Ion exchangers NHX1 and NHX2 mediate active potassium uptake into vacuoles to regulate cell turgor and stomatal function in Arabidopsis

16 pages, 12 figures

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.

This work was supported by grants from Ministerio de Ciencia e Innovacio ́ n (cofinanced by the European Regional Develop- ment Fund) to J.M.P. (BIO2009-08641 and CSD2007-00057) and J.A.F. (CTM2011-30356). Z.A. was supported by the Junta de Ampliacio ́ n de Estudios–Consejo Superior de Investigaciones Cientificas Fellowship Program. Maria A. Parrado and A.D.L. were supported by the Programa Nacional de Potenciacion de Recursos Humanos del Plan Nacional de Investigacion Cientifica.

Peer reviewed