Agrobacterium-mediated transformation of lavandin (Lavandula x intermedia Emeric ex Loiseleur)
Résumé
Lavandin (Lavandula x intermedia Emeric ex Loiseleur) is an aromatic plant, the essential oil of which is widely used in the perfume, cosmetic, flavouring and pharmaceutical industries. The qualitative or quantitative modification of its terpenes-containing essential oil by genetic engineering could have important scientific and commercial applications. In this study, we report the first Agrobacterium tumefaciens-mediated gene transfer into lavandin. The transformation protocol was optimized by lengthening precultivation and cocultivation periods and by testing five different bacterial strains. We obtained transformed callus lines at a frequency of 40-70% with strains AGL1/GI, EHA105/GI and C58/GI. Transgenic shoots were regenerated from these kanamycin resistant calli and rooted on selective medium with 150 mg l −1 kanamycin. The final percentage of transgenic plants obtained varied from 3 to 9%, according to the Agrobacterium strain used, within 6 months of culture. The presence of the introduced β-glucuronidase and neomycin phosphotransferase II genes was shown both by PCR and Southern blot analysis. Transgene expression was investigated using histoenzymatic β-glucuronidase assays, leaf callus assays and RT-PCR. Results showed that both β-glucuronidase and neomycin phosphotransferase II genes were expressed at a high level in at least 41% of the transgenic plants regenerated. This efficient transformation strategy could be used to modify some genetic traits of lavandin (flower colour, pathogens resistance) and to study the biosynthesis of the major monoterpene components of its essential oil (linalool, linalyl acetate, camphor and 1,8-cineole).
Origine : Accord explicite pour ce dépôt
Loading...