Microscopic contributions to the pressure-volume diagram of the cell-water relations as demonstrated with tissue cells

Summary Continuing microscopic studies on cell-water relations with single cells (Gerdenitsch 1979) based on the relationship between the osmotic potential and the cell water volume, tissue cells were investigated. The experiments were done with inner epidermis of the bulb scale of onion (Allium cepa) which seemed to be most suitable for those investigations. Using a diagram described byRichter (1978) pressure volume curves of cells at the margin of the epidermis pieces neighboured by a various amount of dead cells and of cells in the center of tissue were worked out. They were contributed to one of three groups according to their amount of free surface (group 1: >1/3 free surface, group 2: <1/3 free surface, group 3: cells surrounded only by living-cells). Curves of the mean values of each of the three groups were compared as well as the mean -values, calculated as a measure for the elasticity of the cell wall.It was found that cells surrounded only by living cells had -values less than with both other groups. Assuming from observations during the experiments that all cells had very similar properties, this difference could be attributed to the expression of the effect of tissue counterpressure.     

Incorporation of [14C]cinnamate into hydrolase-resistant components of the primary cell wall of spinach

[¹⁴C]Cinnamate was taken up very rapidly by cultured spinach cells and completely incorporated into low-MW conjugates within 20 min. The ¹⁴C-labelled products were similar whether the [¹⁴C]cinnamate was supplied continuously over a period of hours via a peristaltic pump or instantaneously. Radioactivity was slowly recruited from the low-MW pool into aromatic components of the cell-wall fraction. Saponification of the radioactive wall fraction yielded, in addition to radioactive ferulate and p-coumarate, large amounts of ethyl acetate-soluble radioactive material with the properties of oxidatively coupled phenols. The coupled material was associated with the most highly ‘Driselase’-resistant fractions of the cell wall. In contrast, ‘Driselase’ released most of the wall's ferulate and p-coumarate on disaccharide fragments. It is suggested that the oxidatively coupled phenols are formed from simpler phenols by peroxidase and that they cross-link the polysaccharides to which they are attached, making these polysaccharides relatively ‘Driselase’-resistant.     

Structure and biosynthesis of teichoic acids in the cell walls of Staphylococcus xylosus DSM 20266

The simultaneous occurrence of a N-acetylglucosaminyl poly(ribitolphosphate) (-GlcNAc) and a N-acetylglucosaminyl poly(glycerolphosphate) (-GlcNAc) in the cell walls of Staphylococcus xylosus DSM 20266 was demonstrated by different experimental lines:(1) Fractionation of extracted cell wall teichoic acid on DEAE-cellulose, (2) investigation of the composition of cell walls in the growth cycle, (3) in vitro biosynthesis using crude membranes as the source of enzyme.The polymerization of these polymers starts from CDP-ribitol and CDP-glycerol, respectively. In the presence of UDP-N-acetylglucosamine both polymers are substituted with N-acetylglucosamine at a level and with the identical anomeric configuration found in the native cell wall teichoic acids. The in vitro biosynthesis of poly(glycerolphosphate) was unique in that it was highly stimulated by UDP-N-acetylglucosamine and to a lower extent by other UDP-activated sugars. Kinetic studies have provided evidence that this stimulation is due to an increase of V _max while K _m is unchanged. Competition experiments have indicated that poly(ribitolphosphate) and poly(glycerolphosphate) were synthesized in the in vitro system in a close spatial relationship.Abbreviations ADP adenosine 5-diphospho - CDP cytidine 5-diphospho - GDP guanosine 5-diphospho - GalNAc N-acetyl-galactosamine - Glc glucose, glucosyl - GlcNAc N-acetyl-glucosamine - N acetylglucosaminyl - GlcUA glucuronic acid - Gro glycerol - Man mannose, mannosyl - Rit ribitol - SDS sodium dodecyl sulfate - UDP uridine 5-diphospho     

On the conservative nature of the gill filaments of sharks

Synopsis Gill filaments of one highly active and two less active shark species exhibit a conservative morphological scheme including such features as branchial canopies, marginal lamellar projections, and enlarged, discrete outer marginal lamellar channels and lateral lamellar sinuses. The specific spatial orientation of the secondary lamellae respective to one another, the gill filaments, and the interbranchial septa create what appears as one-way interfilament water channels, suggesting the presence of an efficient branchial countercurrent system. It is proposed that the fortified structure of shark gills allows many shark species to ventilate passively without having evolved gill filament modifications as apparently did some highly active teleosts. This in turn may have expedited the evolution of lamnid shark species through pre-adaptation to a swift oceanic lifestyle.     

Localization of polyphosphate in vacuoles of Saccharomyces cerevisiae

Virtually all of the polyphosphate (PP) present in yeast protoplasts can be recovered in a crude particulate fraction if polybase-induced lysis is used for disrupting the protoplasts. This fraction contains most of the vacuoles, mitochondria and nuclei. Upon the purification of vacuoles the PP is enriched to the same extent as are the vacuolar markers. The amount of PP per vacuole is comparable to the amount of PP per protoplast.The possibility that PP is located in the cell wall is also considered. In the course of the incubation necessary for preparing protoplasts, 20% of the cellular PP is broken down. As this loss of PP occurs to the same extent in the absence of cell wall degrading enzymes, it is inferred that internal PP is metabolically degraded, no PP being located in the cell walls.It is concluded that in Saccharomyces cerevisiae most if not all of the PP is located in the vacuoles, at least under the growth conditions used.Non-Standard Abbreviations PIPES piperazine-N,N-bis-2-ethanolsulfonic acid - DEAE-dextran diethylaminoethyl-dextran     

CALCIFICATION IN THE GREEN ALGA HALIMEDA. I. AN ULTRASTRUCTURE STUDY OF THALLUS DEVELOPMENT1

The ultrastructure of 4 species of the calcareous, siphonaceous alga Halimeda (H. cylindracea Decaisne, H. discoidea Decaisne, H. macroloba Decaisne and H. tuna (Ellis & Solander) Lamour) has been studied, and the observed changes during growth and development are related to changes in the degree of calcification. A distinct gradient in the types and quantities of cell organelles exists in a growing apical filament. As these filaments grow, branch, and eventually develop into a mature segment, changes in the organization of organelles such as mitochondria and chloroplasts are observed. Calcification begins when the chloroplasts reach structural maturity and when the peripheral utricles adhere (fuse). This adhesion of the peripheral utricles isolates the intercellular space (ICS) in which calcification occurs from the external seawater. Calcification begins in the outermost (pilose) cell wall layer of the walls facing into the ICS. The cell walls at the thallus exterior undergo extensive changes after utricular fusion; the pilose layer is lost, the cuticles of adjacent utricles fuse forming a ridge at their junction, and multiple cuticles are formed. The aragonite (CaCO₃) crystals which are initially precipitated within the pilose wall layer, rapidly increase in size and number, eventually filling much of the ICS. Only the initial nucleation of aragonite is associated with the pilose wall layer, the later precipitation of aragonite is totally independent of the pilose layer. In older segments secondary deposition of CaCO₃ also occurs around existing aragonite needles.     

Roles of auxin and gibberellic acid in growth and maturation of epicotyls of Vigna angularis: Cell wall changes

The effects of auxin and gibberellic acid on cell wall composition in various regions of epicotyls of azuki bean ( Vigna angularis Ohwi and Ohashi cv. Takara) were investigated with the following results. (1) Young segments excised from apical regions of the epicotyl elongated in response to added 10⁻⁴ M indole-3-acetic acid (IAA). When the segments were supplied with 50 m M sucrose, the IAA-induced segment growth was accompanied by enhanced overall synthesis of cell wall polysaccharides, such as xyloglucans, polyuronides and cellulose. This IAA effect on the cell wall synthesis is a consequence of extension growth induced by IAA. Gibberellic acid (GA) at 10⁻⁴ M synergistically enhanced the IAA-induced cell wall synthesis as well as IAA-induced extension growth, although GA by itself neither stimulated the cell wall synthesis nor extension growth. In the absence of sucrose, cell wall synthesis was not induced by IAA or GA. (2) In mature segments excised from basal regions of the epicotyl, no extension growth was induced by IAA or GA. GA enhanced the synthesis of xylans and cellulose when the segments were supplied with 50 m M sucrose. IAA had no effect on the cell wall synthesis. These findings indicate that synthesis of polyuronides, xyloglucans and cellulose, which occurs during extension growth of the apical region of the epicotyl, is regulated chiefly by auxin whereas synthesis of xylans and cellulose during cell maturation in the basal region of the epicotyl is regulated by GA.     

Physiological and biochemical changes associated with cotton fibre development. II. Auxin oxidising system

Changes in IAA oxidase, and in cytoplasmic and ionically wall-bound peroxidase activities were studied in the developing fibres of three cotton cultivars ( Gossypium hirsutum L. cv. Gujarat-67, cv. Khandwa-2 and G. herbaceum L. cv. Digvijay), designated as long, medium and short staple cultivars, respectively. In all the three cultivars IAA oxidase activity was low during the fibre elongation phase, while the activity increased significantly during the secondary thickening phase. The increase in IAA oxidase activity in the three cultivars showed close correspondence with their respective total period of elongation. No relationship between cytoplasmic peroxidase activity and fibre development was discernible. The ionically bound wall peroxidase activity, however, recorded low levels during the elongation phase and higher levels during the secondary thickening phase. The role of wall peroxidase in cessation of elongation growth is discussed.     

The case for multinet growth in growing walls of plant cells

The basis of multinet gwoth, the multinet growth hypothesis, is examined in view of recent criticisms. It is shown that the strain across a growing wall may be calculated by simple means and the expected reorientations are deduced (a) for a wall in which the microfibrils of the innermost wall lamella always lie helically with the same pitch and (b) in which the microfibrils lie at random. Calculations are presented both for cells increasing in length only and for cells also increasing in breadth. Both the strains and the reorientations are smaller than commonly implied and are too small to be reliably detectable in wall sections. Observations on wall sections cannot therefore be accepted as proof that microfibril reorientation has not occured and it is concluded that the multinet growth hypothesis still stands as applying both to parenchyma and to collenchyma cells. In view of the wide dispersity in the structure of the walls of growing cells, it is recommended that the qualifying multinet should be dropped and replaced by passive reorientation.Abbreviation MGH multinet growth hypothesis     

Composition of the cell wall of Chlorella fusca

Isolated cell walls of mature Chlorella fusca consisted of about 80% carbohydrate, 7% protein, and 13% unidentified material. Mannose and glucose were present in a ratio of about 2.7:1 and accounted for most of the carbohydrate. Minor components were glucuronic acid, rhamnose, and traces of other sugars; galactose was absent. After treatment with 2 M trifluoroacetic acid or with 80% acetic acid/HNO₃ (10/1, v/v), a residue with a mannose/glucose ratio of 0.3:1 was obtained, probably representing a structural polysaccharide. An X-ray diffraction diagram of the walls showed one diffuse reflection at 0.44 nm and no reflections characteristic of cellulose. Walls from young cells contained about 51% carbohydrate, 12% protein, and 37% unidentified material. Mannose and glucose were also the main sugars; their absolute amounts per wall increased 6–7 fold during cell growth. Walls isolated with omission of a dodecylsulphate/mercaptoethanol/urea extraction step had a higher protein content and, with young walls, a significantly higher glucose and fucose content. These data and other published cell wall analyses show a wide variability in cell wall composition of the members of the genus Chlorella.Abbreviations GLC gas liquid chromatography - TFA trifluoroacetic acid