Genome composition of 'Elatior'-begonias hybrids analyzed by genomic in situ hybridisation

Interspecific hybridization of various tuberous Begonia species hybrids with Begonia socotrana results in so-called 'Elatior'-begonias hybrids (B. x hiemalis Fotsch). In our study, genomic in situ hybridization (GISH) has been employed to assess the genome composition in eleven 'Elatior'-begonias hybrids and their ancestor genotypes. Genomic DNA of tuberous Begonia was sonicated to 1-10-kb fragments, labelled by nick translation with digoxigenin-11-dUTP and used as a probe whereas B. socotrana DNA was autoclaved to 100 bp fragments and used as block. The genome of tuberous Begonia was clearly pronounced in 'Elatior'-begonias when the probe concentration was similar to 3.75 ng/mu l (150 ng/slide), with 30 times the excess of B. socotrana blocking DNA and stringency of post hybridization washings at 73% (0.1x SSC at 42A degrees C). In 'Elatior'-begonias hybrids GISH distinguished two groups comprising short (0.6-1.03 mu m in length) and relatively longer chromosomes (1.87-3.88 mu m) which represent B. socotrana and tuberous Begonia genomes, respectively. The number of chromosomes derived from tuberous Begonia ranged from 14 to 56 and for B. socotrana from 7 to 28 which suggest the presence of different ploidy levels in analyzed 'Elatior'-begonia hybrids. Intergenomic recombination has not been detected through GISH in hybrids analyzed. Genomic in situ hybridization turned out to be useful to identify the genome constitution of 'Elatior'-begonia hybrids and thus gain an insight into the origins of these cultivars. This knowledge on the ploidy level and genome composition is essential for further progress in breeding Begonias