Seed dormancy is crucial for plant survival and prevents seed germination out of season. However, little is known about the regulatory mechanism of morphophysiological seed dormancy. Ginkgo biloba L. is one of the most ancient gymnosperms, and the completion of seed germination in this species requires cold and moist stratification. Here, we observed that at the mature seed stage, the embryo was not fully developed in G. biloba seeds. During dormancy stages, the length and weight of the embryo significantly increased, and nutrients accumulated in cotyledons. We further found that abscisic acid (ABA), gibberellic acid (GA), cytokinin and ethylene were integrated in the seed dormancy induction, maintenance and release processes, and GA biosynthesis and signaling transduction specifically act on dormancy release. Combining mRNA and miRNA analyses, we demonstrated that miRNA156 is involved in the regulation of morphophysiological dormancy. Our analyses revealed that G. biloba seed dormancy belongs to the ancestral morphophysiological dormancy type, which is not only regulated by the balance of ABA/GA, but also by other hormones associated with embryo morphological development, as well as genes related to embryo differentiation and development. These findings helped with elucidating the comprehensive regulatory network of morphophysiological dormancy in tree seeds.