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<p><i>Capparis spinosa</i> L. is a Mediterranean medicinal and aromatic species valued for its flavonoids, glucosinolates, and volatile terpenoids. However, increasing exploitation raises concerns regarding sustainable utilization. An in vitro callus culture system was developed to optimize biomass production and secondary metabolite accumulation. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different combinations of cytokinins (6-benzylaminopurine, BAP; kinetin, KIN) and auxins (2,4-dichlorophenoxyacetic acid, 2,4-D; naphthaleneacetic acid, NAA). Biomass accumulation was evaluated through fresh and dry weight gain. Volatile compounds were analyzed by HS-SPME-GC-MS, whereas non-volatile metabolites were characterized using UPLC-QTOF-MS/MS and semi-quantified by UPLC-TQ-MS/MS. Response surface methodology identified optimal hormonal conditions for callus biomass production after 30 days, corresponding to BAP (1.5-2.0&#xa0;mg/L) combined with 2,4-D (0.5–1.5&#xa0;mg/L), yielding fresh and dry weight gains exceeding 3000&#xa0;mg and 300&#xa0;mg, respectively. Volatile profiling revealed terpenes and sulfur-containing compounds, with 2,4-D treatments inducing safranal, (E)-rose oxide, and menthol, not detected in leaf tissues. NAA treatments mainly influenced relative volatile abundance. Non-volatile analysis indicated enhanced accumulation of glycosylated flavonoids, including rutin, kaempferol-3-<i>O</i>-rutinoside, and quercetin-3-<i>O</i>-glucoside, under KIN/2,4-D or NAA conditions, while BAP/2,4-D favored indolic and phenolic derivatives. These results demonstrate that hormonal balance significantly influences biomass production and metabolic profiles in <i>C. spinosa</i> callus cultures, highlighting their potential for sustainable phytochemical production.</p>

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Biotechnological production of secondary metabolites in Capparis spinosa L. through in vitro culture callus induction

  • 期刊论文

  • 发表日期：

    2026年5月29日

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  • Marouane Mohaddab,
  • Younes EL Goumi,
  • Manon Genva,
  • Louise Vilain,
  • Clément Burgeon,
  • Mouna Belkessam,
  • Malika Fakiri,
  • Marie-Laure Fauconnier

摘要

Capparis spinosa L. is a Mediterranean medicinal and aromatic species valued for its flavonoids, glucosinolates, and volatile terpenoids. However, increasing exploitation raises concerns regarding sustainable utilization. An in vitro callus culture system was developed to optimize biomass production and secondary metabolite accumulation. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different combinations of cytokinins (6-benzylaminopurine, BAP; kinetin, KIN) and auxins (2,4-dichlorophenoxyacetic acid, 2,4-D; naphthaleneacetic acid, NAA). Biomass accumulation was evaluated through fresh and dry weight gain. Volatile compounds were analyzed by HS-SPME-GC-MS, whereas non-volatile metabolites were characterized using UPLC-QTOF-MS/MS and semi-quantified by UPLC-TQ-MS/MS. Response surface methodology identified optimal hormonal conditions for callus biomass production after 30 days, corresponding to BAP (1.5-2.0 mg/L) combined with 2,4-D (0.5–1.5 mg/L), yielding fresh and dry weight gains exceeding 3000 mg and 300 mg, respectively. Volatile profiling revealed terpenes and sulfur-containing compounds, with 2,4-D treatments inducing safranal, (E)-rose oxide, and menthol, not detected in leaf tissues. NAA treatments mainly influenced relative volatile abundance. Non-volatile analysis indicated enhanced accumulation of glycosylated flavonoids, including rutin, kaempferol-3-O-rutinoside, and quercetin-3-O-glucoside, under KIN/2,4-D or NAA conditions, while BAP/2,4-D favored indolic and phenolic derivatives. These results demonstrate that hormonal balance significantly influences biomass production and metabolic profiles in C. spinosa callus cultures, highlighting their potential for sustainable phytochemical production.