Plant Cell, Tissue and Organ Culture

, Volume 81, Issue 3, pp 313–318 | Cite as

Optimisation of growing conditions for the apple rootstock M26 grown in RITA containers using temporary immersion principle



The use of bioreactors may provide an efficient and economic tool for mass clonal propagation of plants if technical problems can be solved. In this paper, we report the results of experiments aimed at optimising conditions for apple rootstock M26 grown in RITA containers using the temporary immersion principle. We tested different types and sizes of explants, different concentrations of plant growth regulators (BAP, kinetin and IBA) in the multiplication and elongation phases, and medium exchange during the shoot elongation period. The results show that the higher concentrations of cytokinins were required during the shoot multiplication phase, while the lower concentrations were better during the shoot elongation phase. Hyperhydricity was increased with increasing concentration in of cytokinins during both shoot multiplication and shoot elongation phases. The best shoot production in terms of shoot number and shoot quality was obtained using 4.4 μmol BAP and 0.5 μmol IBA during the shoot multiplication phase and 1.1 μmol BAP and 0.25 μmol IBA during the shoot elongation phase. Medium exchange twice during the shoot elongation phase resulted in higher shoot production compared with no exchange of the medium. However, it also resulted in increased hyperhydricity. Immersion frequency of 16 times per day gave a higher multiplication rate and longer shoots than 8 times per day. The explant size of 0.5 cm or 1 cm resulted in a significantly higher shoot production rate compared with that of 1.5 cm, but shoot length and hyperhydricity were not affected by the explant size. Shoot cultures from the liquid media rooted normally in the RITA containers with more than 90% rooting and the rooted plantlets acclimatised well in the greenhouse.


apple bioreactor micropropagation RITA temporary immersion 





indole butyric acid


Murashige and Skoog medium (1962)


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  1. Aitken-Christie, J, Jones, C 1987Towards automation: radiata pine shoot hedges in vitroPlant Cell Tiss. Org. Cult.8185196Google Scholar
  2. Cabasson, C, Alvard, D, Dambier, D, Ollitrault, P, Teisson, C 1997Improvement of Citrus somatic embryo development by temporary immersionPlant Cell Tiss. Org. Cult.503337Google Scholar
  3. Chu, I 1995

    Economic analysis of automated micropropagation

    Aitken-Christie, JKozai, TSmith, MAL eds. Automation and Environmental Control in Plant Tissue CultureKluwer Academic PublishersDordrecht1926
    Google Scholar
  4. Escalona, M, Lorenzo, JC, González, B, Daquinta, M, González, JL, Desjardins, Y, Borroto, CG 1999Pineapple (Ananas comosus L. Merr) micropropagation in temporary immersion systemsPlant Cell Rep18743748Google Scholar
  5. Jiménez, E, Pérez, N, Feria, M de, Barbón, R, Capote, A, Chávez, M, Quiala, E, Pérez, JC 1999Improved production of potato microtubers using a temporary immersion systemPlant Cell Tiss. Org. Cult.591923Google Scholar
  6. Lorenzo, JC de los Angeles Blanco M, Peláez, O, González, A, Cid, M, Iglesias, A, González, B, Escalona, M, Espinosa, P, Borroto, C 2001Sugarcane micropropagation and phenolic excretionPlant Cell Tiss. Org. Cult.6518Google Scholar
  7. Maene, L, Debergh, P 1985Liquid medium additions to established tissue cultures to improve elongation and rooting in vitroPlant Cell Tiss. Org. Cult.52333Google Scholar
  8. Murashige, F, Skoog, F 1962A revised medium for rapid growth and bioassys with tobacco tissue culturesPhysiol. Plant.15473492Google Scholar
  9. Quoirin M, Lepoivre P & Boxus P (1977) Un premier bilan de dix années de recherche sur les cultures de méristèmes et la multiplication in vitro de fruitiers ligneux (in French). Compte rendu des recherches, Station des Cultures Fruitières et Marrîchères de Gembloux 1976–1977 (pp. 93–117)Google Scholar
  10. Simonton, W, Robacker, C, Krueger, S 1991A programmable micropropagation apparatus using cycled mediumPlant Cell, Tiss. Org. Cult.27211218Google Scholar
  11. Sluis, CJ, Walker, KA 1985Commercialization of plant tissue culture propagationInternational Association for Plant Tissue Culture. Newsletters47212Google Scholar
  12. Teisson, C, Alvard, D 1995

    A new concept of plant in vitro cultivation liquid medium: temporary immersion

    M, Terzi eds. Current Issues in Plant Molecular and Cellular BiologyKluwer Academic PublishersDordrecht105110
    Google Scholar
  13. Teisson C, Alvard D, Lartaud M, Etienne H, Berthouly M, Escalona M & Lorenzo JC (1999) Temporary immersion for plant tissue culture. In: Plant Biotechnology and In Vitro Biology in the 21st Century, Proceedings of the IXth International Congress of Plant Tissue and Cell Culture, Section H: Novel micropropagation methods (pp. 629–632). JerusalemGoogle Scholar
  14. Tisserat, B, Vandercook, CE 1985Development of an automated plant culture systemPlant Cell Tiss. Org. Cult.5107117Google Scholar
  15. Walkey, DG 1972Production of apple plantlets from axillary bud meristemsCan. J. Plant Sci.5210851087Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  1. 1.Department of Crop ScienceSwedish University of Agricultural SciencesAlnarpSweden

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