Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1992 Oct;100(2):756–761. doi: 10.1104/pp.100.2.756

Purification of Maize Pollen Exines and Analysis of Associated Proteins 1

Christopher H Chay 1, Erich G Buehler 1, Judith M Thorn 1, Thomas M Whelan 1, Patricia A Bedinger 1
PMCID: PMC1075623  PMID: 16653056

Abstract

Zea mays (maize) pollen exines have been purified with the use of differential centrifugation and sucrose gradients, followed by mild detergent and high salt treatment. The final exine fraction is highly purified from other organelles and subcellular structures as assayed by transmission electron microscopy. Using mature maize pollen as the starting material, 0.2 to 0.3% of the total pollen protein remained associated with the exine fraction throughout the purification. Seven abundant sodium dodecyl sulfate-extractable proteins are detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the final fraction. Amino acid analysis reveals that one of the proteins contains a substantial amount of hydroxyproline, a characteristic of some primary cell wall proteins. The amino acid composition of the 25-kD protein strongly implies that it is an arabinogalactan protein. When exines are purified from earlier pollen developmental stages, a subset of the proteins found in the mature pollen exine is seen.

Full text

PDF
756

Images in this article

758Figure 2
on p.758
759Figure 3
on p.759
759Figure 4
on p.759
760Figure 5
on p.760

Selected References

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

  1. Bedinger P. A., Edgerton M. D. Developmental staging of maize microspores reveals a transition in developing microspore proteins. Plant Physiol. 1990 Feb;92(2):474–479. doi: 10.1104/pp.92.2.474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bohlmann H., Clausen S., Behnke S., Giese H., Hiller C., Reimann-Philipp U., Schrader G., Barkholt V., Apel K. Leaf-specific thionins of barley-a novel class of cell wall proteins toxic to plant-pathogenic fungi and possibly involved in the defence mechanism of plants. EMBO J. 1988 Jun;7(6):1559–1565. doi: 10.1002/j.1460-2075.1988.tb02980.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Drózdz M., Kucharz E., Szyja J. A colorimetric micromethod for determination of hydroxyproline in blood serum. Z Med Labortech. 1976 Aug 4;17(4):163–171. [PubMed] [Google Scholar]
  4. Gleeson P. A., McNamara M., Wettenhall R. E., Stone B. A., Fincher G. B. Characterization of the hydroxyproline-rich protein core of an arabinogalactan-protein secreted from suspension-cultured Lolium multiflorum (Italian ryegrass) endosperm cells. Biochem J. 1989 Dec 15;264(3):857–862. doi: 10.1042/bj2640857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Guilford W. J., Schneider D. M., Labovitz J., Opella S. J. High resolution solid state C NMR spectroscopy of sporopollenins from different plant taxa. Plant Physiol. 1988 Jan;86(1):134–136. doi: 10.1104/pp.86.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hood E. E., Shen Q. X., Varner J. E. A developmentally regulated hydroxyproline-rich glycoprotein in maize pericarp cell walls. Plant Physiol. 1988 May;87(1):138–142. doi: 10.1104/pp.87.1.138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kieliszewski M. J., Leykam J. F., Lamport D. T. Structure of the Threonine-Rich Extensin from Zea mays. Plant Physiol. 1990 Feb;92(2):316–326. doi: 10.1104/pp.92.2.316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kieliszewski M., Lamport D. T. Purification and Partial Characterization of a Hydroxyproline-Rich Glycoprotein in a Graminaceous Monocot, Zea mays. Plant Physiol. 1987 Nov;85(3):823–827. doi: 10.1104/pp.85.3.823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  10. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  11. Sheldon J. M., Dickinson H. G. Determination of patterning in the pollen wall of Lilium henryi. J Cell Sci. 1983 Sep;63:191–208. doi: 10.1242/jcs.63.1.191. [DOI] [PubMed] [Google Scholar]
  12. Tsumuraya Y., Ogura K., Hashimoto Y., Mukoyama H., Yamamoto S. Arabinogalactan-Proteins from Primary and Mature Roots of Radish (Raphanus sativus L.). Plant Physiol. 1988 Jan;86(1):155–160. doi: 10.1104/pp.86.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES