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<p>Ascorbic acid (AsA) and reduced glutathione (GSH) function as antioxidants and play crucial roles in somatic embryogenesis (SE). However, the effects of GSH, either alone or in combination with AsA, on the embryogenesis of <i>Fraxinus mandshurica</i> remain unclear. In this study, mature cotyledons of <i>F. mandshurica</i> were used as explants and treated with exogenous AsA and GSH. At later stages, the contents of intermediate metabolites in the AsA-GSH cycle, the activities of key and antioxidant enzymes, as well as the levels of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) in the explants were measured. The results showed that the browning rate of explants decreased with increasing concentrations of AsA and GSH. The highest efficiency in somatic embryo production was observed following treatment with GSH 300&#xa0;mg·L⁻¹ (G300) alone or a combination of AsA 50&#xa0;mg·L⁻¹ and GSH 200&#xa0;mg·L⁻¹ (A50G200). Compared to the control (CK), the application of GSH and AsA significantly increased the levels of antioxidants, including AsA, GSH, and oxidized glutathione (GSSG). It also enhanced the activities of reactive oxygen species (ROS)-scavenging enzymes—such as ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR), and glutathione reductase (GR)—which are integral components of the AsA-GSH cycle. Similarly, treatments with G300 or A50G200 also led to higher activities of the antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as increased H₂O₂ levels. This study demonstrates that exogenous GSH and AsA enhance somatic embryo production in <i>F. mandshurica</i> by activating the AsA-GSH cycle and ROS-scavenging enzymes. This lays a foundation for the large-scale propagation of <i>F. mandshurica</i> using SE technology.</p>

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Enhancing Somatic Embryogenesis in Fraxinus mandshurica with Glutathione and Ascorbic Acid

  • 期刊论文

  • 发表日期：

    2026年4月28日

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  • Xue Cheng,
  • Jianfei Yang,
  • Hailong Shen

摘要

Ascorbic acid (AsA) and reduced glutathione (GSH) function as antioxidants and play crucial roles in somatic embryogenesis (SE). However, the effects of GSH, either alone or in combination with AsA, on the embryogenesis of Fraxinus mandshurica remain unclear. In this study, mature cotyledons of F. mandshurica were used as explants and treated with exogenous AsA and GSH. At later stages, the contents of intermediate metabolites in the AsA-GSH cycle, the activities of key and antioxidant enzymes, as well as the levels of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) in the explants were measured. The results showed that the browning rate of explants decreased with increasing concentrations of AsA and GSH. The highest efficiency in somatic embryo production was observed following treatment with GSH 300 mg·L⁻¹ (G300) alone or a combination of AsA 50 mg·L⁻¹ and GSH 200 mg·L⁻¹ (A50G200). Compared to the control (CK), the application of GSH and AsA significantly increased the levels of antioxidants, including AsA, GSH, and oxidized glutathione (GSSG). It also enhanced the activities of reactive oxygen species (ROS)-scavenging enzymes—such as ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR), and glutathione reductase (GR)—which are integral components of the AsA-GSH cycle. Similarly, treatments with G300 or A50G200 also led to higher activities of the antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as increased H₂O₂ levels. This study demonstrates that exogenous GSH and AsA enhance somatic embryo production in F. mandshurica by activating the AsA-GSH cycle and ROS-scavenging enzymes. This lays a foundation for the large-scale propagation of F. mandshurica using SE technology.