Abstract
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.
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- Carlson P. S., Smith H. H., Dearing R. D. Parasexual interspecific plant hybridization. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2292–2294. doi: 10.1073/pnas.69.8.2292. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kao K. N., Keller W. A., Miller R. A. Cell division in newly formed cells from protoplasts of soybean. Exp Cell Res. 1970 Oct;62(2):338–340. doi: 10.1016/0014-4827(70)90563-x. [DOI] [PubMed] [Google Scholar]
- Lewis L. N., Varner J. E. Synthesis of Cellulase during Abscission of Phaseolus vulgaris Leaf Explants. Plant Physiol. 1970 Aug;46(2):194–199. doi: 10.1104/pp.46.2.194. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Power J. B., Cummins S. E., Cocking E. C. Fusion of isolated plant protoplasts. Nature. 1970 Mar 14;225(5237):1016–1018. doi: 10.1038/2251016a0. [DOI] [PubMed] [Google Scholar]
- Ruesink A. W., Thimann K. V. Protoplasts from the Avena coleoptile. Proc Natl Acad Sci U S A. 1965 Jul;54(1):56–64. doi: 10.1073/pnas.54.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]