Harboured cation/proton antiporters modulate stress response to integrated heat and salt via up-regulating KINI and GOLSI in double transgenic Arabidopsis

Abstract

Recent research has pointed to improved salt tolerance by co-overexpression of Arabidopsis thaliana NHX1 (Na+/H+ antiporter) and SOS1 (Salt Overly Sensitive1). However, functionality under salt stress accompanying heat is less understood in double transgenics. To further advance possible co-operational interactions of AtNHX1 (N) and AtSOS1 (S) under combined stress, modulation of osmolyte, redox, energy, and abscisic acid metabolism genes was analysed. The expression of the target BIP3, KIN1, GOLS1, OHP2, and CYCA3;2 in transgenic Arabidopsis seedlings were significantly regulated towards a dramatic suppression by ionic, osmotic, and heat stresses. AtNHX1 and AtSOS1 co-overexpression (NS) outpaced the single transgenics and control in terms of membrane disorganisation and the electrolyte leakage of the cell damage caused by heat and salt stress in seedlings. While NaCl slightly induced CYCA3;2 in transgenics, combined stress up-regulated KIN1 and GOLS1, not other genes. Single N and S transgenics overexpressing AtNHX1 and AtSOS1 only appeared similar in their growth and development; however, different to WT and NS dual transgenics under heat + salt stress. Seed germination, cotyledon survival, and hypocotyl length were less influenced by combined stress in NS double transgenic lines than in single N and S and wild type. Stress combination caused significant reprogramming of gene expression profiles, mainly towards downregulation, possibly as a trade-off strategy. Analysing phenotypic, cellular, and transcriptional responses regulating growth facets of tolerant transgenic genotypes may support the ongoing efforts to achieve combined salt and heat tolerance.