The development of seedless cultivars is a primary goal in grapevine breeding. Since grapes are stenospermic and tend to abort their embryos before development, traditional breeding methods often yield seedless cultivars at a low frequency. Therefore, embryo rescue has emerged as a promising approach for creating seedless grape cultivars. This study aimed to optimize the ideal sampling time and protocol for efficient embryo rescue in grapes. Ovules from immature berries collected at various days after pollination (20DAP, 30DAP, 40DAP, and 50DAP) were cultured on Nitsch and Nitsch (NN) medium with different concentrations of Benzyl aminopurine (BAP) (0.1, 0.2, and 0.5 mg/L) and Activated Charcoal (AC) (1.5, 2, and 2.5 g/L). Several parameters were assessed, including maximum ovule growth, the percentage of enlarged ovules, percentage of collapsed ovules, callus formation percentage, and embryo germination percentage. The study’s results indicated that berries collected at 40 DAP yielded the best outcomes across all parameters. Regarding treatments, the most favorable results were achieved when ovules were cultured on NN medium supplemented with 0.5 mg/L BAP and 2 g/L AC. In conclusion, the study underscores the significance of choosing the right sampling time and treatments to ensure efficient embryo rescue in grapes. The protocol standardized from this research is recommended for effectively rescuing embryos and developing seedless hybrids.

Activated charcoal, Benzyl aminopurine, Embryo rescue, Grape, Nitsch, Ovule culture