In vitro epiphyllous buds formation in Enicostemma hyssopifolium (willd.) ver. induction and cellular origin

In vitro epiphyllous buds have developed on tender excised leaves of Enicostemma hyssopifolium (Willd.) Ver., cultured on Murashige and Skoog’s (1962) medium supplemented with cytokinins (Benzyladenine and Kinetin). This phenomenon was shown to be due to cytokinins and occurred only when abaxial surface was in contact with the media. A two-day exposure to BA (2.0 mgl-1) was sufficient for epiphyllous buds induction, although an exposure for more than four days yielded maximal shoot development from these buds. Culture of excised leaves on media supplemented with BA (2.0 mgl-1) and subsequent subculture on hormone free medium was insufficient to induce epiphyllous buds formation indicating cytokinin dependence. Buds become microscopically visible after fourteen days. These epiphyllous buds originate from sub-epidermal cells on the adaxial side and consisted of functional meristems producing shoots formed by multicellular processes. The role of cytokinin and other potential factors in the induction and development of epiphyllous buds are discussed.