cinnabarinus BRFM 137 coding regions (NCBI accession numbers AAY40456 and AF152170; Otterbein et al., 2000; Schmitt et al., 2008) and by identifying the eukaryotic consensus splicing sites (5′-GT and 3′-AG nucleotides). The nucleotide sequences (only exons for β-tubulin and laccase gene fragments) were aligned using the clustalw algorithm (Higgins et al., 1991). The alignments were then hand-refined. Phylogenetic analyses

were performed from single genes according to the method developed for the figenix platform (Gouret et al., 2005) using the heuristic search for maximum likelihood trees. Bootstrap values were calculated over 1000 replicates to assess branch topology. Phylogenetic trees were rooted with T. suaveolens as an outgroup. The filamentous fungi, among MLN0128 manufacturer which the genus Pycnoporus is considered a strong contender for white biotechnology processes, form a huge worldwide source of biological diversity that needs to be explored. In the present work, the phylogenetic relationships of a large sample of Pycnoporus strains of different geographical origins were analysed

using three complementary DNA markers. The nuclear rDNA region, ITS1-5.8S-ITS2, was often used to infer phylogenetic relationships Metabolism inhibition among wood decay basidiomycetes species within a particular genus such as Phanerochaete (de Koker et al., 2003) or a species complex such as Postia caesia (Yao Cyclic nucleotide phosphodiesterase et al., 2005) but it often fails to provide robust phylogenetic resolution among

the fungal species (Wang et al., 2004). The β-tubulin gene sequences were shown to resolve phylogenetic relationships within ascomycetes genera that could not be distinguished on the basis of morphology, especially in Aspergillus or Pestalotiopsis genera (Giraud et al., 2007; Hu et al., 2007). The genus Pycnoporus is described to overproduce laccase (encoded by lac3-1 gene) as an extracellular ligninolytic enzyme in induced culture conditions (Eggert et al., 1996; Lomascolo et al., 2003). To date, genes encoding laccases have not been used to gain phylogenetic information within a fungal genus. In this study, amplification of the ITS1-5.8S-ITS2 region yielded fragments 550–650 bp in length. After clean-up, the 36 sequences of Pycnoporus strains were aligned in 467 nucleotide positions (see Supporting Information, File S1). The sequencing analysis showed that the ITS1 and ITS2 regions were different in the strains studied, due to nt-insertions/deletions or substitutions, whereas the 5.8S rRNA gene sequences (157 bp long) were conserved for all the taxa. Within the ITS1 sequences, 44 of the 131 aligned positions (33.6%) varied among the strains of Pycnoporus. Within the ITS2 sequences, 36 of the 177 aligned positions (20.3%) varied among the strains of Pycnoporus.