Exogenous 2,4-Epibrassinolide Alleviates Alkaline Stress in Cucumber by Modulating Photosynthetic Performance

Brassinosteroids (BRs) are recognized for their ability to enhance plant salt tolerance. While considerable research has focused on their effects under neutral salt conditions, the mechanisms through which BRs regulate photosynthesis under alkaline salt stress are less well understood. This study investigates these mechanisms, examining plant growth, photosynthetic electron transport, gas exchange parameters, Calvin cycle dynamics, and the expression of key antioxidant and Calvin cycle genes under alkaline stress conditions induced by NaHCO₃. The findings indicate that NaHCO₃ stress substantially impairs cucumber growth and photosynthesis, significantly reducing chlorophyll content, net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (E), maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ΦPSII), antenna conversion efficiency (Fv'/Fm'), and photochemical quenching coefficient (qP). This disruption suggests a severe dysregulation of the photosynthetic electron transport system, impairing electron transfer from photosystem II (PSII) to photosystem I (PSI) and subsequently the Calvin cycle. Application of exogenous 24-epibrassinolide (EBR) alleviated these effects, reducing leaf chlorosis and growth inhibition and significantly enhancing the expression of key genes within the antioxidant system (AsA-GSH cycle) and the Calvin cycle. This intervention also led to a reduction in reactive oxygen species (ROS) accumulation and improved photosynthetic performance, as evidenced by enhancements in Pn, Gs, E, Fv/Fm, ΦPSII, Fv'/Fm', and qP. Moreover, NaHCO₃ stress hindered chlorophyll synthesis, primarily by blocking the conversion from porphobilinogen (PBG) to uroporphyrinogen III (UroIII) and by increasing chlorophyllase (Chlase) and decreasing porphobilinogen deaminase (PBGD) activity. Exogenous EBR countered these effects by enhancing PBGD activity and reducing Chlase activity, thereby increasing chlorophyll content under stress conditions. In summary, EBR markedly mitigated the adverse effects of alkaline stress on cucumber leaf photosynthesis by stabilizing the photosynthetic electron transport system, accelerating photosynthetic electron transport, and promoting the Calvin cycle. This study provides valuable insights into the regulatory roles of BRs in enhancing plant resilience to alkaline stress.