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The first whole genome sequencing of historical lichen specimens enables genome-wide analysis of fungal and algal symbionts
  • Mieko Kono,
  • Yoshihito Ohmura,
  • Yohey Terai
Mieko Kono
SOKENDAI

Corresponding Author:mieko.kono10001@gmail.com

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Yoshihito Ohmura
National Museum of Nature and Science
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Yohey Terai
SOKENDAI
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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.