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Taxa in the genus  Melanelia  (Parmeliaceae, Ascomycota) belong to a group of saxicolous lichens with brown to black foliose thalli, which have recently undergone extensive changes in circumscription. Taxa belonging to Parmeliaceae are prolific producers of bioactive compounds, which have also been traditionally used for chemotaxonomic purposes. However, the chemical diversity of the genus  Melanelia  and the use of chemical data for species discrimination in this genus are largely unexplored. In addition, identification based on morphological characters is challenging due to few taxonomically informative characters. Molecular identification methods, such as DNA barcoding, have rarely been applied to this genus. This study aimed to identify the  Melanelia  species from Iceland using DNA barcoding approach, and to explore their chemical diversity using chemical profiling. Chemometric tools were used to see if lichen metabolite profiles determined by LC-MS could be used for the identification of Icelandic  Melanelia  species. Barcoding using the fungal nuclear ribosomal internal transcribed spacer region (nrITS) successfully identified three  Melalenlia  species occurring in Iceland, together with  Montanelia disjuncta  (Basionym:  Melanelia disjuncta ). All species formed monophyletic clades in the neighbor-joining nrITS gene tree. However, high intraspecific genetic distance of  M .  stygia  suggests the potential of unrecognized species lineages. Principal component analysis (PCA) of metabolite data gave a holistic overview showing that  M .  hepatizon  and  M .  disjuncta  were distinct from the rest, without the power to separate  M .  agnata  and  M .  stygia  due to their chemical similarity. Orthogonal partial least–squares to latent structures–discriminate analysis (OPLS-DA), however, successfully distinguished  M .  agnata  and  M .  stygia  by identifying statistically significant metabolites, which lead to class differentiation. This work has demonstrated the potential of DNA barcoding, chemical profiling and chemometrics in identification of  Melanelia  species.

plos one

DNA barcoding and LC-MS metabolite profiling of the lichen-forming genus Melanelia: Specimen identification and discrimination focusing on Icelandic taxa