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<p><i>Microchirita involucrata</i> is a promising source of bioactive secondary metabolites. This study aimed to establish an efficient in vitro root culture system for <i>M. involucrata</i> through optimization of culture conditions and to evaluate the effect of β-cyclodextrin (CD) elicitation on secondary metabolite production and biological activities. Semi-solid 0.5 MS medium was optimal for root induction, yielding the highest biomass (3.37 ± 0.20&#xa0;g), while supplementation with 30&#xa0;g/L sucrose further enhanced root elongation and fresh biomass (4.75 ± 0.32&#xa0;g). The integration of optimized culture conditions with CD elicitation significantly enhanced the production of bioactive metabolites. The culture medium extract obtained from CD-elicited roots grown on semi-solid 0.5 MS medium supplemented with 4&#xa0;mg/L indole-3-butyric acid (IBA) and 30&#xa0;g/L sucrose exhibited strong antioxidant activity (ABTS: 25.31 ± 4.56 µmol Trolox/L; FRAP: 6.86 ± 2.21&#xa0;mg ascorbic acid/L), together with elevated total phenolic (9.41 ± 1.35&#xa0;mg gallic acid/L) and flavonoid contents (14.37 ± 1.43&#xa0;mg quercetin/L). HPLC analysis revealed enhanced accumulation of anthraquinone derivatives following CD elicitation. Notably, this study provides the first evidence of cytotoxic activity associated with culture medium extracts of <i>M. involucrata</i>. The CD-elicited extract demonstrated lower IC₅₀ values in colorectal cancer cells (HCT116; 182.23 ± 44.33&#xa0;µg/mL) and lung cancer cells (A549; 239.43 ± 15.82&#xa0;µg/mL) compared to normal keratinocytes (HaCaT; 400.77 ± 36.79&#xa0;µg/mL), suggesting greater sensitivity of cancer cells than normal keratinocytes under the tested conditions. These findings demonstrate the effectiveness of combining culture optimization with CD elicitation to enhance biomass accumulation, bioactive metabolite production, and biological activities in <i>M. involucrata</i>.</p> Graphical Abstract <p></p>

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Integrated root culture optimization and cyclodextrin elicitation enhance bioactive metabolite production and biological activities in Microchirita involucrata

  • 期刊论文

  • 发表日期：

    2026年7月30日

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  • Adsadayu Thonnondang,
  • Onrut Sapatee,
  • Anupan Kongbangkerd,
  • Runglatda Srisawang,
  • Suchada Phimsen,
  • Wipaporn Chuaymaung,
  • Apinun Limmongkon

摘要

Microchirita involucrata is a promising source of bioactive secondary metabolites. This study aimed to establish an efficient in vitro root culture system for M. involucrata through optimization of culture conditions and to evaluate the effect of β-cyclodextrin (CD) elicitation on secondary metabolite production and biological activities. Semi-solid 0.5 MS medium was optimal for root induction, yielding the highest biomass (3.37 ± 0.20 g), while supplementation with 30 g/L sucrose further enhanced root elongation and fresh biomass (4.75 ± 0.32 g). The integration of optimized culture conditions with CD elicitation significantly enhanced the production of bioactive metabolites. The culture medium extract obtained from CD-elicited roots grown on semi-solid 0.5 MS medium supplemented with 4 mg/L indole-3-butyric acid (IBA) and 30 g/L sucrose exhibited strong antioxidant activity (ABTS: 25.31 ± 4.56 µmol Trolox/L; FRAP: 6.86 ± 2.21 mg ascorbic acid/L), together with elevated total phenolic (9.41 ± 1.35 mg gallic acid/L) and flavonoid contents (14.37 ± 1.43 mg quercetin/L). HPLC analysis revealed enhanced accumulation of anthraquinone derivatives following CD elicitation. Notably, this study provides the first evidence of cytotoxic activity associated with culture medium extracts of M. involucrata. The CD-elicited extract demonstrated lower IC₅₀ values in colorectal cancer cells (HCT116; 182.23 ± 44.33 µg/mL) and lung cancer cells (A549; 239.43 ± 15.82 µg/mL) compared to normal keratinocytes (HaCaT; 400.77 ± 36.79 µg/mL), suggesting greater sensitivity of cancer cells than normal keratinocytes under the tested conditions. These findings demonstrate the effectiveness of combining culture optimization with CD elicitation to enhance biomass accumulation, bioactive metabolite production, and biological activities in M. involucrata.

Graphical Abstract