Haplotype Threading: Accurate Polyploid Phasing from Long Reads

Abstract

Resolving genomes at haplotype level is crucial for understanding the evolutionary history of polyploid species and for designing advanced breeding strategies. As a highly complex computational problem, polyploid phasing still presents considerable challenges, especially in regions of collapsing haplotypes. We present WO_SCPLOWHATSC_SCPLOWHO_SCPLOWAPC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWPOLYPHASEC_SCPLOW, a novel two-stage approach that addresses these challenges by (i) clustering reads using a position-dependent scoring function and (ii) threading the haplotypes through the clusters by dynamic programming. We demonstrate on a simulated data set that this results in accurate haplotypes with switch error rates that are around three times lower than those obtainable by the current state-of-the-art and even around seven times lower in regions of collapsing haplotypes. Using a real data set comprising long and short read tetraploid potato sequencing data we show that WO_SCPLOWHATSC_SCPLOWHO_SCPLOWAPC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWPOLYPHASEC_SCPLOW is able to phase the majority of the potato genes after error correction, which enables the assembly of local genomic regions of interest at haplotype level. Our algorithm is implemented as part of the widely used open source tool WhatsHap and ready to be included in production settings.