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<p>Phenolic oxidation and browning pose significant challenges in coconut cell suspension culture. Although activated charcoal is commonly used to mitigate phenolic interference, its powder form reduces medium clarity, thereby hindering precise quantification. The present study aimed to address this issue by developing an alternative method of incorporating activated charcoal that maintains its functional role while preserving culture transparency. For this, calcium alginate spherules of two sizes (4 and 6&#xa0;mm) were prepared using varying concentrations of sodium alginate and calcium chloride, and their stability in suspension culture conditions was evaluated. Selected spherules were incorporated with activated charcoal and were further tested for polyphenol adsorption capacity in gallic acid solutions and coconut cell suspension cultures. While the spherules of treatment S<sub>1</sub>T<sub>20</sub> (6&#xa0;mm diameter, 3.0% sodium alginate with 0.4&#xa0;M calcium chloride), incorporated with 2.0&#xa0;g L<sup>−1</sup> of activated charcoal, adsorbed 68 to 80% of the polyphenols in coconut cell suspension, those devoid of activated charcoal exhibited no phenol adsorption. Proliferation of the cell and formation of cell clumps occurred in media with activated charcoal (either in the powder form or as incorporated in calcium alginate spherules), whereas the cells elongated and vacuolated in media without activated charcoal. This study introduced, for the first time, an effective alternative to the conventional approach of incorporating activated charcoal powder in suspension cultures using calcium alginate spherules. This approach effectively addressed the challenge of phenolic interference while simultaneously enhancing medium clarity facilitating accurate visualization and quantification of cultured cells. The proposed technology holds promise not only for coconut suspension cultures but also for application across a wide range of plant species where activated charcoal is essential during various stages of <i>in vitro</i> culture. The study provided important insights that can guide future research efforts to enhance the somatic embryogenesis protocol in coconut.</p>

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Effect of charcoal-impregnated calcium alginate spherules in mitigating polyphenolic interference in coconut (Cocos nucifera L.) suspension cultures

  • 期刊论文

  • 发表日期：

    2026年3月11日

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  • Mohamed Neema,
  • Veluru Aparna,
  • Raju Sudha,
  • K. P. Chandran,
  • Fathimath Zaeema,
  • Padmanabhan Sugatha,
  • S. V. Ramesh

摘要

Phenolic oxidation and browning pose significant challenges in coconut cell suspension culture. Although activated charcoal is commonly used to mitigate phenolic interference, its powder form reduces medium clarity, thereby hindering precise quantification. The present study aimed to address this issue by developing an alternative method of incorporating activated charcoal that maintains its functional role while preserving culture transparency. For this, calcium alginate spherules of two sizes (4 and 6 mm) were prepared using varying concentrations of sodium alginate and calcium chloride, and their stability in suspension culture conditions was evaluated. Selected spherules were incorporated with activated charcoal and were further tested for polyphenol adsorption capacity in gallic acid solutions and coconut cell suspension cultures. While the spherules of treatment S1T20 (6 mm diameter, 3.0% sodium alginate with 0.4 M calcium chloride), incorporated with 2.0 g L−1 of activated charcoal, adsorbed 68 to 80% of the polyphenols in coconut cell suspension, those devoid of activated charcoal exhibited no phenol adsorption. Proliferation of the cell and formation of cell clumps occurred in media with activated charcoal (either in the powder form or as incorporated in calcium alginate spherules), whereas the cells elongated and vacuolated in media without activated charcoal. This study introduced, for the first time, an effective alternative to the conventional approach of incorporating activated charcoal powder in suspension cultures using calcium alginate spherules. This approach effectively addressed the challenge of phenolic interference while simultaneously enhancing medium clarity facilitating accurate visualization and quantification of cultured cells. The proposed technology holds promise not only for coconut suspension cultures but also for application across a wide range of plant species where activated charcoal is essential during various stages of in vitro culture. The study provided important insights that can guide future research efforts to enhance the somatic embryogenesis protocol in coconut.