Planta

, Volume 198, Issue 3, pp 356–364 | Cite as

An analysis of vacuole development in oat aleurone protoplasts

  • T. G. Emyr Davies
  • Susan H. Steele
  • David J. Walker
  • Roger A. Leigh
Article

Abstract

Cultured oat (Avena sativa L. — naked form) aleurone protoplasts were employed as a model system for following changes which accompany the development of vacuoles during in-vitro incubation. Over a 5-d period, the aleurone grains progressively grew and fused to form a large central vacuole and the volume of the protoplasts increased sevenfold. The growth of the vacuole was accompanied by a progressive acidification of the vacuolar sap. Vacuolation was inhibited by high concentrations of mannitol and by cycloheximide and cordycepin applied at various times during the incubation period. Neither cycloheximide nor cordycepin affected the initial phases of vacuolation but cycloheximide retarded subsequent stages, particularly if added early in the incubation. Cordycepin inhibited only the later stages of vacuolation. Radiolabelling studies identified at least three novel microsomal proteins, with relative molecular masses of approximately 34, 47 and 48 kDa, which appeared during vacuolation and whose synthesis was markedly affected by these inhibitors.

Key words

Avena (aleurone) Membrane protein (microsomal) Protoplast Tonoplast intrinsic Protein Vacuole 

Abbreviations

CF

carboxyfluorescein

CFDA

6 carboxyfluorescein diacetate

TIP

tonoplast intrinsic protein

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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • T. G. Emyr Davies
    • 1
  • Susan H. Steele
    • 1
  • David J. Walker
    • 1
  • Roger A. Leigh
    • 1
  1. 1.Biochemistry and Physiology DepartmentIACR-RothamstedHertsUK

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