Protoplast Isolation and Culture

  • Stephen C. Gleddie
Part of the Springer Lab Manual book series (SLM)


Protoplasts can be isolated from plant tissues or cultured cells by enzymatic digestion to remove the cell walls. The enzymes for this purpose are preparations which are commercially available. The success of protoplast isolation depends especially on the condition of the tissue and the combination of enzymes being used. There is no standard method for the isolation and culture of protoplasts. However, the procedure follows a general pattern. The individual cell or tissue source may require special conditions for successful isolation or for culture. Protoplasts from cell suspension cultures generally are the most readily obtained and usually regenerate into dividing cells at a reasonable frequency. Moreover, if plant regeneration is possible from the cells of the suspension culture, the same is often true for the cells regenerated from the protoplasts derived from the culture. Leaf mesophyll cells from a wide range of plants also have been used as protoplast sources with success.


Plant Regeneration Somatic Hybrid Protoplast Fusion Donor Plant Protoplast Isolation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References and Advanced Readings

  1. Gleddie S, Keller WA (1989) Protoplast fusion technology. J Tiss Cult Meth 12: 157–161CrossRefGoogle Scholar
  2. Gleddie S, Keller WA, Poysa V (1989) Plant regeneration from stem cortex protoplasts of a tomato hybrid. Plant Cell Rep 8: 21–24CrossRefGoogle Scholar
  3. Keller WA, Setterfield G, Douglas G, Gleddie S, Nakamura C (1982) Production, characterization, and utilization of somatic hybrids of higher plants. In: Tomes D, Ellis B, Harney P, Kasha K, Peterson R (eds) Application of plant cell and tissue culture to agriculture and industry. University of Guelph, Guelph, ON, pp 81–114Google Scholar
  4. Klimaszewska K, Keller WA (1987) Plant regeneration from stem cortex protoplasts of Brassica napus. Plant Cell Tissue Organ Cult 8: 225–233CrossRefGoogle Scholar
  5. Pelletier G, Primard C, Vedel F, Chetrit P, Remy R, Rouselle, Renard M (1983) IntergenericGoogle Scholar
  6. cytoplasmic hybridization in cruciferae by protoplast fusion. Mol Gen Genet 191:244–250 Rashid A (1988) Cell physiology and genetics of higher plants, vol II. CRC Press, Boca Raton, FLGoogle Scholar
  7. pp 85–117Google Scholar
  8. Shillito RD, Paszkowski J, Potrykus I (1983) Agarose plating and a bead type technique enable and stimulate development of protoplast-derived colonies in a number of plant species. Plant Cell Rep 2: 244–247CrossRefGoogle Scholar
  9. Tan ML, Boerrigter HS, Kool AJ (1987) A rapid procedure for plant regeneration from protoplasts isolated from suspension cultures and leaf mesophyll cells of wild Solanum species and Lycopersicon pennellii. Plant Sci 49: 63–72CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1995

Authors and Affiliations

  • Stephen C. Gleddie

There are no affiliations available

Personalised recommendations