ISHS Acta Horticulturae 738: International Symposium on Biotechnology of Temperate Fruit Crops and Tropical Species
TRANSGENIC PLUM (PRUNUS DOMESTICA L.) PLANTS OBTAINED BY AGROBACTERIUM-MEDIATED TRANSFORMATION OF LEAF EXPLANTS WITH VARIOUS SELECTIVE AGENTS |
| Authors: | R.V. Mikhailov, S.V. Dolgov |
| Keywords: | GFP, nptII, hpt, transformation, plum, Prunus domestica |
| Abstract:
Plum (P. domestica L.) scion cv. ‘Startovaya’ was transformed using Agrobacterium tumefaciens strain CBE21:pCamGFP carrying the hygromycin phosphotransferase gene (hpt) and a synthetic codon optimized red-shifted gfp gene; pBin-mGFP5-ER carrying neomycin phosphotransferase gene (nptII) and gfp gene with a leader sequence for expression in endoplasmic reticulum. Whole leaf explants were co-cultivated with A. tumefaciens, and selection and regeneration of transformed cells and shoots was carried out during 6-8 months on selection medium containing 35-50 mg/L kanamycin (Km) or 8 mg/L hygromycin. One of 1207 (0% early / 0.23% delayed selection) explants on kanamycin selection and 10 of 793 (0.27% early / 2.2% delayed selection) explants on hygromycin selection cv. ‘Startovaya’ produced 10/11 GFP and 11/11 PCR-positive independent transgenic lines from the cut midribs via an intermediate callus stage. The selected transformants had a normal phenotype in vitro. | |
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