Effect of Culture Medium Concentration Times of Overlay and Cytokinins on Development of New Shoot Buds Purplish Red Leaf and Callus Formation

Abstract

In order to study development of mangosteen leaves for callus formation subsequently a high frequency of plantlet regeneration, preparation of the leaves was carried out using two-phase culture media. Lower phase medium was woody plant (WPM) medium supplemented with 3% sucrose, 500 mg/l polyvinylpyroridone (PVP) and 0.1 mg/l benzyladenine (BA). Upper phase medium or overlay medium was liquid Murashige and Skoog (MS) reduced the compon to various concentrations and overlayed at various periods of culture. Moreover, we also study the effect of kinds of cytokinins on development of new shoots, shoot length, purplish red leaf and callus formation. It is found that liquid 1/2MSNB (MS which was reduced the components to a half strength of origin and supplemented with 3% sucrose, 0.06 mg/l naphthaleneacetic acid (NAA) and 0.03 mg /l BA) gave the best results in increment of new shoot buds and purplish red leaf formation. Period of culture shoot clump on WPM for 2 weeks followed by overlay the liquid medium gave the best results in increment of new shoot buds, shoot length and purplish red leaf formation. Between cytokinin tested 1/2MSNBT (both BA and thidiazuron, TDZ at the same concentration of 0.03 mg/l) provided the best formation of both purplish red leaves and callus.