De novo assembly and phasing of dikaryotic genomes from two isolates of puccinia coronata f. Sp. avenae, the causal agent of oat crown rust

Marisa E. Miller, Ying Zhang, Vahid Omidvar, Jana Sperschneider, Benjamin Schwessinger, Castle Raley, Jonathan M. Palmer, Diana Garnica, Narayana Upadhyaya, John Rathjen, Jennifer M. Taylor, Robert F. Park, Peter N. Dodds, Cory D. Hirsch, Shahryar F. Kianian, Melania Figueroa

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Oat crown rust, caused by the fungus Pucinnia coronata f. sp. avenae, is a devastating disease that impacts worldwide oat production. For much of its life cycle, P. coronata f. sp. avenae is dikaryotic, with two separate haploid nuclei that may vary in virulence genotype, highlighting the importance of understanding haplotype diversity in this species. We generated highly contiguous de novo genome assemblies of two P. coronata f. sp. avenae isolates, 12SD80 and 12NC29, from long-read sequences. In total, we assembled 603 primary contigs for 12SD80, for a total assembly length of 99.16 Mbp, and 777 primary contigs for 12NC29, for a total length of 105.25 Mbp; approximately 52% of each genome was assembled into alternate haplotypes. This revealed structural variation between haplotypes in each isolate equivalent to more than 2% of the genome size, in addition to about 260,000 and 380,000 heterozygous single-nucleotide polymorphisms in 12SD80 and 12NC29, respectively. Transcript-based annotation identified 26,796 and 28,801 coding sequences for isolates 12SD80 and 12NC29, respectively, including about 7,000 allele pairs in haplotype-phased regions. Furthermore, expression profiling revealed clusters of coexpressed secreted effector candidates, and the majority of orthologous effectors between isolates showed conservation of expression patterns. However, a small subset of orthologs showed divergence in expression, which may contribute to differences in virulence between 12SD80 and 12NC29. This study provides the first haplotype-phased reference genome for a dikaryotic rust fungus as a foundation for future studies into virulence mechanisms in P. coronata f. sp. avenae. IMPORTANCE Disease management strategies for oat crown rust are challenged by the rapid evolution of Puccinia coronata f. sp. avenae, which renders resistance genes in oat varieties ineffective. Despite the economic importance of understanding P. coronata f. sp. avenae, resources to study the molecular mechanisms under- pinning pathogenicity and the emergence of new virulence traits are lacking. Such limitations are partly due to the obligate biotrophic lifestyle of P. coronata f. sp. avenae as well as the dikaryotic nature of the genome, features that are also shared with other important rust pathogens. This study reports the first release of a haplotype-phased genome assembly for a dikaryotic fungal species and demonstrates the amenability of using emerging technologies to investigate genetic diversity in populations of P. coronata f. sp. avenae.

Original languageEnglish (US)
Article numbere01650-17
JournalmBio
Volume9
Issue number1
DOIs
StatePublished - Jan 1 2018

Bibliographical note

Funding Information:
This work was funded by the USDA-ARS and the University of Minnesota Standard Cooperative Agreement (grant 3002-11031-00053115 shared by S.F.K. and M.F.), the University of Minnesota Experimental Station USDA-NIFA Hatch/Figueroa project MIN-22-058, and an Organization for Economic Cooperation and Development Fellowship to M.F. M.E.M. was partially supported by a USDA-NIFA Postdoctoral Fellowship Award (2017-67012-26117). J.S. was supported by an OCE Postdoctoral Fellowship. R.F.P. receives funding from the Australian Grains Research Development Corporation (grant US00067). J.M.P. was supported by the Northern Research Station of the USDA Forest Service. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Publisher Copyright:
© 2018 Miller et al.

Keywords

  • Effectors
  • Genomics
  • Oat
  • Rust fungi
  • Virulence

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