A system to induce the deletion of gemomic sequences using R/RS site-specific recombination and the Ac transposon in transgenic rice plants

Abstract 

Recombinase encoded by the R gene of Zygosaccharomyces rouxii mediates reciprocal recombination between two specific recombination sites (RSs) to induce deletion or inversion of the DNA segment that is flanked by the RSs. The R gene under the control of the CaMV 35 S promoter was introduced into rice (Oryza sativa L.). R/RS-specific deletion was first demonstrated in transgenic rice callus carrying the R gene by transient introduction of a cryptic reporter gene that was designed to confer β-glucuronidase (GUS) expression once deletion between two RSs took place. The rice containing the R gene was subsequently crossed with transgenic rice carrying (I-RS/dAc-I-RS) T-DNA that contained RS sequences within the T-DNA and another RS in a modified Ac element that had been transposed to a new locus by Ac transposase. Deletion of the gemomic sequences flanked by the two RSs was detected by PCR analysis in somatic cells of F₂ plants. These results demonstrate a technical advance in that the R/RS recombination system, in combination with the Ac transposable element, can be used to generate deletion in rice chromosomes.