Assembly collapsing versus heterozygosity oversizing: detection of homokaryotic and heterokaryotic Laccaria trichodermophora strains by hybrid genome assembly.

Genome assembly and annotation using short-paired reads is challenging for eukaryotic organisms due to their large size, variable ploidy and large number of repetitive elements. However, the use of single-molecule long reads improves assembly quality (completeness and contiguity), but haplotype duplications still pose assembly challenges. To address the effect of read length on genome assembly quality, gene prediction and annotation, we compared genome assemblers and sequencing technologies with four strains of the ectomycorrhizal fungus Laccaria trichodermophora. By analysing the predicted repertoire of carbohydrate enzymes, we investigated the effects of assembly quality on functional inferences. Libraries were generated using three different sequencing platforms (Illumina Next-Seq, Mi-Seq and PacBio Sequel), and genomes were assembled using single and hybrid assemblies/libraries. Long reads or hybrid assemby resolved the collapsing of repeated regions, but the nuclear heterozygous versions remained unresolved. In dikaryotic fungi, each cell includes two nuclei and each nucleus has differences not only in allelic gene version but also in gene composition and synteny. These heterokaryotic cells produce fragmentation and size overestimation of the genome assembly of each nucleus. Hybrid assembly revealed a wider functional diversity of genomes. Here, several predicted oxidizing activities on glycosyl residues of oligosaccharides and several chitooligosaccharide acetylase activities would have passed unnoticed in short-read assemblies. Also, the size and fragmentation of the genome assembly, in combination with heterozygosity analysis, allowed us to distinguish homokaryotic and heterokaryotic strains isolated from L. trichodermophora fruit bodies.

Hemiaustroboletus, a new genus in the subfamily Austroboletoideae (Boletaceae, Boletales).

The present study describes Hemiaustroboletus gen. nov. in the subfamily Austroboletoideae (Boletaceae). Hemiaustroboletus is supported by morphological and molecular data using LSU and RPB2 regions. Additionally, its geographic distribution and intraspecific variation were inferred using ITS sequences. The genus is characterised by pileate-stipitate basidiomata; purple, brown, reddish-brown, orange-brown to dark brown vinaceous pileus; whitish or lilac to vinaceous context and a subclavate stipe. Microscopically, it is characterised by ornamented, slightly verrucose, cracked to perforated brown basidiospores. Two species are described within the genus, Hemiaustroboletusvinaceobrunneus sp. nov. and H.vinaceus sp. nov. Hemiaustroboletusvinaceus sp. nov. is morphologically similar to Austroboletusgracilis, which suggests they may have been confused in the past. This study presents the phylogenetic placement, microscopic structures, detailed morphological descriptions and illustrations of both new species.