Evidence of Coordinated and Adjustable Osmolytes Movements Following Hyposmotic Swelling in Rainbow Trout Red Blood Cells

BACKGROUND/AIMS: The osmolytes involved in the volume regulation of hyposmotically-swollen fish cells are well identified. However, if a coordination and adjustments of their fluxes are obvious, few studies have clearly illustrated these aspects. METHODS: Trout red blood cells volume variations were estimated from water contents obtained by a gravimetric method. Intracellular K+ and Na+ contents, and Cl- content of haemolysed cells were determined by photometry and colorimetry, respectively. The taurine contribution to cell volume regulation was calculated from the net changes of water, K+, Cl- and Na+ contents. The intracellular pH was calculated from the chloride distribution across the cells membranes according to the Donnan equilibrium. RESULTS: Cells responses to a rapid change (from 296 to 176 mOsm.kg-1) of the saline osmolality were examined in three conditions designed to not impact (Hypo. I) or to reduce the K+ (Hypo. II) and Cl- (Hypo. III) contributions to the volume regulation. Hypo. I condition caused an immediate increase in water content, followed by a 90 min. full regulation, concomitant with gradual lowering of K+ and Cl- contents and a surprising increase in Na+ content. Hypo. II and III conditions showed a partial and complete volume regulation, respectively. This was made possible by an increase in the taurine involvement. These experiments allowed to confirm that K+ and Cl- were released via KCl cotransport and by separate channels. The comparison of Hypo. I and III conditions led to the observation that the partially amiloride-sensitive Na+ influx is proportional to the taurine efflux; the latter being sustained mainly by a Na+/taurine cotransport. The Hypo. II condition was suitable for the (Na+/K+)ATPase activity inhibition. This effect could explain the observed lack of Na+ uptake, the consecutive depletion of intracellular taurine stock and the incomplete volume regulation. Finally, the results support the importance of taurine in pH control under Hypo. I (physiologic) condition. The alkalosis observed in Hypo. II and III conditions were the consequences of changes in the salines compositions, not of physiologic adjustments. CONCLUSION: The regulatory volume decrease process of trout RBCs is complex and adjustable through coordinated osmolytes movements. The obliged decrease in K+ and/or Cl- contributions stimulates taurine and Na+ pathways. This study highlights the importance of taurine as a compensatory variable in cell volume regulation and explains for the first time the significance of the Na+ uptake during this process

Growth, Yield, and Bunch Quality of “Superior Seedless” Vines Grown on Different Rootstocks Change in Response to Salt Stress

The growth and quality of vines are negatively affected by soil salinity if enough salts accumulate in the root zone. As part of the current study, we estimated the remediating effects of rootstocks under salinity. For this reason, “superior seedless” vines were grafted onto three different rootstocks, such as SO4, 1103 Paulson, and own-root (“superior seedless” with their own-root). The experiment was conducted in the 2019 and 2020 seasons. This study examines the effects of different rootstocks on vine growth, yield, and quality using “superior seedless” vines grown in sandy soil with salinity. Four stages of berry development were examined (flowering, fruit set, veraison, and harvest time). At harvest, yield characteristics (clusters per vine and cluster weight) were also assessed. Each parameter of the growth season was influenced separately. The K+ and Na+ ratios were also significantly increased, as were the salinity symptoms index and bunch yield per vine and quality. Rootstock 1103 Paulson improved photosynthetic pigments, K+ accumulation, Na+ uptake, and cell membrane damage in “superior seedless” vines compared to other rootstocks, according to the study results. As determined in the arid regions of northwestern Egypt, the 1103 Paulson can mitigate salinity issues when planting “superior seedless” vines on sandy soil.

Vacuolar H+-ATPase and Na+/K+-ATPase energize Na+ uptake mechanisms in the nuchal organ of the hyperregulating freshwater crustacean Daphnia magna.

The nuchal organ of the embryos and neonates of the cladoceran, Daphnia magna, has been shown to be a site of Na+ influx and H+, NH4+ and Cl− efflux. This study combines the scanning-ion selective electrode technique with application of inhibitors of specific transporters to assess the mechanisms of Na+ transport across the nuchal organ. Na+ influx across the nuchal organ was inhibited both by inhibitors of the Na+/K+-ATPase (ouabain, bufalin) and by inhibitors of the vacuolar H+-ATPase (bafilomycin, N-ethylmaleimde, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, KM91104, S-nitrosoglutathione). Na+ influx was unaffected by the epithelial Na+ channel blocker benzamil, but was sensitive to ethylisopropyl amiloride and elevated external ammonium concentrations, consistent with roles for Na+/H+ and Na+/NH4+ exchangers in the apical membrane but not Na+ channels. Transport across the basolateral membrane into the hemolymph is proposed to involve the Na+/K+-ATPase and a thiazide-sensitive Na+:Cl− cotransporter. Keywords: Daphnia magna, Na+/K+-ATPase, V-ATPase, Iionoregulation, Nnuchal organ

Downregulation of high-affinity potassium and sodium symporter gene, EcHKT1;1, in Eucalyptus roots enhances salt tolerance

Engineering for restricted root Na+ uptake could potentially enhance salt tolerance in Eucalyptus. High-affinity K+ transporters (HKTs) have been implicated in Na+ uptake from the external medium as in the case of TaHKT2;1 or in the unloading of Na+ from xylem like in AtHKT1;1. To rapidly determine the in planta role of EcHKT1:1, composite transgenics in which EcHKT1:1 was specifically downregulated via RNAi in the roots were generated. Compared to the controls that failed to survive at 350 mM NaCl, 33 % of the composite transgenic plantlets generated using the EcHKT1;1 silencing construct were able to tolerate up to 400 mM NaCl. In these composite transgenics, EcHKT1;1 downregulation ranged from 37 % to 74 %. The average shoot to root ratio of sodium was 4.9 folds lower than the controls indicating restricted translocation of Na+ to the shoots. Relative expression analysis in the leaves of two non-transgenic genotypes contrasting for their salt tolerance also showed downregulated EcHKT1;1 expression in the tolerant clone. The study thus determined that EcHKT1;1 is a major gene determining Na+ transport from the roots to shoots. This study also demonstrated the utility of the composite transgenic approach for screening genes conferring salt tolerance in tree species.

Plant HKT Channels: An Updated View on Structure, Function and Gene Regulation

HKT channels are a plant protein family involved in sodium (Na+) and potassium (K+) uptake and Na+-K+ homeostasis. Some HKTs underlie salt tolerance responses in plants, while others provide a mechanism to cope with short-term K+ shortage by allowing increased Na+ uptake under K+ starvation conditions. HKT channels present a functionally versatile family divided into two classes, mainly based on a sequence polymorphism found in the sequences underlying the selectivity filter of the first pore loop. Physiologically, most class I members function as sodium uniporters, and class II members as Na+/K+ symporters. Nevertheless, even within these two classes, there is a high functional diversity that, to date, cannot be explained at the molecular level. The high complexity is also reflected at the regulatory level. HKT expression is modulated at the level of transcription, translation, and functionality of the protein. Here, we summarize and discuss the structure and conservation of the HKT channel family from algae to angiosperms. We also outline the latest findings on gene expression and the regulation of HKT channels.

Effects of single and binary applications of bisphenol A and NaCl on Ceratopyllum demersum

Determining the physiological effects of single and binary applications of BPA and NaCl on the Ceratophyllum demersum L., which is a rootless submerged aquatic macrophyte, is the aim of the present study. For this purpose, the macrophyte was collected from a local pond that was not exposed to any pollution. Macrophytes were acclimatized in 10% nutrient solution for 14 days, than they were treated with 17.2 mg/L BPA; 10 and 100 mM NaCl; 17.2 mg/L BPA plus 10 mM NaCl and 17.2 mg/L BPA plus 100 mM NaCl. The macrophyte without added BPA or/and NaCl served as control (without treatment). Increasing concentrations of NaCl induced a progressive accumulation of Na in the macrophyte tissues. On the other hand, BPA application partially reduced Na uptake. Nutrient uptake was affected differently by the applications. The contents of some nutrients such as Cu, Zn, Mn, K and Mg were generally reduced, whereas the Fe and Ca contents were increased. In general, increases in contents of total carbohydrate, total phenolic and non-protein sulfhydryl groups were found, when compared to control. Protein and photosynthetic pigment contents, on the contrary to these, were decreased. According to findings, the increase in H2O2 and MDA levels showed that single and combined applications of BPA and NaCl in C. demersum tissues induced oxidative stress.