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<p>This study presents the first reproducible vitrification-based cryopreservation protocol for shoot tips of sycamore maple (<i>Acer pseudoplatanus</i> L.), a tree of high ecological and economic significance in Central Europe. By safeguarding genetically verified material, including rare wood traits, this work supports the integration of the species into long-term <i>ex situ</i> conservation programmes. We investigated the influence of explant size, hardening duration, daytime temperature during hardening, and pre-culture across four genotypes. The protocol involved 4 wk of cold hardening under short-day conditions, a pre-culture at low temperature, vitrification with a plant vitrification solution, storage in liquid nitrogen, rapid rewarming, and gradual recovery under controlled light. Explant size proved decisive: shoot tips of 1–2&#xa0;mm in length regrew at substantially higher rates and developed more vigorous shoots than larger explants. Four weeks of hardening with daytime temperatures between 3 and 7°C followed by pre-culture at 5°C produced the most reliable outcomes. The most responsive genotype reached regrowth in up to 90% of the explants, and for the remaining three genotypes, 50–64% regrowth was achieved. Post-cryopreservation, multiplication factors were comparable to non-cryopreserved control. Hyperhydricity occurred in a genotype-dependent manner but remained low overall. Thus, the protocol enables cryopreservation of sycamore maple genotypes, ensuring both survival and propagation potential.</p>

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First successful vitrification-based cryopreservation of in vitro shoot tips of Sycamore Maple (Acer pseudoplatanus L.): Influence of explant size, hardening, and pre-culture

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  • 出版模式：

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  • 发表日期：

    2026年4月23日

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  • Vitalina Karfik,
  • Andreas Meier-Dinkel,
  • Traud Winkelmann

摘要

This study presents the first reproducible vitrification-based cryopreservation protocol for shoot tips of sycamore maple (Acer pseudoplatanus L.), a tree of high ecological and economic significance in Central Europe. By safeguarding genetically verified material, including rare wood traits, this work supports the integration of the species into long-term ex situ conservation programmes. We investigated the influence of explant size, hardening duration, daytime temperature during hardening, and pre-culture across four genotypes. The protocol involved 4 wk of cold hardening under short-day conditions, a pre-culture at low temperature, vitrification with a plant vitrification solution, storage in liquid nitrogen, rapid rewarming, and gradual recovery under controlled light. Explant size proved decisive: shoot tips of 1–2 mm in length regrew at substantially higher rates and developed more vigorous shoots than larger explants. Four weeks of hardening with daytime temperatures between 3 and 7°C followed by pre-culture at 5°C produced the most reliable outcomes. The most responsive genotype reached regrowth in up to 90% of the explants, and for the remaining three genotypes, 50–64% regrowth was achieved. Post-cryopreservation, multiplication factors were comparable to non-cryopreserved control. Hyperhydricity occurred in a genotype-dependent manner but remained low overall. Thus, the protocol enables cryopreservation of sycamore maple genotypes, ensuring both survival and propagation potential.