Effects of methyl viologen on the antioxidant system in cultured Salvia miltiorrhiza cells

doi:10.3724/SP.J.1258.2014.00047

Abstract

Abstract:

Aims We studied the effects of the methyl viologen (MV) in the culture medium on the antioxidant enzyme system in Salvia miltiorrhiza cells and investigated changes in the contents of H₂O₂, malonaldehyde (MDA), glutathione (GSH), and antioxidant enzyme. The differential expressions of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were discussed. Our objective was to explore if the MV could significantly affect the antioxidant system in the cultured S. miltiorrhiza cells.
Methods We treated the S. miltiorrhiza cells in suspension cultures with MV as an elicitor on the accumulation of H₂O₂, and used the diuron (DCMU) as an electron transporting antagonist. The biosynthesis of H₂O₂, MDA, and GSH, and the activities of SOD, CAT, and POD were determined by ultraviolet spectrophotometry. The specific isoenzymes bands of SOD, CAT, and POD were examined by acrylamide gel electrophoresis technique.
Important findings We found that MV significantly increased the accumulation of H₂O₂, MDA, and GSH, along with increased activities of CAT and POD as well as enhanced expressions and increased number of isoenzymes bands of CAT and POD; whereas DCMU inhibited the effects of MV. Results in this study suggest that MV could significantly enhance H₂O₂ accumulation in chloroplasts and regulate the antioxidant system in S. miltiorrhiza cells in suspension cultures, in order to maintain normal physiological conditions.

Key words: antioxidant system, chloroplast, DCMU, methyl viologen, polyacrylamide gel electrophoresis, Salvia miltiorrhiza

XING Bing-Yu,ZHU Nan,ZHANG Hong-Pei,YANG Xi-Ling,DONG Juan-E. Effects of methyl viologen on the antioxidant system in cultured Salvia miltiorrhiza cells[J]. Chin J Plant Ecol, 2014, 38(5): 507-514.

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URL: https://www.plant-ecology.com/EN/10.3724/SP.J.1258.2014.00047

https://www.plant-ecology.com/EN/Y2014/V38/I5/507

Figures/Tables 6

Fig. 1 Effects of methyl viologen on the accumulation of H2O2 in cultured Salvia miltiorrhiza cells (mean ± SE). Test materials are S. miltiorrhiza cells in suspension cultures for six days. Different small letters indicate significant differences (p < 0.05).

Fig. 2 Effects of diuron (DCMU) and methyl viologen (MV) on the accumulation of H2O2 in cultured Salvia miltiorrhiza cells (mean ± SE). Test materials are S. miltiorrhiza cells in suspension cultures for six days. DCMU is added 30 min prior to the MV treatments. Different small letters indicate significant differences (p < 0.05).

Fig. 3 Effects of different treatments on the accumulation of malondialdehyde (MDA) in cultured Salvia miltiorrhiza cells (mean ± SE). DCMU, diuron; MV, methyl viologen. Test materials are S. miltiorrhiza cells in suspension cultures for six days. DCMU is added 30 min prior to the MV treatments. Different small letters indicate significant differences (p < 0.05).

Fig. 4 Effects of different treatments on the accumulation of glutathione (GSH) in cultured Salvia miltiorrhiza cells (mean ± SE). DCMU, diuron; MV, methyl viologen. Test materials are S. miltiorrhiza cells in suspension cultures for six days. DCMU is added 30 min prior to the MV treatments. Different small letters indicate significant differences (p < 0.05).

Fig. 5 Effects of different treatments on the activities of superoxide dismutase (SOD) (A), catalase (CAT) (B), and peroxidase (POD) (C) in cultured Salvia miltiorrhiza cells (mean ± SE). DCMU, diuron; MV, methyl viologen. Test materials are S. miltiorrhiza cells in suspension cultures for six days. DCMU is added 30 min prior to the MV treatments. Different small letters indicate significant differences (p < 0.05).

Fig. 6 Effects of different treatments on the isoenzymes spectrum of superoxide dismutase (SOD) (A), catalase (CAT) (B), and peroxidase (POD) (C) in culture Salvia miltiorrhiza cells. 1, distilled water treatment; 2, methyl viologen treatment; 3, diuron treatment; 4, methyl viologen + diuron treatment. a-e, soenzyme bands of SOD, CAT and POD.

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