A simple method for the isolation of plant protoplasts

Sankara Rao, K. ; Prakash, A. H. (1995) A simple method for the isolation of plant protoplasts Journal of Biosciences, 20 (5). pp. 645-655. ISSN 0250-5991

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Official URL: http://www.ias.ac.in/jarch/jbiosci/20/645-655.pdf

Related URL: http://dx.doi.org/10.1007/BF02703304


A simple protoplast isolation protocol that was designed to recover totipotent plant protoplasts with relative ease has been described. The key elements of the protocol are, tissue digestion at slightly elevated temperatures and use of protoplast-releasing enzymes that are stable and efficient at higher temperatures. Besides enzymes, the protoplast isolation cocktail consisted of an osmoticum (mannitol or MgSO4), and a protectant (CaCl2 2H2O), all dissolved in distilled water. The protocol has ensured reproducibility, higher yields and is gentle on protoplasts as the protoplasts obtained were amenable to cell wall regeneration and cell division. Plant regeneration was demonstrated for Nicotiana tabacum cv. Thompson from protoplasts isolated by this method. Wall regeneration and cell division were obtained in other species. The merits of the protocol are, simple and easy-to-handle procedure, non-requirement of preconditioning of donor plant and explants, incubation without agitation, satisfactory yields, culturability of the protoplasts isolated and applicability of the protocol to a large number of species including mucilage-containing plants.

Item Type:Article
Source:Copyright of this article belongs to Indian Academy of Sciences.
Keywords:Plant Protoplasts; Isolation; Simple Protocol; Totipotency
ID Code:89769
Deposited On:30 Apr 2012 14:33
Last Modified:30 Apr 2012 14:33

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