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Journal of Experimental Botany, Vol. 51, No. 351, pp. 1671-1677, October 2000
© 2000 Oxford University Press

Original Papers

A plasma membrane-enriched fraction isolated from the coats of developing pea seeds contains H+-symporters for amino acids and sucrose

A. de Jong1 and A.C. Borstlap2

Transport Physiology Research Group, Department of Plant Ecology and Evolutionary Biology, Utrecht University, Sorbonnelaan 16, NL-3584 CA Utrecht, The Netherlands

Aqueous polymer two-phase partitioning was used to obtain a plasma membrane-enriched fraction from coats of developing pea (Pisum sativum L.) seeds in the filling stage. Uptake of amino acids and sucrose by vesicles from this fraction was determined after imposition of gradients of proton concentration ({Delta}pH, inside alkaline) and electrical potential ({Delta}{psi}, inside negative) across the vesicle membrane. The uptake of sucrose and the amino acids L-valine, L-lysine, and L-glutamic acid was stimulated by the imposition of {Delta}pH. The imposition of {Delta}{psi}, either in the presence or in the absence of {Delta}pH, stimulated the uptake of L-valine and L-lysine, but had no detectable effect on the uptake of sucrose and L-glutamic acid. The proton-motive-force-driven uptake of all four substrates was abolished by the protonophore carbonylcyanide m-chlorophenyl-hydrazone (CCCP). The results demonstrate the presence of H+-symporters for sucrose and amino acids in pea seed coats. This is running counter to the previously reported finding that their uptake by isolated pea seed coats was insensitive to CCCP, and that the uptake of sucrose, L-valine, and L-glutamic acid displayed linear kinetics. Possible causes of this discrepancy will be discussed.

Key words: Amino acids, plasma membrane, proton symport, seed coat, sucrose.


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