Synergistic effect of IBA, BAP, and salicylic acid on the in vitro regeneration, total polyphenols, and antioxidant activity of Ruta graveolens L

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发表日期：

2026年5月23日

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Aysar Mohammed Salim,
Ziyad Shihab Ahmed,
Bashar Zeki Ameen

摘要

Ruta graveolens L. is a medicinal plant currently cultured in most countries of the world. A factorial design was employed using three concentrations each of: IBA: 0.5, 1.0, 1.5 mg/L, BAP: 0.5, 1.0, 2.0 mg/L, and SA: 4, 8, 12 mg/L, along with a control (no hormone), resulting in 27 treatment combinations to optimizing hormonal in tissue culture of this plant. Results exhibited a progressively enhancement for callus induction, organogenesis and phenolic compounds with increasing hormonal levels. Application of IBA 1.5 mg/L + BAP 2.0 mg/l + SA 4 mg/L induced the highest callus formation (65.33%), IBA 1.5 mg/L + BAP 2.0 mg/L, with SA 8 or 12 lead to highest callus weight (0.86%). The highest survival rate was observed in the plantlets treated with IBA 1.5 mg/L + BAP 2.0 mg/L + SA 12 mg/L (81.5%), the highest root length (42.2 mm) and root number (15.3 n), respectively were observed in plantlets treated with 1.5 mg/L IBA + 2.0 mg/L BAP + 12/ 8 mg/L SA. Furthermore, the highest shoot number (7.7) and shoot length (34.4 mm), respectively were recorded in response to 1.5 mg/L IBA + 2.0 mg/L BAP + 8/12 mg/L SA. The highest flavonoid accumulation was observed in cultures treated with IBA 1.5 mg/L + BAP 2.0 + SA 12 mg/L (42.5 mg quercetin/g extract). Finally, the highest phenolic compounds (63.18 mg/g gallic acid equivalent) and antioxidant activity (63.18%) were obtained with application of 1.5 mg/L IBA + 2.0 mg/L BAP + 12 mg/L SA. A comprehensive univariate and multivariate (PCA) analysis were revealed IBA at 1.5 mg/L in combination with BAP at 2.0 mg/L and SA at 8–12 mg/L significantly enhanced callus induction, shoot and root traits, and levels of flavonoids, phenols, and antioxidants compared with other treatments. An optimized synergistic combination of IBA, BAP, and salicylic acid establishes an efficient and reproducible protocol for enhanced morphogenesis, metabolite accumulation, and large-scale in vitro propagation of Ruta graveolens.