Evidence for active Phloem loading in the minor veins of sugar beet.

Phloem loading in source leaves of sugar beet (Beta vulgaris, L.) was studied to determine the extent of dependence on energy metabolism and the involvement of a carrier system. Dinitrophenol at a concentration of 4 mm uncoupled respiration, lowered source leaf ATP to approximately 40% of the level in the control leaf and inhibited translocation of exogenously supplied (14)C-sucrose to approximately 20% of the control. Dinitrophenol at a concentration of 8 mm inhibited rather than promoted CO(2) production, indicating a mechanism of inhibition other than uncoupling of respiration. The 8 mm dinitrophenol also reduced ATP to approximately 40% of the level in the control source leaf and reduced translocation of exogenous sucrose to approximately 10% of the control. Application of 4 mm ATP to an untreated source leaf promoted the translocation rate by approximately 80% over the control, while in leaves treated with 4 mm dinitrophenol, 4 mm ATP restored translocation to the control level. No recovery of translocation was observed when ATP was applied to leaves treated with 8 mm dinitrophenol. The results indicate an energy-requiring process for both phloem loading and translocation in the source leaf.Application of (14)C-sucrose solutions in a series of concentrations through the upper surface of a source leaf produced a biphasic isotherm for translocation out of the fed region. A similar dual isotherm was obtained for phloem loading with leaf discs floated on (14)C-sucrose solutions. The first and possibly the second phases were attributed to active, carrier-mediated accumulation in the minor vein phloem. Autoradiography of the tissue confirmed that most of the sucrose was localized in the minor veins. Data from uptake through the abraded surface of intact leaves, the most reliable method, were analyzed by the Hofstee method. Kinetic parameters, analogous to Km and V(max) of enzyme studies, were calculated to be: K(j) = 16 mm and J(max) = 70 mug C/min dm(2) or 490 nmoles sucrose/min.dm(2). Rates for phloem loading and translocation of exogenous sucrose are equal to or greater than those observed for compounds derived from photosynthetically fixed CO(2). The data indicate that a free space sucrose concentration in the region of the minor vein phloem of approximately 20 mm can support translocation at the rates commonly observed for photosynthetically produced sugars.