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A large genetic screen for sos (for salt overly sensitive) mutants was performed in an attempt to isolate mutations in any gene with an sos phenotype. Our search yielded 28 new alleles of sos1, nine mutant alleles of a newly identified locus, SOS2, and one allele of a third salt tolerance locus, SOS3. The sos2 mutations, which are recessive, were mapped to the lower arm of chromosome V, approximately 2.3 centimorgans away from the marker PHYC. Growth measurements demonstrated that sos2 mutants are specifically hypersensitive to inhibition by Na+ or Li+ and not hypersensitive to general osmotic stresses. Interestingly, the SOS2 locus is also necessary for K+ nutrition because sos2 mutants were unable to grow on a culture medium with a low level of K+. The expression of several salt-inducible genes was superinduced in sos2 plants. The salt tolerance of sos1, sos2, and sos3 mutants correlated with their K+ tissue content but not their Na+ tissue content. Double mutant analysis indicated that the SOS genes function in the same pathway. Based on these results, a genetic model for salt tolerance mechanisms in Arabidopsis is presented in which SOS1, SOS2, and SOS3 are postulated to encode regulatory components controlling plant K+ nutrition that in turn is essential for salt tolerance

Plant roots contain both high- and low-affinity transport systems for uptake of K+ from the soil. In this study, we characterize a K+ transporter that functions in both high- and low-affinity uptake. Using yeast complementation analysis, we isolated a cDNA for a functional K+ transporter from Arabidopsis (referred to as AtKUP1 for Arabidopsis thaliana K+ uptake). When expressed in a yeast mutant, AtKUP1 dramatically increased K+ uptake capacity at both a low and high [K+] range. Kinetic analyses showed that AtKUP1-mediated K+ uptake displays a \"biphasic\" pattern similar to that observed in plant roots. The transition from the high-affinity phase (Km of 44 micromolars) to the low-affinity phase (Km of 11 mM) occurred at 100 to 200 micromolars external K+. Both low- and high-affinity K+ uptake via AtKUP1 were inhibited by 5 mM or higher concentrations of NaCl. In addition, AtKUP1-mediated K+ uptake was inhibited by K+ channel blockers, including tetraethylammonium, Cs+, and Ba2+. Consistent with a possible function in K+ uptake from the soil, the AtKUP1 gene is primarily expressed in roots. We conclude that the AtKUP1 gene product may function as a K+ transporter in Arabidopsis roots over a broad range of [K+] in the soil

KAT1 is a voltage-dependent inward rectifying K+ channel cloned from the higher plant Arabidopsis thaliana [Anderson, J. A., Huprikar, S. S., Kochian, L. V., Lucas, W. J. and Gaber, R. F. (1992) Proc. Natl. Acad. Sci. USA 89, 3736-3740]. It is related to the Shaker superfamily of K+ channels characterized by six transmembrane spanning domains (S1-S6) and a putative pore-forming region between S5 and S6 (H5). The H5 region between Pro-247 and Pro-271 in KAT1 contains 14 additional amino acids when compared with Shaker [Aldrich, R. W. (1993) Nature (London) 362, 107-108]. We studied various point mutations introduced into H5 to determine whether voltage-dependent plant and animal K+ channels share similar pore structure. Through heterologous expression in Xenopus oocytes and voltage-clamp analysis combined with phenotypic analysis involving a potassium transport-defective Saccharomyces cerevisiae strain, we investigated the selectivity filter of the mutants and their susceptibility toward inhibition by cesium and calcium ions. With respect to electrophysiological properties, KAT1 mutants segregated into three groups: (i) wild-type-like channels, (ii) channels modified in selectivity and Cs+ or Ca2+ sensitivity, and (iii) a group that was additionally affected in its voltage dependence. Despite the additional 14 amino acids in H5, this motif in KAT1 is also involved in the formation of the ion-conducting pore because amino acid substitutions at Leu-251, Thr-256, Thr-259, and Thr-260 resulted in functional channels with modified ionic selectivity and inhibition. Creation of Ca2+ sensitivity and an increased susceptibility to Cs + block through mutations within the narrow pore might indicate that both blockers move deeply into the channel. Furthermore, mutations close to the rim of the pore affecting the half-activation potential (U1/2) indicate that amino acids within the pore either interact with the voltage sensor or ion permeation feeds back on gating

Inward-rectifying K+ (K+in) channels in the guard cell plasma membrane have been suggested to function as a major pathway for K+ influx into guard cells during stomatal opening. When K+in channels were blocked with external Cs+in wild-type Arabidopsis guard cells, light-induced stomatal opening was reduced. Transgenic Arabidopsis plants were generated that expressed a mutant of the guard cell K+in channel, KAT1, which shows enhanced resistance to the Cs+ block. Stomata in these transgenic lines opened in the presence of external Cs+. Patch-clamp experiments with transgenic guard cells showed that inward K+in currents were blocked less by Cs+ than were K+ currents in controls. These data provide direct evidence that KAT1 functions as a plasma membrane K+ channel in vivo and that K+in channels constitute an important mechanism for light-induced stomatal opening. In addition, biophysical properties of K+in channels in guard cells indicate that components in addition to KAT1 may contribute to the formation of K+in channels in vivo

Low and intermediate level radioactive wastes are generated from daily applications of radioisotopes in various medical, industrial, agricultural and research fields. The release of these wastes to the surrounding environment represents a major complicated ecological crisis. Treatment of that radwastes is the most essential process in the radioactive waste management scheme. For simplicity and low running costs, phytoremediation technique has been candidate for processing some selected hazardous liquid waste streams. The submitted work aims at evaluating the capability of one of the aquatic plant, namely Lemna gibba, to biosorb Cs-137 and Co-60 from aqueous radioactive waste simulate. The study discusses, in batchwise laboratory scale experiments, the parameters that may affect the efficiency of Lemna gibba to bioremove and bioaccumulate the two radionuclides (e.g: contact time, pH value and the initial activity content of the waste simulate, light effect, biomass used, ...). The uptake values, biosorption efficiency percentages, rate constant and isotherm factors were evaluated for the process. The uptakes values for Co-60 and Cs-137 respectively recorded 1213 Bq/gm and 872 Bq/gm from the waste simulate solution containing 6100 Bq and at pH=6.9 after 24 hours contact time. The results obtained exhibit the potential of the wide aquatic plant Lemna gibba to be used as a biological sorber for cesium-137 and cobalt-60 from their low and intermediate level aqueous radioactive waste stream successfully and efficiently.

Uptake by roots from contaminated soil is one of the key steps in the entry of radiocaesium into the food chain. We have measured the uptake by roots of radiocaesium and its transfer to shoots of a heathland grass, sheep fescue (Festuca ovina L.) from two contrasting agricultural soils, a sandy podzol and a clayey calcareous soil. A culture device which keeps the roots separate from the soil was used thus allowing rhizosphere soil to be obtained easily and enhancing the effect of root action. Biomass production and ¹³⁷Cs in shoots and roots were recorded. Cs adsorption was studied on both the initial, nonrhizosphere soil and on rhizosphere soil in dilute soil suspension. Cs desorption was measured by resuspending subsamples of contaminated soil in solutions containing various concentrations of stable Cs. The proportion of Cs fixed, i.e. not readily desorbable, was calculated by comparison of the adsorption and desorption isotherms. Uptake by roots varied considerably between soils and did not appear to be diffusion limited. Root-to-shoot transfer did not differ for the two soils studied. Root action considerably enhanced Cs adsorption on the soils, particularly the in sandy podzol with a low Cs affinity. The value of Kd was increased by up to an order of magnitude. A large proportion of adsorbed Cs was found to be fixed, the Kd was up to seven times greater on desorption than adsorption, indicating that up to 80% of adsorbed Cs was not readily exchangeable. Root action had little effect on the fixed fraction.

A novel effect of ammonium ions on root growth was investigated to understand how environmental signals affect organ development. Ammonium ions (3-12 mM) were found to dramatically inhibit Arabidopsis thaliana seedling root growth in the absence of potassium even if nitrate was present. This inhibition could be reversed by including in the growth medium low levels (20-100 micromole) of potassium or alkali ions Rb+ and Cs+ but not alkali ions Na+ and Li+. The protective effect of low concentrations of potassium is not due to an inhibition of ammonium uptake. Ammonium inhibition is reversible, because root growth was restored in ammonium-treated seedlings if they were subsequently transferred to medium containing potassium. It is known that plant hormones can inhibit root growth. We found that mutants of Arabidopsis resistant to high levels of auxin and other hormones (aux1, axr1, and axr2) are also resistant to the ammonium inhibition and produce roots in the absence of potassium. Thus, the mechanisms that mediate the ammonium inhibition of root development are linked to hormone metabolic or signaling pathways. These findings have important implications for understanding how environmental signals, especially mineral nutrients, affect plant root development

Biotite is a potassium rich mineral, which is used as a fertilizer in organic farming and as a soil amendment in conventional farming. Its ability to reduce ^sup 134^Cs uptake by ryegrass from peat soil was studied in pot experiments and compared with zeolite, heavy clay, bentonite and apatite. In addition, the long-term effect of biotite on ^sup 137^Cs uptake from peat soil was studied in the peat field. In the pot experiments in the first cut of ryegrass, the minerals decreased ^sup 134^Cs uptake by plants in the following order: zeolite > heavy clay > bentonite > biotite > apatite. Apatite did not have any effect on the plant ^sup 134^Cs level. In the later cuts, the uptake of ^sup 134^Cs from biotite-treated soil decreased further while that from soils treated with other minerals remained unchanged or even increased. In general, ^sup 134^Cs uptake by plants decreased with increasing mineral level. The decrease of ^sup 134^Cs uptake became more efficient, especially at the early growth stage, by mixing small amounts of zeolite in biotite. The results of the field experiment indicated the long-term effect of biotite on reducing ^sup 134^Cs uptake by plants. Biotite application rate was 30 t ha-1. The five-year mean of the plant/soil concentration ratio of ^sup 137^Cs was 0.05 for biotite-treated soil, in contrast to 0.14 for the control soil. On the whole, biotite reduced considerably the ^sup 137^Cs level of plants on peat soil and this effect was long-lasting. For an effective reduction of plant radiocesium a great quantity of biotite is needed and therefore it is most suitable for greenhouse cultivation where contaminated slightly decomposed peat is used as a growing medium.[PUBLICATION ABSTRACT]

Nowadays, there is still the need to improve the quantification of parameters that affect radionuclide mobility. With this aim, radiocesium and radiostrontium soil-to-plant transfer was measured in lysimeters in a Calcic Luvisol, loamy soil and in a Fluvisol, loam-sandy soil, using lettuce [Lactuca sativa L. cv. Kinemontepas] and pea plants [Pisum sativum L. cv. Kelvedon Wonder]. Weighted Concentration Ratios (WCR), expressed as kg soil/kg plant, were calculated for different growth stages. Weighted Concentration Ratios were in general higher for 85Sr than for 134Cs, and also higher in the loam-sandy than in the loamy soil. To predict plant uptake, we evaluated a set of soil properties to define a prediction factor for the relative transfer in the two soils using cation exchange capacity (CEC) and radionuclide available fraction (fav) for radiostrontium, and soil solution composition, solid-liquid distribution coefficient, and radionuclide available fraction for radiocesium. The ratios of WCR in the loam-sandy and loamy soil were compared with the prediction factor. There was good agreement in lettuce for 85Sr (ratio of WCR was 5.4 for seedling and 3.9 for commercial samples, whereas prediction factor was 3.1) and for 134Cs (ratio of WCR was 5.1 for seedling and 5.5 for commercial samples, the prediction factor being 5.1), although for pea only the relative root uptake of radiocesium in seedling pea was well predicted (the ratio of WCR was 8.8, the prediction factor being 9.1). These soil parameters improved former predictions based solely on the fav, although factors depending on plant physiology should be better evaluated