Multiple protocorm like body (MPLB) formation and plant regeneration from the PLB culture of a rare orchid Aerides crispum Lindl
Ashok N. Pyati*
Plant tissue culture laboratory, Department of PG Botany, Maharani’s Science College for Women, JLB Road, Mysore-570005 (India) Email: drashoknpyati@gmail.com
ABSTRACT
An efficient protocol for micropropagation of Aerides crispum using axenic protocorm like bodies (PLBs) derived from in vitro seed germination was developed. These PLBs were used to induce multiple protocorm like bodies (MPLBs) on different PGRs through the formation of primary and secondary PLBs. The effect of α-Naphthalene acetic acid (NAA), 6- Benzyl amino purine (BAP) and Thidiazuron (TDZ) singly and in combinations were investigated. The formation of primary and secondary PLBs leading to the formation of MPLBs was assessed. The MPLB formation was optimum in ½ MS medium supplemented with TDZ (0.5 mg/l) where 70.3% MPLBs were induced with an average number of 38.5 MPLBs/explants was recorded. To increase the frequency of MPLB formation various combinations of PGRs were also tested. The MPLB formation was greater from the PLB explants cultured on ½ MS medium containing TDZ with NAA, followed by BAP with NAA. The medium fortified with TDZ (1.0 mg/l) + NAA (0.5 mg/l) the highest frequency of MPLBs (91.8%) were recorded with an average number of (68.0) MPLBs/explant. Medium containing BAP (1.0 mg/l) + NAA (0.5 mg/l) stimulated the formation of MPLBs (84.3%) and an average number of MPLBs/explant (51.9) was recorded. On subculturing the MPLBs on to the same media further developed into plantlets. A survival rate of more than 89% under green house conditions was achieved.