Agrobacterium - mediated transformation and transgenic plant regeneration from embryogenic calli derived from an immatureseed produced from miniature rose cultivar 'Shortcake'

摘要

The immature seeds were harvested from self - pollinated miniature rose 'Shortcake'. The pale yellow and friable cmbryogcnic calli were induced from an immature seed. The calli showed high frequency (50-60 percent) of transient GUS expression in Agrobacterium - mediated transformation. The population density of Agrobacterium and the sonication assistance treatment had slight effects on the transformation frequency. Among 65 putative transgenic shoots grown in vitro, seven transgenic rose plants havebeen established and flowered in a contained greenhouse. The flowers on the primary shoots showed unstable and quite different characteristics from those on the 2nd or later emerged shoots, which were very similar to the flowers of non - transgenic regenerated plants. The integration of T - DNA fragment in transgenic plants was confirmed by both PCR - Southern blot analysis with a GUS probe and Southern blot analysis with a hygromycin phosphotransferase RNA probe.