Highly efficient transformation protocol for plum (Prunus domestica L.)

Abstract

A high-throughput transformation system for plum has been developed using hypocotyl slices excised from zygotic embryos as the source of explants. The hypocotyl slices are infected in an Agrobacterium tumefaciens suspension and then cocultivated for 3 days in shoot regeneration ¾ MS basal medium supplemented with 9 μM 2,4-dichlorophenoxyacetic acid. Transgenic shoots are regenerated in a medium containing 7.5 μM thidiazuron and elongated in a medium containing 3 μM benzyladenine in the presence of 80 mg/L kanamycin in both media. Transformed shoots are rooted in ½ MS basal medium supplemented with 5 μM NAA and 40 mg/L kanamycin. The transgenic plants are acclimatized in a growth chamber and transferred to a temperature-controlled greenhouse. This protocol has allowed transformation efficiencies up to 42% and enabled the production of self-rooted transgenic plum plants within 6 months of transformation.