High frequency of protocorm like bodies (PLBs) induction and plant regeneration from protocorm and leaf sections of Aerides crispum

An efficient and reproducible method for the large-scale propagation of Aerides crispum L. using protocorm and leaf sections has been developed. Protocorm and leaf sections were cultured on Murashige and Skoog (MS) medium supplemented with cytokinins [N6-benzyl adenine (BA), thidiazuron (TDZ), and kinetin (KN), 0.5, 1.0, 2.0 and 5.0 μM], auxins [α-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), 0.5, 1.0, 2.0 and 5.0 μM] and coconut liquid endosperm (CW: 5, 10 and 15%). The explants developed protocorm like bodies (PLBs) within 5-8 weeks on the growth medium. BA supplemented medium was found best for the induction of PLBs and an optimum of 49.1 and 22.0 PLBs developed from protocorm and leaf sections on medium supplemented with 1 and 2 μM BA, respectively. Upon subculture on basal MS medium, the PLBs differentiated plantlets within 6-8 weeks. The resulting plantlets were successfully transferred to potting mixture and 85% of plantlets survived after green house transplantation. This simple protocol will be useful for large-scale propagation of A. crispum L.