PepsiCo OT3098 Hexaploid Oat Version 2 Genome Assembly Release in collaboration with GrainGenes

PepsiCo releases version 2 of OT3098 reference genome [April 2021]
By: Dr. Mandy Waters, PepsiCo

Method comparison between OT3098 v1 and v2:
Written by Dr. Kevin Fengler, Corteva Agrisciences

The v1 contig assembly was built from 35x PacBio HiFi (read N50 = 17.3 kb, min predicted accuracy > .992) using the HiCanu assembler which resulted in 1,890 contigs with a contig N50= 30.3 Mb and a total assembly size of 10.97 Gb. The contigs were then scaffolded with BioNano genome maps (450 maps, map N50= 98.5, total length = 11.2 Gb) on the BioNano Access platform to create hybrid scaffolds (n= 100, scaffold N50 = 293.9 Mb, total length = 10.84 Gb). Hybrid scaffolds were tentatively placed into pseudomolecules using oat consensus markers. The final ordering of scaffolds was refined using HiC data and contact map visualization using JuiceBox and the Juicer tools.

The v2 release is based on an improved contig assembly using the same PacBio HiFi dataset, but with the hifiasm assembler. The v2 assembly exhibited a significant improvement in contiguity (1,343 contigs, contig N50 = 71 Mb, total size= 10.75 Gb). This assembly was similarly scaffolded with the same BioNano maps as v1, however, the improved contigs significantly improved the contiguity of the scaffolds as well (84 scaffolds, scaffold N50= 374 kb, total size = 10.83 Gb). The v2 assembly was manually curated to resolve overlapping contigs that were not addressed during BioNano hybrid scaffolding (this was not done for v1). Scaffolds were placed into pseudomolecules using the v1 chromosomes as a guide and were validated by HiC contact map visualization as before.

Nomenclature, orientation, and stats:
Chromosomes in v1 were oriented such that the short arm was on top. The OT3098 v2 pseudomolecules use chromosome names and orientations adopted by PanOat in an effort to maintain consistency with future assemblies. The PanOat consortium is currently constructing chromosome-scale reference for multiple oat (Avena sativa) genotypes and has therefore adopted a chromosome nomenclature (naming and orientation). Chromosome names specify homologous group (1 to 7) and sub genome (A/C/D). Chromosomes were oriented in such a way that proximal regions of homoeologous chromosomes are collinear. PepsiCo would like to thank Dr. Martin Mascher for updating the v2 pseudomolecule orientation and naming in accordance with PanOat.

Below are the names and lengths of chromosomes in v1 and v2. Chromosomes highlighted in bold were swapped between v1 and v2 (please see the attached document for a better formatted version).

v1_chr_name v1_chr_length v2_chr_name v2_chr_length
1A 542795238 chr1A 540897063
2A 454026946 chr2A 449127287
3A 426317889 chr3A 425675180
4A 462057589 chr4A 463192880
5A 485535456 chr5A 485323027
6A 431567647 chr6A 448461343
7A 493489733 chr7A 493511962
1C 463431985 chr1C 462796039
2C 585391692 chr2C 589118817
3C 636099650 chr3C 638425132
7C 731989224 chr4C 716105986
5C 612252875 chr5C 613160974
6C 624915216 chr6C 626220839
4C 552251759 chr7C 551718542
1D 485732902 chr1D 484215583
2D 532459853 chr2D 532103454
3D 467934025 chr3D 480949782
4D 424978419 chr4D 455353809
5D 502323219 chr5D 499214392
6D 301592285 chr6D 298028472
7D 529301501 chr7D 528225653
Chr00 235316112 ChrUn 58832055

Future release of data:
PepsiCo is currently working to annotate a set of PacBio transcripts. After annotation is complete researchers will be able to access these data as a track on the genome browser. Additional files including a GFF, ORF nucleotide fasta, and ORF translated peptide fasta will also be available via the download site on GrainGenes. These data are anticipated to be available mid-May 2021. Additionally, Dr. Nick Tinker has mapped relevant SNP markers to v2, which will be available as a track on the genome browser, GFF for download, and visualized as CMaps physical maps.

Researchers are free to use and publish with all OT3098 genomic resources shared on GrainGenes. Given that no direct publication will be submitted for this individual genome assembly we choose to opt out of the Toronto Agreement, so researchers can freely use these resources as they become available:
• Genome Browser:
• BLAST: (select “PepsiCo OT3098 Hexaploid Oat v2 pseudomolecules (2021)” under the “Oat Selections”)
• Data Download:

If you use these resources, please cite:
"Avena sativa – OT3098 v2, PepsiCo,

Associated File: