Esterase activity and isoenzymes in relation to morphogenesis in Mammillaria gracillis Pfeiff. tissue culture

Cactus Mammillaria gracillis Pfeiff. (Cactaceae), cultivated in vitro, spontaneously switches from an organised to unorganised way of growth, producing a habituated organogenic callus which regenerates normal and hyperhydric shoots without the addition of any growth regulators. Tumour tissues, induced by A. tumefaciens wild strain B6S3 (tumour TW) and rooty mutant GV3101 (tumour TR), do not express any organogenic potential. The esterase (arylesterase EC 3.1.1.2 and carboxylesterase EC 3.1.1.1) activity and isoenzyme pattern of morphologically different cactus tissues: shoot, callus, hyperhydric regenerant, tumours TW and TR, were compared. Tissue samples were frozen at –80 °C and lyophilized before protein extraction. Two esterase substrates, 1and 2-naphthylacetate, were used. Esterase activity of all tissues varied during the period of one subculture. In shoots and tumours, the highest esterase activity for both substrates was measured on the 21st day, while in the callus and hyperhydric regenerants the highest activity was on the 7th day. Esterases were separated electrophoretically in polyacrylamide gradient gels under non-denaturating conditions. In total, 13 isoesterases, reacting with both substrates, were resolved. No differences in isoenzyme profile were noticed in correlation with the age of tissues, but the esterase activity varied among tissues. The significance of these results is discussed.