0 program. Branch lengths are proportional to the number of changes. Seven different intron sequence types (bolded) identified from 57 B. bassiana isolates were aligned with 24 representative intron sequences from Metarhizium anisopliae (Ma), Beauveria bassiana (Bb) and Cordyceps profilica (Csp), and an intron sequence from Emricasan nmr Naegleria sp. (Nsp) was used as outgroup. The four group I intron insertion positions are shown as Ec1921 (position 4), Ec2066 (position 3), Ec2449 (position 2) and Ec2563 (position 1). EF1-α gene analysis With the

exception of isolate Bb49, where no amplification was observed, all isolates afforded PCR products of 1.1 kb for the EF1-α gene with the primers tef1fw and 1750-R. Eleven different EF1-α gene sequences were identified among the 56 isolates. The alignment and comparison of these 11 sequences and another 18 GenBank-deposited sequences, representing different lineages from B. bassiana s.s. (sensu stricto), B. brongniartii and B. bassiana clade C [7, 8, 12], produced 1757 aligned positions, with 1542 constant characters and 114 parsimony-informative characters. The MP tree is shown

in Figure 2. Of the 56 isolates analyzed, 94.6% (53 isolates) were located in the B. bassiana s.s. clade, and 5.4% (3 isolates) in clade C, which includes B. cf. (uncertain taxonomy) bassiana isolates. Within B. bassiana s.s., the 53 isolates analyzed in this study were separated in five subgroups (Eu-7, Eu-8 and Eu-9 with isolates from Spain and Portugal; Eu-3 from Spain, France and Denmark; and Wd-2 with world-wide distribution), selleck chemicals supported by bootstrap values Selleck PD-L1 inhibitor higher than 50%. Figure 2 Phylogenetic analysis based on EF1- a sequences from Beauveria bassiana. The MP tree was generated by parsimony analysis after heuristic searches (TBR option). A bootstrap full heuristic analysis, with bootstrap intervals from 1000 replications and nodes supported in >50% of bootstrap replicates, was generated using the PAUP 4.0 program. Branch lengths are proportional to the number of changes. Eleven sequence types identified from 56 B. bassiana isolates, of which 52 were sampled

in Spain (bolded), were aligned with 18 GenBank B. bassiana s.s., B. brongniartii and B. cf. bassiana (clade C) sequences, indicated by accession numbers as in previous works [7, 8]. B. bassiana s.s. EF1-α sequences representing European subgroups learn more [7] are marked with an asterisk. Reference isolates from countries different to Spain, are referred to as: Eu-1 (KVL0376 from Denmark and ARSEF1628 from Hungary), Eu-3 (KVL0373 from Denmark and ARSEF1185 from France), Eu-4 (KVL03114 from Denmark and ARSEF1848 from Belgium), Eu-5 (KVL0392 and KVL03112 from Denmark), Eu-6 (KVL0384 from Denmark and 815 from France), Eu-7 (Bb45 from Portugal), Wd-1 (296 and 344 from USA), Wd-2 (681 from Romania, 792 from USA, Bb55 from Georgia and Bb56 from Greece), C1 (4933 from France and Bb57 from Poland), C2 (812 from France) and B. brogniartii (KVL0392 from Denmark and 4384 from China). Cordyceps cf.