Inhibition of agrobacterial oncogene expression by means of antisense RNAs

Infection of plants by soil bacterium Agrobacterium tumefaciens induces tumors referred to as crown galls. Tumor development is determined by the introduction of agrobacterial genes governing phytohormone (auxin and cytokinin) production into the plant genome. The most important of these genes are iaaM and ipt. Development of transgenic plants inhibiting the expression of these genes allows a raise of varieties resistant to crown gall disease. For this purpose, single and double tobacco transformants with antisense copies of iaaM and ipt fused with single and double promoters for the 35S RNA of the cauliflower mosaic virus (CaMV 35S and CaMV 35SS) were obtained. Inoculation of transgenic plants harboring the antisense oncogene copies with virulent A. tumefaciens strains C58 (pTiC58) and A6 (pTiA6) revealed significant, but still incomplete, inhibition of these genes. Agrobacterium-mediated transformation of transgenic plants gave rise to weakened tumors, which varied in morphology and allowed regeneration of whole plants. Analysis of the inhibition of the iaaM and ipt expression in tumor cells demonstrated that the RNA interference strategy is promising for developing plant varieties resistant to agrobacterial infection.