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(European beech) is a dominant hardwood forest tree species across Central Europe, supporting diverse ecosystem services and forming the basis of a significant market for high-value timber. However, climate change increasingly threatens beech vitality and productivity, making molecular insights into its stress resilience and functional validation of underlying genes urgently needed. Here, we report a protocol for protoplast isolation from seedling leaves and demonstrate transient genetic transformation and CRISPR\/Cas-mediated genome editing in\n                    <jats:italic>F. sylvatica<\/jats:italic>\n                    . PEG-mediated transformation was sequentially optimized, achieving efficiencies of 59\u2009\u00b1\u20096.19% within distinct seasonal windows. Protoplast yield and transformation efficiency showed pronounced temporal variation throughout the year, indicating a strong seasonal influence on reproducibility of the workflow despite controlled growth conditions. A basic molecular toolkit for functional genomics and future biotechnological applications was established by testing a set of promoters and reporters. For proof-of-concept genome editing, we achieved 4.75 to 32.69% editing efficiencies in the\n                    <jats:italic>PHYTOENE DESATURASE<\/jats:italic>\n                    gene (\n                    <jats:italic>FsPDS<\/jats:italic>\n                    ) using temperature-tolerant\n                    <jats:italic>Lb<\/jats:italic>\n                    Cas12a (tt\n                    <jats:italic>Lb<\/jats:italic>\n                    Cas12a). Although further optimization of transformation reproducibility and regeneration systems remains necessary, the presented protoplast platform provides a valuable foundation for transient functional assays and genome editing studies in this non-model tree species.\n                  <\/jats:p>","DOI":"10.1038\/s42003-026-10805-9","type":"journal-article","created":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T09:46:04Z","timestamp":1787737564000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Genetic transformation and CRISPR\/Cas12a-mediated gene editing of European beech (Fagus sylvatica L.) employing a transient protoplast system"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-1473-9311","authenticated-orcid":false,"given":"Virginia","family":"Zahn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-0591-7176","authenticated-orcid":false,"given":"Alice-Jeannine","family":"Sievers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9900-9133","authenticated-orcid":false,"given":"Birgit","family":"Kersten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9301-8581","authenticated-orcid":false,"given":"Matthias","family":"Fladung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9823-5630","authenticated-orcid":false,"given":"Tobias","family":"Bruegmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,26]]},"reference":[{"key":"10805_CR1","first-page":"62","volume":"70","author":"Z Fuchs","year":"2024","unstructured":"Fuchs, Z. et al. 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