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Plant Physiology 77:963-968 (1985)
© 1985 American Society of Plant Biologists

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Articles

Translocation of Photosynthates into Vacuoles in Spinach Leaf Protoplasts

Sumio Asami1, Ikuko Hara-Nishimura, Mikio Nishimura and Takashi Akazawa

Research Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Chikusa, Nagoya 464, Japan

A method was developed for the isolation of vacuoles from the mesophyll protoplasts of spinach leaf, employing the discontinuous Ficoll density gradient centrifugation technique. Isolated vacuole preparations were judged to be free from other organellar fractions based on the assays of marker enzyme activities of individual organelles.

Using this isolation method, a time-dependent translocation of 14C-labeled photosynthates into vacuoles was determined. In contrast to a significant transport of 14C organic acids such as malate and citrate within 10 to 15 minutes 14C neutral sugars and amino acids were barely transported into vacuoles during 40 minutes incubation, in spite of the fact that a relatively large amount of these compounds are found in the vacuoles. It was also found that a majority of [14C]sucrose remains in the cytosol, apparently not actively moving into the vacuoles. Overall results appear to suggest that vacuoles are not actively engaged in photosynthetic carbon metabolism in spinach leaf protoplasts.

1 Present address: Central Research Laboratory, Suntory Brewery Co. Ltd., Mishima, Osaka, Japan.






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Copyright © 1985 by the American Society of Plant Biologists