NUCLEAR GENOMIC COMPOSITION OF ASYMMETRIC FUSION PRODUCTS BETWEEN IRRADIATED TRANSGENIC SOLANUM-BREVIDENS AND SOLANUM-TUBEROSUM - LIMITED ELIMINATION OF DONOR CHROMOSOMES AND POLYPLOIDIZATION OF THE RECIPIENT GENOME

The production of asymmetric somatic hybrid calli after fusion between gamma-irradiated protoplasts from transgenic Solanum brevidens and pr otoplasts from S. tuberosum are reported. Transgenic (kanamycin-resist ant, GUS-positive) S. brevidens plants and hairy root clones were obta ined after transformation with Agrobacterium tumefaciens LBA 1060 (pRi 1855) (PBI121) and LBA 4404 (pRAL4404) (pBI121), and A. rhizogenes LBA 9402 (pRi1855) (pBI121), respectively. Leaf protoplasts isolated from the transgenic plants or root protoplasts from the hairy root clones were fused with S. tuberosum leaf protoplasts, and several calli were selected on kanamycin-containing medium. The relative nuclear DNA cont ent of the hybrid calli was measured by flow cytometry (FCM), and the percentages of DNA of the S. brevidens and S. tuberosum genomes in the calli were determined by dot blot analysis using species-specific DNA probes. Chromosome-specific restriction fragment length polymorphism (RFLP) markers were used to investigate the elimination of specific S. brevidens chromosomes in the hybrids. The combined data on FCM, dot b lot and RFLP analysis revealed that 18-62% of the S. brevidens DNA was eliminated in the hybrid calli and that the RFLP marker for chromosom e 7 was absent in seven out of ten calli. The absence of RFLP markers for chromosomes 5 and 11 hardly ever occurred. In most of the hybrids the ploidy level of the S. tuberosum genome had increased considerably .