Molecular Physiological Evidence for the Role of Na+-Cl- Co-Transporter in Branchial Na+ Uptake in Freshwater Teleosts.

The gills are the major organ for Na+ uptake in teleosts. It was proposed that freshwater (FW) teleosts adopt Na+/H+ exchanger 3 (Nhe3) as the primary transporter for Na+ uptake and Na+-Cl- co-transporter (Ncc) as the backup transporter. However, convincing molecular physiological evidence to support the role of Ncc in branchial Na+ uptake is still lacking due to the limitations of functional assays in the gills. Thus, this study aimed to reveal the role of branchial Ncc in Na+ uptake with an in vivo detection platform (scanning ion-selective electrode technique, SIET) that has been recently established in fish gills. First, we identified that Ncc2-expressing cells in zebrafish gills are a specific subtype of ionocyte (NCC ionocytes) by using single-cell transcriptome analysis and immunofluorescence. After a long-term low-Na+ FW exposure, zebrafish increased branchial Ncc2 expression and the number of NCC ionocytes and enhanced gill Na+ uptake capacity. Pharmacological treatments further suggested that Na+ is indeed taken up by Ncc, in addition to Nhe, in the gills. These findings reveal the uptake roles of both branchial Ncc and Nhe under FW and shed light on osmoregulatory physiology in adult fish.

Optimization of eco-friendly amendments as sustainable asset for salt-tolerant plant growth-promoting bacteria mediated maize (Zea Mays L.) plant growth, Na uptake reduction and saline soil restoration.

Soil salinity is progressively affecting global agriculture area, and act as a brutal environmental factor for the productivity of plants, therefore, sustainable remediation of the saline soil is urgently required. In this study, we tested the effectiveness of PM (poultry manure), SMS (spent mushroom substrate), and CD (cow dung) for the recovery of salt soil and the optimization of the productivity of the maize plant. PM and SMS showed the valuable source of OC, N, P, K as the CD. The HCA analysis showed that 47% of the bacterial population from PM, SMS, and CD survived at 6% NaCl (w/v), which had PGP attributes such as IAA, P-solubilizers, and siderophore activity. The results from pot experiments of plant growth and PCA analysis of bacterial PGP attributes reveled re formulation of PM, SMS, and CD, which were further optimized at the saline field level. T-2 treated plant increased their shoot length, chlorophyll content, reducing sugar, nitrogen, phosphorus, and potassium levels significantly after 30 and 60 days, followed by T-4 and T-3 as the control. A significant (P < 0.01) increase in particle density and decrease in bulk density was observed for all combinations treated (T-2 to T-7). A two-year field study revealed that the T-2 combination increased 43% OC, 57% N, 66% P, 48% K, 32% DHA, 76% PPO in the soil than the control after 60 days. T2-combination decreased ≈50% of Na content in root and shoot, and increased 27% of maize crop yield. The dose of 10% PM + 10% SMS can significantly induce the growth of maize plants and the restoration of saline soil health.

Respirometry and cutaneous oxygen flux measurements reveal a negligible aerobic cost of ion regulation in larval zebrafish (Danio rerio).

Fishes living in fresh water counter the passive loss of salts by actively absorbing ions through specialized cells termed ionocytes. Ionocytes contain ATP-dependent transporters and are enriched with mitochondria; therefore ionic regulation is an energy-consuming process. The purpose of this study was to assess the aerobic costs of ion transport in larval zebrafish (Danio rerio). We hypothesized that changes in rates of Na+ uptake evoked by acidic or low Na+ rearing conditions would result in corresponding changes in whole-body oxygen consumption (MO2 ) and/or cutaneous oxygen flux (JO2 ), measured at the ionocyte-expressing yolk sac epithelium using the scanning micro-optrode technique (SMOT). Larvae at 4 days post-fertilization (dpf) that were reared under low pH (pH 4) conditions exhibited a higher rate of Na+ uptake compared with fish reared under control conditions (pH 7.6), yet they displayed a lower MO2  and no difference in cutaneous JO2 Despite a higher Na+ uptake capacity in larvae reared under low Na+ conditions, there were no differences in MO2  and JO2  at 4 dpf. Furthermore, although Na+ uptake was nearly abolished in 2 dpf larvae lacking ionocytes after morpholino knockdown of the ionocyte proliferation regulating transcription factor foxi3a, MO2 and JO2  were unaffected. Finally, laser ablation of ionocytes did not affect cutaneous JO2 Thus, we conclude that the aerobic costs of ion uptake by ionocytes in larval zebrafish, at least in the case of Na+, are below detection using whole-body respirometry or cutaneous SMOT scans, providing evidence that ion regulation in zebrafish larvae incurs a low aerobic cost.

Use of gene knockout to examine serotonergic control of ion uptake in zebrafish reveals the importance of controlling for genetic background: A cautionary tale.

Freshwater (FW) fishes inhabit dilute environments and must actively absorb ions in order to counteract diffusive salt loss. Neuroendocrine control of ion uptake in FW fishes is an important feature of ion homeostasis and several important neuroendocrine factors have been identified. The role of serotonin (5-HT), however, has received less attention despite several studies pointing to a role for 5-HT in the control of ion balance. Here, we used a gene knockout approach to elucidate the role of 5-HT in regulating Na+ and Ca2+ uptake rates in larval zebrafish. Tryptophan hydroxylase (TPH) is the rate-limiting step in 5-HT synthesis and we therefore hypothesized that ion uptake rates would be altered in zebrafish larvae carrying knockout mutations in tph genes. We first examined the effect of tph1b knockout (KO) and found that tph1bKO larvae, obtained from Harvard University, had reduced rates of Na+ and Ca2+ uptake compared to wild-type (WT) larvae from our institution (uOttawa WT), lending support to our hypothesis. However, further experiments controlling for differences in genetic background demonstrated that WT larvae from Harvard University (Harvard WT) had lower ion uptake rates than those of uOttawa WT, and that ion uptake rate between Harvard WT and tph1bKO larvae were not significantly different. Therefore, our initial observation that tph1bKO larvae (Harvard source) had reduced ion uptake rates relative to uOttawa WT was a function of genetic background and not of knockout itself. These data provide a cautionary tale of the importance of controlling for genetic background in gene knockout experiments.

Characterization of Na+ transport to gain insight into the mechanism of acid-base and ion regulation in white sturgeon (Acipenser transmontanus).

Freshwater fish actively take up ions via specific transporters to counter diffusive losses to their hypotonic environment. While much is known about the specific mechanisms employed by teleosts, almost nothing is known about the basal fishes, such as white sturgeon (Acipenser transmontanus) which may offer insight into the evolution of osmo- and ionoregulation in fishes. We investigated Na+ uptake in juvenile white sturgeon in the presence and absence of transporter inhibitors. We found that sturgeon acclimated to 100µmoll-1 Na+ have Na+ uptake kinetics typical of teleosts and that a Na+/H+ exchanger (NHE) is the predominant transporter for Na+ uptake. White sturgeon are tolerant to hypercarbia-induced respiratory acidoses and recover blood pH (pHe) at 1.5kPa PCO2 but not at higher PCO2 (6kPa PCO2) where they preferentially regulate intracellular pH (pHi). It was hypothesized that during exposure to hypercarbia Na+ uptake would increase at CO2 tensions at which fish were capable of pHe regulation but decrease at higher tensions when they were preferentially regulating pHi. We found that Na+ uptake did not increase at 1.5kPa PCO2, but at 6kPa PCO2 Na+ uptake was reduced by 95% while low water pH equivalent to 6kPa PCO2 reduced Na+ uptake by 71%. Lastly, we measured net acid flux during hypercarbia, which indicates that net acid flux is not associated with Na+ uptake. These findings indicate Na+ uptake in sturgeon is not different from freshwater teleosts but is sensitive to hypercarbia and is not associated with pHe compensation during hypercarbia.

Spermidine and Ca(2+), but not Na(+), can permeate NMDA receptors consisting of GluN1 and GluN2A or GluN2B in the presence of Mg(2+).

N-Methyl-D-aspartate receptors (NMDA receptors) are known to be permeable to Na(+) and Ca(2+) ions. In this study, we tested whether polyamines (putrescine, spermidine, spermine), organic cations found in cells, can permeate NMDA receptors expressed in Xenopus laevis oocytes and HEK293 cells. It was found that polyamines, especially spermidine, can permeate NMDA channels expressed from GluN1/GluN2A or GluN1/GluN2B activated by glycine and glutamate. Furthermore, spermidine and Ca(2+) influx through NMDA receptors was observed in the presence of Mg(2+), although Na(+) influx was strongly inhibited by Mg(2+). The Km values for spermidine influx through GluN1/GluN2A and GluN1/GluN2B were 2.2 mM and 2.7 mM, respectively in the presence of isotonic extracellular ion solutions. Spermidine uptake by NMDA receptors was dependent on the presence of glycine and glutamate, and inhibited by Ca(2+) and by memantine, an NMDA receptor channel blocker. The Km values for Ca(2+) influx through GluN1/GluN2A and GluN1/GluN2B were 4.6 mM and 3.3 mM, respectively, under the same ionic conditions. The results indicate that spermidine and Ca(2+), but not Na(+), can permeate NMDA receptors in the presence of Mg(2+). Spermidine, if released locally from presynaptic terminals (where its concentration is high in synaptosomes and synaptic vesicles) could permeate NMDA receptors and play a role in synaptic plasticity mediated by NMDA receptors together with Ca(2+).

Ammonia first? The transition from cutaneous to branchial ammonia excretion in developing rainbow trout is not altered by exposure to chronically high NaCl.

Larval rainbow trout (Oncorhynchus mykiss) were reared from hatch under control ([Na(+)]=0.60 mmol l(-1)) or high NaCl ([Na(+)]=60 mmol l(-1)) conditions to elucidate the driving force for the ontogeny of branchial Na(+)/NH4 (+) exchange, one of the earliest gill functions. We hypothesized that if Na(+) uptake is the driving force, then in high NaCl there would be a delay in the skin-to-gill shift in ammonia excretion (Jamm) and/or an elevation in whole-body total ammonia (Tamm). In both groups, however, the skin-to-gill shift for Jamm, determined using divided chambers, occurred at the same time (13 days post-hatch; dph) and whole-body Tamm was unchanged. Moreover, high NaCl larvae displayed elevated whole-body [Na(+)] relative to controls by 18 dph, suggesting that maintaining branchial Jamm occurs at the expense of Na(+) balance. Overall, these results support the 'ammonia hypothesis', which posits that ammonia excretion, probably as Na(+)/NH4 (+) exchange, is the primary function of the early fish gill.

(22)Na influx is significantly lower in salt tolerant groundnut (Arachis hypogaea) varieties.

Distinct varieties differing in salt tolerance were initially identified from two separate green house experiments using two systems; solution as well as soil culture. The first screening involved a diverse group of 27 cultivars. Several physiological traits; Chlorophyll Stability Index (CSI), Salt Tolerance Index (STI) and ion content were determined to screen the cultivars for differences in salt tolerance using solution culture in the first experiment. A set of six varieties (three tolerant and three susceptible) were selected from this experiment and then subjected again to salt stress adopting a natural soil system in the second experiment which involved a screening approach essentially similar to that of the first experiment. In the third experiment using two distinct cultivars differing in salt tolerance selected from experiment II, (22)Na influx rate was determined in the root and shoot at the end of a 24 h salt imposition in Hoagland's nutrient system containing 180 KBq of (22)Na. The results suggested that there were distinct differences in (22)Na influx rate into root and concurrently in the shoot. The salt tolerant Spanish improved and one of the moderately tolerant Trombay variety TAG 24, showed good regulation of (22)Na influx resulting in low (22)Na concentration. The salt susceptible variety JSP39 had nearly 7-8 fold higher root (22)Na content as compared to the tolerant and moderately tolerant cultivars. The results have highlighted the importance of Na exclusion as an important determinant of salt tolerance in groundnut.

Effect of low pH exposure on Na(+) regulation in two cichlid fish species of the Amazon.

We evaluated the effects of acute exposure to low pH on Na(+) regulation in two Amazon cichlids collected from natural ion-poor "blackwaters", angelfish (Pterophyllum scalare) and discus (Symphysodon discus). Na(+) uptake kinetic parameters, unidirectional Na(+) fluxes, and net Cl(-) fluxes were determined at pH6.0 and 3.6. At pH6.0, both species presented low unidirectional Na(+) flux rates, with kinetics showing a relatively low affinity for Na(+) (angelfish Km=79, discus Km=268µmolL(-1)), with similar maximum transport capacities (Jmax~535nmolg(-1)h(-1)). Overall, there appeared to be high sensitivity to inhibition by low pH, yet low intrinsic branchial permeability limiting diffusive ion effluxes, resulting in relatively low net loss rates of Na(+), the same strategy as seen previously in other blackwater cichlids, and very different from the strategy of blackwater characids. At low pH, Na(+) uptake in angelfish was inhibited competitively (increased Km=166µmolL(-1)) and non-competitively (decreased Jmax=106nmolg(-1)h(-1)), whereas in discus, only a decrease in Jmax (112nmolg(-1)h(-1)) was statistically significant. An acute reduction in H(+)-ATPase activity, but not in Na(+)/K(+)-ATPase activity, in the gills of angelfish suggests a possible mechanism for this non-competitive inhibition at low pH. Discus fish were more tolerant to low pH than angelfish, as seen by lesser effects of exposure to pH3.6 on unidirectional Na(+) uptake and efflux rates and net Na(+) and Cl(-) loss rates. Overall, discus are better than angelfish in maintaining ionic balance under acidic, ion-poor conditions.

Overexpression of OsGF14C enhances salinity tolerance but reduces blast resistance in rice.

High-salinity and blast disease are two major stresses that cause dramatic yield loss in rice production. GF14 (14-3-3) genes have been reported to play important role in biotic and abiotic stresses in plants. However, the roles of OsGF14C remain unknown. To understand the functions and regulatory mechanisms of OsGF14C in regulating salinity tolerance and blast resistance in rice, we have conducted OsGF14C-overexpressing transgenic experiments in the present study. Our results showed that overexpression of OsGF14C enhanced salinity tolerance but reduced blast resistance in rice. The enhanced salinity tolerance is related to the reduction of methylglyoxal and Na+ uptake instead of exclusion or compartmentation and the negative role of OsGF14C in blast resistance is associated with the suppression of OsGF14E, OsGF14F and PR genes. Our results together with the results from the previous studies suggest that the lipoxygenase gene LOX2 which is regulated by OsGF14C may play roles in coordinating salinity tolerance and blast resistance in rice. The current study for the first time revealed the possible roles of OsGF14C in regulating salinity tolerance and blast resistance in rice, and laid down a foundation for further functional study and crosstalk regulation between salinity and blast resistance in rice.

Teleostean fishes may have developed an efficient Na+ uptake for adaptation to the freshwater system.

Understanding Na+ uptake mechanisms in vertebrates has been a research priority since vertebrate ancestors were thought to originate from hyperosmotic marine habitats to the hypoosmotic freshwater system. Given the evolutionary success of osmoregulator teleosts, these freshwater conquerors from the marine habitats are reasonably considered to develop the traits of absorbing Na+ from the Na+-poor circumstances for ionic homeostasis. However, in teleosts, the loss of epithelial Na+ channel (ENaC) has long been a mystery and an issue under debate in the evolution of vertebrates. In this study, we evaluate the idea that energetic efficiency in teleosts may have been improved by selection for ENaC loss and an evolved energy-saving alternative, the Na+/H+ exchangers (NHE3)-mediated Na+ uptake/NH4 + excretion machinery. The present study approaches this question from the lamprey, a pioneer invader of freshwater habitats, initially developed ENaC-mediated Na+ uptake driven by energy-consuming apical H+-ATPase (VHA) in the gills, similar to amphibian skin and external gills. Later, teleosts may have intensified ammonotelism to generate larger NH4 + outward gradients that facilitate NHE3-mediated Na+ uptake against an unfavorable Na+ gradient in freshwater without consuming additional ATP. Therefore, this study provides a fresh starting point for expanding our understanding of vertebrate ion regulation and environmental adaptation within the framework of the energy constraint concept.

Frequent SLC12A3 mutations in Chinese Gitelman syndrome patients: structure and function disorder.

Purposes: This study was conducted to identify the frequent mutations from reported Chinese Gitelman syndrome (GS) patients, to predict the three-dimensional structure change of human Na-Cl co-transporter (hNCC), and to test the activity of these mutations and some novel mutations in vitro and in vivo. Methods: SLC12A3 gene mutations in Chinese GS patients previously reported in the PubMed, China National Knowledge Infrastructure, and Wanfang database were summarized. Predicted configurations of wild type (WT) and mutant proteins were achieved using the I-TASSER workplace. Six missense mutations (T60M, L215F, D486N, N534K, Q617R, and R928C) were generated by site-directed mutagenesis. 22Na+ uptake experiment was carried out in the Xenopus laevisoocyte expression system. In the study, 35 GS patients and 20 healthy volunteers underwent the thiazide test. Results: T60M, T163M, D486N, R913Q, R928C, and R959frameshift were frequent SLC12A3 gene mutations (mutated frequency >3%) in 310 Chinese GS families. The protein's three-dimensional structure was predicted to be altered in all mutations. Compared with WT hNCC, the thiazide-sensitive 22Na+ uptake was significantly diminished for all six mutations: T60M 22 ± 9.2%, R928C 29 ± 12%, L215F 38 ± 14%, N534K 41 ± 15.5%, Q617R 63 ± 22.1%, and D486N 77 ± 20.4%. In thiazide test, the net increase in chloride fractional excretion in 20 healthy controls was significantly higher than GS patients with or without T60M or D486N mutations. Conclusions: Frequent mutations (T60M, D486N, and R928C) and novel mutations (L215F, N534K, and Q617R) lead to protein structure alternation and protein dysfunction verified by 22Na+ uptake experiment in vitro and thiazide test on the patients.

High Pseudocapacitance Boosts Ultrafast, High-Capacity Sodium Storage of 3D Graphene Foam-Encapsulated TiO2 Architecture.

Anatase TiO2 is an attractive anode for Li-ion batteries and Na-ion batteries because of its structural stability. However, the electrochemical capability of anatase TiO2 is unsatisfactory due to its intrinsically low electrical conductivity and poor ion diffusivity at the electrode/electrolyte interface. We prepared 3D lightweight graphene aerogel-encapsulated anatase TiO2, which exhibits a high reversible capacity (390 mA h g-1 at 50 mA g-1), a superior rate performance (164.9 mA h g-1 at 5 A g-1), and a long-term cycling capability (capacity retention of 86.8% after 7800 cycles). The major energy-storage mechanism is surface capacitance dominated, which favors a high capacity and fast Na+ uptake. The inherent features of 3D porous aerogels provide additional active reaction sites and facilitate fast charge diffusion and easy ion access. This will enable the development of 3D interconnected, graphene-based, high-capacity active materials for the development of next-generation energy-storage applications.

Ion uptake pathways in European sea bass Dicentrarchus labrax.

Ion uptake mechanisms are diverse in fish species, certainly linked to duplication events that have led to the presence of a multitude of paralogous genes. In fish, Na+ uptake involves several ion transporters expressed in different ionocyte subtypes. In the European sea bass Dicentrarchus labrax, several key transporters potentially involved in Na+ uptake have been investigated in seawater (SW) and following a 2 weeks freshwater (FW) acclimation. Using gel electrophoresis, we have shown that the Na+/H+-exchanger 3 (nhe3, slc9a3) is expressed in gills and kidney at both salinities. Quantitative realtime PCR analysis showed a significantly higher nhe3 expression in fresh water (FW) compared to SW. Its apical localization in a subset of gill ionocytes in freshwater-acclimated fish supports the role of NHE3 in Na+ uptake. Interestingly, NHE3-immunopositive cells also express basolateral Na+/K+/2Cl- cotransporter 1 (NKCC1) and are mainly localized in gill lamella. Among the three nhe2 (slc9a2) paralogs, only nhe2c shows differential branchial expression levels with higher mRNA levels in SW than in FW. The increased branchial expression of the ammonia transporter rhcg1 (Rhesus protein), nhe3 and cytoplasmic carbonic anhydrase (cac) in FW could indicate the presence of a functional coupling between ion transporters to form a Na+/NH4+ exchange complex. Acid-sensing ion channel 4 (asic4) seems not to be expressed in sea bass gills. Na+/Cl- cotransporter (ncc2a or ncc-like) is about three times more expressed in FW compared to SW suggesting coupled Na+ and Cl- uptake in a subset of gill ionocytes. Besides the main pump Na+/K+-ATPase, branchial NCC2a and NHE3 may be key players in ion uptake in sea bass following a long-term freshwater challenge.

Physiological protective action of dissolved organic carbon on ion regulation and nitrogenous waste excretion of zebrafish (Danio rerio) exposed to low pH in ion-poor water.

Dissolved organic carbon (DOC) represents a heterogeneous group of naturally-occurring molecules in aquatic environments, and recent studies have evidenced that optically dark DOCs can exert some positive effects on ionoregulatory homeostasis of aquatic organisms in acidic waters. We investigated the effects of Luther Marsh DOC, a dark allochthonous DOC, on ion regulation and N-waste excretion of zebrafish acutely exposed to either neutral or low pH in ion-poor water. In the first experiment, simultaneous exposure to pH 4.0 and DOC greatly attenuated the stimulation of Na+ diffusive losses (J outNa ), and prevented the blockade of Na+ uptake (J inNa ) seen in zebrafish exposed to pH 4.0 alone, resulting in much smaller disturbances in Na+ net losses (J netNa ). DOC also attenuated the stimulation of net Cl- losses (J netCl ) and ammonia excretion (J netAmm ) during acidic challenge. In the second experiment, zebrafish acclimated to DOC displayed similar regulation of J inNa and J outNa , and, therefore, reduced J netNa at pH 4.0, effects which persisted even when DOC was no longer present. Protective effects of prior acclimation to DOC on J netCl and J netAmm at pH 4.0 also occurred, but were less marked than those on Na+ balance. Urea fluxes were unaffected by the experimental treatments. Overall, these effects were clearly beneficial to the ionoregulatory homeostasis of zebrafish at low pH, and were quite similar to those seen in a recent parallel study using darker DOC from the upper Rio Negro. This suggests that dark allochthonous DOCs share some chemical properties that render fish tolerant to ionoregulatory disturbances during acidic challenge.

High waterborne Mg does not attenuate the toxic effects of Fe, Mn, and Ba on Na+ regulation of Amazonian armored catfish tamoatá (Hoplosternum litoralle).

Formation water (FoW) is a by-product from oil and gas production and usually has high concentrations of soluble salts and metals. Calcium (Ca) and magnesium (Mg) have been shown to reduce the toxicity of metals to aquatic animals, and previous study showed that high waterborne Ca exerts mild effect against disturbances on Na+ regulation in Amazonian armored catfish tamoatá (Hoplosternum littorale) acutely exposed to high Fe, Mn, and Ba levels. Here, we hypothesized that high Mg levels might also reduce the toxic effects of these metals on Na+ regulation of tamoatá. The exposure to 5% FoW promoted an increase in Na+ uptake and a rapid accumulation of Na+ in all tissues analyzed (kidney

External potassium (K(+)) application improves salinity tolerance by promoting Na(+)-exclusion, K(+)-accumulation and osmotic adjustment in contrasting peanut cultivars.

Achieving salt-tolerance is highly desirable in today's agricultural context. Apart from developing salt-tolerant cultivars, possibility lies with management options, which can improve crop yield and have significant impact on crop physiology as well. Thus present study was aimed to evaluate the ameliorative role of potassium (K(+)) in salinity tolerance of peanut. A field experiment was conducted using two differentially salt-responsive cultivars and three levels of salinity treatment (control, 2.0 dS m(-1), 4.0 dS m(-1)) along with two levels (with and without) of potassium fertilizer (0 and 30 kg K2O ha(-1)). Salinity treatment incurred significant changes in overall physiology in two peanut cultivars, though the responses varied between the tolerant and the susceptible one. External K(+) application resulted in improved salinity tolerance in terms of plant water status, biomass produced under stress, osmotic adjustment and better ionic balance. Tolerant cv. GG 2 showed better salt tolerance by excluding Na(+) from uptake and lesser accumulation in leaf tissue and relied more on organic osmolyte for osmotic adjustment. On the contrary, susceptible cv. TG 37A allowed more Na(+) to accumulate in the leaf tissue and relied more on inorganic solute for osmotic adjustment under saline condition, hence showed more susceptibility to salinity stress. Application of K(+) resulted in nullifying the negative effect of salinity stress with slightly better response in the susceptible cultivar (TG 37A). The present study identified Na(+)-exclusion as a key strategy for salt-tolerance in tolerant cv. GG 2 and also showed the ameliorating role of K(+) in salt-tolerance with varying degree of response amongst tolerant and susceptible cultivars.

Wheat responses to sodium vary with potassium use efficiency of cultivars.

The role of varied sodium (Na) supply in K nutrition of wheat (Triticum aestivum L.) is not well understood especially among cultivars differing in K efficiency. We examined the response of K-efficient and K-inefficient Australian wheat cultivars to Na supply (low to high Na) under K-deficient and K-adequate conditions. In a pot experiment, wheat cvv Wyalkatchem, Cranbrook (K-efficient), and cvv Gutha, Gamenya (K-inefficient) were grown for 8 weeks in a sandy soil containing 40 or 100 mg K/kg in combination with nil, 25, 50, 100, or 200 mg Na/kg. High soil Na levels (100, 200 mg Na/kg) greatly reduced plant growth in all four cultivars especially at low soil K (40 mg K/kg). By contrast, low to moderate soil Na levels (25, 50 mg Na/kg) stimulated root dry weight at low K supply, particularly in K-efficient cultivars compared with K-inefficient cultivars. At low K supply, low to moderate Na failed to increase shoot Na to a concentration where substitution of K would be feasible. However, low to moderate Na supply increased shoot K concentration and content in all four wheat cultivars, and it increased leaf photosynthesis and stomatal conductance to measured values similar to those under adequate K and nil Na conditions. The results showed that low to moderate Na stimulated K uptake by wheat particularly in K-efficient cultivars and through increased shoot K enhanced the photosynthesis. We conclude that increased photosynthesis supplied more assimilates that led to increased root growth and that greater root growth response of K-efficient cultivars is related to their greater K-utilization efficiency. However, the process by which low to moderate Na increased shoot K content warrants further investigation.

Exposure to waterborne Cu inhibits cutaneous Na⁺ uptake in post-hatch larval rainbow trout (Oncorhynchus mykiss).

In freshwater rainbow trout (Oncorhynchus mykiss), two common responses to acute waterborne copper (Cu) exposure are reductions in ammonia excretion and Na(+) uptake at the gills, with the latter representing the likely lethal mechanism of action for Cu in adult fish. Larval fish, however, lack a functional gill following hatch and rely predominantly on cutaneous exchange, yet represent the most Cu-sensitive life stage. It is not known if Cu toxicity in larval fish occurs via the skin or gills. The present study utilized divided chambers to assess cutaneous and branchial Cu toxicity over larval development, using disruptions in ammonia excretion (Jamm) and Na(+) uptake (Jin(Na)) as toxicological endpoints. Early in development (early; 3 days post-hatch; dph), approximately 95% of Jamm and 78% of Jin(Na) occurred cutaneously, while in the late developmental stage (late; 25 dph), the gills were the dominant site of exchange (83 and 87% of Jamm and Jin(Na), respectively). Exposure to 50 µg/l Cu led to a 49% inhibition of Jamm in the late developmental stage only, while in the early and middle developmental (mid; 17 dph) stages, Cu had no effect on Jamm. Jin(Na), however, was significantly inhibited by Cu exposure at the early (53% reduction) and late (47% reduction) stages. Inhibition at the early stage of development was mediated by a reduction in cutaneous uptake, representing the first evidence of cutaneous metal toxicity in an intact aquatic organism. The inhibitions of both Jamm and Jin(Na) in the late developmental stage occurred via a reduction in branchial exchange only. The differential responses of the skin and gills to Cu exposure suggest that the mechanisms of Jamm and Jin(Na) and/or Cu toxicity differ between these tissues. Exposure to 20µg/l Cu revealed that Jamm is the more Cu-sensitive process. The results presented here have important implications in predicting metal toxicity in larval fish. The Biotic Ligand Model (BLM) is currently used to predict metal toxicity in aquatic organisms. However, for rainbow trout this is based on gill binding constants from juvenile fish. This may not be appropriate for post-hatch larval fish where the skin is the site of toxic action of Cu. Determining Cu binding constants and lethal accumulation concentrations for both skin and gills in larval fish may aid in developing a larval fish-specific BLM. Overall, the changing site of toxic action and physiology of developing larval fish present an interesting and exciting avenue for future research.