Overexpression of SlMDHAR in transgenic tobacco increased salt stress tolerance involving S-nitrosylation regulation

Abstract

Protein S-nitrosylation, which refers to the redox-based posttranslational modification of a cysteine thiol by the attachment of a nitric oxide (NO) group, modulates a variety of enzyme activities. Monodehydroascorbate reductase (MDHAR) is essential for ascorbic acid (AsA) regeneration, which protects plant cells against damage by detoxifying reactive oxygen species (ROS). However, the relationship between S-nitrosylation and the role of tomato MDHAR (SlMDHAR) under salt stress remains unclear. In this paper, we show that the SlMDHAR mRNA expression, enzyme activity, protein level, total S-nitrosylated proteins and S-nitrosylated SlMDHAR protein level in tomato leaves significantly increase after NaCl treatment. To further evaluate the function of SlMDHAR under salt stress, overexpressed transgenic tobacco plants were used. The germination rate and root length of the overexpressed plants under NaCl stress were significantly higher than those of wild-type (WT) plants. Meanwhile, the transgenic plants had lower ROS accumulation, higher antioxidant enzyme activities and AsA-DHA ratio, more proline and soluble sugar contents than those in WT plants under salt stress. With a higher expression of stress-related genes, the transgenic plants demonstrated lower Na⁺ and higher K⁺ accumulation compared with WT plants. The NO accumulation and S-nitrosylated MDHAR level were higher in transgenic plants than in WT plants after NaCl treatment. In contrast, virus-induced gene silencing (VIGS) of SlMDHAR tomato plants showed enhanced sensitivity to salt stress and have lower S-nitrosylated MDHAR protein. These results suggested that SlMDHAR confers salt stress tolerance by alleviating oxidative damage probably involving the S-nitrosylation of MDHAR.

Keywords: AsA-GSH; MDHAR; NaCl; ROS; Tomato.