The SbMT-2 Gene from a Halophyte Confers Abiotic Stress Tolerance and Modulates ROS Scavenging in Transgenic Tobacco

Abstract

Heavy metals are common pollutants of the coastal saline area and Salicornia brachiata an extreme halophyte is frequently exposed to various abiotic stresses including heavy metals. The SbMT-2 gene was cloned and transformed to tobacco for the functional validation. Transgenic tobacco lines (L2, L4, L6 and L13) showed significantly enhanced salt (NaCl), osmotic (PEG) and metals (Zn ⁺⁺, Cu ⁺⁺ and Cd ⁺⁺) tolerance compared to WT plants. Transgenic lines did not show any morphological variation and had enhanced growth parameters viz. shoot length, root length, fresh weight and dry weight. High seed germination percentage, chlorophyll content, relative water content, electrolytic leakage and membrane stability index confirmed that transgenic lines performed better under salt (NaCl), osmotic (PEG) and metals (Zn ⁺⁺, Cu ⁺⁺ and Cd ⁺⁺) stress conditions compared to WT plants. Proline, H ₂O ₂ and lipid peroxidation (MDA) analyses suggested the role of SbMT-2 in cellular homeostasis and H ₂O ₂ detoxification. Furthermore in vivo localization of H ₂O ₂ and O ₂ ⁻; and elevated expression of key antioxidant enzyme encoding genes, SOD, POD and APX evident the possible role of SbMT-2 in ROS scavenging/detoxification mechanism. Transgenic lines showed accumulation of Cu ⁺⁺ and Cd ⁺⁺ in root while Zn ⁺⁺ in stem under stress condition. Under control (unstressed) condition, Zn ⁺⁺ was accumulated more in root but accumulation of Zn ⁺⁺ in stem under stress condition suggested that SbMT-2 may involve in the selective translocation of Zn ⁺⁺ from root to stem. This observation was further supported by the up-regulation of zinc transporter encoding genes NtZIP1 and NtHMA-A under metal ion stress condition. The study suggested that SbMT-2 modulates ROS scavenging and is a potential candidate to be used for phytoremediation and imparting stress tolerance.