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Mascher, Martin; Wicker, Thomas; Jenkins, Jerry; Plott, Christopher; Lux, Thomas; Koh, Chu Shin; Ens, Jennifer; Gundlach, Heidrun; Boston, Lori B; Tulpová, Zuzana; Holden, Samuel; Hernández-Pinzón, Inmaculada; Scholz, Uwe; Mayer, Klaus F X; Spannagl, Manuel; Pozniak, Curtis J; Sharpe, Andrew G; Šimková, Hana; Moscou, Matthew J; Grimwood, Jane; Schmutz, Jeremy; Stein, Nils
The Plant cell, 07/2021, Volume: 33, Issue: 6Journal Article
Sequence assembly of large and repeat-rich plant genomes has been challenging, requiring substantial computational resources and often several complementary sequence assembly and genome mapping approaches. The recent development of fast and accurate long-read sequencing by circular consensus sequencing (CCS) on the PacBio platform may greatly increase the scope of plant pan-genome projects. Here, we compare current long-read sequencing platforms regarding their ability to rapidly generate contiguous sequence assemblies in pan-genome studies of barley (Hordeum vulgare). Most long-read assemblies are clearly superior to the current barley reference sequence based on short-reads. Assemblies derived from accurate long reads excel in most metrics, but the CCS approach was the most cost-effective strategy for assembling tens of barley genomes. A downsampling analysis indicated that 20-fold CCS coverage can yield very good sequence assemblies, while even five-fold CCS data may capture the complete sequence of most genes. We present an updated reference genome assembly for barley with near-complete representation of the repeat-rich intergenic space. Long-read assembly can underpin the construction of accurate and complete sequences of multiple genomes of a species to build pan-genome infrastructures in Triticeae crops and their wild relatives.
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