Changes in the expression pattern of the plasma membrane H+-ATPase in developing Ricinus communis cotyledons...

The plasma membrane (PM) H+-ATPase is thought to play a key role in generating the proton motive force used to drive the uptake and accumulation of solutes in plant cells. Changes in its expression pattern were studied in the Ricinus communis L. cotyledon as it changed from a sink to a source organ. Expression was monitored in 3-, 10- and 14-day-old cotyledons using an antibody to the maize PM H+-ATPase. The antibody labelled a 100-kDa protein in membrane fractions prepared from cotyledons and this protein occurred at higher levels in the PM-enriched fractions compared to those enriched in intracellular membranes. Immunostaining of tissue sections of 3-day-old Ricinus cotyledons (sinks) with this antibody demonstrated that the PM H+-ATPase was highly expressed in the lower epidermal cells and also in the vascular bundles, particularly the phloem. The high expression in the epidermis suggests that these cells may be important in the initial active uptake of solutes from the endosperm. A similar distribution was observed in the 10-day-old seedlings but, in addition, larger, more spherical cells (idioblasts) had developed in the lower and upper epidermal layers and these were also labelled. In 14-day-old seedlings the cotyledons are no longer reliant on nutrients from the endosperm (which has totally degraded) and they are functioning as source organs. This is reflected in a decrease in PM H+-ATPase expression in the lower epidermal cells, apart from idioblasts and stomatal guard cells. The latter were also observed in the upper epidermis. Expression remained high in the vascular bundles of 14-day-old seedlings with strong staining in the phloem.