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The effects of carbon source and concentration and of seedcoat were tested on the in vitro germination of peach seeds derived from crosses performed in the field. Seeds were extracted from the fruit and cultured in Woody Plant Medium (WPM) supplemented with sucrose, glucose, or sorbitol at concentrations of 15, 30, and 45 g·L−1. The percentage of germination as well as the root and hypocotyl lengths were measured after the stratification process and before acclimatization. Seedcoat did not have any influence on seed germination in any tested media and genotype. Glucose at a concentration of 15 g·L−1 and sucrose at 15, 30, and 45 g·L−1 resulted in greater stem seedling growth. The root developed the most when seeds were cultured in media with 15 or 30 g·L−1 of sucrose.