|Petri, Cesar -|
Submitted to: Transgenic Plants: Methods and Protocols
Publication Type: Book / Chapter
Publication Acceptance Date: May 26, 2011
Publication Date: January 9, 2012
Citation: Petri, C., Scorza, R., Srinivasan, C. 2012. Highly efficient transformation protocol for plum (Prunus domestica L.). In: Dunwell, J. M. and Wetten, A. C., editors. Transgenic Plants: Methods and Protocols. New York, NY:Humana Press. p. 191-200. Technical 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 co-cultivated for 3 days in shoot regeneration three-quarter MS basal medium supplemented with 9 micrometer 2.4-dichlorophenoxyacetic acid. Transgenic shoots are regenerated in a medium containing 7.5 micrometer thidiazuron and elongated in a medium containing 3 micrometer benzyladenine in the presence of 80 mg l-1 kanamycin in both media. Transformed shoots are rooted in one-half MS basal medium supplemented with 5 micrometer NAA and 40 mg l-1 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 percent and enabled the production of self-rooted transgenic plum plants within 6 months of transformation.