Strawberries, as one of the small fruits of temperate regions, have been a popular delicious fruit for its flavor, taste, fresh use and become the most important and commercial horticultural produces in recent decades. Strawberry cultivars are highly heterozygote, so every seedling in the population is genetically unique. Thus, the use of achenes is important as a true representative of the plant populations in order to produce seedlings. Since germination of seeds of strawberry cultivars is not uniform, achieving an efficient techniques for uniform seedling production in short-term along with increasing the branch crown numbers are important for the mass production. In this study, in separate experiments, germinability of stratified and Non-stratified achenes of two strawberry cultivars ( Fragaria x ananassa Duch. ‘Selva’ and ‘Camarosa’) by using the cut achenes and embryos as explants, and utilizing Prohexadione – Calcium (Pro - Ca) in various concentrations (0.5, 1.5, 3, 4.5, 6 mM) in order to enhance branch crowns were examined in vitro . in factorial experiments in completely randomized design with four replications, the excised embryos and the cut achene were cultured in MS basal medium lacking hormones. the cultures were stored up to 8 weeks at 27°C. The results indicated that the maximum germination percentage (90%-100%), germination rate (two-three days) and the rate of normal seedling regeneration (80-90 percent) in explants of cut achenes and embryo culture comparing to whole seeds were high and it was clearly observed in two cultivars. Utilizing Prohexadione – Calcium at the concentrations of 1.5, 4.5 and 6 mM obviously increased crown branching in Camarosa cultivar and no effect on Selva cultivar. The number of induced branch crowns in Camarosa cultivar at 4.5 and 6 mM was twice more than of 1.5 mM of applied Pro - Ca . Exogenously applied Pro-Ca had significant effect on developmental indices and decreased the main stem length, main stem diameter and enhanced number of leaves and roots, and wet and dry weight of regenerated seedlings. In conclusion, the main results of this study was confirmed that in vitro application of Pro-Ca as an anti-GA chemical has the potential to differentiate and develop the branch crowns in strawberry that is genotype-dependent, absence of physiological dormancy in strawberry seeds, and identifying the hard seed coats as a preventing agent of uniform seed germination.