Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1994 Jun;105(2):659–670. doi: 10.1104/pp.105.2.659

Uridine Diphosphate Glucose Metabolism and Callose Synthesis in Cultured Pollen Tubes of Nicotiana alata Link et Otto.

H Schlupmann 1, A Bacic 1, S M Read 1
PMCID: PMC159407  PMID: 12232233

Abstract

Membrane preparations from cultured pollen tubes of Nicotiana alata Link et Otto contain a Ca2+ -independent (1-3)-[beta]-D-glucan (callose) synthase activity that has a low affinity for UDP-glucose, even when activated by treatment with trypsin (H. Schlupmann, A. Basic, S.M. Read [1993] Planta 191: 470-481). Therefore, we investigated whether UDP-glucose was a likely substrate for callose synthesis in actively growing pollen tubes. Deposition of (1-3)-[beta]-glucan occurred at a constant rate, 1.4 to 1.7 nmol glucose min-1, in tubes from 1 mg of pollen from 3 h after germination; however, the rate of incorporation of radioactivity from exogenous [14C]-sucrose into wall polymers was not constant, but increased until at least 8 h after germination, probably due to decreasing use of internal reserves. UDP-glucose was a prominent ultraviolet-absorbing metabolite in pollen-tube extracts, with 1.6 nmol present in tubes from 1 mg of pollen, giving a calculated cytoplasmic concentration of approximately 3.5 mM. Radioactivity from [14C]-sucrose was rapidly incorporated into sugar monophosphates and UDP-glucose by the growing tubes, consistent with a turnover time for UDP-glucose of less than 1 min; the specific radioactivity of extracted UDP-[14C]glucose was equal to that calculated from the rate of incorporation of [14C]sucrose into wall glucans. Large amounts of less metabolically active neutral sugars were also present. The rate of synthesis of (1-3)-[beta]-glucan by nontrypsin-treated pollen-tube membrane preparations incubated with 3.5 mM UDP-glucose and a [beta]-glucoside activator was slightly greater than the rate of deposition of (1-3)-[beta]-glucan by intact pollen tubes. These data are used to assess the physiological significance of proteolytic activation of pollen-tube callose synthase.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

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

  1. Chen M., Loewus F. A. myo-Inositol Metabolism in Lilium longiflorum Pollen: Uptake and Incorporation of myo-Inositol-2-H. Plant Physiol. 1977 Apr;59(4):653–657. doi: 10.1104/pp.59.4.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dancer J., Neuhaus H. E., Stitt M. Subcellular compartmentation of uridine nucleotides and nucleoside-5' -diphosphate kinase in leaves. Plant Physiol. 1990 Mar;92(3):637–641. doi: 10.1104/pp.92.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Deshusses J., Gumber S. C., Loewus F. A. Sugar Uptake in Lily Pollen : A PROTON SYMPORT. Plant Physiol. 1981 Apr;67(4):793–796. doi: 10.1104/pp.67.4.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Duran A., Cabib E. Solubilization and partial purification of yeast chitin synthetase. Confirmation of the zymogenic nature of the enzyme. J Biol Chem. 1978 Jun 25;253(12):4419–4425. [PubMed] [Google Scholar]
  5. Fry S. C., Northcote D. H. Sugar-nucleotide precursors of arabinopyranosyl, arabinofuranosyl, and xylopyranosyl residues in spinach polysaccharides. Plant Physiol. 1983 Dec;73(4):1055–1061. doi: 10.1104/pp.73.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hayashi T., Read S. M., Bussell J., Thelen M., Lin F. C., Brown R. M., Delmer D. P. UDP-Glucose: (1-->3)-beta-Glucan Synthases from Mung Bean and Cotton: Differential Effects of Ca and Mg on Enzyme Properties and on Macromolecular Structure of the Glucan Product. Plant Physiol. 1987 Apr;83(4):1054–1062. doi: 10.1104/pp.83.4.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lee Y. C. High-performance anion-exchange chromatography for carbohydrate analysis. Anal Biochem. 1990 Sep;189(2):151–162. doi: 10.1016/0003-2697(90)90099-u. [DOI] [PubMed] [Google Scholar]
  8. Li L., Brown R. M., Jr [beta]-Glucan Synthesis in the Cotton Fiber (II. Regulation and Kinetic Properties of [beta]-Glucan Synthases. Plant Physiol. 1993 Apr;101(4):1143–1148. doi: 10.1104/pp.101.4.1143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Meinert M. C., Delmer D. P. Changes in biochemical composition of the cell wall of the cotton fiber during development. Plant Physiol. 1977 Jun;59(6):1088–1097. doi: 10.1104/pp.59.6.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Meyer R., Wagner K. G. Determination of nucleotide pools in plant tissue by high-performance liquid chromatography. Anal Biochem. 1985 Aug 1;148(2):269–276. doi: 10.1016/0003-2697(85)90228-3. [DOI] [PubMed] [Google Scholar]
  11. Ohana P., Benziman M., Delmer D. P. Stimulation of Callose Synthesis in Vivo Correlates with Changes in Intracellular Distribution of the Callose Synthase Activator [beta]-Furfuryl-[beta]-Glucoside. Plant Physiol. 1993 Jan;101(1):187–191. doi: 10.1104/pp.101.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Olempska-Beer Z., Freese E. B. Optimal extraction conditions for high-performance liquid chromatographic determination of nucleotides in yeast. Anal Biochem. 1984 Jul;140(1):236–245. doi: 10.1016/0003-2697(84)90159-3. [DOI] [PubMed] [Google Scholar]
  13. Shaw J. A., Mol P. C., Bowers B., Silverman S. J., Valdivieso M. H., Durán A., Cabib E. The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. J Cell Biol. 1991 Jul;114(1):111–123. doi: 10.1083/jcb.114.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ulane R. E., Cabib E. The activating system of chitin synthetase from Saccharomyces cerevisiae. Purification and properties of an inhibitor of the activating factor. J Biol Chem. 1974 Jun 10;249(11):3418–3422. [PubMed] [Google Scholar]

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

RESOURCES