Genetic transformation of korean chrysanthemum (Dendranthema zawadskii x D. x grandiflorum) has been successfully achieved by co-cultivation with Agrobacterium tumefaciens strain EHA101 carrying the plasmid pIGP3 containing the maize transposable element Ac and the nptII gene as selectable marker. Selection with 100 mg l-1 kanamycin led to a transformation frequency of 7.8% with respect to co-cultivated explants. The nptII gene was detected by PCR analysis in transgenic regenerants and in the 4 progenies achieved. Southern blot analysis confirmed the presence of Ac in both the primary transformants and in their progeny. The restriction analysis of DNA with the methylation sensitive enzyme PvuII showed that the transposon was in a non-methylated status at the PvuII sites. We hypothesize that the Ac element is inserted in an active form and Ac might be successfully employed as gene tagging system in korean chrysanthemum and related diploid species.

Transformation of Korean chrysanthemum (Dendranthema zawadskii x grandiflorum) and insertion of the maize autonomous element Ac using Agrobacterium tumefaciens

DELLEDONNE, Massimo;FURINI, Antonella;
2000-01-01

Abstract

Genetic transformation of korean chrysanthemum (Dendranthema zawadskii x D. x grandiflorum) has been successfully achieved by co-cultivation with Agrobacterium tumefaciens strain EHA101 carrying the plasmid pIGP3 containing the maize transposable element Ac and the nptII gene as selectable marker. Selection with 100 mg l-1 kanamycin led to a transformation frequency of 7.8% with respect to co-cultivated explants. The nptII gene was detected by PCR analysis in transgenic regenerants and in the 4 progenies achieved. Southern blot analysis confirmed the presence of Ac in both the primary transformants and in their progeny. The restriction analysis of DNA with the methylation sensitive enzyme PvuII showed that the transposon was in a non-methylated status at the PvuII sites. We hypothesize that the Ac element is inserted in an active form and Ac might be successfully employed as gene tagging system in korean chrysanthemum and related diploid species.
genetic transformation; korean chrysanthemum
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/305523
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