Subcellular Alterations Induced by Cyanotoxins in Vascular Plants—A Review

Phytotoxicity of cyanobacterial toxins has been confirmed at the subcellular level with consequences on whole plant physiological parameters and thus growth and productivity. Most of the data are available for two groups of these toxins: microcystins (MCs) and cylindrospermopsins (CYNs). Thus, in this review we present a timely survey of subcellular cyanotoxin effects with the main focus on these two cyanotoxins. We provide comparative insights into how peculiar plant cellular structures are affected. We review structural changes and their physiological consequences induced in the plastid system, peculiar plant cytoskeletal organization and chromatin structure, the plant cell wall, the vacuolar system, and in general, endomembrane structures. The cyanotoxins have characteristic dose-and plant genotype-dependent effects on all these structures. Alterations in chloroplast structure will influence the efficiency of photosynthesis and thus plant productivity. Changing of cell wall composition, disruption of the vacuolar membrane (tonoplast) and cytoskeleton, and alterations of chromatin structure (including DNA strand breaks) can ultimately lead to cell death. Finally, we present an integrated view of subcellular alterations. Knowledge on these changes will certainly contribute to a better understanding of cyanotoxin–plant interactions.

Tradescantia bracteata pollen in vitro: pollen tubes development and mitosis

About 90 per cent of <i>Tradescantia bracteata</i> pollen germinates <i>in vitro</i> after 15 min. Mitosis starts in the pollen tube after about 3 h. The mitotic trans-formations of chromosomes within the generative nucleus are not synchronized. They involve succesively the linearly arranged chromosomes in the elongated generative nucleus. In metaphase the chromosomes are arranged tandem-like linearly along the pollen tube. The chromatides translocate in anaphase from various distances to the poles in a plane parallel to the metaphase plate. This suggests that chromosomes have individual mitotic spindles and that coordination of the chromosome transformations in the generative cell is much less strict than in a typical somatic mitosis. Starch is the storage material of pollen grains. In the vegetative cytoplasm of mature pollen grains minute reddish-orange vesicular structures are visible after staining with neutral red. They do not fuse with the vacuoles proper arising in germinating pollen grains to form the vacuolar system of the pollen tube.

Translocation of the generative cell towards the centre of the pollen grain due to changes in the vacuolar system in Ranunculus repens L.

On the basis of observations performed under the light microscope on the development of the male gametophyte of <i>Ranunculus repens</i> L., the hypothesis is advanced that the translocation of the generative cell from a position adjacent to the wall towards the centre of the pollen grain, associated with change in the shape of this cell, is the result of vacuolization of the generative and vegetative cells and the interaction of these two vacuole systems.

Nuclear DNA content and ultrastructure of secretory cells of Vicia faba L. stigma

The object of study was the level of nuclear DNA and the ultrastructural transformations in the secretory cells of the stigma in <i>Vicia faba</i> L. It has been found that the stigmal cells which are active in biogenesis and exudate secretion are diploid cells whose differentiation starts from 2C DNA level. The presence of a population of nuclei with an amount DNA of about 2.5 C suggests that the metabolic activity of those cells may be regulated through supplementary incomplete replication. The ultrastructural transformations of secretory cells point to three stages of biogenesis and secretion of exudate. Stage I, before the start of the cell's secretory functions, is characterized by the development of the protein synthesizing apparatus and the activity of dictyosomes. In development stage II vesicular electron-transparent exudate is secreted. Stage III of exudate biogenesis is production of lipids. They form mainly in the plastids and are secreted with the involvement of the cell's vacuolar system.

Darkness descends with two Rabs

Over 60 distinct Rab GTPases regulate specific vesicular transport steps in the mammalian central vacuolar system. Wasmeier et al. (this issue, p. 271) reveal a redundant role for two tissue-specific Rab proteins in regulating transport to a tissue-specific lysosome-related organelle, the melanosome.

Vacuolar localization of phosphorus in hyphae of Phialocephala fortinii, a dark septate fungal root endophyte

Phialocephala fortinii is a dark septate fungal endophyte that colonizes roots of many host species. Its effect on plant growth varies from being pathogenic to beneficial. The basic biology of this species has received little research, and thus the main objectives of this study were to determine cytological features of hyphae, including the nature of the vacuolar system, and whether polyphosphate was present in vacuoles. Both living hyphae and hyphae that had been rapidly frozen and freeze substituted before embedding were studied. A complex system of vacuoles, including a motile tubular vacuolar system, elongated vacuoles, and spherical vacuoles, was demonstrated in living hyphae by the fluorescent probe Oregon Green 488 carboxylic acid diacetate, using laser scanning confocal microscopy. The motile tubular vacuolar system was more prevalent at the hyphal tip than in more distal regions, whereas elongated vacuoles and spherical vacuoles were more abundant distal to the tip. All vacuoles contained polyphosphate as shown by labelling embedded samples with recombinant polyphosphate binding domain of Escherichia coli exopolyphosphatase, containing Xpress tag at the N-terminal end, followed by anti-Xpress antibody and a secondary antibody conjugated either to a fluorescent probe for laser scanning confocal microscopy or colloidal gold for transmission electron microscopy. The polyphosphate was dispersed in vacuoles. This was confirmed by staining embedded samples with 4′,6-diamidino-2-phenylindole and viewing with UV light using epifluorescence microscopy. These cytological methods showed that the tubular vacuolar system had lower concentrations of polyphosphate than the spherical vacuoles. Lipid bodies were present around vacuoles.Key words: Phialocephala, motile tubular vacuoles, vacuoles, polyphosphate, lipid.

Ubiquitin-mediated Targeting of a Mutant Plasma Membrane ATPase, Pma1-7, to the Endosomal/Vacuolar System in Yeast

Pma1-7 is a mutant plasma membrane ATPase that is impaired in targeting to the cell surface at 37°C and is delivered instead to the endosomal/vacuolar pathway for degradation. We have proposed that Pma1-7 is a substrate for a Golgibased quality control mechanism. By contrast with wild-type Pma1, Pma1-7 is ubiquitinated. Ubiquitination and endosomal targeting of Pma1-7 is dependent on the Rsp5-Bul1-Bul2 ubiquitin ligase protein complex but not the transmembrane ubiquitin ligase Tul1. Analysis of Pma1-7 ubiquitination in mutants blocked in protein transport at various steps of the secretory pathway suggests that ubiquitination occurs after ER exit but before endosomal entry. In the absence of ubiquitination in rsp5-1 cells, Pma1-7 is delivered to the cell surface and remains stable. Nevertheless, Pma1-7 remains impaired in association with detergent-insoluble glycolipid-enriched complexes in rsp5-1 cells, suggesting that ubiquitination is not the cause of Pma1-7 exclusion from rafts. In vps1 cells in which protein transport into the endosomal pathway is blocked, Pma1-7 is routed to the cell surface. On arrival at the plasma membrane in vps1 cells, Pma1-7 remains stable and its ubiquitination disappears, suggesting deubiquitination activity at the cell surface. We suggest that Pma1-7 sorting and fate are regulated by ubiquitination.