Changes in protein profiles during shoot differentiation in callus cultures from Tylophora Indica Wight & ARN. (Antamul)

Leaf explants were cultured on MS medium supplemented with 2.0 mg/l of 2,4-D + 0.5 mg/l KIN + 200 mg/l CH. Callus obtained on this initiation medium was sub cultured for further growth on same medium and thus it acted as maintenance medium also. For differentiation of shoots medium containing MS + 1.0 mg/l KIN+ 0.5 mg/l IAA was used. The growth and shoot-formation of the calli on maintenance and differentiation media was respectively followed (0, 5, 10, 15 and 20 days after) and the protein content and profile were determined. Comparison of protein profiles of the callus on maintenance medium and during differentiation of shoots showed that additional polypeptides of MW 107. 109 and 114 kDa appeared only at the time of shoots initiation and are specific to shoots formation. Similarly, the accumulation of total soluble proteins and acid-protease activity were greater during sequential stages of the differentiation than during the callus growth.