Characterization of mating type genes in heterothallic Neonectria species, with emphasis on N. coccinea, N. ditissima, and N. faginata.

Neonectria ditissima and N. faginata are canker pathogens involved in an insect-fungus disease complex of American beech (Fagus grandifolia) in North America commonly known as beech bark disease (BBD). In Europe, both N. ditissima and N. coccinea are involved in BBD on European beech (Fagus sylvatica). Field observations across the range of BBD indicate ascospores to be the dominant spore type in the environment. Several studies report a heterothallic (self-sterile) mating strategy for Neonectria fungi, but one study reported homothallism (self-fertility) for N. ditissima. As such, investigations into mating strategy are important for understanding both the disease cycle and population genetics of Neonectria. This is particularly important in the United States given that over time N. faginata dominates the BBD pathosystem despite high densities of nonbeech hosts for N. ditissima. This study utilized whole-genome sequences of BBD-associated Neonectria spp. along with other publicly available Neonectria and Corinectria genomes and in vitro mating assays to characterize mating type (MAT) locus and confirm thallism for select members of Neonectria and Corinectria. MAT gene-specific primer pairs were developed to efficiently characterize the mating types of additional single-ascospore strains of N. ditissima, N. faginata, and N. coccinea and several other related species lacking genomic data. These assays also confirmed the sexual compatibility among N. ditissima strains from different plant hosts. Maximum likelihood phylogenetic analyses of both MAT1-1-1 and MAT1-2-1 sequences recovered trees with similar topology to previously published phylogenies of Neonectria and Corinectria. The results of this study indicate that all Neonectria and Corinectria tested are heterothallic based on our limited sampling and, as such, thallism cannot help explain the inevitable dominance of N. faginata in the BBD pathosystem.

Genome Resources for Thelonectria rubi, the Causal Agent of Nectria Canker of Caneberry.

Thelonectria rubi is the causal agent of Nectria canker of Rubus spp. Here, we report a high-quality draft genome sequence for this pathogen, which also represents the first genome sequence for a Thelonectria species. The genome assembly was 44.6 Mb in size, assembled into 669 scaffolds and consisting of 12,973 predicted protein-coding genes. The availability of genome data for T. rubi provides a critical additional resource for an important plant pathogen and will be useful for fungal biology, comparative genomic, taxonomic and population studies of this and related species.

Isolation of Nectria galligena from Cankers on Sweet Birch.

The fungus Nectria galligena infects many species of hardwood trees, causing "perennial canker," "European canker," or "target canker." Cultures from ascospores produced in stromata on the cankers have confirmed the pathogen species, but the fungus has rarely been successfully isolated from inside the cankers. Concern for the sweet birch trees affected by this disease in Connecticut prompted us to try isolations, using a method successful in isolating chestnut tree pathogens. Small pieces of cankered bark were stabbed into Granny Smith apples and, after incubation in boxes in the laboratory, pure cultures of N. galligena were easily removed as the pathogen slowly grew out, unchallenged, into the tissue of the apples. This technique will now be used to sample the N. galligena population in the birch populations being studied.