Fragaria vesca L. has become a model species for genomic studies relevant to important crop plant species in the Rosaceae family, but generating large numbers of plants from non-runner-producing genotypes is slow. To develop a protocol for the rapid generation of plants, leaf explants were compared to single axillary bud shoot explants, both from in vitro-grown Fragaria vesca seedlings, as sources of shoots for new plant production in response to benzyladenine (BA) or thidiazuron (TDZ) combined with indolebutyric acid (IBA) on Murashige and Skoog’s Basal Salt (MS) medium. BA at 2.0 and 4.0 mg L−1 and TDZ at 1.5 mg L−1 promoted the greatest number of shoots produced per shoot explant. There were no IBA effects or IBA interactions with BA or TDZ. Significant interactions between BA and IBA, but not TDZ and IBA, occurred in leaf explant callus formation and % explants with callus at 6 and 9 weeks of culture and on shoots per leaf explant at 9 weeks. TDZ treatments produced uniformly high levels of callus but low numbers of shoots. The treatment generating the most shoot production was BA at 4.0 mg L−1 plus IBA at 0.50 mg L−1. After 9 weeks of culture, leaf explants of the non-runner-producing genotype Baron Solemacher had generated 4.6 shoots per explant with the best treatment, while axillary bud explants had generated 30.8 shoots with the best treatment. Thus, in vitro culture of shoot axillary bud explants can generate high numbers of clonal shoots from a single seedling plant in vitro
