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.
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References
Bajaj YPS (1974) Potentials of protoplast culture work in agriculture. Euphytica 23:633–649
Binding H, Behls R, Schieder O, Sopory SK, Wenzel G (1978) Regeneration of mesophyll protoplasts isolated from dihaploid clones of Solanum tuberosum. Physiol Plant 43:52–54
Cocking EC (1960) A method for the isolation of plant protoplasts and vacuoles. Nature (London) 187:927–929
Cocking EC (1972) Plant cell protoplasts—isolation and development. A Rev Plant Physiol 23:29–50
Cocking EC, Pojnar E (1969) An electron microscopic study of the infection of isolated tomato fruit protoplasts by tobacco mosaic virus. J Gen Virol 4:305–312
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 437–444
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 99:261–270
Evans DA, Crocomo OJ, De Carvalho MTV (1980a) Protoplast isolation and subsequent callus regeneration in sugarcane. Z Pflanzenphysiol 98:355–358
Evans DA, Wetter LR, Gamborg OL (1980b) Somatic hybrid plants of Nicotiana glauca and Nicotiana tabacum obtained by protoplast fusion. Physiol Plant 48:225–230
Frearson EM, Power JB, Cocking EC (1973) The isolation, culture, and regeneration of Petunia leaf protoplasts. Dev Biol 33:130–137
Gregory DW, Cocking EC (1965) The large scale isolation of protoplasts from immature tomato fruit. J Cell Biol 24:143–146
Grun P, Chu LJ (1978) Development of plants from protoplasts of Solanum (Solanaceae). Am J Bot 65:538–543
Harada H (1973) A new method for obtaining protoplasts from mesophyll cells. Z Pflanzen-physiol 69:77–80
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 343–351
Power JB, Cocking EC (1969) A simple method for the isolation of very large numbers of leaf protoplasts. Biochem J 111:33
Schenk RU, Hilderbrandt AC (1969) Production of protoplasts from plant cells in liquid culture using purified commercial cellulases. Crop Sci 9:629–631
Sink KC, Power JB (1977) The isolation, culture, and regeneration of leaf protoplasts of Petunia parviflora Juss. Plant Sci Lett 10:335–340
Takebe I, Labib G, Melchers G (1971) Regeneration of whole plants from isolated mesophyll protoplasts of tobacco. Naturwissenschaften 58:318–320
Vasil V, Vasil IK (1974) Regeneration of tobacco and Petunia plants from protoplasts and culture of corn protoplasts. In Vitro 10:83–96
Vasil V, Vasil IK (1980) Isolation and culture of cereal protoplasts II. Embryogenesis and plantlet formation from protoplasts of Pennisetum americanum. Theor Appl Genet 56:97–99
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 301–307
Wilson HM, Styer DJ, Conard PL, Durbin RD, Helgeson JP (1980) Isolation of sterile protoplasts from unsterilized leaves. Plant Sci Lett 18:151–154
Zieg RG, Outka DE (1980) The isolation, culture and callus formation of soybean pod protoplasts. Plant Sci Lett 18:105–114
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Kumari, I. (2020). A Simple Procedure for Isolation, Culture of Protoplast, and Plant Regeneration. In: Kumar, M., Kumar, V., Prasad, R. (eds) Phyto-Microbiome in Stress Regulation. Environmental and Microbial Biotechnology. Springer, Singapore. https://doi.org/10.1007/978-981-15-2576-6_13
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DOI: https://doi.org/10.1007/978-981-15-2576-6_13
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