Poster: Emerging Technologies

Abs # 886: Genetic transformation of soutooth oak (Quercus acutissima) somatic embryo

Presenter:	Ohmiya, Yasunori , yohmiya@affrc.go.jp
Authors	Ohmiya, Yasunori  (A)   Taniguchi, Toru  (A)   Okamura, Masanori  (A)
Affiliations:	(A): Forest Tree Breeding Center

Soutooth oak tree(Quercus acutissima Carruth.) is one of the important breeding species that is commonly available for fuels(charcoal), or bed logs for the cultivation of Shiitake mushroom, and also value as furniture materials because of their superior timber quality. However, their growth is relatively slow and difficult to proliferate by cutting. Introducing foreign genes of interest might be excellent and advanced techniques to improve their growth. In this study, we showed Quercus acutissima somatic embryos are infectable by Agrobacterium. Quercus acutissima somatic embryo was infected for 15 min with a suspension of A. tumefaciens strain EHA105 containing the binary vector pBIN19-sgfp, which harbors the green fluorescent protein driven by the CaMV35S promoter, and NPTII gene as a selectable marker, and co-cultivated for 2 days on 1/2 MS, 100mM acetosyringone. Transgenic embryos were isolated after 3 months cultivation on 1/2 MS supplemented with 50mg/l kanamycin and 300mg/l carbenicillin, with green fluorescence as a landmark. Integration of their genes was confirmed by PCR and Southern hybridization analyses, and 16 transgenic lines were identified from 300 of infected embryos. They were maintained and multiplied by secondary embryogenesis for additional 3 months and then transferred to the germination medium (1/2 WPM, 1% sucrose) . Regenerated transgenic plants were obtained after 2 months of further cultivation and finally confirmed by their fluorescence of GFP proteins in each organ.