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Plant and Cell Physiology, 2000, Vol. 41, No. 7 857-863
© 2000 Oxford University Press

Demonstration of Cl– Requirement for Inhibition of Vacuolar Acidification by Cycloprodigiosin in Situ

Tsuyoshi Nakayasu, Keiko Kawauchi, Hajime Hirata and Teruo Shimmen1

Department of Life Science, Faculty of Science, Himeji Institute of Technology, Harima Science Park City, Hyogo, 678-1297 Japan

Applying vacuole-perfusion and plasma membrane per-meabilization techniques to internodal cells of Chara, we analyzed the requirement of Cl– for the action of cycloprodigiosin (cPrG) to inhibit vacuole acidification in situ. By combining the two techniques, the Cl– concentration on both sides of the tonoplast could be controlled. In permeabilized cell fragments lacking Cl– in the vacuole, the inhibitory effect of cPrG on vacuole acidification was cancelled. On the other hand, Cl– in the cytoplasm was not needed for the cPrG action. These results supported the function of cPrG as a H+/Cl– symporter. Requirement of Cl– for the cPrG action was also demonstrated in vacuole-perfused living cells. This is the first report on the mechanism of cPrG action in situ.

1 Corresponding author: E-mail, shimmen@sci.himeji-tech.ac.jp; Fax, +81-791-58-0175.


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