title: Interspecific somatic hybrids between Cyclamen persicum and C. coum, two sexually incompatible species.
abstract: By applying polyethylene glycol (PEG)-mediated protoplast fusion, the first somatic hybrids were obtained between Cyclamen persicum (2n = 2x = 48) and C. coum (2n = 2x = 30)-two species that cannot be combined by cross breeding. Heterofusion was detected by double fluorescent staining with fluorescein diacetate and scopoletin. The highest heterofusion frequencies (of about 5%) resulted from a protocol using a protoplast density of 1 × 10(6)/mL and 40% PEG. The DNA content of C. coum was estimated for the first time by propidium iodide staining to be 14.7 pg/2C and was 4.6 times higher than that of C. persicum. Among 200 in vitro plantlets regenerated from fusion experiments, most resembled the C. coum parent, whereas only 5 plants showed typical C. persicum phenotypes and 46 had a deviating morphology. By flow cytometry, six putative somatic hybrids were identified. A species-specific DNA marker was developed based on the sequence of the 5.8S gene in the ribosomal nuclear DNA and its flanking internal transcribed spacers ITS1 and ITS2. The hybrid status of only one plant could be verified by the species-specific DNA marker as well as sequencing of the amplification product. RAPD markers turned out to be less informative and applicable for hybrid identification, as no clear additivity of the parental marker bands was observed. Chromosome counting in root tips of four hybrids revealed the presence of the 30 C. coum chromosomes and 2-41 additional ones indicating elimination of C. persicum chromosomes.
authors: Institute of Floriculture and Woody Plant Science, Gottfried Wilhelm Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419 Hannover, Germany.
authors: Anika Nadja Sabine
authors: AN
authors: Prange
authors: Melanie
authors: M
authors: Bartsch
authors: Julia
authors: J
authors: Meiners
authors: Margrethe
authors: M
authors: Serek
authors: Traud
authors: T
authors: Winkelmann
mesh_terms: D001483:Base Sequence; D002459:Cell Fusion; D032461:Chromosomes, Plant / Q000235:genetics*; D031824:Cyclamen / Q000196:embryology* / Q000235:genetics*; D020732:Cytogenetic Analysis; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005434:Flow Cytometry; D005452:Fluoresceins; D005819:Genetic Markers / Q000235:genetics; D018745:Genome, Plant / Q000235:genetics*; D006824:Hybridization, Genetic / Q000235:genetics*; D018519:Meristem / Q000235:genetics; D008969:Molecular Sequence Data; D010641:Phenotype; D058439:Plant Somatic Embryogenesis Techniques / Q000379:methods*; D011003:Ploidies; D011092:Polyethylene Glycols; D011523:Protoplasts; D019105:Random Amplified Polymorphic DNA Technique; D012603:Scopoletin; D016415:Sequence Alignment; D013045:Species Specificity
chemical_list: D018744:DNA, Plant; D004275:DNA, Ribosomal; D005452:Fluoresceins; D005819:Genetic Markers; D011092:Polyethylene Glycols; D012603:Scopoletin; C018506:diacetylfluorescein