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Liquid Culture Systems for in vitro Plant Propagation

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Chapter 38<br />

Development and validation of an efficient low cost<br />

bioreactor <strong>for</strong> furanocoumar<strong>in</strong> production with<br />

Ruta graveolens shoot cultures<br />

E. Gontier (1) , S. Piutti (1) , A. Gravot (1) , S. Milesi (1) , A. Grabner (1) , B. Massot (1) ,<br />

K. Lievre (1) , M. Tran (1) , J.L. Goergen (2) & F. Bourgaud (1)<br />

(1) UMR 1121 INPL-ENSAIA-INRA, (2) UPR CNRS 6811<br />

Ecole Nationale Supérieure d’Agronomie et des Industries Alimentaires, Institut National<br />

Polytechnique de Lorra<strong>in</strong>e, 2 avenue de la <strong>for</strong>êt de Haye, 54505 Vandoeuvre les Nancy,<br />

France. Correspondence: Eric.Gontier@ensaia.<strong>in</strong>pl-nancy.fr; Tel: (+33)3 83 59 57 81<br />

Fax: (+33)3 83 59 57 99<br />

Abstract: Despite ef<strong>for</strong>ts made to produce plant secondary metabolites from cell<br />

suspensions, only a few <strong>in</strong>dustrial applications have been successful. Generally, higher yields<br />

are obta<strong>in</strong>ed when cultivat<strong>in</strong>g organs (roots or leafy stems) <strong>in</strong>stead of undifferentiated cells.<br />

In this case, new problems arise because of the structure of the plant material, and special<br />

bioreactors have to be built <strong>for</strong> such applications. Furthermore, the high cost of commercial<br />

bioreactors may limit the number available <strong>for</strong> the researcher to carry out many experiments<br />

<strong>in</strong> parallel. Because of this, we developed a very low cost system (i.e; bioreactors) that allows<br />

good growth of Ruta graveolens L. shoots and production of secondary metabolites (i.e.<br />

furanocoumar<strong>in</strong>s). The development of a very simple auto-prim<strong>in</strong>g siphon allows the use of<br />

common jars rang<strong>in</strong>g from 3 to 20 litres <strong>for</strong> temporary immersion cultures. The very low cost<br />

of such a home-made bioreactor allows scientists to run many different experiments at the<br />

same time. It thus saves time <strong>in</strong> optimis<strong>in</strong>g the culture medium parameters and <strong>in</strong> replicat<strong>in</strong>g<br />

trials be<strong>for</strong>e reach<strong>in</strong>g the step of f<strong>in</strong>al culture system development with highly equipped<br />

(costly) bioreactors.<br />

Key words: psoralen, temporary immersion, Tween20, permeabilisation, secondary<br />

metabolites, simplified bioreactor system<br />

Abbreviations: FC - furanocoumar<strong>in</strong>; FW - fresh weight; DW - dry weight; TIS -<br />

temporary immersion system; 5Mop - 5-methoxypsoralen; 8Mop - 8-methoxypsoralen;<br />

5,8Mop -5,8-methoxypsoralen; Pso - psoralen; vvm - volume of gas per volume of liquid<br />

per m<strong>in</strong>ute<br />

509<br />

A.K. Hvoslef-Eide and W. Preil (eds.), <strong>Liquid</strong> <strong>Culture</strong> <strong>Systems</strong> <strong>for</strong> <strong>in</strong> <strong>vitro</strong> <strong>Plant</strong> <strong>Propagation</strong>, 509–524.<br />

© 2005 Spr<strong>in</strong>ger. Pr<strong>in</strong>ted <strong>in</strong> the Netherlands.

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