T he genus Armillaria causes root rot disease in both gymnoand angiosperms, in forests, parks, and even vineyards in more than 500 host plant species 1 across the world. Most Armillaria species are facultative necrotrophs, which, after colonizing and killing the root cambium, transition to a saprobic phase, decomposing dead woody tissues of the host. As saprotrophs, Armillaria spp. are white rot (WR) fungi, which can efficiently decompose all components of plant cell walls, including lignin, (hemi-)cellulose and pectin 2 . They produce fleshy fruiting bodies (honey mushrooms) that appear in large clumps around infected plants and produce sexual spores. The vegetative phase of Armillaria is predominantly diploid rather than dikaryotic like most basidiomycetes.Individuals of Armillaria can reach immense sizes and include the 'humongous fungus' , one of the largest terrestrial organisms on Earth 3 , measuring up to 965 hectares and 600 tons 4 , and can display a mutation rate ≅ 3 orders of magnitude lower than most filamentous fungi 5 . Individuals reach this immense size via growing rhizomorphs, dark mycelial strings 1-4 mm wide that allow the fungus to bridge gaps between food sources or host plants 1,6 (hence the name shoestring root rot). Rhizomorphs develop through the aggregation and coordinated parallel growth of hyphae, similar to some fruiting body tissues 7,8 . As migratory and exploratory organs, rhizomorphs can grow approximately 1 m yr −1 and cross several metres underground in search for new hosts, although roles in uptake and longrange translocation of nutrients have also been proposed 1,9,10 . Root contact by rhizomorphs is the main mode of infection by the fungus, which makes the prevention of recurrent infection in Armillariacontaminated areas particularly difficult 1 . Despite their huge impact on forestry, horticulture and agriculture, the genetics of the pathogenicity of Armillaria species is poorly understood. The only -omics data published so far have highlighted a substantial repertoire of plant cell wall degrading enzymes (PCWDE) and secreted proteins, among others, in A. mellea and A. solidipes 11,12 , while analyses of the genomes of other pathogenic basidiomycetes (such as Moniliophthora 13,14 , Heterobasidion 15 and Rhizoctonia 16 ) identified genes coding for PCWDEs, secreted and effector proteins or secondary metabolism (SM) as putative pathogenicity factors. However, the lifecycle and unique dispersal strategy of Armillaria prefigure other evolutionary routes to pathogenicity, which, along with other potential genomic factors (such as transposable elements 17 ) are not yet known.Here, we investigate genome evolution and the origin of pathogenicity in Armillaria using comparative genomics, transcriptomics

Dermatophagoides pteronyssinus is the European dust mite and a major source of human allergens. Here, we present the first draft genome sequence of the mite, as well as the ab initio gene prediction and functional analyses that will facilitate comparative genomic analyses with other mite species.

The European house dust mite Dermatophagoides pteronyssinus is of significant medical importance as it is a major elicitor of allergic illnesses. In this analysis we have undertaken comprehensive bioinformatic and proteomic examination of Dermatophagoides pteronyssinus airmid, identified 12,530 predicted proteins and validated the expression of 4,002 proteins. Examination of homology between predicted proteins and allergens from other species revealed as much as 2.6% of the D . pteronyssinus airmid proteins may cause an allergenic response. Many of the potential allergens have evidence for expression ( n = 259) and excretion ( n = 161) making them interesting targets for future allergen studies. Comparative proteomic analysis of mite body and spent growth medium facilitated qualitative assessment of mite group allergen localisation. Protein extracts from house dust contain a substantial number of uncharacterised D . pteronyssinus proteins in addition to known and putative allergens. Novel D . pteronyssinus proteins were identified to be highly abundant both in house dust and laboratory cultures and included numerous carbohydrate active enzymes that may be involved in cuticle remodelling, bacteriophagy or mycophagy. These data may have clinical applications in the development of allergen-specific immunotherapy that mimic natural exposure. Using a phylogenomic approach utilising a supermatrix and supertree methodologies we also show that D . pteronyssinus is more closely related to Euroglyphus maynei than Dermatophagoides farinae .

Between 1954 and 1963 cone production of individual trees and seedfall were measured in a mature white spruce stand at the Riding Mountain Forest Experimental Area in Manitoba.Results showed that dominant and co-dominant trees produced heavier cone crops and produced them more frequently than intermediates; intermediates produced heavier and more frequent cone crops than suppressed trees.Over the ten-year period 11.7 million white spruce seed fell per acre, of which 6.7 million were sound. Total annual recorded seedfall varied with a high of 5,625,000 seeds per acre in 1960 and a low of 10,000 per acre in 1958. Seed soundness was highest in years of heavy seedfall and lowest in years of nil and light seedfall.Seedfall generally began in early August with peak seedfall occuring in late August or early September. However, in several years below average air temperatures and sunshine and above average precipitation delayed peak seedfall until late September. Early- and late-falling seed was not as sound as that which fell during the period of peak seedfall.

The genus Armillaria includes some of the most devastating forest pathogens worldwide. 58Armillaria causes root rot disease in both gymno-and angiosperms, in forests, parks, and even 59 vineyards in more than 500 host plant species 1 . Most Armillaria species are facultative 60 necrotrophs, which, after colonizing and killing the root cambium, transition to a saprobic phase, 61 decomposing dead woody tissues of the host. As saprotrophs, Armillaria spp. are white rot (WR) 62 fungi, which can efficiently decompose all components of plant cell walls, including lignin, (hemi-63 )cellulose and pectin 2 . They produce fleshy fruiting bodies (honey mushrooms) that appear in 64 large clumps around infected plants and produce sexual spores. The vegetative phase of 65Armillaria is predominantly diploid rather than dikaryotic like most basidiomycetes. 66 peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/166231 doi: bioRxiv preprint first posted online Jul. 20, 2017; are not yet known. 87Here, we investigate genome evolution and the origins of pathogenicity in Armillaria 88 using comparative genomics, transcriptomics and proteomics. We sequenced the genomes of 89four Armillaria species to combine with that of related saprotrophic, hemibiotrophic and 90 mycorrhizal fungi. Transcript and proteome profiling of invasive and reproductive developmental 91 peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission. Cylindrobasidium as their closest relatives ( Fig. 1/a). We estimated the age of pathogenic 109Armillaria spp. at 21 million years (myr) and their divergence from Guyanagaster at 42 myr 110 ( Supplementary Fig. 4), coincident with decreasing temperatures and the spread of deciduous 111 forests in the Eocene. Reconstruction of genome-wide gene duplication and loss histories in 27 112Agaricales species revealed an early origin for most genes, followed by lineage-specific gene 113 losses in most family and genus level groups, except Armillaria, which showed a net genome 114 expansion: 16,687 protein-coding genes were inferred for the most recent common ancestor 115 peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission. Table 4 20, 2017; 8 buried in sawdust-rich medium and four fruiting body stages (Supplementary Table 6). 185peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission. The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/166231 doi: bioRxiv preprint first posted online Jul.Differential expression analysis identified 1,303 and 1,610 over-and underexpressed genes in 186 rhizomorphs relative to vegetative mycelium grown on the same medium (FC VM >4, p≤0.05), 187 respectively, marking one of the largest expression change in our experiments (Fig. 2). 188Similarly, the highest number of unique proteins (n=729) Figs 13-14). 207We observed signi...

The European house dust mite, Dermatophagoides pteronyssinus is a major source of airborne allergens worldwide and is found in half of European homes. Interactions between microbes and house dust mites (HDM) are considered important factors that allow them to persist in the home. Laboratory studies indicate the European HDM, D. pteronyssinus is a mycophagous mite, capable of utilising a variety of fungi for nutrients, however specific mycolytic digestive enzymes are unknown. Our previous work identified a number of putative glycosyl hydrolases present in the predicted proteome of D. pteronyssinus airmid and validated the expression of 42 of these. Of note, three GH16 proteins with predicted β-1,3 glucanase activity were found to be consistently present in the mite body and excretome. Here, we performed an extensive bioinformatic, proteomic and biochemical study to characterize three-novel β-1,3 glucanases from this medically important house dust mite. The genes encoding novel β-1,3 glucanases designated Glu1, Glu2 and Glu3 were identified in D. pteronyssinus airmid, each exhibited more than 59% amino acid identity to one another. These enzymes are encoded by Glu genes present in a tri-gene cluster and protein homologs are found in other acari. The patchy phyletic distribution of Glu proteins means their evolutionary history remains elusive, however horizontal gene transfer cannot be completely excluded. Recombinant Glu1 and Glu2 exhibit hydrolytic activity toward laminarin, pachyman and barley glucan. Excreted β-1,3 glucanase activity was increased in response to D. pteronyssinus airmid feeding on baker's yeast. Active β-1,3 glucanases are expressed and excreted in the faeces of D. pteronyssinus airmid indicating they are digestive enzymes capable of breaking down β-1,3 glucans of fungi present in house dust.

Treatment of tree seed with fungicides is an accepted technique in nursery practice to control losses due to damping-off; in field sowing, seeds are generally treated with bird and rodent repellents. However, in the province of Manitoba, Captan-SOW3 a fungicide with certain rodent repellent q~a l i t i e s ,~ has been applied to coniferous seed sown both in the field and in the nursery. Excellent germination has normally occurred in the nursery. However, results from field sowing have been variable and even in very favourable years germination failures have occurred. For personal use only.

In the version of this Article originally published, it was incorrectly stated that "16,687 protein-coding genes were inferred for the most recent common ancestor (MRCA) of Armillaria"; the value was incorrect and it should have read "15,787". This has now been corrected.