Optimised Agrobacterium-Mediated Transformation and Application of Developmental Regulators Improve Regeneration Efficiency in Melons

Melon (Cucumis melo L.) is a protected crop in China with high economic value. Agrobacterium-mediated genetic transformation powerful tool to improve agronomic traits and obtain elite germplasm. However, current protocols melons are inefficient highly genotype-dependent. To melon, we tested different infiltration methods for transformation. Among these methods, micro-brushing sonication 20 s, followed by vacuum at −1.0 kPa 90 resulted the strongest green fluorescent protein signal increased proportion of infected explants. We transformed melon developmental regulatory genes AtGRF5, AtPLT5, AtBBM, AtWUS, AtWOX5, AtWIND1 from Arabidopsis estimated regeneration frequencies as number regenerating shoots/total inoculated explants selection medium. The overexpression AtGRF5 AtPLT5 efficiencies 42.3% 33% ZHF 45.6% 32.9% Z12, respectively, which were significantly higher than those control. expression cassettes added CRISPR/Cas9 genome-editing vectors transgenic phytoene desaturase CmPDS knockout mutants. Using or multi-allelic mutations observed target sites recalcitrant genotypes. This strategy enables genotype-flexible promotes precise genome modification technologies melons.