Liquid Culture Systems for in vitro Plant Propagation

Mass propagation of tropical crops 201
A scale up and production facility was established at Centro de
Bioplantas (Ciego de Avila, Cuba) for Ananas shoot multiplication in 10litre
twin flask TIS. Shoots multiplied in TIS are used as starting material in
order to speed up the propagation process in standard semisolid medium and
also for the production of transplants in a two stage system which involves
multiplication and elongation in TIS (Escalona et al., 1999). The same group
also developed a protocol for sugarcane shoot multiplication (Lorenzo et al.,
1999), this technology was transferred to a commercial facility at the
Instituto Nacional de Investigaciones de la Caña de Azúcar (INICA) which
is used for the multiplication of elite varieties and basic seed production.
CIRAD (Montpellier, France) has been using mass propagation by
somatic embryogenesis with the RITA® system to disseminate clones of
selected F1 Coffea arabica hybrids in Central America (in cooperation with
CATIE and PROMECAFE) and in Tanzania (in cooperation with ARTI of
Lyamungu) (Etienne and Berthouly, 2002).
BioPlanta GmbH (Leipzig, Germany) in collaboration with IBP (Cuba)
developed a scale up and production facility for biomass (shoots) and
secondary metabolite production in Cymbopogon citratus, Lavandula
offcinialis, Hypericum perforatum and Fabiana imbricata using TIS 5-litre
twin flasks (Hohe et al., 2002).
The Institute for Ornamental Plant Breeding (Ahrensburg, Germany)
established a procedure for Phalaenopsis propagation in 5-litre twin flasks,
which comprises a first step of adventitious shoot multiplication followed by
rooting in TIS (Hempfling and Preil, 2002).
At the Institute of Plant Biotechnology (Santa Clara, Cuba) a production
facility with 10-litre twin flask TIS was built (Figure 1). Potato microtubers
are produced according to the protocol described by Jiménez et al. (1999).
Tubers are directly transplanted to greenhouse or field for elite seed
production (Pérez et al., 2000). A second application is banana propagation
by shoot multiplication in medium containing growth retardants in 10-litre
TIS and subsequent elongation and rooting in TIS using a hormone free
medium (Albany et al., 2002).
From the above mentioned examples some conclusions may be drawn:
– Regeneration system: Beside the development of methods for producing
somatic embryos in TIS, shoot multiplication still remains the main
alternative for tropical crop propagation.
– TIS design: RITA® is a suitable tool for research and lab scale, however
for commercial application larger vessels are frequently used. Most
important tropical species are commercially propagated in twin flasks
TIS ranging from 5 to 10 litres.