Physiological and proteomic responses of reactive oxygen species metabolism and antioxidant machinery in mulberry (Morus alba L.) seedling leaves to NaCl and NaHCO3 stress

Abstract

In this paper, the effects of 100 mM NaCl and NaHCO₃ stress on reactive oxygen species (ROS) and physiological and proteomic aspects of ROS metabolism in mulberry seedling leaves were studied. The results showed that NaCl stress had little effect on photosynthesis and respiration of mulberry seedling leaves. Superoxide dismutase (SOD) activity and the expression of related proteins in leaves increased by varying degrees, and accumulation of superoxide anion (O₂·^-) not observed. Under NaHCO₃ stress, photosynthesis and respiration were significantly inhibited, while the rate of O₂·^- production rate and H₂O₂ content increased. The activity of catalase (CAT) and the expression of CAT (W9RJ43) increased under NaCl stress. In response to NaHCO₃ stress, the activity and expression of CAT were significantly decreased, but the ability of H₂O₂ scavenging of peroxidase (POD) was enhanced. The ascorbic acid-glutathione (AsA-GSH) cycle in mulberry seedling leaves was enhancement in both NaCl and NaHCO₃ stress. The expression of 2-Cys peroxiredoxin BAS1 (2-Cys Prx BAS1), together with thioredoxin F (TrxF), thioredoxin O1 (TrxO1), thioredoxin-like protein CITRX (Trx CITRX), and thioredoxin-like protein CDSP32 (Trx CDSP32) were significantly increased under NaCl stress. Under NaHCO₃ stress, the expression of the electron donor of ferredoxin-thioredoxin reductase (FTR), together with Trx-related proteins, such as thioredoxin M (TrxM), thioredoxin M4 (TrxM4), thioredoxin X (TrxX), TrxF, and Trx CSDP32 were significantly decreased, suggesting that the thioredoxin-peroxiredoxin (Trx-Prx) pathway's function of scavenging H₂O₂ of in mulberry seedling leaves was inhibited. Taken together, under NaCl stress, excessive production of O₂·^- mulberry seedlings leaves was inhibited, and H₂O₂ was effectively scavenged by CAT, AsA-GSH cycle and Trx-Prx pathway. Under NaHCO₃ stress, despite the enhanced functions of POD and AsA-GSH cycle, the scavenging of O₂·^- by SOD was not effective, and that of H₂O₂ by CAT and Trx-Prx pathway were inhibited; and in turn, the oxidative damage to mulberry seedling leaves could not be reduced.

Keywords: Alkalinity; Antioxidant machinery; Morus alba L.; Proteomics; Reactive oxygen species; Salinity.