3 Biotech

, 8:317 | Cite as

In vitro multiplication and growth improvement of Olea europaea L. cv Canino with temporary immersion system (Plantform™)

  • Carla BenelliEmail author
  • Anna De Carlo
Short Reports


Olea europaea L. cv Canino shoots were micropropagated to test two different culture systems: (1) on conventional semi-solid medium in glass jars and (2) in liquid culture in a Plantform™ bioreactor. The temporary immersion system, Plantform™, is a new propagation approach that uses liquid culture, where shoots undergo periodic immersion in liquid medium alternated with dry periods, avoiding gas accumulation through forced ventilation. This study proposes a protocol to improve in vitro propagation of olive reducing production costs. Our findings revealed that olive shoots propagated in Plantform™, with an immersion frequency of 8 min every 16 h and additional ventilation, showed good adaptability and better growth rates than those cultured in conventional system. Overall, the Plantform™ improves in vitro culture of ‘Canino’, showing higher proliferation, shoot length and better vigour of shoots. Moreover, the study found no significant differences in shoot length when 5 or 10 µM zeatin was applied in Plantform™ (3.04 and 3.13 cm, respectively); it is, therefore, possible to achieve efficient olive proliferation also with half hormone concentration. The positive performance of the bioreactor approach was also confirmed by Relative Growth Rate index. This is the first documented study of the Plantform™ technique for olive propagation.


Bioreactor In vitro propagation Olea europaea Zeatin 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Benelli C, Biricolti S, Calamai L, Dongarrà L, Lambardi M (2002) CO2 and ethylene evolution in the culture vessels during in vitro conservation of olive (cv. Frantoio) at 4 °C. Adv Hort Sci 16:184–187Google Scholar
  2. Benelli C, da Conceicao Moreira F, De Carlo A (2015) ‘Plant Form’, a temporary immersion system, for in vitro propagation of Myrtus communis and Olea europea. In: Proceedings of ‘6th International Symposium on Production and Establishment of Micropropagated Plants (PEMP), 19–24 April 2015, Sanremo, Italy, p 163Google Scholar
  3. Briccoli-Bati C, Fodale A, Mulè R, Trombino T (1999) Trials to increase in vitro rooting of (Olea europaea L.) cuttings. Acta Hort 474:91–94CrossRefGoogle Scholar
  4. Debnath SC (2009) A scale-up system for lowbush blueberry micropropagation using a bioreactor. HortScience 44:1962–1966Google Scholar
  5. Debnath SC (2011) Bioreactors and molecular analysis in berry crop micropropagation—a review. Can J Plant Sci 91:147–157CrossRefGoogle Scholar
  6. Etienne H, Berthouly M (2002) Temporary immersion systems in plant micropropagation. Plant Cell Tissue Organ Cult 69:215–231CrossRefGoogle Scholar
  7. Fabbri A, Bartolini G, Lambardi M, Kailis S (2004) Olive propagation manual. CSIRO Publishing, Collingwood, pp 77–95Google Scholar
  8. Gatti E, Sgarbi E, Ozudogru EA, Lambardi M (2017) The effect of Plantform™ bioreactor on micropropagation of Quercus robur in comparison to a conventional in vitro culture system on gelled medium, and assessment of the microenvironment influence on leaf structure. Plant Biosyst 151:1129–1136CrossRefGoogle Scholar
  9. George EF, Sherrington PD (1984) Plant propagation by tissue culture. Handbook and directory of commercial laboratories. Exegetics, BasingstokeGoogle Scholar
  10. Georgiev V, Schumann A, Pavlov A, Bley T (2014) Temporary immersion systems in plant. Biotechnol Eng Life Sci 14:607–621CrossRefGoogle Scholar
  11. Grigoriadou K, Vasilakakis M, Tzoulis T, Eleftheriou EP (2005) Experimental use of a novel temporary immersion system for liquid culture of olive microshoots. In: Hvolslef-Eide AK, Preil W (eds) Liquid culture systems for in vitro plant propagation. Springer, Netherlands, pp 263–274CrossRefGoogle Scholar
  12. Lambardi M, Rugini E (2003) Micropropagation of Olive (Olea europaea L.). In: Jain SM, Ishii K (eds) Micropropagation of woody trees and fruits. forestry sciences, vol 75. Springer, Dordrecht, pp 621–646CrossRefGoogle Scholar
  13. Lambardi M, Benelli C, Ozden-Tokatli Y, Ozudogru EA, Gumusel F (2006) A novel approach to olive micropropagation: the temporary immersion system. In: Proc. “2nd Int. Seminar OLIVEBIOTEQ 2006”, Vol I. Marsala—Mazara del Vallo, Italy, pp 319–326Google Scholar
  14. Lambardi M, Ozudogru EA, Roncasaglia R (2013) In vitro propagation of olive (Olea europaea L.) by nodal segmentation of elongated shoots. In: Lambardi M, Ozudogru E, Jain S (eds) Protocols for micropropagation of selected economically-important horticultural plants. Methods in molecular biology (methods and protocols), vol 994. Humana Press, Totowa, pp 33–44Google Scholar
  15. Paek KY, Chakrabarty D, Hahn EJ (2005) Application of bioreactor systems for large scale production of horticultural and medicinal plants. Plant Cell Tissue Organ Cult 81:287–300CrossRefGoogle Scholar
  16. Roels S, Noceda C, Escalona M, Sandoval J, Canal MJ, Rodriguez R, Debergh P (2006) The effect of headspace renewal in a temporary immersion bioreactor on plantain (Musa AAB) shoot proliferation and quality. Plant Cell Tissue Organ Cult 84:155–163CrossRefGoogle Scholar
  17. Rugini E (1984) In vitro propagation of some olive (Olea europaea sativa L.) cultivars with different root-ability, and medium development using analytical data from developing shoots and embryos. Sci Hortic 24:123–134CrossRefGoogle Scholar
  18. Rugini E, Cristofori V, Silvestri C (2016) Genetic improvement of olive (Olea europaea L.) by conventional and in vitro biotechnology methods. Biotech Adv 34:687–696CrossRefGoogle Scholar
  19. Welander M, Persson J, Asp H, Zhu LH (2014) Evaluation of a new vessel system based on temporary immersion system for micropropagation. Sci Hortic 179:227–232CrossRefGoogle Scholar
  20. Ziv M (2005) Simple bioreactors for mass propagation of plants. Plant Cell Tissue Organ Cult 81:277–285CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Trees and Timber Institute, National Research Council (CNR-IVALSA)FlorenceItaly

Personalised recommendations