Skip to main content
Log in

Purification and viability determinations of plant protoplasts

  • Published:
Planta Aims and scope Submit manuscript

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.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

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)

    Google Scholar 

  • Böyum, A.: Separation of white blood cells. Nature (Lond.) 21, 793–794 (1964)

    Google Scholar 

  • Böyum, A.: Isolation of leucocytes from human blood. Further observations. Scand. J. Clin. Lab. Invest. 21, Suppl. 97, 31–50 (1968)

    Google Scholar 

  • Brown, G., Greaves, M.F.: Enumeration of absolute numbers of T and B lymphocytes in human blood. Scand. J. Immunol. 3, 161–172 (1974)

    Google Scholar 

  • 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)

    Google Scholar 

  • Cercek, L., Cercek, B.: Effect of centrifugal forces on the structuredness of cytoplasm in growing yeast cells. Biophysik 9, 105–108 (1973a)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • Davey, M.R., Bush, E., Power, J.B.: Cultural studies of a dividing legume leaf protoplast system. Plant Sci. Letters 3, 127–133 (1974)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • Glimelius, K., Wallin, A., Eriksson, T.: Agglutinating effects of concanavalin A on isolated protoplasts of Daucus carota. Physiol. Plant. 31, 225–230 (1974)

    Google Scholar 

  • Gregory, D.W., Cocking, E.C.: The large-scale isolation of protoplasts from immature tomato fruit. J. Cell Biol 24, 143–146 (1965)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • Kameya, T., Uchimiya, H.: Embryoids derived from isolated protoplasts of carrot. Planta (Berl.) 103, 356–360 (1972)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • Pelcher, L.E., Gamborg, O.L., Kao, K.N.: Bean mesophyll protoplasts: production, culture and callus formation. Plant Sci. Letters 3, 107–111 (1974)

    Google Scholar 

  • Power, J.B., Cocking, E.C.: Isolation of leaf protoplasts: macromolecule uptake and growth substance response. J. exp. Bot. 21, 64–70 (1970)

    Google Scholar 

  • Raj, B., Herr, J.M.: Isolation of protoplasts from the placental cells of Lycopersicum pimpinellifolium Mill. Exp. Cell Res. 64, 479–480 (1970)

    Google Scholar 

  • 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)

    Google Scholar 

  • Shepard, J.F., Totten, R.E.: Isolation and regeneration of tobacco mesophyll cell protoplasts under low osmotic conditions. Plant Physiol. 55, 689–694 (1975)

    Google Scholar 

  • 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)

    Google Scholar 

  • Thorsby, E., Bratlie, A.: A rapid method for preparation of pure lymphocyte suspensions. Histocompat. Testing pp. 655–656 Copenhagen: Munksgaard 1970

    Google Scholar 

  • Ting, A., Morris, P.J.: A technique for lymphocyte preparation from stored heparinized blood. Vox Sang. (Basel) 20, 561–563 (1971)

    Google Scholar 

  • 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)

    Google Scholar 

  • Widholm, J.M.: The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol. 47, 189–194 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints 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

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00393231

Keywords

Navigation