Abstract
We have established a stable genetic transformation method for Hinoki cypress by Agrobacterium--mediated gene transfer system. Embryogenic tissues are co-cultivated with disarmed Agrobacterium tumefaciens strain C58/pMP90 containing a binary vector pBIN19-sgfp, and the infected tissues were screened by both nptII selectable marker and GFP fluorescence. The transformation frequency varied up to 22.5 independent transgenic lines isolated per petri dish that include 250 mg of embryogenic tissues. Eight independent transgenic lines were regenerated through maturation and germination steps of developing somatic embryo and their intensity of GFP fluorescence was observed under a fluorescence microscope. The 35S promoter induced strong GFP fluorescence in embryogenic tissues and roots, but showed quite low intensity in leaves and stems. In searching for constitutive promoters, we are producing transgenic lines expressing a gfp gene under the control of maize ubiquitin promoter.
