A cDNA fragment comprising 80% of complete cDNA nucleotide sequence was identified in barley (Hordeum vulgare L.); it encoded a polypeptide of 401 amino acids and had a high (83%) homology to the tomato K+/H+antiporter, LeNHX2. We supposed that the identified cDNA fragment encodes a new isoform of Na+,K+/H++ antiporter and designated it HvNHX4. The transcripts of the HvNHX4 gene were detected in roots and leaves of barley seedlings; their content did not change under salt stress conditions. Antibodies to C-terminal fragment of HvNHX4 were raised. Western blot analysis of membrane proteins showed that HvNHX4 was located in the vacuolar membranes and probably in the endoplasmic reticulum and Golgi membranes; tracing amounts of this protein were present in plasma membranes. A comparative Western blot analysis was performed for two isoforms (HvNHX4 and HvNHX2) in vacuolar membranes isolated from roots and leaves of four barley cultivars with different resistances to salt stress. The HvNHX4 protein was detected in roots of a salt-susceptible cv. Belogorskii grown under control conditions; its content in roots of a salt-tolerant cv. Moskovskii 121 was found to increase after salt stress. The content of HvNHX2 protein in roots and leaves increased during salt stress in the salt-tolerant cv. Elo but did not change in the susceptible cv. Belogorskii. The results provide evidence for the organ-and cultivar-specific expression of different isoforms of Na+,K+/H+ antiporter in barley and indicate that the expression of these isoforms is regulated at the posttranscriptional level during salt stress.
Russian Journal of Plant Physiology – Springer Journals
Published: Jan 6, 2007
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