This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Uchimiya, H. Articles by Murashige, T. Search for Related Content PubMed PubMed Citation Articles by Uchimiya, H. Articles by Murashige, T. Agricola Articles by Uchimiya, H. Articles by Murashige, T.

Articles

Evaluation of Parameters in the Isolation of Viable Protoplasts from Cultured Tobacco Cells ¹

^a Department of Plant Sciences, University of California, Riverside, California 92502

A systematic evaluation disclosed the following conditions to be optimum for the isolation of viable protoplasts from cultured cells of Nicotiana tabacum L. `Bright Yellow' grown in liquid suspensions: (a) the cell culture in the early phase of cell number increase, (b) an enzyme mixture of 1% cellulase "Onozuka" and 0.2% Macerozyme, (c) an enzyme solution pH of 4.7 or 5.7, (d) a 2- to 3-hr incubation period, (e) 5 ml of enzyme solution per 500 mg cells and contained in a 50-ml Delong flask, (f) agitation on a gyrotory shaker at 50 rpm, and (g) 0.3 to 0.8 M mannitol as osmoticum in the cell enzyme mixture. The incubation temperature may be varied from 22 to 37 C. The procedure enabled 30% of the tobacco cells to form protoplasts, 80% of which regenerated cell walls in 4 days and 40% resumed cell division activity when returned to cell culture medium.

This article has been cited by other articles:

				T. Kunik, T. Tzfira, Y. Kapulnik, Y. Gafni, C. Dingwall, and V. Citovsky Genetic transformation of HeLa cells by Agrobacterium PNAS, February 13, 2001; 98(4): 1871 - 1876. [Abstract] [Full Text] [PDF]