Agrobacterium-mediated genetic transformation of Prunus salicina

Abstract

We report Agrobacterium tumefaciens-mediated transformation of two Prunus salicina varieties, ‘Angeleno’ and ‘Larry Anne’, using a modification of the hypocotyl slice technique previously described for P. domestica. Regeneration rates on thidiazuron (TDZ) and indole-3-butyric acid (IBA) supplemented Murashige and Skoog (MS) media reached 11% for ‘Angeleno’ and 19% for ‘Larry Anne’ hypocotyl slices. Transformation using Agrobacterium tumefaciens GV3101 harboring a plasmid with the neomycin phosphotransferase II (nptII) and the green fluorescent protein (gfp) genes produced ten independent lines, six from ‘Angeleno’ and four from ‘Larry Anne’, representing transformation efficiencies of 0.8 and 0.3%, respectively, relative to the initial number of hypocotyl slices. Plants of six lines were found to produce the transgene encoded mRNAs. DNA blotting demonstrated the presence of transgene sequences in trees from five lines after 18 months of growth in the greenhouse.