Prospects of telomere-to-telomere assembly in barley: Analysis of sequence gaps in the MorexV3 reference genome

Pavla Navrátilová, Helena Toegelová, Zuzana Tulpová, Yi Tzu Kuo, Nils Stein, Jaroslav Doležel, Andreas Houben, Hana Šimková*, Martin Mascher*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


The first gapless, telomere-to-telomere (T2T) sequence assemblies of plant chromosomes were reported recently. However, sequence assemblies of most plant genomes remain fragmented. Only recent breakthroughs in accurate long-read sequencing have made it possible to achieve highly contiguous sequence assemblies with a few tens of contigs per chromosome, that is a number small enough to allow for a systematic inquiry into the causes of the remaining sequence gaps and the approaches and resources needed to close them. Here, we analyse sequence gaps in the current reference genome sequence of barley cv. Morex (MorexV3). Optical map and sequence raw data, complemented by ChIP-seq data for centromeric histone variant CENH3, were used to estimate the abundance of centromeric, ribosomal DNA, and subtelomeric repeats in the barley genome. These estimates were compared with copy numbers in the MorexV3 pseudomolecule sequence. We found that almost all centromeric sequences and 45S ribosomal DNA repeat arrays were absent from the MorexV3 pseudomolecules and that the majority of sequence gaps can be attributed to assembly breakdown in long stretches of satellite repeats. However, missing sequences cannot fully account for the difference between assembly size and flow cytometric genome size estimates. We discuss the prospects of gap closure with ultra-long sequence reads.

Original languageEnglish
Pages (from-to)1373-1386
Number of pages14
JournalPlant Biotechnology Journal
Issue number7
StatePublished - Jul 2022
Externally publishedYes


  • CenH3
  • Cereba
  • ChIP-seq
  • PacBio HiFi reads
  • flow cytometry
  • nanopore
  • ribosomal DNA
  • satellite
  • telomeric repeats


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