Cloning and characterization of two repeated sequences in the symbiotic fungus Tuber melanosporum Vitt.
Article first published online: 5 JAN 2006
FEMS Microbiology Ecology
Volume 34, Issue 2, pages 139–146, December 2000
How to Cite
Paolocci, F., Rubini, A., Riccioni, C., Granetti, B. and Arcioni, S. (2000), Cloning and characterization of two repeated sequences in the symbiotic fungus Tuber melanosporum Vitt. FEMS Microbiology Ecology, 34: 139–146. doi: 10.1111/j.1574-6941.2000.tb00763.x
- Issue published online: 5 JAN 2006
- Article first published online: 5 JAN 2006
- Received 6 March 2000, Revised 1 August 2000, Accepted 21 August 2000
- Tuber spp.;
- Species-specific probe;
- Mycorrhizal root;
- Repeated genomic fragment;
- Multiplex PCR;
Two repeated DNA sequences of European strains of the symbiotic fungus Tuber melanosporum were isolated and characterized. One of these, SS14, representing about 0.05% of the fungal genome, was shown to be a T. melanosporum-specific sequence by Southern and dot-blot hybridization. The second one, named SS15, is about 0.0025% of the entire genome, and it is specific not only to T. melanosporum but also to the Asian black truffle Tuber indicum. Neither of these two fragments hybridizes with any of the other European truffle species tested. By sequence analysis of these two fragments, PCR primers were designed and used to selectively amplify DNA from T. melanosporum ascocarps and ectomycorrhizae by simple and multiplex PCR. No amplification products were obtained with DNA from either mycorrhizal roots or fruit bodies of other ectosymbiotic fungi. The two identified genomic traits also provided useful information for a better understanding of the phylogenetic relationships among black truffle species and for testing T. melanosporum intraspecific variability.