Haplotype-phased and chromosome-level assembly of Puccinia polysora, a
giga-scale fungal pathogen causing southern corn rust
Abstract
Rust fungi are characterized by large genomes with high repeat content,
and have two haploid nuclei in most life stages, which makes achieving
high-quality genome assemblies challenging. Here, we describe a pipeline
using HiFi reads and Hi-C data to assemble a gigabase-sized fungal
pathogen, Puccinia polysora f.sp. zeae, to haplotype-phased and
chromosome-scale. The final assembled genome is 1.71 Gbp, with
~850 Mbp and 18 chromosomes in each haplotype, being
currently the largest fungal genome assembled to chromosome scale.
Transcript-based annotation identified 47,512 genes with a similar
number for each haplotype. A high level of interhaplotype variation was
found with 10% haplotype-specific BUSCO genes, 5.8 SNPs/kbp, and
structural variation accounting for 3% of the genome size. The P.
polysora genome displayed over 85% repeat content, with genome-size
expansion, gene losses and gene family expansions suggested by multiple
copies of species-specific orthogroups. Interestingly, these features
did not affect overall synteny with other Puccinia species with smaller
genomes. Fine-time-point transcriptomics revealed seven clusters of
co-expressed secreted proteins that are conserved between two
haplotypes. The fact that candidate effectors interspersed with all
genes indicated the absence of a “two-speed genome” evolution in P.
polysora. Genome resequencing of 79 additional isolates revealed a
clonal population structure of P. polysora in China with low geographic
differentiation. Nevertheless, a minor population drifted from the major
population by having mutations on secreted proteins including AvrRppC,
indicating the ongoing evolution and population differentiation. The
high-quality assembly provides valuable genomic resources for future
studies on the evolution of P. polysora.