Potassium physiology from Archean to Holocene: A higher-plant perspective.

In this paper, we discuss biological potassium acquisition and utilization processes over an evolutionary timescale, with emphasis on modern vascular plants. The quintessential osmotic and electrical functions of the K+ ion are shown to be intimately tied to K+-transport systems and membrane energization. Several prominent themes in plant K+-transport physiology are explored in greater detail, including: (1) channel mediated K+ acquisition by roots at low external [K+]; (2) K+ loading of root xylem elements by active transport; (3) variations on the theme of K+ efflux from root cells to the extracellular environment; (4) the veracity and utility of the "affinity" concept in relation to transport systems. We close with a discussion of the importance of plant-potassium relations to our human world, and current trends in potassium nutrition from farm to table.

Plasma-membrane electrical responses to salt and osmotic gradients contradict radiotracer kinetics, and reveal Na+-transport dynamics in rice (Oryza sativa L.).

MAIN CONCLUSION: A systematic analysis of NaCl-dependent, plasma-membrane depolarization (∆∆Ψ) in rice roots calls into question the current leading model of rapid membrane cycling of Na+ under salt stress. To investigate the character and mechanisms of Na+ influx into roots, Na+-dependent changes in plasma-membrane electrical potentials (∆∆Ψ) were measured in root cells of intact rice (Oryza sativa L., cv. Pokkali) seedlings. As external sodium concentrations ([Na+]ext) were increased in a step gradient from 0 to 100 mM, membrane potentials depolarized in a saturable manner, fitting a Michaelis-Menten model and contradicting the linear (non-saturating) models developed from radiotracer studies. Clear differences in saturation patterns were found between plants grown under low- and high-nutrient (LN and HN) conditions, with LN plants showing greater depolarization and higher affinity for Na+ (i.e., higher Vmax and lower Km) than HN plants. In addition, counterion effects on ∆∆Ψ were pronounced in LN plants (with ∆∆Ψ decreasing in the order: Cl- > SO42- > HPO 4 2- ), but not seen in HN plants. When effects of osmotic strength, Cl- influx, K+ efflux, and H+-ATPase activity on ∆∆Ψ were accounted for, resultant Km and Vmax values suggested that a single, dominant Na+-transport mechanism was operating under each nutritional condition, with Km values of 1.2 and 16 mM for LN and HN plants, respectively. Comparing saturating patterns of depolarization to linear patterns of 24Na+ radiotracer influx leads to the conclusion that electrophysiological and tracer methods do not report the same phenomena and that the current model of rapid transmembrane sodium cycling may require revision.

From aquaporin to ecosystem: Plants in the water cycle.

Vascular plants are major intermediaries in the global water cycle, and are highly adapted to both facilitate and resist water fluxes, such as during root uptake, translocation in the xylem, and transpiration by leaves. Here, we summarize the contributions to a Special Issue on water in the Journal of Plant Physiology, which cluster around the theme of control and facilitation of water movement in plants. We conclude with an editorial view of the need for plant physiologists to consider larger cultural issues surrounding water use, especially in terms of the increasing agricultural demand for water to produce animal feed, with its associated trophic nutritive losses and environmental damage.

Membrane fluxes, bypass flows, and sodium stress in rice: the influence of silicon.

Provision of silicon (Si) to roots of rice (Oryza sativa L.) can alleviate salt stress by blocking apoplastic, transpirational bypass flow of Na+ from root to shoot. However, little is known about how Si affects Na+ fluxes across cell membranes. Here, we measured radiotracer fluxes of 24Na+, plasma membrane depolarization, tissue ion accumulation, and transpirational bypass flow, to examine the influence of Si on Na+ transport patterns in hydroponically grown, salt-sensitive (cv. IR29) and salt-tolerant (cv. Pokkali) rice. Si increased growth and lowered [Na+] in shoots of both cultivars, with minor effects in roots; neither root nor shoot [K+] were affected. In IR29, Si lowered shoot [Na+] via a large reduction in bypass flow, while in Pokkali, where bypass flow was small and not affected by Si, this was achieved mainly via a growth dilution of shoot Na+. Si had no effect on unidirectional 24Na+ fluxes (influx and efflux), or on Na+-stimulated plasma-membrane depolarization, in either IR29 or Pokkali. We conclude that, while Si can reduce Na+ translocation via bypass flow in some (but not all) rice cultivars, it does not affect unidirectional Na+ transport or Na+ cycling in roots, either across root cell membranes or within the bulk root apoplast.

How Plant Root Exudates Shape the Nitrogen Cycle.

Although the global nitrogen (N) cycle is largely driven by soil microbes, plant root exudates can profoundly modify soil microbial communities and influence their N transformations. A detailed understanding is now beginning to emerge regarding the control that root exudates exert over two major soil N processes - nitrification and N2 fixation. We discuss recent breakthroughs in this area, including the identification of root exudates as nitrification inhibitors and as signaling compounds facilitating N-acquisition symbioses. We indicate gaps in current knowledge, including questions of how root exudates affect newly discovered microbial players and N-cycle components. A better understanding of these processes is urgent given the widespread inefficiencies in agricultural N use and their links to N pollution and climate change.

Nitrogen transformations in modern agriculture and the role of biological nitrification inhibition.

The nitrogen (N)-use efficiency of agricultural plants is notoriously poor. Globally, about 50% of the N fertilizer applied to cropping systems is not absorbed by plants, but lost to the environment as ammonia (NH3), nitrate (NO3-), and nitrous oxide (N2O, a greenhouse gas with 300 times the heat-trapping capacity of carbon dioxide), raising agricultural production costs and contributing to pollution and climate change. These losses are driven by volatilization of NH3 and by a matrix of nitrification and denitrification reactions catalysed by soil microorganisms (chiefly bacteria and archaea). Here, we discuss mitigation of the harmful and wasteful process of agricultural N loss via biological nitrification inhibitors (BNIs) exuded by plant roots. We examine key recent discoveries in the emerging field of BNI research, focusing on BNI compounds and their specificity and transport, and discuss prospects for their role in improving agriculture while reducing its environmental impact.

The nitrogen-potassium intersection: membranes, metabolism, and mechanism.

Nitrogen (N) and potassium (K) are the two most abundantly acquired mineral elements by plants, and their acquisition pathways interact in complex ways. Here, we review pivotal interactions with respect to root acquisition, storage, translocation and metabolism, between the K+ ion and the two major N sources, ammonium (NH4+ ) and nitrate (NO3- ). The intersections between N and K physiology are explored at a number of organizational levels, from molecular-genetic processes, to compartmentation, to whole plant physiology, and discussed in the context of both N-K cooperation and antagonism. Nutritional regulation and optimization of plant growth, yield, metabolism and water-use efficiency are also discussed.

The Role of Silicon in Higher Plants under Salinity and Drought Stress.

Although deemed a "non-essential" mineral nutrient, silicon (Si) is clearly beneficial to plant growth and development, particularly under stress conditions, including salinity and drought. Here, we review recent research on the physiological, biochemical, and molecular mechanisms underlying Si-induced alleviation of osmotic and ionic stresses associated with salinity and drought. We distinguish between changes observed in the apoplast (i.e., suberization, lignification, and silicification of the extracellular matrix; transpirational bypass flow of solutes and water), and those of the symplast (i.e., transmembrane transport of solutes and water; gene expression; oxidative stress; metabolism), and discuss these features in the context of Si biogeochemistry and bioavailability in agricultural soils, evaluating the prospect of using Si fertilization to increase crop yield and stress tolerance under salinity and drought conditions.

Nutrient constraints on terrestrial carbon fixation: The role of nitrogen.

Carbon dioxide (CO2) concentrations in the earth's atmosphere are projected to rise from current levels near 400ppm to over 700ppm by the end of the 21st century. Projections over this time frame must take into account the increases in total net primary production (NPP) expected from terrestrial plants, which result from elevated CO2 (eCO2) and have the potential to mitigate the impact of anthropogenic CO2 emissions. However, a growing body of evidence indicates that limitations in soil nutrients, particularly nitrogen (N), the soil nutrient most limiting to plant growth, may greatly constrain future carbon fixation. Here, we review recent studies about the relationships between soil N supply, plant N nutrition, and carbon fixation in higher plants under eCO2, highlighting key discoveries made in the field, particularly from free-air CO2 enrichment (FACE) technology, and relate these findings to physiological and ecological mechanisms.

Measurement of Differential Na(+) Efflux from Apical and Bulk Root Zones of Intact Barley and Arabidopsis Plants.

Rapid sodium cycling across the plasma membrane of root cells is widely thought to be associated with Na(+) toxicity in plants. However, the efflux component of this cycling is not well understood. Efflux of Na(+) from root cells is believed to be mediated by Salt Overly-Sensitive-1, although expression of this Na(+)/H(+) antiporter has been localized to the vascular tissue and root meristem. Here, we used a chambered cuvette system in which the distal root of intact salinized barley and Arabidopsis thaliana plants (wild-type and sos1) were isolated from the bulk of the root by a silicone-acrylic barrier, so that we could compare patterns of (24)Na(+) efflux in these two regions of root. In barley, steady-state release of (24)Na(+) was about four times higher from the distal root than from the bulk roots. In the distal root, (24)Na(+) release was pronouncedly decreased by elevated pH (9.2), while the bulk-root release was not significantly affected. In A. thaliana, tracer efflux was about three times higher from the wild-type distal root than from the wild-type bulk root and also three to four times higher than both distal- and bulk-root fluxes of Atsos1 mutants. Elevated pH also greatly reduced the efflux from wild-type roots. These findings support a significant role of SOS1-mediated Na(+) efflux in the distal root, but not in the bulk root.

How high do ion fluxes go? A re-evaluation of the two-mechanism model of K(+) transport in plant roots.

Potassium (K(+)) acquisition in roots is generally described by a two-mechanism model, consisting of a saturable, high-affinity transport system (HATS) operating via H(+)/K(+) symport at low (<1mM) external [K(+)] ([K(+)]ext), and a linear, low-affinity system (LATS) operating via ion channels at high (>1mM) [K(+)]ext. Radiotracer measurements in the LATS range indicate that the linear rise in influx continues well beyond nutritionally relevant concentrations (>10mM), suggesting K(+) transport may be pushed to extraordinary, and seemingly limitless, capacity. Here, we assess this rise, asking whether LATS measurements faithfully report transmembrane fluxes. Using (42)K(+)-isotope and electrophysiological methods in barley, we show that this flux is part of a K(+)-transport cycle through the apoplast, and masks a genuine plasma-membrane influx that displays Michaelis-Menten kinetics. Rapid apoplastic cycling of K(+) is corroborated by an absence of transmembrane (42)K(+) efflux above 1mM, and by the efflux kinetics of PTS, an apoplastic tracer. A linear apoplastic influx, masking a saturating transmembrane influx, was also found in Arabidopsis mutants lacking the K(+) transporters AtHAK5 and AtAKT1. Our work significantly revises the model of K(+) transport by demonstrating a surprisingly modest upper limit for plasma-membrane influx, and offers insight into sodium transport under salt stress.

Measuring fluxes of mineral nutrients and toxicants in plants with radioactive tracers.

Unidirectional influx and efflux of nutrients and toxicants, and their resultant net fluxes, are central to the nutrition and toxicology of plants. Radioisotope tracing is a major technique used to measure such fluxes, both within plants, and between plants and their environments. Flux data obtained with radiotracer protocols can help elucidate the capacity, mechanism, regulation, and energetics of transport systems for specific mineral nutrients or toxicants, and can provide insight into compartmentation and turnover rates of subcellular mineral and metabolite pools. Here, we describe two major radioisotope protocols used in plant biology: direct influx (DI) and compartmental analysis by tracer efflux (CATE). We focus on flux measurement of potassium (K(+)) as a nutrient, and ammonia/ammonium (NH3/NH4(+)) as a toxicant, in intact seedlings of the model species barley (Hordeum vulgare L.). These protocols can be readily adapted to other experimental systems (e.g., different species, excised plant material, and other nutrients/toxicants). Advantages and limitations of these protocols are discussed.

The physiology of channel-mediated K+ acquisition in roots of higher plants.

K(+) channels are among the best-characterized classes of membrane protein in plants. Nevertheless, in-planta demonstrations of traits emerging from molecular characterizations have often been insufficient or lacking altogether. Such linkages are, however, critical to our basic understanding of plant nutrition and to addressing 'real-world' issues that are faced in environmental and agricultural settings. Here, we cover some of the recent advances in K(+) acquisition with particular focus on voltage-gated K(+) channel functioning and regulation in roots, and highlight where linkages to in-planta behavior have been successfully made and, conversely, where such linkages are yet to be made.

Rapid ammonia gas transport accounts for futile transmembrane cycling under NH3/NH4+ toxicity in plant roots.

Futile transmembrane NH3/NH4(+) cycling in plant root cells, characterized by extremely rapid fluxes and high efflux to influx ratios, has been successfully linked to NH3/NH4(+) toxicity. Surprisingly, the fundamental question of which species of the conjugate pair (NH3 or NH4(+)) participates in such fluxes is unresolved. Using flux analyses with the short-lived radioisotope (13)N and electrophysiological, respiratory, and histochemical measurements, we show that futile cycling in roots of barley (Hordeum vulgare) seedlings is predominately of the gaseous NH3 species, rather than the NH4(+) ion. Influx of (13)NH3/(13)NH4(+), which exceeded 200 µmol g(-1) h(-1), was not commensurate with membrane depolarization or increases in root respiration, suggesting electroneutral NH3 transport. Influx followed Michaelis-Menten kinetics for NH3 (but not NH4(+)), as a function of external concentration (Km = 152 µm, Vmax = 205 µmol g(-1) h(-1)). Efflux of (13)NH3/(13)NH4(+) responded with a nearly identical Km. Pharmacological characterization of influx and efflux suggests mediation by aquaporins. Our study fundamentally revises the futile-cycling model by demonstrating that NH3 is the major permeating species across both plasmalemma and tonoplast of root cells under toxicity conditions.

Ecological significance and complexity of N-source preference in plants.

BACKGROUND: Plants can utilize two major forms of inorganic N: NO3(-) (nitrate) and NH4(+) (ammonium). In some cases, the preference of one form over another (denoted as ß) can appear to be quite pronounced for a plant species, and can be an important determinant and predictor of its distribution and interactions with other species. In many other cases, however, assignment of preference is not so straightforward and must take into account a wide array of complex physiological and environmental features, which interact in ways that are still not well understood. SCOPE: This Viewpoint presents a discussion of the key, and often co-occurring, factors that join to produce the complex phenotypic composite referred to by the deceptively simple term 'N-source preference'. CONCLUSIONS: N-source preference is much more complex a biological phenomenon than is often assumed, and general models predicting how it will influence ecological processes will need to be much more sophisticated than those that have been so far developed.

Capacity and plasticity of potassium channels and high-affinity transporters in roots of barley and Arabidopsis.

The role of potassium (K(+)) transporters in high- and low-affinity K(+) uptake was examined in roots of intact barley (Hordeum vulgare) and Arabidopsis (Arabidopsis thaliana) plants by use of (42)K radiotracing, electrophysiology, pharmacology, and mutant analysis. Comparisons were made between results from barley and five genotypes of Arabidopsis, including single and double knockout mutants for the high-affinity transporter, AtHAK5, and the Shaker-type channel, AtAKT1. In Arabidopsis, steady-state K(+) influx at low external K(+) concentration ([K(+)]ext = 22.5 µm) was predominantly mediated by AtAKT1 when high-affinity transport was inhibited by ammonium, whereas in barley, by contrast, K(+) channels could not operate below 100 µm. Withdrawal of ammonium resulted in an immediate and dramatic stimulation of K(+) influx in barley, indicating a shift from active to passive K(+) uptake at low [K(+)]ext and yielding fluxes as high as 36 µmol g (root fresh weight)(-1) h(-1) at 5 mm [K(+)]ext, among the highest transporter-mediated K(+) fluxes hitherto reported. This ammonium-withdrawal effect was also established in all Arabidopsis lines (the wild types, atakt1, athak5, and athak5 atakt1) at low [K(+)]ext, revealing the concerted involvement of several transport systems. The ammonium-withdrawal effect coincided with a suppression of K(+) efflux and a significant hyperpolarization of the plasma membrane in all genotypes except athak5 atakt1, could be sustained over 24 h, and resulted in increased tissue K(+) accumulation. We discuss key differences and similarities in K(+) acquisition between two important model systems and reveal novel aspects of K(+) transport in planta.

K+ efflux and retention in response to NaCl stress do not predict salt tolerance in contrasting genotypes of rice (Oryza sativa L.).

Sudden elevations in external sodium chloride (NaCl) accelerate potassium (K(+)) efflux across the plasma membrane of plant root cells. It has been proposed that the extent of this acceleration can predict salt tolerance among contrasting cultivars. However, this proposal has not been considered in the context of plant nutritional history, nor has it been explored in rice (Oryza sativa L.), which stands among the world's most important and salt-sensitive crop species. Using efflux analysis with (42)K, coupled with growth and tissue K(+) analyses, we examined the short- and long-term effects of NaCl exposure to plant performance within a nutritional matrix that significantly altered tissue-K(+) set points in three rice cultivars that differ in salt tolerance: IR29 (sensitive), IR72 (moderate), and Pokkali (tolerant). We show that total short-term K(+) release from roots in response to NaCl stress is small (no more than 26% over 45 min) in rice. Despite strong varietal differences, the extent of efflux is shown to be a poor predictor of plant performance on long-term NaCl stress. In fact, no measure of K(+) status was found to correlate with plant performance among cultivars either in the presence or absence of NaCl stress. By contrast, shoot Na(+) accumulation showed the strongest correlation (a negative one) with biomass, under long-term salinity. Pharmacological evidence suggests that NaCl-induced K(+) efflux is a result of membrane disintegrity, possibly as result of osmotic shock, and not due to ion-channel mediation. Taken together, we conclude that, in rice, K(+) status (including efflux) is a poor predictor of salt tolerance and overall plant performance and, instead, shoot Na(+) accumulation is the key factor in performance decline on NaCl stress.

Isotope techniques to study kinetics of Na+ and K+ transport under salinity conditions.

Radioisotopes (particularly (22)Na, (24)Na, (42)K, and (86)Rb) have been used for many decades to trace the fluxes and accumulation of sodium and potassium ions in plant tissues. In this article, standard procedures for the tracing of ion fluxes are described, with emphasis on special problems encountered when examining K(+) and Na(+) transport under salinity conditions. We focus in particular on unidirectional influx measurements, while also providing a brief introduction to compartmental analysis by tracer efflux.

A pharmacological analysis of high-affinity sodium transport in barley (Hordeum vulgare L.): a 24Na+/42K+ study.

Soil sodium, while toxic to most plants at high concentrations, can be beneficial at low concentrations, particularly when potassium is limiting. However, little is known about Na(+) uptake in this 'high-affinity' range. New information is provided here with an insight into the transport characteristics, mechanism, and ecological significance of this phenomenon. High-affinity Na(+) and K(+) fluxes were investigated using the short-lived radiotracers (24)Na and (42)K, under an extensive range of measuring conditions (variations in external sodium, and in nutritional and pharmacological agents). This work was supported by electrophysiological, compartmental, and growth analyses. Na(+) uptake was extremely sensitive to all treatments, displaying properties of high-affinity K(+) transporters, K(+) channels, animal Na(+) channels, and non-selective cation channels. K(+), NH(4)(+), and Ca(2+) suppressed Na(+) transport biphasically, yielding IC(50) values of 30, 10, and <5 µM, respectively. Reciprocal experiments showed that K(+) influx is neither inhibited nor stimulated by Na(+). Sodium efflux constituted 65% of influx, indicating a futile cycle. The thermodynamic feasibility of passive channel mediation is supported by compartmentation and electrophysiological data. Our study complements recent advances in the molecular biology of high-affinity Na(+) transport by uncovering new physiological foundations for this transport phenomenon, while questioning its ecological relevance.

Silver ions disrupt K⁺ homeostasis and cellular integrity in intact barley (Hordeum vulgare L.) roots.

The heavy metals silver, gold, and mercury can strongly inhibit aquaporin-mediated water flow across plant cell membranes, but critical examinations of their side effects are rare. Here, the short-lived radiotracer (42)K is used to demonstrate that these metals, especially silver, profoundly change potassium homeostasis in roots of intact barley (Hordeum vulgare L.) plants, by altering unidirectional K(+) fluxes. Doses as low as 5 µM AgNO(3) rapidly reduced K(+) influx to 5% that of controls, and brought about pronounced and immediate increases in K(+) efflux, while higher doses of Au(3+) and Hg(2+) were required to produce similar responses. Reduced influx and enhanced efflux of K(+) resulted in a net loss of >40% of root tissue K(+) during a 15 min application of 500 µM AgNO(3), comprising the entire cytosolic potassium pool and about a third of the vacuolar pool. Silver also brought about major losses of UV-absorbing compounds, total electrolytes, and NH(4)(+). Co-application, with silver, of the channel blockers Cs(+), TEA(+), or Ca(2+), did not affect the enhanced efflux, ruling out the involvement of outwardly rectifying ion channels. Taken together with an examination of propidium iodide staining under confocal microscopy, the results indicate that silver ions affect K(+) homeostasis by directly inhibiting K(+) influx at lower concentrations, and indirectly inhibiting K(+) influx and enhancing K(+) efflux, via membrane destruction, at higher concentrations. Ni(2+), Cd(2+), and Pb(2+), three heavy metals not generally known to affect aquaporins, did not enhance K(+) efflux or cause propidium iodide incorporation. The study reveals strong and previously unknown effects of major aquaporin inhibitors and recommends caution in their application.