Callus-mediated Somatic Embryogenesis and Plant Regeneration in Vanda tricolor Lindl. var. Pallida

Abstract

In this study, a protocol to induce indirect somatic embryogenesis from the basal leaf segments of Vanda tricolor Lindl. var. Pallida has been developed. 2 0 T The experiments consisted of two stages، i.e. induction of SEs from calli and regeneration of plantlet from SEs. 2 0 T Embryogenic calli obtained from previous experiment (0.05 mg LP -1 P NAA + 0.01 mg LP -1 P BAP) were used to induce somatic embryos (SEs) on half-strength Murashige and Skoog (MS) medium supplemented with (0.05 mg LP -1 P toP P 0.20 mg LP -1 P ) 6-benzylaminopurine (BAP) alone or in combination with 0.01 mg LP -1 P α-naphtalene acetic acidP P (NAA). Embryogenic calli, those cultured on 0.05 mg LP -1 P BAP and 0.01 mg LP -1 P NAA resulted in 90 % induction of SE structures at 30 d of culture period. Histological observation exhibited development of pro-embryo to form completed embryo. The pattern of SEs development started from embryogenic callus to form pro-embryo, followed by globular phase at 10 d of culture. Globular embryo elongated to form suspensor at 20 d of incubation period, and completed embryo. Regeneration of SEs into complete plantlets was attained on the half-strength MS medium without addition of any plant growth regulator (PGR). Based on the results of the present study, it can be concluded that 2 0 T half-strength MS medium supplemented with 2 0 T NAA 0.01 mg LP -1