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. 1995 Aug;108(4):1679–1689. doi: 10.1104/pp.108.4.1679

The Use of Nonaqueous Fractionation to Assess the Ionic Composition of the Apoplast during Fruit Ripening.

A J MacDougall 1, R Parker 1, R R Selvendran 1
PMCID: PMC157550  PMID: 12228573

Abstract

We have examined the possibility that pectin solubilization and cell separation in fruit may be due to organic acids disrupting calcium bridges between pectic polysaccharides. With fruit from a wild tomato (Lycopersicon pimpinellifolium [Dunal]) we demonstrated the validity of a nonaqueous fractionation method to obtain reliable estimates of the ionic content of the apoplast. In unripe fruit no organic acids were associated with the cell wall, which contained 67% of the total calcium and 47% of the magnesium. In ripe fruit 4% of the malate, 10% of the citrate, and 15% of the oxalate were estimated to be in the cell wall, together with 84% of the calcium and 52% of the magnesium. In contrast to the cultivated tomato, we did not find a consistent decrease in the degree of methyl esterification between unripe and ripe fruit, and an overall average of 75% was observed. In the cell walls of ripe fruit the ratio of calcium:magnesium:organic acid:unesterified uronic acid, on the basis of charge, was 15:4:4:16. The use of a computer program to predict the proportions of different ionic species in complex mixtures suggested that in ripe fruit 70% of the unesterified uronic acid would be complexed with calcium. Our results show that organic acids do not accumulate in the cell wall sufficiently to disrupt calcium cross-linking, nor is the calcium removed from the wall into the cell. We therefore conclude that organic acids do not contribute to cell separation during the ripening of tomato fruit.

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Selected References

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  1. Boller T., Kende H. Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 1979 Jun;63(6):1123–1132. doi: 10.1104/pp.63.6.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cooper T. G., Beevers H. Mitochondria and glyoxysomes from castor bean endosperm. Enzyme constitutents and catalytic capacity. J Biol Chem. 1969 Jul 10;244(13):3507–3513. [PubMed] [Google Scholar]
  3. De Ruiter G. A., Schols H. A., Voragen A. G., Rombouts F. M. Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods. Anal Biochem. 1992 Nov 15;207(1):176–185. doi: 10.1016/0003-2697(92)90520-h. [DOI] [PubMed] [Google Scholar]
  4. Gerhardt R., Heldt H. W. Measurement of subcellular metabolite levels in leaves by fractionation of freeze-stopped material in nonaqueous media. Plant Physiol. 1984 Jul;75(3):542–547. doi: 10.1104/pp.75.3.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gordon A. H., Bacon J. S. Fractionation of cell-wall preparations from grass leaves by centrifuging in non-aqueous density gradients. Biochem J. 1981 Mar 1;193(3):765–771. doi: 10.1042/bj1930765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kim J. B., Carpita N. C. Changes in Esterification of the Uronic Acid Groups of Cell Wall Polysaccharides during Elongation of Maize Coleoptiles. Plant Physiol. 1992 Feb;98(2):646–653. doi: 10.1104/pp.98.2.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Koch J. L., Nevins D. J. Tomato fruit cell wall : I. Use of purified tomato polygalacturonase and pectinmethylesterase to identify developmental changes in pectins. Plant Physiol. 1989 Nov;91(3):816–822. doi: 10.1104/pp.91.3.816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Legge R. L., Cheng K. H., Lepock J. R., Thompson J. E. Differential effects of senescence on the molecular organization of membranes in ripening tomato fruit. Plant Physiol. 1986 Aug;81(4):954–959. doi: 10.1104/pp.81.4.954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lurie S., Ben-Arie R. Microsomal Membrane Changes during the Ripening of Apple Fruit. Plant Physiol. 1983 Nov;73(3):636–638. doi: 10.1104/pp.73.3.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Poovaiah B. W., Mizrahi Y., Dostal H. C., Cherry J. H., Leopold A. C. Water Permeability during Tomato Fruit Development in Normal and rin Nonripening Mutant. Plant Physiol. 1975 Dec;56(6):813–815. doi: 10.1104/pp.56.6.813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Redgwell R. J., Melton L. D., Brasch D. J. Cell Wall Dissolution in Ripening Kiwifruit (Actinidia deliciosa) : Solubilization of the Pectic Polymers. Plant Physiol. 1992 Jan;98(1):71–81. doi: 10.1104/pp.98.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Soboll S., Akerboom T. P., Schwenke W. D., Haase R., Sies H. Mitochondrial and cytosolic ATP/ADP ratios in isolated hepatocytes. A comparison of the digitonin method and the non-aqueous fractionation procedure. Biochem J. 1980 Dec 15;192(3):951–954. doi: 10.1042/bj1920951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Tieman D. M., Handa A. K. Reduction in Pectin Methylesterase Activity Modifies Tissue Integrity and Cation Levels in Ripening Tomato (Lycopersicon esculentum Mill.) Fruits. Plant Physiol. 1994 Oct;106(2):429–436. doi: 10.1104/pp.106.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tieman D. M., Harriman R. W., Ramamohan G., Handa A. K. An Antisense Pectin Methylesterase Gene Alters Pectin Chemistry and Soluble Solids in Tomato Fruit. Plant Cell. 1992 Jun;4(6):667–679. doi: 10.1105/tpc.4.6.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Van Der Wilden W., Chrispeels M. J. Characterization of the Isozymes of alpha-Mannosidase Located in the Cell Wall, Protein Bodies, and Endoplasmic Reticulum of Phaseolus vulgaris Cotyledons. Plant Physiol. 1983 Jan;71(1):82–87. doi: 10.1104/pp.71.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wu Q., Szakacs-Dobozi M., Hemmat M., Hrazdina G. Endopolygalacturonase in Apples (Malus domestica) and Its Expression during Fruit Ripening. Plant Physiol. 1993 May;102(1):219–225. doi: 10.1104/pp.102.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]

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