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Journal of Cell Science, Vol 102, 133-139, Copyright © 1992 by Company of Biologists

Submitted on December 16, 1991
Accepted on February 3, 1992

Development of a probenecid-sensitive Lucifer Yellow transport system in vacuolating oat aleurone protoplasts

K. M. WRIGHT 1, T. G. E. DAVIES 2, S. H. STEELE 2, R. A. LEIGH 2, and K. J. OPARKA 1

1 Department of Cellular and Environmental Physiology, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK
2 Biochemistry and Physiology Department, AFRC Institute of Arable Crops Research, Rothamsted Experimental Station, Harpenden, Herts AL5 2JQ, UK

Oat aleurone protoplasts, maintained in liquid culture over a period of five days, have been shown to develop an uptake system capable of transporting the membrane-impermeant probe Lucifer Yellow CH (LYCH). The dye was completely excluded from the densely cytoplasmic, early developmental stages but its uptake increased exponentially after the protoplasts had been cultured for between 2 and 5 days. Culturing induced vacuolation and uptake of the dye was largely restricted to highly vacuolate protoplasts. No cytoplasmic staining was evident. In vacuolate protoplasts uptake was linear with time and saturated with increasing substrate concentration. Low temperature, and addition of the drug probenecid to the incubation medium, completely eliminated LYCH uptake. In contrast to unconjugated LYCH, LYCH-dextrans (Mr 10,000 and 40,000) were excluded from the protoplasts.

The data negate simple diffusion and fluid-phase endocytosis as possible candidates for dye uptake and suggest, instead, the development of highly co-ordinated membrane transport systems on both plasmalemma and tonoplast.

Key words: aleurone protoplasts, oat (Avena sativa), Lucifer Yellow transport, vacuolation

Submitted on December 16, 1991
Accepted on February 3, 1992




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© The Company of Biologists Ltd 1992