the simple pits of a wall adjacent to a vessel can be transformed into corresponding pit-pairs. In this case the residues of the protuberances within the pit chamber can be transformed into incrustations similar to the vestures of bordered pits described byBailey (1933). Structural similarities between the brush-like protuberances in the half-bordered pits of theValeriana transfer cells and the ingrowths found inLauraceae (Castro 1982, 1985) are evident. Supposedly, all the cambial derivatives inValerianaceae can develop protuberances at least within their pits. Thus, it appears possible to interpret the vestures of the bordered pits as rudimentary protuberances, and to suggest that they have a specific function in the selective transport of solutes.

---

Transferzellen im Xylem derValerianaceae

     

Unusual transfer cells in the epithelium of the nectaries ofAsclepias curassavica L.

Summary The secretory cells of the nectaries ofAsclepias curassavica form a glandular epithelium in the inner parts of the stigmatic chambers. They resemble transfer cells in having many infoldings of the plasmalemma. The wall protuberances, however, are poorly developed and often lacking. The plasmalemma is highly convoluted and forms, in places, external compound membranes where the extracytoplasmic space is collapsed completely. Active glands contain dilated cisternae of the ER and large vesicles which are mainly associated with the cis face of the dictyosomes. In addition, small vesicles are observed in high number. It is discussed whether the secretion is granulocrine or eccrine and whether the enlargement of the plasmalemma is the cause or the consequence of the high secretory activity. After the secretory phase the outer peripheral part of the cytoplasm disintegrates. The remaining part of the protoplast is covered by a new plasmalemma.     

Microbiology in the 1990s: reflections about open questions and trends

The optimal conditions for protoplast formation ofCandida apicola were by using an enzyme fromArthrobacter sp. in combination with 2-mercaptoethanol. The kinetic data support the two-layered structure model of cell wall for this yeast but the structure of the cell wall depended on the age of cells and culture conditions. To regenerate the protoplasts, the type of osmotic stabilizer was important: sorbitol gave 16 to 30% regeneration. Electron microscopy revealed the presence of vesicles in the sections of protoplasts and whole cells ofCandida apicola grown in production medium and producing glycolipids. In sections of whole cells, vesicle-like structures are located in the periplasmic space and in protoplasts they can either be attached to, or released from, the cell surface. These vesicles are thought to be involved in the transport of the surface-active glycolipids and in the protection of the cell against denaturing effects.     

Transfer cells in the vascular parenchyma of roots

Structural adaptations to increased transport activities were investigated in the cells of vascular parenchyma at the site of the lateral root junction, in non-stressed plant roots. Typical transfer cells were differentiated in dicotyledonousHelianthus tuberosus and in two different genotypes ofH. annuus, the cv. IBH166 and a decorative form. In the representatives of monocotyledonous, no structural adaptations occurred in the roots ofHordeum vulgare but small and rare cell wall protuberances were found in xylem and phloem ofZea mays inbred line VIR17. Some degree of cell wall labyrinth differentiation was seen in xylem and typical transfer cells were found in phloem of the roots of the maize hybrid CE380. The capability of vascular parenchyma to differentiate transfer cells did not depend on species, genotype, or on the growing conditions withHelianthus. On the other hand, the development of the structural adaptations in monocotyledonous representatives depended on both the species and the genotype. This capability may be linked with the taxonomic and evolutionary position of plant species.     

Spores of microorganisms

Spores ofBacillus cereus (strain NCIB 8122) were germinated in a synthetic germination limited medium (GL-medium), which permitted germination but did not make the termination of post-germinative development possible. Incorporation of¹⁴C-diaminopimelic acid into the newly formed cell wall was followed in this culture. Morphological changes were studied by optical and electron microscopy. Germination was associated with the usual germination changes,i.e. depolymerization of the “bulky” cortex, differentiation of nuclear structure and mesosomes and ribosomes in the cytoplasm. At this stage the spore protoplast is surrounded by several layers: exosporium, laminated coat with four layers, residual spore wall and the protoplast membrane. During incubation in this limited medium the residual wall layer thickens and the nuclear structure, mesosomes and ribosomes were not more detectable. After enrichment of the GL medium (shift up) the thick-walled cells can form additional cell wall material, elongate and an atypical septum formation can occur. The cell wall material forms local thickenings. On long-term cultivation in the GL medium some of the cells in the GL medium lyze. If, in the course of 3–6 h the cells are transferred from the GL-medium to a solid complex medium (Difco Nutrient Agar) the thickwalled cells are transformed into dividing cells. When the cells are transferred later, their colony-forming ability rapidly decreases. The decrease of viability of the thick-walled cells derived directly from spores after their germination in the limited medium indicates that these cellular forms probably do not represent more stable cellular types that would be of considerable importance for survival of the populat ion of bacilli.     

Variations of the Surface Sculpture and Cell Wall Ultrastructure of the Zygospores in Chlamydomonas geitleri (Chlorophyta)

The superficial cell wall ornamentation in the zygospores of the alga Chlamydomonas geitleri Ettl (Chlorophyta) is formed by thickenings of the cell wall which are shaped into a network of anastomosing ribs, sometimes with local wart-like protuberances. Clearly different sculpture patterns (given by presence, arrangement and/or morphological modification of sculpture elements) were accompanied by many transient forms. Sculpture variations occurred even in clonal cultures. In the zygospore cell wall of C. geitleri, the inner, outer and middle layer can be distinguished from the morphological point of view. The relatively thin outer (sporopollenin) layer covers the whole surface of the zygospore wall. The thicker inner layer adhering to the zygospore protoplast forms, either solely or together with the middle layer (possessing a fine meshwork substructure), variously shaped thickening of the zygospore cell wall. Discussed are the ultrastructural morphology of the cell wall in Chlamydomonas zygospores, the striking similarity of the cell wall ultrastructure of zygospores in C. geitleri to the ultrastructure of the cell wall of vegetative cells in some green algae (subfamily Scotiellocystoideae), as well as the extensive morphological variability of the zygospore wall sculpture in C geitleri and its species specificity.     

Uptake of protoplasts from the filamentous fungiAspergillus nidulans andFusarium oxysporum into protoplasts from celery (Apium graveolens L.)

Summary Protoplasts isolated from celery cell suspension cultures, were mixed with fungal protoplasts, from either the saprophytic speciesAspergillus nidulans or the pathogenic speciesFusarium oxysporum. The incubation of protoplast mixtures with PEG caused close adhesion between plant and fungal protoplasts. Subsequent dilution of PEG resulted in the uptake of protoplasts from either fungal species into the plant protoplast cytoplasm. A range of PEG concentrations, incubation times and dilution rates were tested to maximise adhesion and uptake frequencies. Identification of uptake was achieved either by fluorescent staining of nuclei or by electron-microscopy. A maximum of 10% celery protoplasts had taken upA. nidulans protoplasts after PEG treatment. Fungal protoplasts were taken up into celery protoplast cytoplasm by endocytosis, and were maintained within vesicles; two bounding membranes were observed by electron microscopy. Plant protoplast viability was determined during prolonged incubation following fungal protoplast uptake. The presence ofA. nidulans protoplasts tended to maintain celery protoplast viability and although some morphological disintegration occurred intact celery protoplasts remained for at least 92 h after uptake. The uptake ofF. oxysporum protoplasts markedly depressed celery protoplast viability after 24 h incubation and greater celery protoplast disintegration occurred.Abbreviations PEG Polyethylene glycol - DAPI 4,6-diaminido-2-phenylindole - 2,4-D 2,4-dichlorophenoxyacetic acid     

Intercellular protuberances in the ground tissue ofCocos nucifera L.

Summary Prolific filamentous intercellular protuberances have been observed in the intercellular spaces of the ground parenchyma tissue in the stems ofCocos nucifera. They are visually similar to some intercellular material reported in several other plant tissues but their chemical composition is unknown. Tests for lignin, cellulose, callose, suberin and waxes have proved negative and those for pectin inconclusive. The amount of intercellular material is closely related to the thickness of the parenchyma cell wall and the protuberances appear to be produced continuously by an active cytoplasm.     

Fine structure of the host-fungus interface in orchid mycorrhiza

Summary Electron microscopy of protocorms of Dactylorhiza purpurella infected with a symbiotic Rhizoctonia sp. showed that the intracellular hyphae examined did not penetrate the plasmalemma of the host cell. Walls of hyphae within cells bore many hemispherical protuberances over which the host plasmalemma was closely pressed. we estimate that these protuberances would increase the area of contact between hyphae and host plasmalemma by about 15%. They were not found on hyphae growing on agar. Except for these protuberances, and some vesicles or tubules which invaginated the fungus plasmalemma, no other structures were seen which could be suggested to be adaptations to transport across the living fungus-host interface.     

Cell cycle related changes in the quantity of TMV virions bound to protoplasts ofNicotiana sylvestris

Summary Attachment of virions of tobacco mosaic virus to protoplasts isolated from dividing suspension cultured cells ofNicotiana sylvestris was estimated using quantitative autoradiography of individual protoplasts. Additionally, the position of each protoplast in the cell cycle was assessed by Feulgen microspectrophotometry. At pH 5.6, after preincubation with 4 g 1^–1 poly-L-ornithine, protoplasts in the G₁ and G₂ phases bound more virions than protoplasts in the S-phase. The possibility that such differential binding was caused by cyclical variation in the net charge on the protoplast membrane has been investigated. It was found that S-phase protoplasts ofN. sylvestris can be separarated from protoplasts of other cycle stages by partition in aqueous, two-phase, immiscible polymer systems, presumably because they differ in charge. Also, electrophoretic studies suggest that G₁ phase protoplasts bear higher surface charge than some non-G₁ protoplasts.     

Beeinflussung der Blattzellen vonFontinalis antipyretica durch den Bewuchs vonCocconeis placentula

In a special habitat near Lunz (Mittersee) long time epiphytism ofCocconeis placentula var.tenuistriata and var.klinoraphis onFontinalis antipyretica leaves causes characteristic, strongly localized alterations in the walls of affected cells. They turn brown, may thicken, and may produce cystolithe-like protuberances into the cell lumina. These reactions obviously are brought about by a specific property ofCocconeis, for other epiphytes do not cause any change. The comparison of theCocconeis vegetation on younger and olderFontinalis leaves shows a considerable variability of the reactions mentioned, and in another habitat none could be found. Apparently several factors cause these reactions. Altogether,Cocconeis placentula is not a completely harmless epiphyte under certain circumstances. The alterations of the moss cells may be interpreted partly as a premature aging process, partly as a defensive action corresponding to alterations induced by some parasitic fungi. Furthermore, the reactions ofCocconeis onFontinalis are remarkably parallel to the influence of the dinococcalean algaRaciborskia inermis onSpirogyra grevilleana.

     

Efficient protoplast regeneration ofBacillus thuringiensis andAgrobacterium tumefaciens

Summary Treatment ofBacillus thuringiensis andAgrobacterium tumefaciens taken from the early growth phase (8 h) with lysozyme at 1 mg/ml gave 90–99% protoplast formation and 10–12% protoplast regeneration on the minimal medium in absence of plasma expander (Bovine serum albumin). Enhanced fusion frequency was obtained when protoplasts from 8 h grown cells were used for fusion experiments.     

The ultrastructure of the salt gland of Spartina foliosa

Summary The salt gland in Spartina foliosa is composed of two cells, a large basal cell and a smaller, dome-shaped cap cell which is located on a neck-like protrusion of the basal cell. There is no cuticular layer separating the salt gland from the mesophyll tissue. The basal cell has dense cytoplasm which contains numerous mitochondria, rod-like wall protuberances, and infoldings of the plasmalemma which extend into the basal cell and partition the basal cell cytoplasm. The protuberances originate on the wall between the basal and the cap cells and are isolated from the basal-cell cytoplasm by the infoldings of the plasmalemma. While the cap cell has no partitioning membrane system or wall protuberances, it resembles the basal cell by having dense cytoplasm and numerous mitochondria.The basal cell seems to be designed for efficient movement of ions toward the cap cell. The long, dead-end extracellular channels in the basal cell of Spartina appear comparable to surface specializations seen in the secreting epithelium of animal cells which carry out solute-linked water transport. The number of mitochondria and their close association with the plasmalemma extensions suggest that they have an important role in the transfer of ions through the basal cell.The accumulated ions would move into the extracellular spaces along an osomotic gradient where the accompanying passive flow of water would move the ions into the cap-cell wall and from there the solution would pass out through the pores in the cuticle.     

ANALYSIS OF GAMETIC PROTOPLAST RELEASE IN THE CLOSTERIUM PERACEROSUM-STRIGOSUM-LITTORALE COMPLEX (CHLOROPHYTA)1

A biologically active glycoprotein (protoplast-release-inducing protein; PR-IP), which induces the release of gametic protoplasts from mating type minus (mt^-) cells of the Closterium peracerosum-strigosum-littorale complex, was prepared from a medium in which mt^- and mt⁺ cells had been previously incubated together. The process of PR-IP-inducing protoplast release was analyzed. Induction of protoplast release was dependent upon the duration of both PR-IP treatment and preincubation in nitrogen-deficient mating medium before PR-IP treatment. Low cell density in the preculture stage had a significant stimulative effect upon the induction of protoplast release. Light was necessary for protoplast release, especially just before PR-IP treatment. Chloramphenicol and 3-(4-chlorophenyl)-1,1-dimethylurea (CMU) exerted inhibitory effects on protoplast release, especially when they were applied to the preculture stage but not when they were applied to the protoplast-releasing stage after the PR-IP treatment. We suggest that preculture at a low cell density under continuous light conditions that may cause metabolic changes in the chloroplast is a very important stage for gametic protoplast release in this Closterium.     

Experimental inhibition of cell wall formation and of reversion inNadsonia elongata andSchizosaccharomyces pombe protoplasts

Summary The growth, cell wall regeneration, and the reversion of the protoplasts ofNadsonia elongata andSchizosaccbaromyces pombe cultivated in nutrient media containing snail enzyme was studied by light and electron microscopy. The protoplasts grew in the presence of snail enzyme and an incomplete cell wall composed of fibrils was formed on their surface. Thus, the presence of snail enzyme inhibited the completion of cell wall structure and, consequently, the reversion of the protoplasts to normal cells. The transfer of these protoplasts to medium free from snail enzyme led first to the completion of the cell wall and then to the reversion of the protoplasts to normal cells. The reported experiments confirmed that the regeneration of the complete cell wall preceded the protoplast reversion.     

Determination of malate levels during the swelling of vacuoles isolated from guard-cell protoplasts

A method for the preparation of vacuoles from guard cells ofVicia faba L. is described. Vacuoles were released from guard-cell protoplasts by osmotic shock and purified on a Ficoll gradient. Contamination of the vacuoles was examined by assaying marker enzymes, such as fumarase, glucose-6-phosphate dehydrogenase, phosphofructokinase, acid phosphatase and mannosidase. Potassium ions in the incubation medium caused increases in the volume of the vacuoles by a factor of about 2.6, while the malate level remained unchanged. In contrast, malate synthesis was stimulated during the swelling phase when complete guard-cell protoplasts were exposed to K⁺. The possible role of K⁺ as an efficient osmotic effector is discussed.Abbreviations DEAE diethylaminoethyl - GCP guard-cell protoplast(s) - GCV guard-cell vacuoles(s) - MCP mesophyll cell protoplast(s) - MCV mesophyll cell vacuole(s)     

Multinucleate transfer cells induced in coleus roots by the root-knot nematode,Meloidogyne arenaria

Summary The occurrence and position of wall protuberances in giant cells induced in coleus roots by the root-knot nematodeMeloidogyne arenaria is described, and the structure and function of giant cells is compared with that of syncytia induced by cyst-nematodes.Extensive protuberance development occurs on walls of giant cells adjacent to xylem vessels. Protuberances are less well developed next to sieve elements, and almost absent next to parenchyma cells. On walls between giant cells they occur on both sides or only one side. The formation of protuberances indicates that giant cells are multinucleate transfer cells. The position of protuberances marks the wall area where solutes enter the cell. Solutes are obtained from xylem and phloem elements, and the position of protuberances at the junction between giant cells and vascular elements indicates an extensive flow of solutes along cell walls. The observations support the hypothesis that wall protuberances form as a result of selective solute flow across the plasmalemma.No cell wall dissolution was observed, although wall gaps may occur between giant cells as a result of breakage during rapid cell expansion.     

Transfer cells and lipid droplets of someValerianaceae

Development, structure and the axial distribution of transfer cells and their lignification were investigated inValerianella locusta, Valeriana officinalis, andV. tuberosa (Valerianaceae). Fundamental new results are: (1) Transfer cells often contain numerous lipid droplets. Within the stem the distribution of cells containing lipid droplets correlates to that of transfer cells. (2) InValeriana officinalis persisting protuberances are frequently found on pit membranes of xylem transfer cells. Lignified transfer cells can undergo a second modification: a layer covering the secondary wall forms wall ingrowths similar to those of transfer cells. (3) Peripheral pith cells, abuting transfer cells, are able to modify into transfer cells. Cambial derivatives are only temporarily developed as transfer cells. (4) Phloem transfer cells are found in vascular bundles of the whole axis. (5) In roots, xylem transfer cells are poorly developed or absent. (6) Oil cells with oil bodies are present in the rape ofValeriana tuberosa. They are absent however in the stem of the species investigated. (7) Tannins occur in elements of the primary cortex, phloem and secondary xylem ofValeriana officinalis.     

Organization of mature external glands on the trap and other organs of the bladderwortUtricularia monanthos

Summary The mature dome-shaped glands which cover the outer surfaces of the trap, leaves, anchor and runner stolons inU. monanthos are described using conventional and some high voltage transmission electron microscopy. The glands occur as scattered ordinary external glands and as a compact clump of vestibule glands at the entrance to the doorway. Each gland rests on a basal epidermal cell and consists of a single pedestal and terminal cell. Vestibule and leaf glands differ slightly from the other glands mainly in the structure of the outer wall of the terminal cell. Nuclear crystals are prominent and the cytoplasm of the pedestal and terminal cells contains tubular structures usually aggregated near the nucleus. The pedestal cell is a transfer cell with short wall protuberances on the outer wall, conspicuous mitochondria and a heavily impregnated lateral wall.The terminal cell often has an outer wall that is greatly thickened and a protoplast that may degenerate early. In the most developed cells the protoplast remains active for a long period and the outer wall is differentiated into several layers. The outermost layer is cuticularized consisting of an open meshwork of deposits. In leaf glands a local polysaccharide mass is usually developed within the cuticularized region. The inner non-impregnated region of the outer wall may show four layers. In vestibule glands fewer layers are present and the wall shows prominent lamellations. Some ordinary external glands differentiate a sponge-like substructure within the inner wall.The ultrastructure and function of the glands are discussed. We support the concept that mature external glands are responsible for secreting water, with those on traps being particularly active during the resetting of the organ. Our work provides a structural basis for recent suggestions by other workers that the mechanism of secretion probably involves establishing a standing osmotic gradient within the gland.     

Use of protoplasts in the improvement of filamentous fungi I. Mutagenization of protoplasts ofOudemansiella mucida

Summary The mutagenic action of ultraviolet light and of N-methyl-N-nitro-N-nitrosoguanidine (NTG) on protoplasts of the basidiomyceteOudemansiella mucida has been evaluated. Since the fungus does not form imperfect spores, we tested the possibilities of using protoplast mutagenesis for improving mucidin-producing strains ofOudemansiella mucida. Protoplasts ofOudemansiella mucida proved to be a convenient starting material for strain improvement and their mutagenization enabled us to obtain not only higher-producing strains, but also auxotrophic mutants, which were used for protoplast fusion experiments.