Direct introduction of foreign gene in higher plants

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In order to introduce conventionally foreign gene to higher plants, I tried graft method using transgenic tobacco plants which contained kanasycin resistant and nopaline-synthase genes as stock. Non-transgenic normal tobacco plants were grafted on the transgenic tobacco ones. 6 km resistant plants in more than 33000 (0.08%) from the selfed seeds of scion capsules were selected on kanamycin medium (100ug/ml). Four graft transformants had one km gene in the respective one of two homologous chromesomes, one had km genes in homozygous state and the other one had more than 2 km genes (probably 5) in independent state on its chromosomes from the results of test cross experiment. Southern blot analysis of the graft transformants and the progenies show the existence of the maker km and Nos genes and complicated band patterns.It is concluded that the graft transformation is hopeful as a conventional method of gene introduction in higher plant. However, it is further necessary to make clear the mechanism and increase the efficiency.