Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1989 Apr;89(4):1049–1052. doi: 10.1104/pp.89.4.1049

Measurement of the Cytoplasmic and Vacuolar Buffer Capacities in Chara corallina1

Kazuhiko Takeshige 1, Masashi Tazawa 1
PMCID: PMC1055973  PMID: 16666662

Abstract

The cytoplasm and the vacuole were isolated from internodal cells of Chara corallina by using the intracellular perfusion technique, and their buffer capacities (βi) were determined from the titration curves. The pH of the isolated vacuolar sap was 5.19 ± 0.029 (mean ± standard error). At this pH, βi was minimal and amounted to 0.933 ± 0.11 millimoles H+/pH unit/liter vacuolar sap. The pH of isolated cytoplasm was 7.22 ± 0.028. βi was minimal in this pH region and amounted to 14.2 ± 0.80 millimoles H+/pH unit/liter cytoplasm. When 1% (volume/volume) Triton X-100 was added to the cytoplasmic solution to permeabilize the subcellular organelles, the cytoplasmic pH increased to 7.32 ± 0.026, where βi was 20.35 ± 2.66 millimoles H+/pH unit/liter cytoplasm. This shows that alkaline subcellular compartments exist in the cytoplasm and also that the cytoplasmic pH before adding Triton X-100 may represent the cytosolic pH. These data indicate that the pH values of the cytoplasm and the vacuole are regulated at the values where the βi values are minimal. This suggests that ATP- and inorganic pyrophosphate-dependent H+ pumps in the plasma membrane and the tonoplast could efficiently regulate the pH of both cytoplasm and vacuole in Chara internodal cells.

Full text

PDF
1051

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Kitasato H. The influence of H+ on the membrane potential and ion fluxes of Nitella. J Gen Physiol. 1968 Jul;52(1):60–87. doi: 10.1085/jgp.52.1.60. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Roos A., Boron W. F. Intracellular pH. Physiol Rev. 1981 Apr;61(2):296–434. doi: 10.1152/physrev.1981.61.2.296. [DOI] [PubMed] [Google Scholar]
  3. Sanders D., Slayman C. L. Control of intracellular pH. Predominant role of oxidative metabolism, not proton transport, in the eukaryotic microorganism Neurospora. J Gen Physiol. 1982 Sep;80(3):377–402. doi: 10.1085/jgp.80.3.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Takeshige K., Tazawa M., Hager A. Characterization of the H Translocating Adenosine Triphosphatase and Pyrophosphatase of Vacuolar Membranes Isolated by Means of a Perfusion Technique from Chara corallina. Plant Physiol. 1988 Apr;86(4):1168–1173. doi: 10.1104/pp.86.4.1168. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES