Abstract

Hydrophobins—secreted small cysteine-rich, amphipathic proteins—foster interactions of fungal hyphae with hydrophobic surfaces, and are involved in the formation of aerial hyphae. Phylogenetic analyses of Tricholoma vaccinum hydrophobins showed a grouping with hydrophobins of other ectomycorrhizal fungi, which might be a result of co-evolution. Further analyses indicate angiosperms as likely host trees for the last common ancestor of the genus Tricholoma. The nine hydrophobin genes in the T. vaccinum genome were investigated to infer their individual roles in different stages of the life cycle, host interaction, asexual and sexual development, and with respect to different stresses. In aerial mycelium, hyd8 was up-regulated. In silico analysis predicted three packing arrangements, i.e., ring-like, plus-like and sheet-like structure for Hyd8; the first two may assemble to rodlets of hydrophobin covering aerial hyphae, whereas the third is expected to be involved in forming a two-dimensional network of hydrophobins. Metal stress induced hydrophobin gene hyd5. In early steps of mycorrhization, induction of hyd4 and hyd5 by plant root exudates and root volatiles could be shown, followed by hyd5 up-regulation during formation of mantle, Hartig’ net, and rhizomorphs with concomitant repression of hyd8 and hyd9. During fruiting body formation, mainly hyd3, but also hyd8 were induced. Host preference between the compatible host Picea abies and the low compatibility host Pinus sylvestris could be linked to a stronger induction of hyd4 and hyd5 by the preferred host and a stronger repression of hyd8, whereas the repression of hyd9 was comparable between the two hosts.

Highlights

  • The mutualistic ectomycorrhizal symbiosis of basidiomycetes with tree roots plays an important role in forest ecosystems, where trees are connected via fungal mycelia leading to better nutrient and water distribution [1]

  • Nine hydrophobin genes were identified from a genomic sequence of T. vaccinum GK6514 based on the lenght of the probable gene, position of introns and the cysteine motif in encoded protein (Table 1)

  • The hydrophobicity pattern of all hydrophobins from T. vaccinum was consistent with class I hydrophobins (Supporting Figure 3 in S1 File)

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Summary

Introduction

The mutualistic ectomycorrhizal symbiosis of basidiomycetes with tree roots plays an important role in forest ecosystems, where trees are connected via fungal mycelia leading to better nutrient and water distribution [1]. The plant benefits from mineral supply, resistance against pathogens, growth promotion and increased metal tolerance, PLOS ONE | DOI:10.1371/journal.pone.0167773. Hydrophobins in Ectomycorrhiza whereas the fungus is supported with growth promoting factors, vitamins and up to 30% of net photosynthetic carbohydrates (for review [2]). More than 50% of the mRNAs were differentially expressed upon host interaction in Pisolithus tinctorius [3]. Knowledge on ectomycorrhiza (ECM) increased significantly, when studies of gene expression, carbohydrate metabolism or nitrogen transport became available for different ectomycorrhizal fungi. Signaling involved in host recognition, e.g. by abietic acid, oligopeptides, phytohormones, volatiles, and hydrophobins, has been discussed (for reviews, see [2, 4, 5] and citations therein)

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