Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1985 Apr;77(4):963–968. doi: 10.1104/pp.77.4.963

Translocation of Photosynthates into Vacuoles in Spinach Leaf Protoplasts

Sumio Asami 1,1, Ikuko Hara-Nishimura 1, Mikio Nishimura 1, Takashi Akazawa 1
PMCID: PMC1064641  PMID: 16664172

Abstract

A method was developed for the isolation of vacuoles from the mesophyll protoplasts of spinach leaf, employing the discontinuous Ficoll density gradient centrifugation technique. Isolated vacuole preparations were judged to be free from other organellar fractions based on the assays of marker enzyme activities of individual organelles.

Using this isolation method, a time-dependent translocation of 14C-labeled photosynthates into vacuoles was determined. In contrast to a significant transport of 14C organic acids such as malate and citrate within 10 to 15 minutes 14C neutral sugars and amino acids were barely transported into vacuoles during 40 minutes incubation, in spite of the fact that a relatively large amount of these compounds are found in the vacuoles. It was also found that a majority of [14C]sucrose remains in the cytosol, apparently not actively moving into the vacuoles. Overall results appear to suggest that vacuoles are not actively engaged in photosynthetic carbon metabolism in spinach leaf protoplasts.

Full text

PDF
963

Images in this article

964Fig. 2
on p.964

Selected References

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

  1. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Hatch M. D. A simple spectrophotometric assay for fumarate hydratase in crude tissue extracts. Anal Biochem. 1978 Mar;85(1):271–275. doi: 10.1016/0003-2697(78)90299-3. [DOI] [PubMed] [Google Scholar]
  3. Jones M. G., Outlaw W. H., Lowry O. H. Enzymic assay of 10 to 10 moles of sucrose in plant tissues. Plant Physiol. 1977 Sep;60(3):379–383. doi: 10.1104/pp.60.3.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  5. Lin W., Wagner G. J., Siegelman H. W., Hind G. Membrane-bound ATPase of intact vacuoles and tonoplasts isolated from mature plant tissue. Biochim Biophys Acta. 1977 Feb 14;465(1):110–117. doi: 10.1016/0005-2736(77)90359-5. [DOI] [PubMed] [Google Scholar]
  6. Maruyama H., Easterday R. L., Chang H. C., Lane M. D. The enzymatic carboxylation of phosphoenolpyruvate. I. Purification and properties of phosphoenolpyruvate carboxylase. J Biol Chem. 1966 May 25;241(10):2405–2412. [PubMed] [Google Scholar]
  7. Nawa Y., Asahi T. Rapid Development of Mitochondria in Pea Cotyledons during the Early Stage of Germination. Plant Physiol. 1971 Dec;48(6):671–674. doi: 10.1104/pp.48.6.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nishimura M., Akazawa T. Photosynthetic activities of spinach leaf protoplasts. Plant Physiol. 1975 Apr;55(4):712–716. doi: 10.1104/pp.55.4.712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nishimura M., Beevers H. Hydrolases in vacuoles from castor bean endosperm. Plant Physiol. 1978 Jul;62(1):44–48. doi: 10.1104/pp.62.1.44. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nishimura M., Takabe T., Sugiyama T., Akazawa T. Structure and function of chloroplast proteins. XIX. Dissociation of spinach leaf ribulose-1,5-diphosphate carboxylase by p-mercuribenzoate. J Biochem. 1973 Nov;74(5):945–954. [PubMed] [Google Scholar]
  11. Nishimura M. pH in Vacuoles Isolated from Castor Bean Endosperm. Plant Physiol. 1982 Sep;70(3):742–744. doi: 10.1104/pp.70.3.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Thom M., Komor E., Maretzki A. Vacuoles from Sugarcane Suspension Cultures : II. CHARACTERIZATION OF SUGAR UPTAKE. Plant Physiol. 1982 Jun;69(6):1320–1325. doi: 10.1104/pp.69.6.1320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wagner G. J. Content and vacuole/extravacuole distribution of neutral sugars, free amino acids, and anthocyanin in protoplasts. Plant Physiol. 1979 Jul;64(1):88–93. doi: 10.1104/pp.64.1.88. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wittenbach V. A., Lin W., Hebert R. R. Vacuolar localization of proteases and degradation of chloroplasts in mesophyll protoplasts from senescing primary wheat leaves. Plant Physiol. 1982 Jan;69(1):98–102. doi: 10.1104/pp.69.1.98. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES