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Mycelial interactions of Laccaria bicolor strains were studied in pure culture and after inoculation onto mycorrhizal hosts. Monokaryon–monokaryon and dikaryon–monokaryon crosses were performed on an agar medium that enhanced nuclear migration to study mating events. The first observance of clamped hyphae, their location, evidence of nuclear migration, and the occurrence of dikaryon–monokaryon matings varied among crosses. One monokaryon–monokaryon and two dikaryon–monokaryon combinations were selected for seedling inoculation to compare their mycorrhizosphere interactions with those observed on agar medium. Seedlings of Pinus banksiana were grown for 20 weeks in a mycelium inoculated soil medium. Three seedlings from each treatment were selected at harvest and ectomycorrhizae reisolates were subject to randomly amplified polymorphic DNA analysis to identify genotypes. This analysis was also done for mycelial samples of the same crosses on agar medium. Variation in the mycobiont genotype was observed for different root isolates from the same seedling, which had been initially inoculated with a compatible monokaryon–monokaryon or a dikaryon–monokaryon strain combination. Root isolates from seedlings receiving the latter treatment included a new dikaryotic genotype produced by a dikaryon–monokaryon mating. Seedling growth was reduced with ectomycorrhizal colonization, most likely because of the photosynthate requirements of the mycobiont during this study. The nature of mycelial interactions and the potential value of a genetically variable mycobiont are discussed. Key words: Buller phenomenon, ectomycorrhizae, intraspecific variability, mycelial interactions, RAPD analysis.

The Ascomycete Ophiostoma novo-ulmi threatens elm populations worldwide. The molecular mechanisms underlying its pathogenicity and virulence are still largely uncharacterized. As part of a collaborative study of the O. novo-ulmi-elm interactome, we analyzed the O. novo-ulmi ssp. americana transcriptomes obtained by deep sequencing of messenger RNAs recovered from Ulmus americana saplings from one resistant (Valley Forge, VF) and one susceptible (S) elm genotypes at 0 and 96 h post-inoculation (hpi). Transcripts were identified for 6424 of the 8640 protein-coding genes annotated in the O. novo-ulmi nuclear genome. A total of 1439 genes expressed in planta had orthologs in the PHI-base curated database of genes involved in host-pathogen interactions, whereas 472 genes were considered differentially expressed (DEG) in S elms (370 genes) and VF elms (102 genes) at 96 hpi. Gene ontology (GO) terms for processes and activities associated with transport and transmembrane transport accounted for half (27/55) of GO terms that were significantly enriched in fungal genes upregulated in S elms, whereas the 22 GO terms enriched in genes overexpressed in VF elms included nine GO terms associated with metabolism, catabolism and transport of carbohydrates. Weighted gene co-expression network analysis identified three modules that were significantly associated with higher gene expression in S elms. The three modules accounted for 727 genes expressed in planta and included 103 DEGs upregulated in S elms. Knockdown- and knockout mutants were obtained for eight O. novo-ulmi genes. Although mutants remained virulent towards U. americana saplings, we identified a large repertoire of additional candidate O. novo-ulmi pathogenicity genes for functional validation by loss-of-function approaches.

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