The Early Oxidative Stress Induced by Mercury and Cadmium Is Modulated by Ethylene in Medicago sativa Seedlings.

Cadmium (Cd) and mercury (Hg) are ubiquitous soil pollutants that promote the accumulation of reactive oxygen species, causing oxidative stress. Tolerance depends on signalling processes that activate different defence barriers, such as accumulation of small heat sock proteins (sHSPs), activation of antioxidant enzymes, and the synthesis of phytochelatins (PCs) from the fundamental antioxidant peptide glutathione (GSH), which is probably modulated by ethylene. We studied the early responses of alfalfa seedlings after short exposure (3, 6, and 24 h) to moderate to severe concentration of Cd and Hg (ranging from 3 to 30 µM), to characterize in detail several oxidative stress parameters and biothiol (i.e., GSH and PCs) accumulation, in combination with the ethylene signalling blocker 1-methylcyclopropene (1-MCP). Most changes occurred in roots of alfalfa, with strong induction of cellular oxidative stress, H2O2 generation, and a quick accumulation of sHSPs 17.6 and 17.7. Mercury caused the specific inhibition of glutathione reductase activity, while both metals led to the accumulation of PCs. These responses were attenuated in seedlings incubated with 1-MCP. Interestingly, 1-MCP also decreased the amount of PCs and homophytochelatins generated under metal stress, implying that the overall early response to metals was controlled at least partially by ethylene.

Specific mechanisms of tolerance to copper and cadmium are compromised by a limited concentration of glutathione in alfalfa plants.

The induction of oxidative stress is a characteristic symptom of metal phytotoxicity and is counteracted by antioxidants such as glutathione (GSH) or homoglutathione (hGSH). The depletion of GSH│hGSH in fifteen-day-old alfalfa (Medicago sativa) plants pre-incubated with 1mM buthionine sulfoximine (BSO) affected antioxidant responses in a metal-specific manner under exposure to copper (Cu; 0, 6, 30 and 100µM) or cadmium (Cd; 0, 6 and 30µM) for 7 days. The phytotoxic symptoms observed with excess Cu were accompanied by an inhibition of root glutathione reductase (GR) activity, a response that was augmented in Cd-treated plants but reverted when combined with BSO. The synthesis of phytochelatins (PCs) was induced by Cd, whereas the biothiol concentration decreased in Cu-treated plants, which did not accumulate PCs. The depletion of GSH│hGSH by BSO also produced a strong induction of oxidative stress under excess Cu stress, primarily due to impaired GSH│hGSH-dependent redox homeostasis. In addition, the synthesis of PCs was required for Cd detoxification, apparently also determining the distribution of Cd in plants, as less metal was translocated to the shoots in BSO-incubated plants. Therefore, specific GSH│hGSH-associated mechanisms of tolerance were triggered by stress due to each metal.