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Arabidopsis thaliana is a glycophyte capable to tolerate mild salinity. Although salt sensitivity of this species, a variability of this characteristic was revealed between different ecotypes. This study presents the physiological and molecular characteristics of salt response of two ecotypes, NOK2 and Columbia (Col). Seedlings were cultivated in hydroponics in the presence of 0 or 50 mM NaCl during 25 days. Rosette leaf samples were collected after 19, 22, and 25 days for determination of physiological parameters, and after 18 days for study of DNA polymorphism. Salt treatment decreased rosette dry matter, leaf number, leaf hydration, and leaf surface area. All these effects were significantly more visible in Col than in NOK2. Moreover, the NOK2 leaves accumulated less Na⁺ and more K⁺ than those of Col. DNA polymorphism between the two ecotypes was analyzed with codominant molecular markers based on PCR amplification, namely, microsatellites, cleaved amplified polymorphism sequence (CAPS), and single nucleotide polymorphism markers (SNP). Among the 35 tested markers, 17 showed a clear polymorphism and were distributed on the five Arabidopsis chromosomes ending with a genetic map construction. These results could play an important role in the future establishment of cartography of candidate gene controlling the K⁺/Na⁺ selectivity of ion transport in leaves, a component of plant salt tolerance.
The Arabidopsis thaliana NOK2 accession displays salt tolerance compared to more commonly known A. thaliana accessions, such as Col-0, but the basis of this phenotypic feature is unknown. This work was focused on determining whether salt tolerance in NOK2 plants is affected by calcium supplementation to the growth medium. A. thaliana seedlings were grown in pots containing a mixture of sand and peat under controlled conditions in a low-level Ca(NO₃)₂ medium supplemented with 0 or 50 mM NaCl with and without amendment with two higher levels of Ca(NO₃)₂. Calcium amendment was beneficial for salt-treated NOK2 plants, as shown by the increase in dry weight of NOK2 plants with and without NaCl, but had no impact on Col-0 biomass. Sodium accumulation decreased as a function of calcium amendment in NOK2, while Col-0 maintained its high Na levels under these conditions. Leaf K⁺ content, K⁺ uptake, and Ca content decreased in NOK2 and Col-0 plants growing in the low-level Ca medium when NaCl was added, but rose in leaves of both accessions with calcium amendment, although K remained low in both accessions in the absence of NaCl. K⁺/Na⁺ selectivity increased preferentially in NOK2 with increasing calcium in the presence of NaCl, but when Na was restricted and not under any conditions in Col-0. Preferential effects of calcium were not observed on the transcript accumulation of seven Na⁺, K⁺ or Ca²⁺ transport genes for either of the accessions, except for increased transcription of the CAX4 gene in NOK2 leaves at the highest calcium concentration used (5 mM). Leaf membrane leakage, which increased two-fold higher in Col-0 under salt application compared with the increase in NOK2, declined for both accessions in response to calcium supplementation, and in NOK2 this decline reached no salt levels when Ca²⁺ amendments were highest. Chlorophyll and carotenoid content dropped two-fold in Col-0 in response to salt, but were unchanged in NOK2 under these conditions. In contrast, leaf anthocyanins, which were normally tenfold higher in Col-0 than in NOK2 in the lowlevel Ca²⁺ medium, declined in Col-0 plants as a function of Ca²⁺ supplementation, but were maintained at low levels in NOK2 leaves regardless of salinity and calcium. In conclusion, NOK2 plants responded positively to calcium supplementation by improving biomass yield during salinity treatment, whereas this amendment only affected Col-0 by reducing its permeability and anthocyanin titre. K⁺/Na⁺ selectivity appeared to be an important characteristic of NOK2 response to calcium. The regulation of this response may involve the CAX4 Ca²⁺/H⁺ vacuolar transport gene, but does not appear to involve six other common ion transporters.
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