Phylogenomics and effector analysis of Plasmodiophora brassicae genomes unveils a unique and highly divergent clade in Australia

Clubroot, caused by the protist pathogen Plasmodiophora brassicae , is a soilborne disease that leads to significant yield losses in a broad range of brassica crops worldwide. Despite its significant impact on agriculture, genomic and molecular studies of clubroot disease are hindered due to the complex life cycle and the obligate biotrophic nature of the pathogen, which prevents its cultivation in vitro . In addition, several genotypes of P. brassicae are often present as a mixture in a single field sample, making it challenging to resolve high-quality assemblies of individual genomes. Recently, several genomes were assembled for some strains of P. brassicae , however, in-depth genomic analysis of this devastating pathogen remains limited.

In this study, we generated complete telomere-to-telomere genome assemblies for three Australian P. brassicae field isolates using PacBio Revio HiFi sequencing combined with chromatin conformation capture technology (Hi-C). Importantly, we could generate two nearly fully phased haplotype genome assemblies from one of the field isolates. Additionally, we sequenced 14 isolates of P. brassicae, sampled from a wide geographical range across Australia using Illumina technology and performed an in-depth global comparative phylogenetic analysis. We revealed that Australian isolates are classified into three different clades, including a highly divergent and unique clade present only in Australia. Comparative analysis showed that the predicted effector profile of the Australian-unique clade is distinct from other clades, further supporting phylogenetic divergence. Altogether, we demonstrate the first haplotype-resolved genome of Australian P. brassicae isolates from a single clubroot field sample, and provide an in-depth phylogenetic analysis of global and Australian isolates. These findings, together with core effector profile analysis, will further advance the research to combat this destructive pathogen.