<i style="">In vitro </i>propagation of threatened terrestrial orchid, <i style="">Malaxis khasiana</i> Soland ex. Swartz through immature seed culture

Abstract

Rapid <i style="">in vitro </i>propagation of the terrestrial orchid, <i style="">M. khasiana </i>through immature seed culture was achieved. Immature seeds of 8-9 week after pollination (WAP) cultured on MS medium (2% sucrose) supplemented with 500 mgl^-1 casein-hydrolysate and 1 <img src='/image/spc_char/micro.gif'><i style="">M</i> N⁶-benzyladenine (BA) exhibited germination of 75% seeds after 107 days of culture and subsequently supported the development of PLBs. Subsequent culture on MS medium enriched with 6 <img src='/image/spc_char/micro.gif'><i style="">M</i> of indole-3-acetic acid (IAA), 18 <img src='/image/spc_char/micro.gif'><i style="">M</i> each of BA and kinetin induced multiple shoots and plantlets. Transfer of PLBs to MS medium with 0.1% activated charcoal (AC) facilitated rapid proliferation of PLBs, while AC at 0.2% favored shoot bud induction and rhizome enlargement. The plantlets, developed on medium with IAA, BA and kinetin, after hardening <i style="">in vitro </i>for 8-10 weeks were planted in community pots and transferred to poly-house. The plantlets showed 65% survival under field conditions.