Genetic and biochemical characterization of somatic hybrids between Bupleurum scorzonerifolium and Gentianopsis paludosa.

Ultraviolet-irradiated protoplasts of Gentianopsis paludosa were fused with those of Bupleurum scorzonerifolium and 28 independent hybrid calli were identified, five of which later differentiated into plants. A genetic analysis of these calli and regenerated plants based on chromosome number, esterase, random amplified polymorphic DNA, and 5S rDNA spacer profiling showed that the majority of their nuclear genomes were represented by the recipient biparent B. scorzonerifolium. A restriction fragment length polymorphism analysis of the plastidial genomes confirmed that DNA from both biparents was present in some of the hybrids. The secondary metabolite composition of the hybrids was analyzed by a combination of high-performance liquid chromatography and gas chromatography-mass spectrometry analysis. The content of oleanolic acid in two of the hybrid lines was substantially higher than in the donor G. paludosa, while that of swertiamarin was equal to that in G. paludosa in two of the six hybrids analyzed. A number of both G. paludosa and B. scorzonerifolium specific compounds were detected in the three hybrids analyzed by GC-MS as were several not present in either of the biparents.