Pretreatment of seeds with hydrogen peroxide improves deep-sowing tolerance of wheat seedlings

Drought is a prevalent natural factor limiting crop production in arid regions across the world. To overcome this limitation, seeds are sown much deeper to boost germination by soil moisture produced by underground water. Seed pretreatment can effectively induce deep-sowing tolerance in plants. In the present study, we evaluated whether H₂O₂ pretreatment of seeds can initiate metabolic changes and lead to improved deep-sowing tolerance in wheat. Pretreatment with 0.05 μM H₂O₂ promoted first internode elongation by 13% in the deep-sowing tolerant wheat cultivar "Tir" and by 32% in the sensitive cultivar "Kıraç-66" under deep-sowing conditions, whereas internode elongation was inhibited by diphenyleneiodonium chloride. In contrast to Tir seedlings, H₂O₂ levels in the first internode of Kıraç-66 seedlings increased under deep-sowing condition in the H₂O₂-treated group compared to controls. Moreover, these seedlings had significantly lower catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX) activities but higher NADPH oxidase (NOX) and superoxide dismutase (SOD) activities under the same conditions, which consequently induced greater H₂O₂ accumulation. Contrary to Tir, both total glutathione and glutathione S-transferase (GST) activity decreased in Kıraç-66 after deep-sowing at 10 cm. However, H₂O₂ treatment increased the total glutathione amounts and the activities of glutathione-related enzymes (except GST and GPX) in the first internode of Kıraç-66. Taken together, these data support that H₂O₂ acts as a signaling molecule in the activation of antioxidant enzymes (specifically NOX, SOD, and CAT), regulation of both glutathione-related enzymes and total glutathione content, and upregulation of the cell wall-loosening protein gene TaEXPB23.