Salinity-induced stress is widely considered a main plant-growth-limiting factor. The positive effects of melatonin in modulating abiotic stresses have led this hormone to be referred to as a growth regulator in plants. This study aims to show how melatonin protects fenugreek against the negative effects of salt stress. Different amounts of melatonin (30, 60, and 90 ppm), salinity stress (150 mM and 300 mM), and the use of both salinity and melatonin were used as treatments. The results showed that applying different melatonin levels to salinity-treated fenugreek plants effectively prevented the degradation of chlorophyll a, chlorophyll b, total chlorophyll, and carotenoid contents compared with salinity treatment without melatonin application. Besides, melatonin increases the biosynthesis of enzymatic and non-enzymatic antioxidants, thereby adjusting the content of reactive oxygen species, free radicals, electrolyte leakage, and malondialdehyde content. It was observed that applying melatonin increased the activity of potassium-carrying channels leading to the maintenance of ionic homeostasis and increased intracellular water content under salinity stress. The results revealed that melatonin activates the defense signaling pathways in fenugreek through the nitric oxide, auxin, and abscisic acid-dependent pathways. Melatonin, in a similar vein, increased the expression of genes involved in the biosynthesis pathway of diosgenin, a highly important steroidal sapogenin in medical and food industries, and hence the diosgenin content. When 150 mM salinity stress and 60 ppm melatonin were coupled, the diosgenin concentration rose by more than 5.5 times compared to the control condition. In conclusion, our findings demonstrate the potential of melatonin to enhance the plant tolerance to salinity stress by stimulating biochemical and physiological changes.

No abstract

High-temperature stress is widely considered a main plant-growth-limiting factor. The positive effects of 24-epibrassinolide (EBR) as analogs of brassinosteroids (BRs) in modulating abiotic stresses have led this hormone to be referred to as a growth regulator in plants. The current study highlights the influence of EBR on enhancing tolerance to high-temperature and altering the diosgenin content in fenugreek. Different amounts of EBR (4, 8, and 16 μM), harvesting times (6, and 24 h), as well as temperature regimes (23 °C, and 42 °C) were, used as treatments. EBR application under normal temperature and high-temperature stress resulted in decreased malondialdehyde content and electrolyte leakage percentage, while the activity of antioxidant enzymes improved significantly. Exogenous EBR application possibly contributes to activating the nitric oxide, H2O2, and ABA-dependent pathways, enhancing the biosynthesis of abscisic acid and auxin, and regulating the signal transduction pathways, which raises fenugreek tolerance to high-temperature. The SQS (eightfold), SEP (2.8-fold), CAS (11-fold), SMT (17-fold), and SQS (sixfold) expression, considerably increased following EBR application (8 μM) compared to the control. Compared to the control, when the short-term (6 h) high-temperature stress was accompanied by EBR (8 μM), a sixfold increase in diosgenin content was achieved. Our findings highlight the potential role of exogenous 24-epibrassinolide in mitigating the high-temperature stress in fenugreek by stimulating the biosynthesis processes of enzymatic and non-enzymatic antioxidants, chlorophylls, and diosgenin. In conclusion, the current results could be of utmost importance in breeding or biotechnology-based programs of fenugreek and also in the researches related to the engineering of the biosynthesis pathway of diosgenin in this valuable plant.

The current study highlights the influence of 24-epibrassinosteroid on enhancing tolerance to high-temperature and altering the diosgenin content in fenugreek. The EBR application under high-temperature stress, resulted in decreased malondialdehyde content and electrolyte leakage percentage compared to plants treated with high-temperature stress. The antioxidant activities rose under high-temperature stress and applying EBR compared to control. Exogenous EBR application possibly contributes in activating the NO, H2O2 and ABA-dependent pathway, enhancing the biosynthesis of abscisic acid and auxin, and regulating the signal transduction pathways, which raises fenugreek tolerance to high-temperature. The SQS (8-fold), SEP (2.8-fold), CAS (11-fold), SMT (17-fold) and SQS (6-fold) expression, considerably increased following EBR application (8 μM) compared to control. Compared to control, when the short-term (6 h) high-temperature stress was accompanied by EBR (8 μM), a 6-fold increase in diosgenin content was achieved. Our findings highlight the potential role of exogenous 24-epibrassinolide in mitigating the high-temperature stress in fenugreek by stimulating the biosynthesis processes of enzymatic and non-enzymatic antioxidants, chlorophylls and diosgenin. In conclusion, the current results could be of utmost importance in breeding or biotechnology-based programs of fenugreek and also in the researches related to the engineering of the biosynthesis pathway of diosgenin in this valuable plant.