2011a, b), its genetic transformation (Wójcik and Rybczyński 2015), and its somatic hybridization (Tomiczak et al. kurroo is particularly obvious in cell suspension cultures (Fiuk and Rybczyński 2008) and in cultures of cell suspension-derived protoplasts (Fiuk and Rybczyński 2007), which provide the basis for successful cryopreservation of this species (Mikuła et al. 2006 Behera and Raina 2012 Baba and Malik 2014). This species is an endangered medicinal herb from western and northwestern Himalaya, which has been overexploited in its natural habitats owing to the extensive collection of its leaves, roots, rhizomes, and flower heads, which are highly valued in traditional Indian medicine (Latif et al. The results of our experiments revealed the tremendous embryogenic potential possessed by Gentiana kurroo Royle. 1996, 2011 Mikuła and Rybczyński 2001 Fiuk and Rybczyński 2008 Tomiczak et al. 2016).įor over 20 years, we have studied the morphogenic abilities of various gentian species in vitro, aimed at conserving and broadening their genetic diversity (Mikuła et al. Thus, detailed investigation is needed, not only to confirm the hybrid status of regenerants, but also to characterize precisely their genome size and composition, which is essential for their further efficient exploitation (Smyda-Dajmund et al. However, regeneration of plants from fused protoplasts is often accompanied by polyploidization and/or the elimination of genome parts of either one or both fusion parents (Babiychuk et al. In contrast to sexual hybridization, following protoplast fusion, all nuclear and cytoplasmic DNA from both parents are united in one individual. In the family Gentianaceae, which contains many valuable medicinal and ornamental species, somatic hybrids representing different nucleocytoplasmic combinations would be very useful as new horticultural cultivars and as a valuable source of secondary metabolites (Wang et al. In a breeding program, somatic cell fusion is usually used for transferring of polygenic traits and those encoded by organellar genomes (Waara and Glimelius 1995). It is used to overcome sexual incompatibility between species and may lead to the production of novel genetic combinations as a consequence of recombination within either nuclear or cytoplasmic genomes. Somatic hybridization by protoplast fusion offers several possibilities for increasing genetic variability in higher plants. In consequence, gradual elimination of nuclear DNA, mixoploidy, and high genetic instability were observed in most hybrid plants and calli during the subsequent 4 years of in vitro culture. Flow cytometry revealed that polyploidization occurred, and probably it took place at the early stage of post-fusion culture. According to the results of chloroplast DNA analysis with the use of CAPS markers, all somatic hybrids inherited chloroplasts from “mesophyll” fusion partner G. AFLP and ISSR analyses revealed that all 21 hybrid plants and 3 lines of hybrid callus were genetically closer to G. were characterized using various molecular markers and flow cytometry.
Here, genome composition and stability of interspecific gentian somatic hybrids obtained following electrofusion of cell suspension-derived protoplasts of diploid Gentiana kurroo Royle with leaf mesophyll-derived protoplasts of tetraploid G. However, in order to evaluate its further usefulness, detailed characterization of the newly created hybrid is essential. Somatic hybrids of the genus Gentiana could provide a useful source of new ornamental cultivars and of secondary metabolites. Somatic hybridization by protoplast fusion is used in breeding programs to obtain plant material that has inherited valuable traits from two different species, and in order to broaden plant genetic diversity.
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