Liquid Culture Systems for in vitro Plant Propagation

Chapter 18
Experimental use of a novel temporary immersion system
for liquid culture of olive microshoots
Katerina Grigoriadou 1 , Miltiadis Vasilakakis 2 , Theofilos Tzoulis 3 &
Eleftherios P. Eleftheriou 4
1
VITRO HELLAS S.A., Nisseli Imathias, 593 00, Greece, tel:+302333024888,
fax: +302333026417, e-mail: vitro2@otenet.gr (author for correspondence).
2
Pomology Department, School of Agriculture, Aristotle Univ. of Thessaloniki, 541 24, Greece.
3 4
B. Hirs 11, 546 41, Thessaloniki, Greece. Department of Botany, School of Biology,
Aristotle University of Thessaloniki, 541 24, Greece
Abstract: A novel temporary immersion system (TIS), designed at the laboratory of VITRO
HELLAS S.A., was used for the liquid culture of olive microshoots during their proliferation
phase. The results were compared with those obtained in agar solidified medium, which was
used as control, and with other bioreactor systems including the LifeReactor © , liquid culture in
Erlenmeyer flasks under agitation and liquid culture on filter paper bridges. After 30 days of
culture, results derived from the novel TIS (1.93 new microshoots per explant, 0.95 cm shoot
length) were statistically the same as the control (1.75 microshoots per explant, 1.22 cm shoot
length respectively). Equally high results were obtained with the agitated Erlenmeyer flasks,
while LifeReactor © gave the poorest ones (0.35 new microshoots per explant, 0.40 cm shoot
length). Cultures on filter paper bridges developed large masses of callus at the explant bases.
However, all the liquid cultures suffered from hyperhydricity, the most serious was observed in
the Erlenmeyer flasks and the least in the novel TIS. In order to reduce hyperhydricity, the TIS
was combined with cultures on agar solidified medium, which eliminated the problem but
shoot proliferation was also reduced. It seems that the novel TIS provides a promising device,
although some technical problems need to be overcome for large scale production.
Key words: bioreactor, in vitro culture, liquid culture, Olea europaea L.
Abbreviations: GA - gibberellic acid; NAA - naphthaleneacetic acid
1. Introduction
The use of liquid media for in vitro culture is considered an ideal
technique for mass propagation. However, the main disadvantage of liquid
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A.K. Hvoslef-Eide and W. Preil (eds.), Liquid Culture Systems for in vitro Plant Propagation, 263–274.
© 2005 Springer. Printed in the Netherlands.