Alleviation of rapid, futile ammonium cycling at the plasma membrane by potassium reveals K+-sensitive and -insensitive components of NH4+ transport.
Futile plasma membrane cycling of ammonium (NH4+) is characteristic of low-affinity NH4+ transport, and has been proposed to be a critical factor in NH4+ toxicity. Using unidirectional flux analysis with the positron-emitting tracer 13N in intact seedlings of barley (Hordeum vulgare L.), it is shown that rapid, futile NH4+ cycling is alleviated by elevated K+ supply, and that low-affinity NH4+ transport is mediated by a K+-sensitive component, and by a second component that is independent of K+. At low external [K+] (0.1 mM), NH4+ influx (at an external [NH4+] of 10 mM) of 92 micromol g(-1) h(-1) was observed, with an efflux:influx ratio of 0.75, indicative of rapid, futile NH4+ cycling. Elevating K+ supply into the low-affinity K+ transport range (1.5-40 mM) reduced both influx and efflux of NH4+ by as much as 75%, and substantially reduced the efflux:influx ratio. The reduction of NH4+ fluxes was achieved rapidly upon exposure to elevated K+, within 1 min for influx and within 5 min for efflux. The channel inhibitor La3+ decreased high-capacity NH4+ influx only at low K+ concentrations, suggesting that the K+-sensitive component of NH4+ influx may be mediated by non-selective cation channels. Using respiratory measurements and current models of ion flux energetics, the energy cost of concomitant NH4+ and K+ transport at the root plasma membrane, and its consequences for plant growth are discussed. The study presents the first demonstration of the parallel operation of K+-sensitive and -insensitive NH4+ flux mechanisms in plants.
Szczerba
Mark W
MW
Department of Biological Sciences, University of Toronto, 1265 Military Trail, Toronto, Ontario, Canada M1C 1A4.
Britto
Dev T
DT
Balkos
Konstantine D
KD
Kronzucker
Herbert J
HJ
D002462:Cell Membrane / Q000378:metabolism*; D001467:Hordeum / Q000378:metabolism*; D017136:Ion Transport / Q000502:physiology; D007811:Lanthanum / Q000378:metabolism; D009590:Nitrogen Radioisotopes / Q000378:metabolism; D011188:Potassium / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism*; D036226:Seedlings / Q000378:metabolism*
D009590:Nitrogen Radioisotopes; D000644:Quaternary Ammonium Compounds; D007811:Lanthanum; D011188:Potassium