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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

on

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).