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PLANT PHYSIOLOGY , Vol 109, Issue 2 481-490, Copyright © 1995 by American Society of Plant Biologists
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WHOLE PLANT, ENVIRONMENTAL, AND STRESS PHYSIOLOGY |
Analysis of 13NH4+ Efflux in Spruce Roots (A Test Case for Phase Identification in Compartmental Analysis)
H. J. Kronzucker, M. Y. Siddiqi and ADM. Glass
Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4
13NH4+-efflux analyses were conducted with roots of intact Picea glauca
(Moench) Voss. seedlings at external NH4+ concentrations of 100 [mu]M and
1.5 mM. Three kinetically distinct phases were identified with half-lives
of exchange of approximately 2 s, 30 s, and 14 min. The presumed identities
of the subcellular compartments corresponding to these phases were
confirmed by several techniques, including pretreatment of roots (a) at
75[deg]C or with SDS, (b) with [alpha]-keto-glutarate or
L-methionine-DL-sulfoximine, (c) at elevated levels of Ca2+, and (d) at low
pH or with Al3+ at low pH. Treatments a and b selectively influenced phase
III without affecting phases I and II. Similarly, treatment c selectively
perturbed phase II, and treatment d affected phases II and III. Based on
these findings and the assumption of an in-series arrangement of root cell
compartments, it was concluded that phase III corresponded to the
cytoplasm, phase II corresponded to the Donnan free space, and phase I
corresponded to a film of solution adhering to the root surface.
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