Growth under Different Trophic Regimes and Synchronization of the Red Microalga Galdieria sulphuraria

The extremophilic unicellular red microalga Galdieria sulphuraria (Cyanidiophyceae) is able to grow autotrophically, or mixo- and heterotrophically with 1% glycerol as a carbon source. The alga divides by multiple fission into more than two cells within one cell cycle. The optimal conditions of light, temperature and pH (500 µmol photons m−2 s−1, 40 °C, and pH 3; respectively) for the strain Galdieria sulphuraria (Galdieri) Merola 002 were determined as a basis for synchronization experiments. For synchronization, the specific light/dark cycle, 16/8 h was identified as the precondition for investigating the cell cycle. The alga was successfully synchronized and the cell cycle was evaluated. G. sulphuraria attained two commitment points with midpoints at 10 and 13 h of the cell cycle, leading to two nuclear divisions, followed subsequently by division into four daughter cells. The daughter cells stayed in the mother cell wall until the beginning of the next light phase, when they were released. Accumulation of glycogen throughout the cell cycle was also described. The findings presented here bring a new contribution to our general understanding of the cell cycle in cyanidialean red algae, and specifically of the biotechnologically important species G. sulphuraria.

Phylogenetic and comparative functional analysis of the cell-separation α-glucanase Agn1p in Schizosaccharomyces

The post-cytokinetic separation of cells in cell-walled organisms involves enzymic processes that degrade a specific layer of the division septum and the region of the mother cell wall that edges the septum. In the fission yeast Schizosaccharomyces pombe, the 1,3-α-glucanase Agn1p, originally identified as a mutanase-like glycoside hydrolase family 71 (GH71) enzyme, dissolves the mother cell wall around the septum edge. Our search in the genomes of completely sequenced fungi identified GH71 hydrolases in Basidiomycota, Taphrinomycotina and Pezizomycotina, but not in Saccharomycotina. The most likely Agn1p orthologues in Pezizomycotina species are not mutanases having mutanase-binding domains, but experimentally non-characterized hypothetical proteins that have no carbohydrate-binding domains. The analysis of the GH71 domains corroborated the phylogenetic relationships of the Schizosaccharomyces species determined by previous studies, but suggested a closer relationship to the Basidiomycota proteins than to the Ascomycota proteins. In the Schizosaccharomyces genus, the Agn1p proteins are structurally conserved: their GH71 domains are flanked by N-terminal secretion signals and C-terminal sequences containing the conserved block YNFNAY/HTG. The inactivation of the agn1Sj gene in Schizosaccharomyces japonicus, the only true dimorphic member of the genus, caused a severe cell-separation defect in its yeast phase, but had no effect on the hyphal growth and yeast-to-mycelium transition. It did not affect the mycelium-to-yeast transition either, only delaying the separation of the yeast cells arising from the fragmenting hyphae. The heterologous expression of agn1Sj partially rescued the separation defect of the agn1Δ cells of Schizosaccharomyces pombe. The results presented indicate that the fission yeast Agn1p 1,3-α-glucanases of Schizosaccharomyces japonicus and Schizosaccharomyces pombe share conserved functions in the yeast phase.

Changes in cell ultrastructure and morphology of Arabidopsis thaliana roots after coumarins treatment

The ultrastructure and morphology of roots treated with coumarin and umbelliferone as well as the reversibility of the coumarins effects caused by exogenous GA, were studied in <em>Arabidopsis thaliana</em>. Both coumarins suppressed root elongation and appreciably stimulated radial expansion of epidermal and cortical cells in the upper part of the meristem and in the elongation zone. The gibberellic acid applied simultaneously with coumarins decreased their inhibitory effect on root elongation and reduced cells swelling.Microscopic observation showed intensive vacuolization of cells and abnormalities in the structure of the Golgi stacks and the nuclear envelope. The detection of active acid phosphatase in the cytosol of swollen cells indicated increased membrane permeability. Significant abnormalities of newly formed cell walls, e.g. the discontinuity of cellulose layer, uncorrect position of walls and the lack of their bonds with the mother cell wall suggest that coumarins affected the cytoskeleton.

The cell wall of the pollen mother cell at the tetrad stage in Convallaria majalis L.

The cell wall at the tetrad stage in <em>Convallaria majalis</em> L. has been studied by light microscope histochemical techniques. The standard PAS reaction has shown the persistence of the primary pecto-cellulosic pollen mother cell wall localized around the callosic special wall (determined both by Bauer's reaction and the fluorescence technique with aniline blue) of individual tetrcds. A PAS-positive spore precursor wall (primexime) is formed while the tetrad of microspores is still enclosed by intact callose and pollen mother cell walls. The pecto-cellulosic wall and callose layer dissolve simultaneously to release the microspores into another loculi.

Haustorial mother cell development by Uromyces vignae on collodion membranes

The development of infection structures by the rust fungus Uromyces vignae was observed on oil-containing collodion membranes. About 40% of infection hyphae formed a haustorial mother cell, but this structure commonly senesced and died more rapidly than the infection hypha to which it was attached. These data suggest that the continued development of the haustorial mother cell requires some component normally provided by the host plant. Before they died, many haustorial mother cells apparently formed the thickened region of the wall which normally is traversed by the penetration peg during haustorium formation. Such a peg was observed in the centre of up to 40% of these thickened regions. However, no pegs protruded beyond the haustorial mother cell far enough to be called a haustorial neck. The thickened region of the haustorial mother cell wall could be differentiated from the rest of the wall by its lack of fluorescence under ultraviolet irradiation when mounted in Calcofluor or SITS (4-acetomido-4′-iso-thiocyanatostilbene-2,2′-disulphonic acid). Treatment with alkali, acid, chloroform–methanol, protease, and laminarinase did not affect this differential fluorescence, and the haustorial mother cell wall stained uniformly for proteins, carbohydrates, and chitin. Since Calcofluor normally binds to chitin, these data suggest that the thickened region of the haustorial mother cell wall may physically exclude the dye or may contain potential binding sites that are masked by other wall components.

Maturation of the megaspore in Marsilea vestita

The course of megasporogenesis was followed from the simultaneous cleavage of the mother cell at the termination of meiosis to the formation of the compound wall around the single surviving megaspore in the maturing sporangium. The favoured tetrad that yielded the surviving megaspore was always that closest to the stalk of the sporangium. Although no asymmetry could be detected in the tetrad, three of the spores regularly degenerated. The beginning of degeneration was sometimes evident before the release of the spores, but only after the spores had acquired distinct walls. Cytochemical tests indicated that separation of the spores was facilitated by the mother cell wall becoming fluid. Growth of the maturing megaspore was accompanied by conspicuous nucleocytoplasmic interaction. Tubular extensions of the nucleus could be followed in the cytoplasm for more than 1 μm, but no connections could be detected with any organelles. The central space into which the megaspore grew was probably generated partly by the expansion of the sporangium as a whole, and partly by the digestion of the plasmodial tapetum. The wall of the mature spore was clearly compound, the unstructured exine of the growing spore eventually meeting and fusing with a chambered component formed at the periphery of the tapetum.