Micropropagation of Bixa orellana using phytohormones and triacontanol

An efficient micropropagation protocol for annatto (Bixa orellana L.) was achieved using nodal shoot tip explants. Shoot buds were obtained on the Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of indole-3-acetic acid (IAA), N-6-benzyladenine (BA) and triacontanol (TRIA). Maximum of 213 shoot buds along with 18 primary shoots were produced on MS medium containing 0.05 A mu M IAA, 8.87 A mu M BA, and 11.2 A mu M TRIA. The primary shoots elongated best on MS medium containing 6.66 A mu M BA and 2.45 A mu M indole-3-butyric acid (IBA). The regenerated shoots rooted best on MS medium supplemented with 4.9 A mu M IBA. The in vitro rooted plantlets were hardened and establishment rate under field conditions was 70 to 80 %.