Identification of mitotic chromosomes of tuberous and non-tuberous Solanum species (Solanum tuberosum and Solanum brevidens) by GISH in their interspecific hybrids

GISH (genomic in situ hybridization) was applied for the analysis of mitotic chromosome constitutions of somatic hybrids and their derivatives between dihaploid clones of cultivated potato (Solanum tuberosum L.) (2n = 2x = 24, AA genome) and the diploid, non-tuberous, wild species Solanum brevidens Phil. (2n = 2x = 24, EE genome). Of the primary somatic hybrids, both tetraploid (2n = 4x) and hexaploid (2n = 6x) plants were found with the genomic constitutions of AAEE and AAEEEE, respectively. Androgenic haploids (somatohaploids) derived from the tetraploid somatic hybrids had the genomic constitutions of AE (2n = 2x = 24) and haploids originating from the hexaploid hybrids were triploid AEE (2n = 3x = 33 and 2n = 3x = 36). As a result of subsequent somatic hybridization from a fusion between dihaploid S. tuberosum (2n = 2x = 24, genome AA) and a triploid somatohaploid (2n = 3x = 33, genome AEE), second-generation somatic hybrids were obtained. These somatic hybrids were pentaploids (2n = 5x, genome AAAEE), but had variable chromosome numbers. GISH analysis revealed that both primary and second-generation somatic hybrids had lost more chromosomes of S. brevidens than of S. tuberosum.Key words: anther culture, genome, haploid, potato, somatic hybridization.

Cytological and molecular characterization of repetitive DNA sequences of Solanum brevidens and Solanum tuberosum

The chromosomal distribution, copy numbers, and nucleotide sequences were determined for four repetitive DNA clones, pSB1 and pSB7 of Solanum brevidens and pST3 and pST10 of Solanum tuberosum. Using fluorescence in situ hybridization (FISH), pSB1 and pSB7 were localized near the telomeres and in some centromeric and interstitial sites of S. brevidens chromosomes, but not in S. tuberosum chromosomes, after high stringency washes. The clone pST3 showed signals in the telomeric areas of a few chromosomes in S. tuberosum, but signals were not detected in S. brevidens. All three repeated sequences (pSB1, pSB7, and pST3) were detected in chromosomal areas that are typically known to contain tandemly repeated sequences. The S. tuberosum clone pST10 did not show signals in either species even at low stringency conditions. The estimated copy numbers of the four clones were 1500, 6750, 300, and 400 for pSB1, pSB7, pST3, and pST10, respectively, in the corresponding haploid genomes (S. brevidens and S. tuberosum). The inserts of the four clones pSB1, pSB7, pST3, and pST10 were 322, 167, 845, and 121 bp, respectively. After sequencing, no significant sequence homologies were found among the four clones. A homology search in sequence data bases showed that pSB7 has variable homology (78-100%) with another repetitive sequence of S. brevidens Sb4/2 depending on its subrepeat. It also showed some homology with one repeat of tomato (pLEG15) and one repeat of Solanum circaeifolium (pSC15).Key words: chromosome, copy number, fluorescence in situ hybridization, FISH, nucleotide sequence, potato.

Differential behavior of Solanum brevidens ribosomal DNA loci in a somatic hybrid and its progeny with potato

Fluorescence in situ hybridization was used to characterize loci encoding ribosomal RNA (rDNA) among parents and progeny of a somatic fusion between tetraploid Solanum tuberosum (potato) and diploid Solanum brevidens. As expected, four major sites of hybridization to rDNA loci were evident in the tetraploid parent species, two in the diploid parent species, and six in the hexaploid somatic fusion plant. Two of the loci in the somatic fusion plant showed differential signals relative to the other four, which were interpreted as a delayed condensation of sites harboring the rDNA loci. This delayed condensation was heritable to the first backcross generation of the somatic hybrid crossed with potato. In the second backcross generation, differential condensation was not evident. However, a heterochromatic isochromosome was observed whose presence was correlated with a S. brevidens specific marker linked with the rDNA locus. It is suggested that the S. brevidens rDNA loci are preferentially affected in the somatic hybrid and its progeny, and that the delayed condensation may have contributed to the formation of the isochromosome.Key words: introgression, recombination, isochromosome, in situ hybridization.