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Barragán, Verónica; Leidi, Eduardo O.; Andrés, Zaida; Rubio, Lourdes; De Luca, Anna; Fernández, José A.; Cubero, Beatriz; Pardo, José M.
The Plant cell, 03/2012, Letnik: 24, Številka: 3Journal Article
Intracellular NHX proteins are Na⁺,K⁺/H⁺ antiporters involved in K⁺ homeostasis, endosomal pH regulation, and salt tolerance. Proteins NHX1 and NHX2 are the two major tonoplast-localized NHX isoforms. Here, we show that NHX1 and NHX2 have similar expression patterns and identical biochemical activity, and together they account for a significant amount of the Na⁺,K⁺/H⁺ antiport activity in tonoplast vesicles. Reverse genetics showed functional redundancy of NHX1 and NHX2 genes. Growth of the double mutant nhx1 nhx2 was severely impaired, and plants were extremely sensitive to external K⁺. By contrast, nhx1 nhx2 mutants showed similar sensitivity to salinity stress and even greater rates of Na⁺ sequestration than the wild type. Double mutants had reduced ability to create the vacuolar K⁺ pool, which in turn provoked greater K⁺ retention in the cytosol, impaired osmoregulation, and compromised turgor generation for cell expansion. Genes NHX1 and NHX2 were highly expressed in guard cells, and stomatal function was defective in mutant plants, further compromising their ability to regulate water relations. Together, these results show that tonoplast-localized NHX proteins are essential for active K⁺ uptake at the tonoplast, for turgor regulation, and for stomatal function.
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