Ewa Szczuka

New ultrastructural features of organelles in leaf cells of Deschampsia antarctica Desv

An electron microscope has been used to investigate the ultrastructure of leaf cells in Deschampsia antarctica Desv. (Poaceae). The leaf anatomy exhibits features typical of xerophytes. New ultrastructural features were found in mesophyll cells. Chloroplasts in mesophyll cells of D. antarctica leaves form small vesicles and pockets. The outer chloroplast membrane forms vesicles, and pockets are invaginations of both membranes. The invaginations contain small vesicles, mitochondria, or lipid droplets. The mitochondria or peroxisomes adhere very tightly to the chloroplasts.

Structural network of arabinogalactan proteins (AGPs) and pectins in apple fruit during ripening and senescence processes

The cell wall is an essential framework determining the overall form of the plant cell. Our study was focused on the distribution of arabinogalactan proteins (AGPs), arabinan, and homogalacturonan in fruit cells during ripening and storage with emphasis on quantitative analysis of their presence in particular regions of the cell wall - plasma membrane. The localization of the examined compounds was determined with immunohistochemistry techniques and immunogold labelling. Spatio-temporal colocalization between AGPs epitopes - [βGlcA(1→3)-αGalA(1→2)Rha] recognized by JIM13 and MAC207 antibodies, and arabinan labelled by the LM16 antibody was detected in the inner cell wall layer, in association with the plasma membrane. The specific arrangement of AGP and arabinan epitopes differentiated them from homogalacturonan epitopes, consisting of GalA residues recognized by LM19 and LM20 antibodies in all the examined fruit maturity stages. The disruption of cell wall - plasma membrane continuum, observed during ripening-associated softening process, was associated with both the substantial decrease of AGPs, pectins content and with remodeling of their arrangement. The results indicate that the textural properties of fruit during growth and postharvest storage, an attribute of fruit quality becoming selection criteria for consumers, depend on the existence of dynamic network organizing polysaccharides and glycoproteins in the extracellular matrix.

Specific ultrastructure of the leaf mesophyll cells of Deschampsia antarctica Desv. (Poaceae)

The ultrastucture of mesophyll cells of Deschampsia antarctica Desv. (Poaceae) leaves was investigated using the standard method of preparing material for examination in transmission electron microscopy (TEM). The investigated leaves were collected from the Antarctic hairgrass growing in a tundra microhabitat and representing xermorphic morphological and anatomical features. The general anatomical features of mesophyll cells are similar to those in cells of another grass leaves. The observations of the ultrastructure of mesophyll cells have shown that the organelles are located close to each other in a relatively small amount of the cytoplasm or closely adhere to each other. Organelles such as mitochondria, peroxisomes, and Golgi apparatus, as well as osmiophilic materials are gathered close to the chloroplasts. The chloroplast of the mesophyll cells of the D. antarctica leaf can form concavities filled with the cytoplasm. Such behaviour and ultrastructure of organelles facilitate exchange/flow of different substances engaged in the metabolic activity of the cell between cooperating organelles.

Optimization of in vitro culture conditions influencing the initiation of raspberry (Rubus idaeus L. cv. Nawojka) cell suspension culture

The purpose of our investigation was to determine appropriate conditions for induction of raspberry (Rubus idaeus cv. Nawojka) cell suspension culture. The established callus culture obtained from leaf explants was used as an inoculum for cell culture initiation. Five combinations of plant growth regulators: 1) 4.0 mg l-1 IAA and 1.0 mg l-1 BAP; 2) 0.25 mg l-1 2,4-D; 3) 0.5 mg l-1 2,4-D; 4) 2.0 mg l-1 NAA and 2.0 mg l-1 BAP; 5) 4.0 mg l-1 NAA and 2.0 mg l-1 BAP, added into modified Murashige and Skoog (1962) medium, were tested in order to get the callus culture suitable for initiation of a cell suspension. The best callus (vigorously growing, healthy and friable) was obtained on the medium supplemented with 4.0 mg l-1 IAA and 1.0 mg l-1 BAP. To find the appropriate culture conditions for dispersing callus tissue in liquid medium into single cells and small aggregates, four combinations of plant hormones (auxins and cytokinins) were tested. The best culture medium for induction of raspberry cv. Nawojka cell suspension appeared to be the one supplemented with 1.0 mg l-1 2,4-D. Also the medium with 8.0 mg l-1 IAA and 1.0 mg l-1 BAP was similarly efficient.

Analysis of antifungal and anticancer effects of the extract from Pelargonium zonale

The extract from Pelargonium zonale stalks exhibits activity against Candida albicans and exerts an effect on the HeLa cell line. The action against C. albicans cells was analysed using light, CLSM, SEM, and TEM microscopes. The observations indicate that the extract influenced fungal cell morphology and cell metabolic activity. The morphological changes include cell wall damage, deformations of cell surfaces, and abnormalities in fungal cell shape and size. Cells of C. albicans treated with the extract exhibited disturbances in the budding pattern and a tendency to form agglomerates and multicellular chains. The P. zonale extract caused a significant decrease in the metabolic activity of C. albicans cells. Cells died via both apoptosis and necrosis. The antitumor activity of the extract was analysed using the MTT assay. The P. zonale extract exhibited minor cytotoxicity against the HeLa cell line but a dose-dependent cytopathic effect was noticed. The P. zonale extract is a promising source for the isolation of antifungal and anticancer compounds.