Seasonal Changes of Plasma Membrane H+-ATPase and Endogenous Ion Current during Cambial Growth in Poplar Plants

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Abstract
The plasma membrane H+-ATPase (PM H+-ATPase), potassium ions, and endogenous ion currents might play a fundamental role in the physiology of cambial growth. Seasonal changes of these parameters were studied in twigs ofPopulus nigra and Populus trichocarpa. Monoclonal and polyclonal antibodies against the PM H+-ATPase, x-ray analysis for K+ localization and a vibrating electrode for measurement of endogenous ion currents were used as probes. In dormant plants during autumn and winter, only a slight immunoreactivity against the PM H+-ATPase was found in cross sections and tissue homogenates, K+ was distributed evenly, and the density of endogenous current was low. In spring during cambial growth, strong immunoreactivity against a PM H+-ATPase was observed in cambial cells and expanding xylem cells using the monoclonal antibody 46 E5 B11 F6 for fluorescence microscopy and transmission electron microscopy. At the same time, K+ accumulated in cells of the cambial region, and strong endogenous current was measured in the cambial and immature xylem zone. Addition of auxin to dormant twigs induced the formation of this PM H+-ATPase in the dormant cambial region within a few days and an increase in density of endogenous current in shoot cuttings within a few hours. The increase in PM H+-ATPase abundance and in current density by auxin indicates that auxin mediates a rise in number and activity of an H+-ATPase in the plasma membrane of cambial cells and their derivatives. This PM H+-ATPase generates the necessary H+-gradient (proton-motive force) for the uptake of K+ and nutrients into cambial and expanding xylem cells.