Pcr Primers for the Amplification of Mitochondrial Small Subunit Ribosomal DNA of Lichen-forming Ascomycetes

TLDR

The primers target conserved U2, U4, and U6 regions of mitochondrial SSU rDNA. Four primers for amplifying mitochondrial DNA in lichen‑forming ascomycetes are introduced. The primers were evaluated by PCR, sequencing an 894‑bp product from *Lobaria pulmonaria*, and comparing it to *Podospora anserine* SSU rDNA. PCR produced single products from eight lichen fungi but not from two algae, and sequencing revealed >76 % identity with *Podospora anserine*, with product sizes 800–1000 bp, indicating the primers amplify mitochondrial SSU rDNA and may aid phylogenetic analyses.

Abstract

Abstract Four primers for the amplification of mitochondrial DNA of lichenforming ascomycetes are presented. The primers match the conserved regions U2, U4, and U6, respectively, of mitochondrial small subunit (SSU) ribosomal DNA (rDNA). Polymerase chain reaction using different combinations of the primers produced single amplification products from DNA of eight lichen-forming fungal species but did not amplify DNA of two axenic cultured algal species. The amplification product obtained from Lobaria pulmonaria was sequenced and the 894-bp sequence was compared with the mitochondrial SSU rDNA sequence of Podospora anserine . The two sequences revealed more than 76% identity in the conserved regions U3 to U5 demonstrating that we amplified mitochondrial DNA. The primers matching U2 and U6 yielded amplification products of 800–1000 bp depending on the species examined. The variation observed suggests that mitochondrial SSU rDNA may be useful for phylogenetic analyses of lichen-forming ascomycetes.