51. Molecular Identification of Tuber Species and Isolates by PCR-Based Techniques
- Author
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P. Nicolas, B. Henrion, Francis Martin, Chantal Dupré, P. Drevet, Gérard Chevalier, D. Gandeboeuf, UR 0874 Unité de recherche Agronomie de Clermont, Institut National de la Recherche Agronomique (INRA)-Environnement et Agronomie (E.A.)-Ecologie des Forêts, Prairies et milieux Aquatiques (EFPA), Institut National de la Recherche Agronomique (INRA)-Unité de recherche Agronomie de Clermont (URAC), Unité de recherche Biogéochimie des Ecosystèmes Forestiers (BEF), Institut National de la Recherche Agronomique (INRA), Unité associée organisation et variabilité des génomes végétaux, V. Stocchi, P. Bonfante, and M. Nuti
- Subjects
0106 biological sciences ,0303 health sciences ,Truffle ,Molecular epidemiology ,[SDV]Life Sciences [q-bio] ,15. Life on land ,Biology ,biology.organism_classification ,01 natural sciences ,030308 mycology & parasitology ,RAPD ,law.invention ,03 medical and health sciences ,law ,Tuber aestivum ,[SDE]Environmental Sciences ,Botany ,Internal transcribed spacer ,Restriction fragment length polymorphism ,Ribosomal DNA ,ComputingMilieux_MISCELLANEOUS ,Polymerase chain reaction ,010606 plant biology & botany - Abstract
Morphological features of fruit bodies and ectomycorrhizas allow the identification of most truffle species. However, economically important truffles (e.g. Tuber aestivum / T. uncinatum; T. borchii/ T. magnatum ) are difficult to distinguish using these criteria. It is therefore required, when morphological traits are unsufficient for discriminating, to call for additional markers (e.g. isozymes and DNA). DNA polymorphisms in truffle genomes have therefore been used to generate molecular markers by PCR amplification of specific regions and by random amplification of polymorphic DNA (RAPD). Amplification of the internal transcribed spacer (ITS) and intergenic spacer (IGS) of the ribosomal DNA combined with endonuclease digestion yielded unique diagnostic patterns for most European truffle species. However, rDNA polymorphism was not high enough to distinguish T. aestivum and T. uncinatum. This approach revealed a low intraspecific variation within T. melanosporum. The PCR amplification of the rDNA spacers can be combined with RFLP, allowing truffle species to be typed in less than a day and is most immediately applicable to molecular epidemiology. DNA polymorphism has been further assessed by RAPD technique. The high interspecific genomic variability already observed for most truffle species using PCR-RFLP was confirmed. In addition, large intraspecific variations have been found in all species, except T. melanosporum and T. brumale. Molecular techniques provide an efficient tool, on the one hand for typing fruiting bodies for the food industry, mycelium for patenting purposes as well as mycorrhiza for checking the inoculated seedlings and on the other hand for tracking the fate of mycorrhizal seedlings within truffieres and forest ecosystems.
- Published
- 1995
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