Applied In Vitro Plant Biotechnology

Prof. dr. ir. Stefaan Werbrouck

For many a crop species vegetative propagation is still very difficult. Although genetic variation is important, clonal plant production has a lot of advantages compared to heterogeneous seedlings. The central motivation of our team is to provide innovative solutions to clone superior herbaceous and principally woody plants for food, feed, fuel, fiber, ornamental and medicinal use. This implies the exploitation of all current botanical, physiological and biotechnological scientific literature. A good scientific network and own research results provide a continuous influx of new research tools. We cooperate worldwide with companies and research groups that are familiar with local plant species and culture conditions to come to applicable long term solutions. Flowering azalea in vitro


The lab focusses mainly on in vitro biotechnology of woody plants. Innovative tissue culture technologies are combined to generate more versatile and efficiency-oriented applied micropropagation systems:

Plant growth regulators:

Phalaenopsis in vitro

Alternative plant growth regulators

Regeneration is a bottle-neck, and therefore a number of small molecules, so called hits from large scale chemical genetic screens are regularly assessed in this regards. These are compounds that interfere with hormonal biosynthesis, catabolism, perception and signal transduction and could stimulate shoot regeneration, normal embryo formation, rooting etc.

GFP expression in an early shoot meristemExpression of LSH4 in Arabidopsis root explants during shoot regeneration on shoot inducing medium (from Motte et al., 2013)

Meta-topolin derivatives

The choice of a particular cytokinin can make the difference between success or failure in the micro propagation system of a plant species. Cytokinins show stimulatory and inhibitory effects during all kinds of developmental processes such as root inhibition, shoot branching, shoot induction and senescence.


In the early years of cytokinin research, only cytokinins with an isoprenoid side chain were thought to be endogenous compounds; however aromatic BA derivatives were identified as natural cytokinins. Also the hydroxylated forms of BA, meta- and ortho-topolin, occur naturally, just like their methoxy derivatives and corresponding 9-ribosides (e.g. MemTR). Our research team experiments with new aromatic cytokinin compounds, as useful additional tools that might solve specific problems in tissue culture of a number of plant species.

MemTR Difficult to micropropagate Petunia Chimera

LEDs as unconventional light sources

LEDs have drawn considerable interest as an alternative light source for in vitro propagation. The advantages of LEDs are their high electrical energy conversion efficiency and specific wavelength, which allows to control growth and development of plants in vitro. In a growth room, 24 growth boxes were installed, in which red, blue and far red LEDs are combined. In another growth room, production LEDs were installed. This allows to study the effect of these monochromatic wave lengths on in vitro plant morphology and development.

Growth room with production LED as light source

Flexible temporary immersion systems (SETIS)

The temporary immersion system SETIS™ consists of an upper recipient for in vitro plants and a connected lower recipient for liquid medium. The combination of periodical short immersion with nutrients and regular aeration of the headspace allows fast in vitro growth of a large number of plant species. Benefits and drawbacks of the system are examined. It offers an exciting new tool for commercial micropropagation. Nevertheless, special attention has to be paid to the setting up and manipulation of the system, in order to avoid contamination. Moreover, the medium in semi-solid medium is not always the best medium in a temporary immersion system. This means that a lot of research is needed to optimize the medium for temporary immersion culture systems.

Temporary immersion systems