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Table 2 Procedure of plantlet regeneration from callus derived protocorm of V. planifolia.

From: Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage

  Stage of plantlet micropropagation
Plant growth regulators in basal medium (BM) PLS germination Embryogenic callus induction on callus induction medium Embryogenic/organogenic callus culture
on callus maintenance medium (A4 medium)
Shoot differentiation, PLB multiplication and elongation on
A10 medium
PLB - derived plantlet rooting
A5 medium
Thidiazuron (TDZ) 0.5 mg l-1 0.5 mg l-1 0.3 mg l-1 0 0
Indole-3-acetic acid (IAA) 0 0.5 mg l-1 0.5 mg l-1 0 0
α-Naphthalene acetic acid (NAA) 0 0 0 0.5 mg l-1 0
Phytagel 7.5 g l-1 7.5 g l-1 7.5 g l-1 7.5 g l-1 7.5 g l-1
Other features      
Container Petri disk Petri disk tube Flask tube
Medium volume 20 ml 20 ml 20 ml 100 ml 20 ml
Subcultures No No Every 21 days No No
Culture time 4 - 6 months 2 months 6 months 4 months 2 months
  1. In the same way as the experiment of shoot differentiation and multiplication on A10 medium with CA10 calli, organogenic calli were cultured on A4 medium (CA4 calli) as control. A4, A5 and A10 media were chosen from results of preliminary experiments (see Additional file 5).