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<p>Vitrification-based cryopreservation holds substantial potential for the large-scale conservation of plant genetic resources. This study investigated the physiological and histological changes in dormant buds of <i>Codonopsis pilosula</i> (Franch.) Nannf. during cryopreservation. At the loading stage, the activity of superoxide dismutase (SOD) and the content of proline were significantly increased, while the activities of catalase (CAT) and peroxidase (POD), as well as the contents of ascorbic acid (AsA) and reduced glutathione (GSH), were markedly decreased. Treatment with Plant Vitrification Solution 2 (PVS2) and freeze-thawing resulted in SOD activity, but had negligible effects on the other aforementioned physiological indicators. Plasmolysis was induced by the loading treatment and was further aggravated after PVS2 exposure. Surviving cells were predominantly distributed in the apical meristem, leaf primordia and the upper segment of the bud axis. Liquid nitrogen (LN) cooling and subsequent freeze-thawing caused extensive cellular damage in the outer and lower segments of the bud axis. In addition, inter-simple sequence repeat (ISSR) markers were employed for the preliminary verification of genetic stability, and no morphological or genomic deoxyribonucleic acid (DNA) variations were detected among the regenerated clones. Collectively, these findings deepen the understanding of the physiological and histological mechanisms underlying the cryopreservation of <i>Codonopsis pilosula</i> dormant buds, and provide a theoretical and technical basis for optimizing cryopreservation protocols for other perennial herbaceous plants.</p>

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Cryopreservation by PVS2-vitrification causes physiological and histology injuries to dormant buds of Codonopsis pilosula (Franch.) Nannf. while cryo-derived plantlets maintain the genetic stability

  • 期刊论文

  • 发表日期：

    2026年3月3日

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  • Shunli Tang,
  • Yanhong Zhang,
  • Sufang Gao,
  • Chunyu He,
  • Qingyi Guo,
  • Jinxiu Wang,
  • Sanbao Shen

摘要

Vitrification-based cryopreservation holds substantial potential for the large-scale conservation of plant genetic resources. This study investigated the physiological and histological changes in dormant buds of Codonopsis pilosula (Franch.) Nannf. during cryopreservation. At the loading stage, the activity of superoxide dismutase (SOD) and the content of proline were significantly increased, while the activities of catalase (CAT) and peroxidase (POD), as well as the contents of ascorbic acid (AsA) and reduced glutathione (GSH), were markedly decreased. Treatment with Plant Vitrification Solution 2 (PVS2) and freeze-thawing resulted in SOD activity, but had negligible effects on the other aforementioned physiological indicators. Plasmolysis was induced by the loading treatment and was further aggravated after PVS2 exposure. Surviving cells were predominantly distributed in the apical meristem, leaf primordia and the upper segment of the bud axis. Liquid nitrogen (LN) cooling and subsequent freeze-thawing caused extensive cellular damage in the outer and lower segments of the bud axis. In addition, inter-simple sequence repeat (ISSR) markers were employed for the preliminary verification of genetic stability, and no morphological or genomic deoxyribonucleic acid (DNA) variations were detected among the regenerated clones. Collectively, these findings deepen the understanding of the physiological and histological mechanisms underlying the cryopreservation of Codonopsis pilosula dormant buds, and provide a theoretical and technical basis for optimizing cryopreservation protocols for other perennial herbaceous plants.