In vitro propagation and conservation of wild bilberry (Vaccinium myrtillus L.) genotypes collected in the Tuscan Apennines (Italy)

BACKGROUND: Vaccinium myrtillus L. is a valuable wild fruit species of conservation concern in the Tuscan Apennines (Italy). OBJECTIVE: The development of a micropropagation protocol for the conservation of this species in in vitro-banks for the year-round mass production of specific selected superior bilberry genotypes, maintaining the genetic variability of natural populations, avoiding genetic erosion. METHODS: Nodal segments from seedlings were cultured on mMS medium with zeatin or 2iP for multiplication and elongation stage, afterwards shoots were rooting in vitro and in vivo. The RAPD analysis was applied to lines obtained and slow growth storage (4 degrees C - darkness) was evaluated for the conservation. RESULTS: Zeatin concentration 9.1 mu M was the most favorable for shoot proliferation. The activated charcoal was essential for in vitro rooting. Both in vitro and ex vitro methods were successfully used to root and acclimatize bilberry shoots. The RAPD analysis indicated a large genetic diversity among the micropropagated genotypes. Shoots were successfully in vitro preserved for up to 6 months with no apparent deterioration effect. CONCLUSIONS: In vitro propagation and conservation are fundamental tools for exploiting the whole potential of this wild crop species for breeding and genetic improvement purposes.