This study was conducted on nine crosses, covering seven female parent and four male parent, to obtain flavorful seedless grapevine seedlings. The hybrid ovules after field hybridization were cultured in vitro to insure the further development of embryo. Then embryos converted into plant in medium, and the hybrid progenies were detected via molecular marker technology. All the seedlings were acclimatized in greenhouse, and then transplanted in field. The study showed: (1) 365 hybrid seedlings were obtained by means of embryo rescue technique, and 182 of them survived after transplanting into field. (2) Two kinds of genetic molecular markers GSLP1569, SCF272000 were selected for detecting the cross parents and some of hybrid progenies. Form 72 hybrid strains, we detected 8, 38 hybrid strains with seedless gene, respectively. (3) The three seedless cultivars (Ruby Seedless, Flame Seedless and Kunxiang Seedless) were suitable as female parent in embryo rescue, with higher seedling rate. Muscat was proved as the best male parent. (4) The optimal sampling times were 43 days and 51 days after pollination (DAP), with the highest developed embryo rate (6.96％-8.54％ and 16.92％-18.13％) and seedling rate (6.45％ and 8.75％-10％), for the crosses of Flame Seedless and Kunxiang Seedless as female parent. For these crosses, Ruby Seedless × Muscat, Melissa Seedless × Muscat, Perlette × Muscat, the suitable sampling times were gained at 59~60 days, 54 days and 52 days after pollination (DAP), respectively. (5) Embryo abortive time of Crimson Seedless × Muscat kyoho and Fantasy Seedless × Kunxiang Seedless were considered at 39 days and 48 days after pollination (DAP), respectively. Cotyledonshape embryo accounted for the highest seedling rate. Besides the percentage of abnormal embryo was found a significant increase with the extension of the culture time. So the research suggests that ovule culture time should be controlled in 8-10 weeks.