Abstract
Mesophyll protoplasts of tobacco (Nicotiana tabacum L. cv. Xanthi) were evacuolated by centrifugation in a density gradient. Evacuolation resulted in the quantitative loss of vacuolar hydrolytic activities. The evacuolated miniprotoplasts were cultivated under different conditions, and the regeneration of the central vacuole was investigated by light and electron microscopy as well as by the determination of activities of vacuolar marker enzymes. Vacuoles and hydrolytic activities, as well as cell wall material reappeared faster when the cells were cultivated at low osmotic strength. A newly synthesized tonoplast polypeptide could be detected using a polyspecific serum raised against tonoplast proteins of barley (Hordeum vulgare L.). Both vacuolar proton pumps, the ATPase as well as the pyrophosphatase appear to be newly synthesized during the regeneration of the vacuole.
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Abbreviations
- GAP-DH:
-
NADP-dependent glyceraldehyde 3-phosphate dehydrogenase
- PEP:
-
phosphoenolpyruvate
- PPi:
-
pyrophosphate
- PPase:
-
pyrophosphatase
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We thank Dr. Ernst Wehrli, Labor für Elektronenmikroskopie I, ETH Zürich, for taking micrographs. Esther Vogt assisted in the determination of the hydrolases. Bafilomycin was kindly provided by Professor Altendorf, Osnabrück FRG. This work was supported by the Swiss National Foundation grant No. 31-25196.88.
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Hörtensteiner, S., Martinoia, E. & Amrhein, N. Reappearance of hydrolytic activities and tonoplast proteins in the regenerated vacuole of evacuolated protoplasts. Planta 187, 113–121 (1992). https://doi.org/10.1007/BF00201632
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DOI: https://doi.org/10.1007/BF00201632