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A Simple Procedure for Isolation, Culture of Protoplast, and Plant Regeneration

  • Indu Kumari
Chapter
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Part of the Environmental and Microbial Biotechnology book series (EMB)

Abstract

This chapter presents different steps involved in isolation and culture of protoplast; discusses basic steps involved in the developmental sequence from protoplast to plant; and overviews a simple procedure for protoplast isolation, culture, and plant regeneration. Protoplasts are naked plant cells without the cell wall, but they possess plasma membrane and other components of cell. Protoplasts are potentially totipotent individuals at the single-cell level. They have the potential to regenerate a cell wall, dedifferentiate, divide mitotically and form unlimited growing cell clones, and differentiate shoot and root meristems or embryos which regenerate whole plant.

Keywords

Isolation Culture Embryos Cell clones Protoplast 

References

  1. Bajaj YPS (1974) Potentials of protoplast culture work in agriculture. Euphytica 23:633–649CrossRefGoogle Scholar
  2. Binding H, Behls R, Schieder O, Sopory SK, Wenzel G (1978) Regeneration of mesophyll protoplasts isolated from dihaploid clones of Solanum tuberosum. Physiol Plant 43:52–54CrossRefGoogle Scholar
  3. Cocking EC (1960) A method for the isolation of plant protoplasts and vacuoles. Nature (London) 187:927–929CrossRefGoogle Scholar
  4. Cocking EC (1972) Plant cell protoplasts—isolation and development. A Rev Plant Physiol 23:29–50CrossRefGoogle Scholar
  5. Cocking EC, Pojnar E (1969) An electron microscopic study of the infection of isolated tomato fruit protoplasts by tobacco mosaic virus. J Gen Virol 4:305–312CrossRefGoogle Scholar
  6. Davey MR, Short KC (1973) The isolation, culture, and fusion of legume and corn flower leaf protoplasts. In: Protoplastes et fusion de cellules somatiques végétales. Colloq Int CNRS, Paris, pp 437–444Google Scholar
  7. Dos Santos AVP, Outka DE, Cocking EC, Davey MR (1980) Organogenesis and somatic embryogenesis in tissues derived from leaf protoplasts and leaf explants of Medicago sativa. Z Pflanzenphysiol 99:261–270CrossRefGoogle Scholar
  8. Evans DA, Crocomo OJ, De Carvalho MTV (1980a) Protoplast isolation and subsequent callus regeneration in sugarcane. Z Pflanzenphysiol 98:355–358CrossRefGoogle Scholar
  9. Evans DA, Wetter LR, Gamborg OL (1980b) Somatic hybrid plants of Nicotiana glauca and Nicotiana tabacum obtained by protoplast fusion. Physiol Plant 48:225–230CrossRefGoogle Scholar
  10. Frearson EM, Power JB, Cocking EC (1973) The isolation, culture, and regeneration of Petunia leaf protoplasts. Dev Biol 33:130–137CrossRefGoogle Scholar
  11. Gregory DW, Cocking EC (1965) The large scale isolation of protoplasts from immature tomato fruit. J Cell Biol 24:143–146CrossRefGoogle Scholar
  12. Grun P, Chu LJ (1978) Development of plants from protoplasts of Solanum (Solanaceae). Am J Bot 65:538–543CrossRefGoogle Scholar
  13. Harada H (1973) A new method for obtaining protoplasts from mesophyll cells. Z Pflanzen-physiol 69:77–80CrossRefGoogle Scholar
  14. Hess D, Potrykus I, Doon G, Durand T, Hoffmann F (1973) Transformation experiments in higher plants: prerequisites for the use of isolated protoplasts (isolation from mesophyll and callus cultures, uptake of proteins and DNA, and regeneration of whole plants). In: Protoplastes et fusion de cellules somatiques végétales. Colloq Int CNRS, Paris, pp 343–351Google Scholar
  15. Power JB, Cocking EC (1969) A simple method for the isolation of very large numbers of leaf protoplasts. Biochem J 111:33Google Scholar
  16. Schenk RU, Hilderbrandt AC (1969) Production of protoplasts from plant cells in liquid culture using purified commercial cellulases. Crop Sci 9:629–631CrossRefGoogle Scholar
  17. Sink KC, Power JB (1977) The isolation, culture, and regeneration of leaf protoplasts of Petunia parviflora Juss. Plant Sci Lett 10:335–340CrossRefGoogle Scholar
  18. Takebe I, Labib G, Melchers G (1971) Regeneration of whole plants from isolated mesophyll protoplasts of tobacco. Naturwissenschaften 58:318–320CrossRefGoogle Scholar
  19. Vasil V, Vasil IK (1974) Regeneration of tobacco and Petunia plants from protoplasts and culture of corn protoplasts. In Vitro 10:83–96CrossRefGoogle Scholar
  20. Vasil V, Vasil IK (1980) Isolation and culture of cereal protoplasts II. Embryogenesis and plantlet formation from protoplasts of Pennisetum americanum. Theor Appl Genet 56:97–99CrossRefGoogle Scholar
  21. Wallin A, Eriksson T (1972) Plating of protoplasts from cell suspension cultures of Daucus carota. In: Protoplastes et fusion de cellules somatiques végétales. Colloq Int. CNRS, Paris, pp 301–307Google Scholar
  22. Wilson HM, Styer DJ, Conard PL, Durbin RD, Helgeson JP (1980) Isolation of sterile protoplasts from unsterilized leaves. Plant Sci Lett 18:151–154CrossRefGoogle Scholar
  23. Zieg RG, Outka DE (1980) The isolation, culture and callus formation of soybean pod protoplasts. Plant Sci Lett 18:105–114CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2020

Authors and Affiliations

  • Indu Kumari
    • 1
  1. 1.Department of BotanyNirmala College, DorandaRanchiIndia

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