ISHS Acta Horticulturae 392: Genetic Improvement of Horticultural Crops by Biotechnology
GENETIC TRANSFORMATION OF EUSTOMA GRANDIFLORUM WITH ROL GENES |
| Authors: | T. Handa, T. Sugimura, E. Kato, H. Kamada, K. Takayanagi |
| Keywords: | Eustoma grandiflorum, rol genes, Agrobacterium, Genetic transformation |
| Abstract:
Hairy roots were obtained from prairie gentian (Eustoma grandiflorum (Griseb.)Schinners) infected with Agrobacterium rhizogenes. Plant regeneration occurred spontaneously on Linsmaier and Skoog (LS) medium lacking phytohormones under the light condition. Phenotypic alterations such as dwarfism, wrinkled leaves and cup-shaped corolla were observed in transformed plants. Southern analysis revealed the introduction of Ri-T-DNA in the plant genome. Transformed plants showed reduced fertility but some plants set viable seeds. The progenies also exhibited the dwarf phenotype and retained the Ri-T-DNA. In addition, some progenies expressed different forms and phyllotaxis in comparison with their parents. Isolated rol genes were reconstructed with promoter sequences and introduced into the binary vector, pBI121. Agrobacterium-mediated transformation was performed to Eustoma cultivars by using different vectors constructed with the combination of promoters and rol genes. Transformed plantlets were regenerated from leaf disks on the selective medium with kanamycin (km). PCR analysis confirmed the transforming events. | |
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