Plant Cell, Tissue and Organ Culture

, Volume 63, Issue 1, pp 67–72 | Cite as

Mass multiplication of protocorm-like bodies using bioreactor system and subsequent plant regeneration in Phalaenopsis

  • Park So Young
  • H.N. Murthy
  • Paek Kee YoeupEmail author


Protocorm-like bodies (PLBs) formed on leaf segmentsin vitro were used as explants for bioreactor cultures. Continuous immersion cultures (air lift column and air lift-balloon bioreactor), and temporary immersion cultures (with or without charcoal filter attached) were used for the culture of PLB sections. A temporary immersion culture with charcoal filter attached was most suitable for PLB culture. About 18,000 PLBs were harvested from 20 g of inoculum (∼1000 PLB sections) in 2 l Hyponex medium after 8 weeks of incubation. Aeration in a bioreactor at 0.5 or 2.0 volume of air per volume of medium min−1 (vvm) yielded similar levels of biomass production. PLBs grown in bioreactors were cultured on solid Murashige and Skoog, Vacin and Went, Knudson C, Lindemann and Hyponex media. Hyponex medium was found to be suitable for conversion of PLBs into plantlets and 83% of PLBs transformed into plantlets on this medium. The feasibility of using PLBs for large-scale micropropagation was evaluated for scaled-up liquid cultures in bioreactors, rate of proliferation, and regeneration.

bioreactor liquid cultures micropropagation orchid 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Aitken-Christie J, Kozai T & Takayama S (1995) Automation in plant tissue culture. General introduction and overview. In: Aitken-Christe J, Kozai T & Smith MAL (eds) Automation and Environmental Control in Plant Tissue Culture (pp 1–18). Kluwer Academic Publishers, DordrechtGoogle Scholar
  2. Akita M & Takayma S (1988) Mass propagation of potato tubers using jar fermentor techniques. Acta Hortic. 230: 55–61Google Scholar
  3. Arditti J & Ernst R (1993) Micropropagation of Orchids. John Wiley & Son, Inc., New YorkGoogle Scholar
  4. Cantliffe DJ, Bieniek ME & Harrell RC (1993) A system approach to developing and automated synthetic seed model. In: Soh WY, Liu JR & Komamine A (eds) Advances in the Developmental Biology and Biotechnology of Higher Plants (pp 160–196). The Korean Society of Plant Tissue Culture, KoreaGoogle Scholar
  5. Griesbach RJ (1983) The use of indoleactylamino acids in the in vitro propagation of Phalaenopsis orchids. Sci. Hortic. 19: 363–366CrossRefGoogle Scholar
  6. Gupta PK, Timmis R & Carlson WC (1993) Somatic embryogenesis: A possible tool for large-scale propagation of forestry species. In: Soh WY, Liu JR & Komamine A (eds) Advances in Developmental Biology and Biotechnology of Higher Plants (pp 18–37). The Korean Society of Tissue Culture, KoreaGoogle Scholar
  7. Ilan A, Ziv m & Halevy AH (1995) Propagation and corm development of Brodiaea in liquid cultures. Sci. Hortic. 63: 101–112CrossRefGoogle Scholar
  8. Kano K (1965) Studies on the media for orchid seed germination. Mem. Fac. Agric. Kagawa Univ. 20: 1–68Google Scholar
  9. Knudson L (1946) A new nutrient solution for germination of orchid seed. Amer. Orchid Soc. Bull. 15: 214–217Google Scholar
  10. Leathers RR, Smith MAL & Aitken-Christie J (1995) Automation of the bioreactor process for mass propagation and secondary metabolites. In: Aitken-Christe J, Kozai T & Smith MAL (eds) Automation and Environmental Control in Plant Tissue Culture (pp 187–214). Kluwer Academic Publishers, DordrechtGoogle Scholar
  11. Lilien-Kipnis H, Ziv M & Kahany S (1990) Proliferation of plantlet regeneration from inflorescence derived Nerine explants cultured in vitro. In: De Jong (ed) Integration of in vitro Techniques in Ornamental Plant Breeding (pp 28–33). CPO Center for Plant Breeding Research, Wageningen, The NetherlandsGoogle Scholar
  12. Lin CC (1986) In vitro culture of flower stalk internodes of Phalaenopsis and Doritaenopsis. Lindleyana 1: 158–163Google Scholar
  13. Lindemann EGP, Gunckel JE & Davidson OW (1970) Meristem cutlure of Cattleya. Amer. Orchid Soc. Bull. 39: 100–127Google Scholar
  14. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497CrossRefGoogle Scholar
  15. Preil W (1991) Application of bioreactors in plant propagation. In: Debergh PC & Zimmerman RH (eds) Micropropagation-Technology and Application (pp 425–445). Kluwer Academic Publishers, DordrechtGoogle Scholar
  16. Preil W & Beck A (1991) Somatic embryogenesis in bioreactor culture. Acta Hortic. 289: 179–192Google Scholar
  17. Sigeoka T, Kozumi Y & Kawamura M (1994) Mass propagation of shoots of Stevia rebuniana using large scale bioreactor. Plant Cell Rep. 13: 180–183Google Scholar
  18. Seon JH, Kim YS, Son SH & Paek KY (2000) The Fed-batch culture system using bioreactor for the bulblet production of oriental lillies. Acta Hortic. 520: 53–59Google Scholar
  19. Son SH, Choi SM, Yun SR, Kwon UW, Lee YH & Paek KY (1999) Large scale culture of plant cell and tissue by bioreactor system. J. Plant Biotech. 1: 1–7Google Scholar
  20. Stuart DA, Strickland SG & Walkar KA (1987) Bioreactor production of alfalfa somatic embryos. HortScience 22: 800–809Google Scholar
  21. Takahashi S, Matsubara K, Yamagata H & Morimoto T (1992) Micropropagation of virus free bulblets of Lilium longiflorum by tank culture 1. Development, culture method and large scale propagation. Acta Hortic. 319: 83–88Google Scholar
  22. Vacin E & Went FW (1949) Some pH changes in nutrient solutions. Bot Gaz. 110: 605–613CrossRefGoogle Scholar
  23. Vanderschaege AM & Debergh PC (1988) Automation of tissue culture manipulations in the final stages. Acta Hortic. 227: 399–401Google Scholar
  24. Vasil IK (1994) Automation of plant propagation. Plant Cell Tiss. Org. Cult. 39: 105–108CrossRefGoogle Scholar
  25. Watad AA, Alper Y, Stav R & Levin R (1999) Mechanization of micropropagation In: Altmann A, Ziv M & Izhar G (eds) Plant Biotechnology and In Vitro Biology in the 21st century (pp. 663–666). Kluwer Academic Publishers, DordrechtGoogle Scholar
  26. Ziv M (1989) Enhanced shoot and cormlet proliferation in liquid cultured gladiolus buds by growth retardants. Plant Cell Tiss. Org. Cult. 17: 101–110CrossRefGoogle Scholar
  27. Ziv M (1992a) The use of growth retardants for the regeneration and acclimatization of in vitro plants. In: Karssen CM, Von Loon LC & Vreugdenhil (eds) Progress in Plant Growth Regulation (pp 809–817). Kluwer Academic Publishers, DordrechtGoogle Scholar
  28. Ziv M (1992b) Morphogenic control of plants micropropagated in bioreactor cultures and its possible impact on acclimatization. Acta Hortic. 319: 119–124Google Scholar
  29. Ziv M (1999) Organogenic plant regeneration in bioreactors. In: Altmann A, Ziv M & Izhar G (eds) Plant Biotechnology and In Vitro Biology in the 21st Century (pp 673–676). Kluwer Academic Publishers, DordrechtGoogle Scholar
  30. Ziv M & Hadar A (1991) Morphogenic pattern of Nephrolepis exalata cv. Bostoniensis in agar or liquid cultures: implications for micropropagation. Isr. J. Bot. 40: 7–16Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  1. 1.Research Center for the Development of Advanced Horticultural TechnologyChungbuk National UniversityKorea

Personalised recommendations