Efficient genetic transformation is of great potential for watermelon research and breeding. By overexpressing a chimeric gene fused by endogenous ClGRF4 and ClGIF1, we were able to achieve highly efficient transformation in watermelon. Mutating the miR396 microRNA targeting site inside ClGRF4 gene can further boost the transformation efficiency up to 67.27% in watermelon in a genotype-independent manner. Chimeric ClGRF4-GIF1 can also be combined with CRISPR/Cas9 genome editing tool to achieve highly efficient gene editing in watermelon, by which we successfully created seedless watermelon at diploid level. Here, we have established a powerful transformation tool for future watermelon research and breeding.

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