Akademik Veri Yönetim Sistemi
8th International Eurasian Conference on Biological and Chemical Sciences (EurasianBioChem 2025), Ankara, Türkiye, 17 - 19 Aralık 2025, ss.59, (Özet Bildiri)
Lettuce (Lactuca sativa L.) is an economically and nutritionally important vegetable cultivated worldwide, yet its susceptibility to biotic and abiotic stresses necessitates genetic improvement through biotechnological tools. Agrobacterium tumefaciens-mediated transformation is a key approach for genetic modification in lettuce. This study evaluated the influence of nine lettuce cultivars (Caipira, Emocion, Festival, Funtime, Garone, Maritima, Rüzgar, Sürpriz, and Vaidosa) and two A. tumefaciens strains (LBA4404 and EHA105) on transformation efficiency using cotyledon explants. A binary vector (pDIRECT_23C, 15,744 bp) carrying CRISPR-Cas9 components and a phosphinothricin (PPT) resistance marker was used as a model to assess DNA delivery, without targeting any specific genomic sequence. Mature seeds were surface sterilized and germinated in vitro. Cotyledon explants from seven-day-old seedlings were pre-cultured, inoculated with Agrobacterium suspensions (OD600 = 0.8–1.0), co-cultured in the dark for two days, and transferred to PPT-based selection and regeneration medium. Regenerated shoots were rooted and acclimatized under controlled greenhouse conditions. PCR analysis confirmed transgene integration using BlpR-specific primers. Statistical analysis revealed that EHA105 significantly outperformed LBA4404 in transformation efficiency (p < 0.05). Among cultivars, Emocion exhibited the highest number of transgenic shoots, with a transformation efficiency of 5%, followed by Caipira. The relatively large size of the pDIRECT_23C vector demonstrates its suitability as a model for future genome editing studies, where guide RNAs can be inserted for targeted modifications. This study optimized an efficient Agrobacterium-mediated transformation protocol for multiple lettuce cultivars, providing a foundation for functional genomics research and biotechnological improvement. The findings contribute to enhancing transformation efficiency in lettuce, facilitating future CRISPR-based genome editing and crop improvement applications.