HAY, JORDAN O.* and ROGER M. SPANSWICK. Department of Biological and Environmental Engineering, Cornell University, Ithaca, NY 14853, USA. - Rice aleurone protoplasts as a system to study sucrose transport.
During post-phloem unloading, sucrose enters the apoplast of the rice
endosperm after it is released from the cells of the nucellus.
Subsequent starch synthesis and accumulation in the endosperm requires
that sucrose be transported across the plasma membranes of starchy
endosperm cells or aleurone cells. We have isolated and purified
protoplasts from the aleurone layer of ten-day-old rice caryopses.
Caryopses were peeled to expose the aleurone layer. The middle
lamellas were digested with pectolyase and the aleurone cells released
by gentle sheering. Digestion of the cell walls with cellulase
isolated protoplasts that were then purified by Percoll
density-gradient centrifugation. Radiotracer and silicone oil
centrifugation techniques are being used to study the kinetics of
sucrose uptake by these protoplasts.
Key words: aleurone, carbon partitioning, rice, sucrose transport