This project aimed at breeding new seedless grape cultivars by embryo rescue through three hybridization methods: 1) using cross-breeding between seedless Vitis vinifera cultivars and wild Chinese Vitis spp; 2) crossing with two seedless cultivars, and 3) hybridization between grapes of different ploidy. Genotype, sampling times, and media were confirmed to play important roles in this system. Among the different genotypes, the productions of hybrid plants were significantly different, ranging from 23.0% (Ruby Seedless x Black Olympia) to only 1.1% (Pink Seedless x Beichun), except for the combinations from which no surviving seedlings were obtained. We got the best sampling times, in days after flowering (DAF), from the following different combinations: ‘Flame Seedless x Beichun’ (39 DAF); ‘Blush Seedless x Shuangyou’ (54 DAF); ‘Pink Seedless x Beichun’ (54 DAF); ‘DA7 x Shuangyou’ (44 DAF); ‘Blush Seedless x Thompson Seedless (54 DAF)’; ‘Pink Seedless x Flame Seedless’ (54 DAF); ‘DA7 x Blush Seedless’ (44 DAF); ‘Ruby Seedless x Black Olympia’ (63 DAF); ‘DA7 x Jingyou’ (44 DAF); ‘Flame Seedless x Fujiminori’ (39 DAF), and ‘Big Black x Kyoho’ (72 DAF). The highest rates of embryo formation (13.2%) and plant development (90.1%) were found when ovules were cultured in MM4 with 500 mg/L mashed banana. Conversely, they were reduced by addition of plant growth regulators. Seven new hybrids were successfully obtained. As a result of early nuclear-free character and ploidy level identification, 11 seedless grape lines, and 3 triploid and 2 haploid grape lines were obtained.

Key words: Grapevine, Embryo rescue, Seedless grapes, Stenospermocarpy, Hybridization.