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Background <p><i>Cyclocarya paliurus</i> (Batal.) Iljinskaja is a medicinal and endangered woody plant, but shoot culture is challenged by higher contamination and lower propagation efficiency.</p> Results <p>This study evaluated the effects of various sterilization and propagation treatments on shoot culture, and further analyzed enzymatic activity and flavonoid metabolomics at 20, 40, 60, and 80 days of culture using UPLC-MS. The optimal disinfection method-cold storage at 4&#xa0;°C for 8 days, followed by immersion in warm water at 37&#xa0;°C, and sterilization with 0.1% mercuric chloride for 5&#xa0;min-resulted in a contamination rate of 12.72%. The highest propagation coefficient (5.91-fold) was achieved on ½ MS medium supplemented with 0.5&#xa0;mg/L 6-BA, 0.01&#xa0;mg/L TDZ, 0.3&#xa0;mg/L GA₃ and 0.02&#xa0;mg/L IBA at 60 days. Enzymatic activity followed the order: SOD &gt; POD &gt; PPO &gt; PAL. A total of 151 flavonoid-related metabolites were identified and classified into 11 categories, including flavonols (28.48%), flavones (18.54%), flavanones (10.60%), benzoic acid and its derivatives (6.62%), flavanols (5.96%), flavanonols (5.30%), isoflavanones (4.64%) and others. Metabolite content varied with culture duration, with catechin (a flavanol) showing the highest content (&gt; 1000&#xa0;µg/g) during shoot culture.</p> Conclusion <p>Pretreatment of “lower-temperature storage and warm-water immersion” efficiently reduced stem contamination, and significant associations existed between enzymatic activity, flavonoid profiles, and the propagation of adventitious shoot. These findings offer a promising strategy to improve shoot propagation, in vitro conservation and utilization in <i>C.</i> species.</p>

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Optimizing disinfection and adventitious shoot propagation in Cyclocarya paliurus stem tissue culture: enzymatic and flavonoid metabolomics insights

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月1日

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  • Ying Feng,
  • Yongshan Chen,
  • Zhixiong Guo,
  • Jinghua Xu

摘要

Background

Cyclocarya paliurus (Batal.) Iljinskaja is a medicinal and endangered woody plant, but shoot culture is challenged by higher contamination and lower propagation efficiency.

Results

This study evaluated the effects of various sterilization and propagation treatments on shoot culture, and further analyzed enzymatic activity and flavonoid metabolomics at 20, 40, 60, and 80 days of culture using UPLC-MS. The optimal disinfection method-cold storage at 4 °C for 8 days, followed by immersion in warm water at 37 °C, and sterilization with 0.1% mercuric chloride for 5 min-resulted in a contamination rate of 12.72%. The highest propagation coefficient (5.91-fold) was achieved on ½ MS medium supplemented with 0.5 mg/L 6-BA, 0.01 mg/L TDZ, 0.3 mg/L GA₃ and 0.02 mg/L IBA at 60 days. Enzymatic activity followed the order: SOD > POD > PPO > PAL. A total of 151 flavonoid-related metabolites were identified and classified into 11 categories, including flavonols (28.48%), flavones (18.54%), flavanones (10.60%), benzoic acid and its derivatives (6.62%), flavanols (5.96%), flavanonols (5.30%), isoflavanones (4.64%) and others. Metabolite content varied with culture duration, with catechin (a flavanol) showing the highest content (> 1000 µg/g) during shoot culture.

Conclusion

Pretreatment of “lower-temperature storage and warm-water immersion” efficiently reduced stem contamination, and significant associations existed between enzymatic activity, flavonoid profiles, and the propagation of adventitious shoot. These findings offer a promising strategy to improve shoot propagation, in vitro conservation and utilization in C. species.