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Plant Physiol, February 2002, Vol. 128, pp. 370-378

Voltage-Dependent Cation Channels Permeable to NH4+, K+, and Ca2+ in the Symbiosome Membrane of the Model Legume Lotus japonicus1

Daniel M. Roberts* and Stephen D. Tyerman2

Department of Biochemistry, Cellular and Molecular Biology, The University of Tennessee, Knoxville, Tennessee, 37996 (D.M.R.); and School of Biological Sciences, The Flinders University of South Australia, G.P.O. Box 2100, Adelaide, South Australia, 5001, Australia (S.D.T.)

The symbiosome of nitrogen fixing root nodules mediates metabolite exchange between endosymbiotic rhizobia bacteria and the legume host. In the present study, the ion currents of the symbiosome membrane of the model legume Lotus japonicus were analyzed by patch-clamp recording. Both excised and symbiosome-attached patches exhibited a large inward (toward the cytosolic side of the membrane) current that is activated in a time-dependent manner by negative (on the cytosolic side) potentials. Based on reversal potential determinations and recordings with the impermeant cation N-methyl-glucamine, this current shows a high permeability for monovalent cations with no apparent permeability for anions. The current also showed a finite Ca2+ permeability. However, the currents were predominantly carried by univalent cations with a slightly greater selectivity for NH4+ over K+. Increased Ca2+ concentration inhibited the current with a K0.5 for inhibition of 0.317 mM. The current showed strong rectification that is mediated by divalent cations (either Mg2+ or Ca2+). The influence of divalent cations is symmetrical in nature, because rectification can be exerted in either direction depending upon which side of the membrane has the highest concentration of divalent cations. However, based on observations with symbiosome-attached patches, the direction of the current in vivo is proposed to be toward the cytosol with cytosolic Mg2+ acting as the putative gating regulator. The findings suggest that L. japonicus possesses a voltage-dependent cation efflux channel that is capable of exporting fixed NH4+, and may also play an additional role in Ca2+ transport.

1 This work was supported by the U.S. Department of Agriculture National Research Initiative Competitive Grants Program (award no. 9703548) and by the National Science Foundation (grant no. MCB-9904978 to D.M.R.), by the Australian Research Council (to S.D.T.), and by a Career Development award to D.M.R. from the University of Tennessee Office of Research Administration.

2 Present address: Department of Horticulture, Viticulture and Oenology, Adelaide University, PMB #1 Glen Osmond, 5042 SA, Australia.

* Corresponding author; e-mail dobert2{at}utk.edu; fax 865- 974-6306.

© 2002 American Society of Plant Physiologists


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