Induction of telomere-mediated chromosomal truncation and stability of truncated chromosomes in Arabidopsis thaliana

Abstract

Minichromosomes possess functional centromeres and telomeres and thus should be stably inherited. They offer an enormous opportunity to plant biotechnology as they have the potential to simultaneously transfer and stably express multiple genes. Segregating independently of host chromosomes, they provide a platform for accelerating plant breeding. Following a top-down approach, we truncated endogenous chromosomes in Arabidopsis thaliana by Agrobacterium-mediated transfer of T-DNA constructs containing telomere sequences. Blocks of A. thaliana telomeric repeats were inserted into a binary vector suitable for stable transformation. After transfer of these constructs into the natural tetraploid A. thaliana accession Wa-1, chromosome truncation by T-DNA-induced de novo formation of telomeres could be confirmed by DNA gel blot analysis, PCR (polymerase chain reaction), and fluorescence in situ hybridisation. The addition of new telomere repeats in this process could start alternatively from within the T-DNA-derived telomere repeats or from adjacent sequences close to the right border of the T-DNA. Truncated chromosomes were transmissible in sexual reproduction, but were inherited at rates lower than expected according to Mendelian rules.