The first whole genome sequencing of historical lichen specimens enables
genome-wide analysis of fungal and algal symbionts
Abstract
With the advent of high-throughput sequencing techniques, natural
history museums and herbaria have become new frontiers for genetic
research. Genomic information from historical specimens has provided
evidence to solve significant questions in broad research areas.
However, access to such valuable genetic resources remains limited in
lichens due to experimental challenges in extracting and amplifying
highly degraded DNA in historical specimens. So far, only a handful of
studies have reported successful sequencing of several short genetic
markers from historical lichen specimens despite the increasing
importance of genetic information in lichenology. Here, we aim to
establish an efficient method for sequencing the whole genome of
historical lichen specimens. We modified a method used in ancient DNA
studies and sequenced the whole genome of 75-year-old Usnea hakonensis
specimens, including the holotype. Our approach shows that 3.9 % and
9.8 % of the total sequenced reads originate from the genomes of U.
hakonensis fungal (mycobiont) and algal (photobiont) symbionts. The
mycobiont- and photobiont-derived reads are comprised of DNA fragments
shorter than 45 bp, covering 77 % and 93 % of the mycobiont and
photobiont reference genomes in average coverage of 27x and 45x,
respectively. We retrieved 792,245 Single Nucleotide Variants (SNVs) to
perform phylogenetic analysis of the mycobiont. We also demonstrated
experimental modifications that improved proportions of
symbionts-derived reads within sequenced data. We believe that our
method is applicable to lichen specimens in a broad range of age and
taxonomic groups, thereby potentially converting historical lichen
specimens into resources of genome-wide studies.