Multilocus Phylogeography of the Tuber mesentericum Complex Unearths Three Highly Divergent Cryptic Species

Tuber mesentericum is an edible European black truffle, apparently easy to recognize, but showing a high degree of genetic variability. In this study, we performed an integrative taxonomic assessment of the T. mesentericum complex, combining a multilocus phylogeographic approach with morphological analyses, and including authentic specimens of Vittadini, and Berkeley and Broome. We performed maximum likelihood phylogenetic analyses, based on single and concatenated gene datasets (ITS rDNA, β-tubulin, elongation factor 1-α), and including all available sequences from previous studies. Phylogenetic analyses consistently recovered three reciprocally monophyletic and well-supported clades: clade I, with a wide range across Europe; clade II, specimens collected mainly in the Iberian, Italian, and Balkan peninsulas; and clade III, specimens collected almost exclusively in central Italy. Genetic distance between clades ranged from 10.4% to 13.1% at the ITS region. We also designed new primer pairs specific for each phylogenetic lineage. Morphology of spores, asci, and peridium were investigated on specimens representing the three lineages. Macro- and micromorphological analyses of ascomata revealed only a few, but not diagnostic, differences between the three phylogenetic lineages, thus, confirming that they are morphologically cryptic. By studying authentic specimens of Vittadini, and Berkeley and Broome, it was possible to identify the three clades as T. mesentericum, Tuber bituminatum, and Tuber suave sp. nov., and to designate an epitype for T. mesentericum s.s. and a lectotype for T. bituminatum. Future investigations on volatile organic compound (VOC) composition are needed to define the aroma repertoires in this species complex.

Tuber mesentericum and Tuber aestivum Truffles: New Insights Based on Morphological and Phylogenetic Analyses

Tuber aestivum, one of the most sought out and marketed truffle species in the world, is morphologically similar to Tuber mesentericum, which is only locally appreciated in south Italy and north-east France. Because T. aestivum and T. mesentericum have very similar ascocarp features, and collection may occur in similar environments and periods, these two species are frequently mistaken for one another. In this study, 43 T. aestivum and T. mesentericum ascocarps were collected in Italy for morphological and molecular characterization. The morphological and aromatic characteristics of the fresh ascocarps were compared with their spore morphology. Afterwards, we amplified and sequenced the elongation factor 1-α (EF1α) locus and built maximum likelihood trees to assess phylogenetic similarities between the two species. Tuber aestivum and T. mesentericum sequences cluster into different clades, with T. mesentericum sequences divided into three different sub-clades. According to their morphological features, three samples (T7, T8 and T12) were classified as T. mesentericum. However, when fresh, these ascocarps lacked the typical phenolic aromatic note. These specimens fall into the sub-clade III of the T. mesentericum phylogeny, which has the lowest genetic distance from the T. aestivum clade.

Tuber mesentericum, an interesting species of black truffles in PoIand

<i>Tuber mesentericum</i> Vitt. is the only member of black truffles collected in Po|and in the calcareous area at the Częstochowa Upland. It was discovered in 1981 in old beech wood at the Zielona Góra nature reserve (Ławrynowicz 1988, 1990). Recently it has been collected in ca 30 years old oak forest situated 13 km NE from the Częstochowa town. Taxonomic notes, description of habitat and distribution are given.