**2.8 DNA isolation and Southern blot analysis**

Genomic DNA was extracted from mycelia using cetyltrimethylammonium bromide (CTAB) isolation buffer (J. J. Doyle & J. L. Doyle, 1987, as cited in Milligan, 1998). To prepare the DIG-labeled probe, genomic DNA was used as a template for PCR with the primer set, LeMnP2En5f2 (5'**-**TCAGGAAAATTCCCGACTAT) and LeMnP2En12r (5'-GAACCTCGATG CCATCAA); this primer set was designed to amplify the region from exon 5 to exon 12 of *lemnp2*, including introns (DDBJ Acc. No. AB306944; Sakamoto et al., 2009), and the resulting probe was named *lemnp2*S. In addition, to examine cross-hybridization between *lemnp2* and a relative of the manganese peroxidases LeMnP1 coded by *lemnp1*, a probe for *lemnp1* was also prepared as described above using the primer set LeMnP1f1 (5'- GATTCCTGAGCCTTTCG) and LeMnP1r (5'-TTCGGGACGGGAATAAC); this primer set was designed to amplify the regions from exon 7 to exon 15 of *lenmp1* including introns (DDBJ Acc. No. AB241061; Nagai et al., 2007) and the resulting probe was named *lemnp1*S. These two probes were then used for Southern blot analysis (Sambrook et al., 1989).
