Regeneration of plantlets from in vitro Root and Leaf culture of Vandaceous orchid, Ascocentrum ampullaceum (Roxb.) Schlter

The paper describes in vitro culture protocol of commercially important Vandaceous orchid from North East India, Ascocentrum ampullaceum using leaves and roots as explants. Leaves and roots of in vitro raised 5–6 months old Ascocentrum ampullaceum were inoculated in Murashige and Skoog (MS) medium supplemented with additive growth hormone (15%) Coconut milk and growth regulators such as NAA, IAA, BAP and Kn (0.5, 1.0, 2.0, 2.5 mgl⁻¹), both singly and in combination. Leaf culture produced highest protocorm like bodies (Plbs) (40%) in the medium supplemented with 0.5 mgl⁻¹ BAP, followed by 30% Plbs supplemented with 15% CM and BAP 2.0 mgl⁻¹, whereas the highest (60%) multiple shoot bud formation was noticed in MS medium supplemented with 15% CM and 0.5 mgl⁻¹ BAP, followed by 30% in 1/2MS medium supplemented with 0.5 mgl⁻¹Kn; Callus formation (30%) was observed in 1/2MS medium supplemented with 2.0 mgl⁻¹ NAA. Roots when cultured in 1/2 MS, only elongation was observed, but in full strength MS medium, it produced both Plbs and shoot buds. The highest Plbs and shoot buds (40%) was observed in the media supplemented in combination of CM (15%) + BAP (0.5 mgl⁻¹) followed by 25% in CM (15%) + BAP (1.0 mgl⁻¹).

• The paper describes in vitro culture protocol of commercially important Vandaceous orchid from North East India, Ascocentrum ampullaceum using leaves and roots as explants.

• Root culture suggested that quality and quantity of growth regulators play an important role for further development of protocorm like bodies (plbs) and shoot bud.

• Results from leaf culture showed best development of Plbs and shoot bud in MS medium supplemented with 15% CM, BAP and NAA.