ASHS 2025 Conference

Cucumber (Cucumis sativus L.) is an economically important crop and is widely cultivated throughout the world. Cucumber plants often suffer from biotic and abiotic stresses during the whole development life cycle, which lead to reduction in yield and quality. Improvement of cucumber for disease, insect, or nematode resistance and other horticultural traits with conventional strategy is limited by long breeding cycle, narrow genetic basis, and severe incompatibility barriers in related species. Emerging plant genome editing techniques provide trait specific breeding for enhancement of plant yield, quality, stress tolerance, and disease resistance. Highly efficient regeneration and transformation system is a prerequisite for cucumber genome editing. We report an efficient Agrobacterium mediated cucumber CRISPR-Cas9 transformation system with the aid of GFP visual selection. Cotyledons from 7 days old in vitro seedlings were harvested, and inoculated with Agrobacterium tumefaciens strain GV3101 contains a binary vector with CRISPR-Cas9 gene, GFP visual selection maker and hygromycin resistance genes. Transgenic callus and shoots obtained with GFP visual selection with high efficiency. PCR double check confirmed transgenes in transgenic plants. Transgenic plants are phenotyping in the greenhouse.