Effective protocol for callus induction from Solanium dubium seeds and its properties to milk clotting activity

Abstract

The aim of this study was to establish an effective protocol for callus induction from the seed explants of the Solanium dubium (Gubbiun) to investigate its properties to milk clotting activity. The effects of growth regulators, basal media strength and sucrose were studied by using different concentrations (0.5, 1.0, 1.5, and 2.0 mglL) indole - 3- acetic acid (IAA), indole 3- butyric acid (IBA), naphthalene acetic acid (NAA) and 2,4- dichlorophenoxy - acetic acid (2,4-D) alone or in combination with 0.5 mglL of 6- bezyladenine (BAP). For milk clotting activity about 50 or 100 u/L extracts of seed callus mixed with 2ml milk held at 55??C for 5 and 10 minutes until milk clotting occurred. The results show that the NAA alone or in combination with BAP possessed a higher callusing percentage and degree compared to the other plant growth regulators at the same concentrations. When an auxin was supplied in combination with BAP a significant increase in callusing percentage or degree of callusing was observed. The time required for callus to be developed was shortened and the quality of the induced callus improved. An increase in callus growth on low sucrose medium concentration was found compared to high sucrose concentration. The effect of crude extracts obtained of S. dubium callusing to milk clotting activity is also included.