RESEARCH CONCERNING THE ACTIVITY AND SPECTRUM OF SOME ENZYMES ON REGENERANTS OF BRASSICA OLERACEA VAR CAPITATA FROM ANTHERES AND OVARIES

Cathalase and peroxidase activity was effected on cabbage leaves (Brassica oleracea) from in vitro regenerants. The study of cathalase and peroxidase specific activity on vitroclones provided by anthers and ovaries pointed out major differences due to genotype, explant type and hormonic balance within the nutritive media. Cathalase activity in most investigated regenerants was higher compared to control plants. Seed-grown plantlets (the same ontogenetic stage as the in vitro plants) were referred to as control plants. Cathalase activity was determined by iodometric method, and peroxidase activity - by spectrophotometry. To evince the isoenzyme spectrum of estherase and peroxidase, enzyme preparations were accomplished in TrisHCl 0.1M, pH= 7.2. Wrigley method was used for electrofocussing. Peroxidasic isoenzymes were evinced by means of McDonald and Smith method, 1972. Cathalase specific activity in most investigated vitroclones was higher related to control plants. In case of ovary- provided regenerants (Z(2)-1(2) genotype) cathalase activity is higher on media supplemented with BAP (benzylaminopurine) and IAA (indole acetic acid): 7.745 mg H2O2/mg proteins, superior to control plants: 1.196 mg H O-2(2) /mg proteins. Peroxidase specific activity in vitroclones was higher than in control plants (0.763 - 1.709 mg H2O2/mg proteins in anther- provided regenerants, and of 1.429 - 4.326 mg H2O2/mg proteins in ovary- provided regenerants, compared to 0.432 mg H-2 O-2 /mg proteins in control). Isoenzymatic spectra of cathalase and of peroxidase evinced some major differences among the regenerants of Brassica oleracea var. capitata provided by anthers and ovaries. Isoestherase spectrum was broad, comprising 9 to 17 electroforetic bands. There was a narrow isoperoxidase spectrum in vitroclones. There were 1 to 4 bands in case of regenerants compared to control (5 bands).