ISHS Acta Horticulturae 673: IX International Symposium on Flower Bulbs
AGROBACTERIUM-MEDIATED TRANSFORMATION OF LILIUM LONGIFLORUM |
| Authors: | Y. Hoshi, M. Kondo, H. Kobayashi, S. Mori, M. Nakano |
| Keywords: | Agrobacterium tumefaciens, Lilium longiflorum, transformed plants, gellan gum, hygromycin |
| Abstract:
We have previously developed a system for Agrobacterium-mediated transformation of the Oriental hybrid lily, Lilium ‘Acapulco’ (Hoshi et al., 2004). Since no transformed plants of L. longiflorum could be obtained with this system, we made some modifications in this system. Following a scratching-treatment of filament-derived calluses of L. longiflorum ‘Georgia’ with sandpaper, they were co-cultivated with A. tumefaciens strain EHA101/pIG121Hm, which harbored the binary vector carrying the neomycin phosphotransferase II (nptII), hygromycin phosphotransferase (hpt), and intron-containing  -glucuronidase (gus-intron) genes in the T-DNA region.
By increasing the concentration of gellan gum in co-cultivation medium from 3 to 10 g L-1, about 3-fold increase was obtained in the number of blue spots resulting from transient expression of the gus gene.
The concentration of hygromycin for selecting putative transformants was decreased from 50 to 35 mg L-1. By using an improved system, 5 hygromycin-resistant (Hygr) culture lines have so far been obtained from 280 co-cultivated calluses of ‘Georgia’. Plantlets were regenerated from all of these Hygr lines, and they were confirmed to be transgenic by inverse PCR analysis and GUS histochemical assay.
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