The origin of Darwin hybrid tulips analyzed by flow cytometry, karyotype analyses and genomic in situ hybridization

Chromosome morphology was studied in diploid cultivars of Tulipa fosteriana and T. gesneriana (2n = 2x = 24) and triploid Darwin hybrids (2n = 3x = 36) developed from interspecific crosses of T. gesneriana and T. fosteriana. Chromosomes were arranged in the karyotype according to decreasing total length. Based on our karyotypic analysis, we propose that median chromosomes may serve as markers for diploid genotypes. Discriminant analysis with respect to total chromosome length and short arm length showed a significant difference between the size of the larger median chromosomes of T. gesneriana and T. fosteriana Comparison of median chromosome length in Darwin hybrid tulips showed that two larger chromosomes and one smaller chromosome were derived from T. gesneriana and T. fosteriana, respectively. This finding was clearly and unambiguously confirmed by simultaneous hybridization of differentially labeled genomic probes of T. fosteriana and T. gesneriana to metaphase chromosomes of the triploid cultivar ‘Yellow Dover’, thereby enabling us to distinguish between the 24 chromosomes derived from T. gesneriana and 12 chromosomes derived from T. fosteriana. Thus, genomic in situ hybridization and median chromosome analyses can be useful to identify the genome constitution of triploid Darwin hybrid tulips. In addition, their hybridity was readily verified by flow cytometry using vegetative tissue of Darwin hybrid tulips. Our results clarify the process of Tulipa cultivar formation and will be useful for interspecific hybridization breeding.The first and second author have contributed equally to this paper