Molecular phylogeny of the tropical lichen family Pyrenulaceae: contribution from dried herbarium specimens and FTA card samples

The Pyrenulaceae is one of the most common lichen families in tropical rainforests. These mostly lichenised ascomycete fungi are relatively well known, as identification tools are available for many members of the family, including the most species-rich genus Pyrenula. However, despite the past boom in DNA sequence generation for most living organisms, molecular data are still mainly lacking for these crustose corticolous lichens, mainly because genomic DNA does not seem to preserve well after specimen desiccation. Because of lengthy customs procedures, newly collected specimens are therefore often too old for molecular work when they reach the laboratory. Here, a new method of DNA extraction and storage using FTA card samples was investigated. New DNA sequences were generated using both DNA extractions from freshly collected material (116 sequences) and FTA card samples (115 sequences) for 100 taxa within Pyrenulaceae. Inferences using three ribosomal genes (nuLSU, mtSSU and ITS) highlighted delimitation problems in few taxa, including the pantropical species P. mamillana and P. quassiicola. Issues related to generic delimitations were also confirmed, with Anthracothecium, Pyrgillus and Lithothelium nested within Pyrenula. Although further taxon and gene sampling will be required to fully revise species and generic concepts within this family, our data allowed us to describe two new species from Brazil (Lithothelium immersum and Pyrenula minutispora) and one from North America (Pyrenula reebiae).