Summary
A method is described for purifying plant protoplasts from cellular and subcellular debris. The procedure utilizes a density buffer containing 9.6% sodium metrizoate and 5.6% Ficoll. The use of fluorescein diacetate for assessing the viability of plant protoplasts is also reported.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- FDA:
-
fluorescein diacetate
References
Bala Bawa, S., Torrey, J.G.: “Budding” and nuclear division in cultured protoplasts of corn, Convolvulus, and onion. Bot. Gaz. 132, 240–245 (1971)
Böyum, A.: Separation of white blood cells. Nature (Lond.) 21, 793–794 (1964)
Böyum, A.: Isolation of leucocytes from human blood. Further observations. Scand. J. Clin. Lab. Invest. 21, Suppl. 97, 31–50 (1968)
Brown, G., Greaves, M.F.: Enumeration of absolute numbers of T and B lymphocytes in human blood. Scand. J. Immunol. 3, 161–172 (1974)
Bui-Dang-Ha, D., Mackenzie, I.A.: The division of protoplasts from Asparagus officinalis L. and their growth and differentiation. Protoplasma 78, 215–221 (1973)
Cercek, L., Cercek, B.: Effect of centrifugal forces on the structuredness of cytoplasm in growing yeast cells. Biophysik 9, 105–108 (1973a)
Cercek, L., Cercek, B.: Relationship between changes in the structuredness of cytoplasm and rate constants for the hydrolysis of FDA in Saccharomyces cerevisiae. Biophysik 9, 109–112 (1973b)
Cercek, L., Cercek, B., Ockey, C.H.: Structuredness of the cytoplasmic matrix and Michaelis-Menten constants for the hydrolysis of FDA during the cell cycle in Chinese Hamster ovary cells. Biophysik 10, 187–194 (1973a)
Cercek, L., Cercek, B., Ockey, C.H.: Cell membrane permeability during the cell generation cycle in Chinese Hamster ovary cells. Biophysik 10, 195–197 (1973b)
Coeking, E.C., Power, J.B., Evans, P.K., Safwat, F., Frearson, E.M., Hayward, C., Berry, S.F., George, D.: Naturally occurring differential drug sensitivities of cultured plant protoplasts. Plant Sci. Letters 3, 341–350 (1974)
Davey, M.R., Bush, E., Power, J.B.: Cultural studies of a dividing legume leaf protoplast system. Plant Sci. Letters 3, 127–133 (1974)
Du Bois, M.J.G.J., Huismans, D.R., Schellekens, P.Th.A., Eijsvoogel, V.P.: Investigation and standardization of the conditions for micro-lymphocyte cultures. Tissue Antigens 3, 402–409 (1973)
Eriksson, T.: Isolation and fusion of plant protoplasts. Les cultures de tissus de plantes, Coll. Int., C.N.R.S. No. 193, 297–302 (1971)
Evans, P.K., Keates, A.G., Cocking, E.C.: Isolation of protoplasts from cereal leaves. Planta (Berl.) 104, 178–181 (1972)
Glimelius, K., Wallin, A., Eriksson, T.: Agglutinating effects of concanavalin A on isolated protoplasts of Daucus carota. Physiol. Plant. 31, 225–230 (1974)
Gregory, D.W., Cocking, E.C.: The large-scale isolation of protoplasts from immature tomato fruit. J. Cell Biol 24, 143–146 (1965)
Grout, B.W.W., Coutts, R.H.A.: Additives for the enhancement of fusion and endocytosis in higher plant protoplasts: An electrophoretic study. Plant Sci. Letters 2, 397–403 (1974)
Heslop-Harrison, J., Heslop-Harrison, Y.: Evaluation of pollen viability by enzymatically induced fluorescence: intracellular hydrolysis of fluorescein diacetate. Stain Technol. 45, 115–120 (1970)
Kameya, T., Uchimiya, H.: Embryoids derived from isolated protoplasts of carrot. Planta (Berl.) 103, 356–360 (1972)
Kanai, R., Edwards, G.E.: Purification of enzymatically isolated mesophyll protoplasts from C3, C4, and crassulacean acid metabolism plants using an aqueous dextran-polyethylene glycol two-phase system. Plant Physiol. 52, 484–490 (1973)
Kartha, K.K., Michayluk, M.R., Kao, K.N., Gamborg, A.L., Constabel, F.: Callus formation and plant regeneration from mesophyll protoplasts of rape plants (Brassica napus L. cv. Zephyr). Plant Sci. Letters 3, 265–271 (1974)
Martel, J.L., Taramillo, S., Allen, F.H. jr., Rubinstein, P.: Serology for automated cytotoxicity assays. Contrast fluorescence test. Vox Sang. (Basel) 27, 13–20 (1974)
Pelcher, L.E., Gamborg, O.L., Kao, K.N.: Bean mesophyll protoplasts: production, culture and callus formation. Plant Sci. Letters 3, 107–111 (1974)
Power, J.B., Cocking, E.C.: Isolation of leaf protoplasts: macromolecule uptake and growth substance response. J. exp. Bot. 21, 64–70 (1970)
Raj, B., Herr, J.M.: Isolation of protoplasts from the placental cells of Lycopersicum pimpinellifolium Mill. Exp. Cell Res. 64, 479–480 (1970)
Rotman, B., Papermaster, B.W.: Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters. Proc. nat. Acad. Sci. (Wash.) 55, 134–141 (1966)
Shepard, J.F., Totten, R.E.: Isolation and regeneration of tobacco mesophyll cell protoplasts under low osmotic conditions. Plant Physiol. 55, 689–694 (1975)
Schenk, R.U., Hildebrandt, A.C.: Production, manipulation and fusion of plant cell protoplasts as steps toward somatic hybridization. Les cultures de tissus de plantes, Coll. Int., C.N.R.S. No. 193, 319–331 (1971)
Taiz, L., Jones, R.L.: The isolation of barley-aleurone protoplasts. Planta (Berl.) 101, 95–100 (1971)
Thorsby, E., Bratlie, A.: A rapid method for preparation of pure lymphocyte suspensions. Histocompat. Testing pp. 655–656 Copenhagen: Munksgaard 1970
Ting, A., Morris, P.J.: A technique for lymphocyte preparation from stored heparinized blood. Vox Sang. (Basel) 20, 561–563 (1971)
Watts, J.W., Motoyoshi, F., King, J.M.: Problems associated with the production of stable protoplasts of cells of tobacco mesophyll. Ann. Bot. 38, 667–671 (1974)
Widholm, J.M.: The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol. 47, 189–194 (1972)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Larkin, P.J. Purification and viability determinations of plant protoplasts. Planta 128, 213–216 (1976). https://doi.org/10.1007/BF00393231
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00393231