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Plant Physiology 69:1315-1319 (1982)
© 1982 American Society of Plant Biologists

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Articles

Vacuoles from Sugarcane Suspension Cultures 1

I. ISOLATION AND PARTIAL CHARACTERIZATION

Margaret Thom, Andrew Maretzki and Ewald Komor2

Experiment Station, Hawaiian Sugar Planters' Association, Aiea, Hawaii 96701

Vacuoles were isolated from suspension cultures of sugarcane (Saccharum sp.) cells by centrifugation of protoplasts at high g force against a 12% (w/v) Ficoll solution. Distribution of marker enzymes and Concanavalin A binding showed an 11% contamination of the vacuole preparation by cytoplasmic components, mitochondria, and endoplasmic reticulum, and 18% contamination by plasma membrane. Acid phosphatase, carboxypeptidase, protease, peroxidase, and ribonuclease activities were enriched in isolated vacuoles. Carboxypeptidase was tonoplast-bound, whereas the other enzymes were soluble. Sucrose, reducing sugars, and free amino acids were measured in protoplasts and vacuoles during growth of cells in suspension culture. Sucrose and reducing sugar content of vacuoles increased as the culture aged, while free amino acids decreased sharply.

2 Permanent address: Botanisches Institut der Universität Bayreuth, Bayreuth, West Germany.

1 This work was supported in part by a grant from the Deutsche Forschungsgemeinschaft (to E. K.) and by a grant from the Governor's Agriculture Coordinating Committee, State of Hawaii (to A. M.). Published with the approval of the Director as Paper No. 519 in the Journal Series of the Experiment Station, Hawaiian Sugar Planters' Association.






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Copyright © 1982 by the American Society of Plant Biologists