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. 2023 Apr 24;13(1):6661.
doi: 10.1038/s41598-023-33913-6.

Exogenous 24-epibrassinolide ameliorates tolerance to high-temperature by adjusting the biosynthesis of pigments, enzymatic, non-enzymatic antioxidants, and diosgenin content in fenugreek

Shahla Sheikhi  1 , Amin Ebrahimi #  2 , Parviz Heidari  1 , Mohamad Reza Amerian  1 , Sajad Rashidi-Monfared  3 , Hadi Alipour #  4
Affiliations

Exogenous 24-epibrassinolide ameliorates tolerance to high-temperature by adjusting the biosynthesis of pigments, enzymatic, non-enzymatic antioxidants, and diosgenin content in fenugreek

Shahla Sheikhi et al. Sci Rep. .

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.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
The effects of temperature, harvesting times, and different levels of EBR on chlorophyll a (a), chlorophyll b (b), total chlorophyll (c), and carotenoid (d). Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
Figure 2
Figure 2
The effects of temperature, harvesting times, and different levels of EBR on electrolyte leakage (a), MDA (b), and total soluble protein (c). Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
Figure 3
Figure 3
The effects of temperature, harvesting times, and different levels of EBR on superoxide dismutase (a), glutathione reductase (b), catalase (c), guaiacol peroxidase (d) and ascorbate peroxidase (e). Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
Figure 4
Figure 4
The effects of temperature, harvesting times, and different levels of EBR on H2O2 (a), and NO (b). Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
Figure 5
Figure 5
The effects of temperature, harvesting times, and different levels of EBR on Abscisic acid (a) and Auxin content (b). Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
Figure 6
Figure 6
The effects of temperature, harvesting times, and different levels of EBR on the expression levels of SQS (a), SEP (b), CAS (c), SMT (d), and SSR (e). Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
Figure 7
Figure 7
The effects of temperature, harvesting times, and different levels of EBR on diosgenin content. Duncan method at a 1% probability level was applied to compare mean values. The columns having similar letters had no significant difference.
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