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Abstract
BackgroundThe wild herb Swertia mussotii is a source of the anti-hepatitis compounds swertiamarin, mangiferin and gentiopicroside. Its over-exploitation has raised the priority of producing these compounds heterologously. Somatic hybridization represents a novel approach for introgressing Swertia mussotii genes into a less endangered species.ResultsProtoplasts derived from calli of Bupleurum scorzonerifolium and S. mussotii were fused to produce 194 putative hybrid cell lines, of which three (all derived from fusions where the S. mussotii protoplasts were pre-treated for 30 s with UV light) later differentiated into green plants. The hybridity of the calli was confirmed by a combination of isozyme, RAPD and chromosomal analysis. The hybrid calli genomes were predominantly B. scorzonerifolium. GISH analysis of mitotic chromosomes confirmed that the irradiation of donor protoplasts increased the frequency of chromosome elimination and fragmentation. RFLP analysis of organellar DNA revealed that mitochondrial and chloroplast DNA of both parents coexisted and recombined in some hybrid cell lines. Some of the hybrid calli contained SmG10H from donor, and produced swertiamarin, mangiferin and certain volatile compounds characteristic of S. mussotii. The expression of SmG10H (geraniol 10-hydroxylase) was associated with the heterologous accumulation of swertiamarin.ConclusionsSomatic hybrids between B. scorzonerifolium and S. mussotii were obtained, hybrids selected all contained introgressed nuclear and cytoplasmic DNA from S. mussotii; and some produced more mangiferin than the donor itself. The introgression of SmG10H was necessary for the accumulation of swertiamarin.
Highlights
The wild herb Swertia mussotii is a source of the anti-hepatitis compounds swertiamarin, mangiferin and gentiopicroside
The medicinal herb Swertia mussotii Franch is native to Tibet, where it has enjoyed a long history of use as a curative for hepatitis [5,6]
We previously reported plant regeneration from cultured B. scorzonerifolium protoplasts [16]
Summary
The wild herb Swertia mussotii is a source of the anti-hepatitis compounds swertiamarin, mangiferin and gentiopicroside. Somatic hybridization represents a novel approach for introgressing Swertia mussotii genes into a less endangered species. The possibility of introgression from exotic sources is of interest not just in the applied field, and because it provides opportunities for the discovery of novel synthetic pathways for secondary metabolites and signalling compounds. Many of the reactions in this pathway are known to be catalyzed by P450 proteins [10,11] Members of this highly diverse protein family are involved in the synthesis of pigments, antioxidants and defense compounds [12], and one of particular importance for the synthesis of swertiamarin is the enzyme geraniol 10-hydroxylase (G10H) [13]. We isolated a full length cDNA clone of S. mussotii
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