Two-dimensional electrophoresis of soluble leaf proteins, isolated from two wheat species (Triticum durum and Triticum aestivum) differing in sensitivity towards NaCl.

Plants of two wheat species (Triticum aestivum cv. Tanit and T. durum cv. Ben Bachir), differing in their sensitivity to NaCl were cultivated in the presence or absence of 100 mM NaCl for 21 days. Soluble proteins extracted from leaves were analyzed by two-dimensional electrophoresis in order to detect NaCl-induced changes in the polypeptide patterns. In all, 500 spots were detected. Results showed species-dependent differences. The greatest alterations in the polypeptide profiles following salt stress were found in the most sensitive cultivar: among the 12 spots (molecular mass, 15-31 kDa) specifically considered in the acidic region of the gel, 11 declined, even disappeared in the NaCl-sensitive leaf profiles, while in the tolerant species only five spots were affected by the salt treatment and five remained untouched; moreover in the latter, two new polypeptides were shown to be induced by NaCl.

Analysis by two-dimensional electrophoresis of the effect of salt stress on the polypeptide patterns in roots of a salt-tolerant and a salt-sensitive cultivar of wheat.

The effect of salt stress on the polypeptide levels in roots of two wheat (Triticum durum) cultivars with different sensitivity to NaCl (cv. Ben Bachir, sensitive; cv. Chili, tolerant), was examined by two-dimensional polyacrylamide gel electrophoresis. Blue-stained gels were analyzed by visual inspection to identify changes that resulted when seedlings were grown in the presence of 200 mM NaCl for four days. Although the protein patterns for control and salt-stressed seedlings were qualitatively similar, the net synthesis of a 26 kDa polypeptide was significantly changed. This observation was mainly noticeable in the more tolerant cultivar. With the intention of identifying its function, the NH2-terminal of this polypeptide was sequenced. A 20 amino acid sequence was obtained and compared to sequences available in different databases. Possible roles of this polypeptide, depending on the homologies of its amino acid sequence with known proteins, in salinity tolerance are discussed.

Phytohormone regulation of isoperoxidases in Catharanthus roseus suspension cultures.

Peroxidase (POD) activity was investigated in Catharanthus roseus cell suspensions cultured under different hormonal conditions. Depletion of 2,4-dichlorophenoxyacetic acid (2,4-D) from the culture medium enhanced POD activity in cells and spent medium. Addition of phytohormones, in particular the auxin 2,4-D, reduced POD activity in medium and cellular compartments and enhanced ionically cell-wall bound POD. The differential modulation of POD is due to hormone effects on synthesis and/or accumulation of POD, rather than on the secretion process. Qualitative analysis showed that 2,4-D, but not cytokinins, regulated the synthesis of a basic isoform. The cytokinin treatment seemed to affect acidic rather than basic isoforms. The presence of basic POD is correlated with the capacity of cells to produce indole alkaloids. The major extracellular basic isoperoxidase was purified to homogeneity from culture medium of Catharanthus roseus cell suspensions. The isolated peroxidase is a haem protein with a M(r) of 33,000 and a pI close to 9. The effect of pH on peroxidase activity was studied using guaiacol as substrate and the optimum pH determined at 25 degrees was 6.0. This enzyme acted on guaiacol, 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), o-dianisidine, o-phenylenediamine (o-PD) and pyrogallol, but had no effect on syringaldazine or coniferyl alcohol substrates.

Cytokinin-enhanced accumulation of indole alkaloids in Catharanthus roseus cell cultures - The factors affecting the cytokinin response.

Cytokinins were found to stimulate the alkaloid synthesis induced by removing auxin from the medium of a cell line of Catharanthus roseus. Diluting the mineral salts of the culture medium decreased the alkaloid production but increased the "sensitivity" of the cells. Addition of high levels of Ca(2+), Mg(2+) or Sr(2+) to B5 media in which the mineral salts were diluted to 5-40%, increased the alkaloid production. The latter effect is related only partially to enhanced osmotic potential.