Production of transgenic lily plants by Agrobacterium -mediated transformation

Abstract

A system for the production of transgenic plants was developed for the Oriental hybrid lily, Lilium cv. Acapulco, by Agrobacterium-mediated genetic transformation. Filament-derived calli were co-cultivated with A. tumefaciens strain EHA101/pIG121Hm, which harbored a binary vector carrying the neomycin phosphotransferase II, hygromycin phosphotransferase, and intron-containing beta-glucuronidase genes in the T-DNA region. Six hygromycin-resistant (Hyg(r)) culture lines were obtained from 200 calli by scratching them with sandpaper prior to inoculation and using NH(4)NO(3)-free medium for co-cultivation and a hygromycin-containing regeneration medium for selection. Hyg(r) culture lines regenerated shoots, which developed into plantlets following transfer to a plant growth regulator-free medium. All of these plantlets were verified to be transgenic by GUS histochemical assay and inverse PCR analysis.

Efficient genetic transformation and regeneration system from hairy root of Origanum vulgare

“…However, more investigations on the effect of these components are needed to determine how gene transfer is influenced by mineral elements. These results are in agreement with the results obtained by other experiments in other plant species (Azadi et al 2010;Hoshi et al 2004;Sharafi et al 2012). The induced callus via hairy root of O. vulgar (Fig.…”

Somatic embryogenesis and plant regeneration of Lilium ledebourii (Baker) Boiss., an endangered species

“…As such, TCL cultures would also appear to be suitable as a valuable target tissue for Agrobacterium transformation. In some Lilium species and cultivars, genetic transformations have already been performed successfully (Watad et al 1998;Hoshi et al 2004;Mori et al 2005;Kim et al 2007). The procedure described here is efficient and can be applied in programs aimed at conserving this valuable genetic resource.…”

Efficient production of transgenic plantls of Vanda through sonication-assisted Agrobacterium-mediated transformation of protocorm-like bodies

“…1) Slicing: The PLBs were wounded by slicing into 3-4 mm thick explants when they were precultured 2 days before inoculation into fresh medium, to which 100 mM acetosyringone (AS) was added. 2) Scratching: The surface of the PLBs was wounded by using sandpaper (grit number 150, Fuchioka, Japan) attached to the inner surface of plastic centrifuge tubes (50 ml; Iwaki, Japan) (Hoshi et al 2004). The PLBs were transferred into the tubes containing 15 ml liquid NDM and stirred using an automatic lab-mixer (Ikeda Scientific Co., Ltd, Japan) for 30 s. 3) Sonication: PLBs were placed in a mixture of 30 ml liquid NDM and Agrobacterium liquid culture (OD 600 ϭ0.6) at 10 : 1 ratio and subjected to sonication treatment for 5 min at 80% ultrasound power (Transsonic Digital S, Elma, Germany).…”