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Detection of the edible ectomycorrhizal fungus Lyophyllum shimeji colonising seedlings of cultivated conifer species in New Zealand

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Abstract

Lyophyllum shimeji is an edible ectomycorrhizal fungus that is widely distributed in East Asia and also present in the northern regions of Europe. In Japan, L. shimeji is a culinary delicacy, considered amongst all edible mushrooms to have the best taste and to be second only to Tricholoma matsutake in price. Traditionally, fruiting bodies of L. shimeji have been collected from the wild but fruiting of L. shimeji is now relatively uncommon and cannot keep up with increasing consumer demand. As a result, methods for its cultivation are being developed for commercial production in Japan and other countries. In this work, techniques were developed to cultivate L. shimeji on coniferous seedlings using a pure culture inoculum. They resulted in successful mycorrhization of Pinus pinaster and Picea abies in only 8 to 10 months. As ectomycorrhizae of L. shimeji are difficult to identify morphologically, mycorrhization was confirmed using an L. shimeji-specific PCR diagnostic, which was designed following a phylogenetic analysis of the Lyophyllum section Difformia using DNA sequences of the internal transcribed spacer (ITS), intergenic spacer (IGS) and elongation factor 1-α (EF1-α) gene. L. shimeji is a member of the Lyophyllum decastes complex in section Difformia, which also includes Lyophyllum fumosum and L. decastes. This analysis confirmed the separation of L. shimeji from closely related Lyophyllum spp. and enabled its unambiguous detection using an IGS-based PCR diagnostic. This is the first report of successful mycorrhization of L. shimeji on P. pinaster and P. abies and provides an opportunity for its commercial cultivation on conifers in New Zealand.

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Acknowledgments

The authors thank Mr Naoki Endo and Mr Keiichi Okada for their help in the field sampling of Honshimeji basidiomata and Simon Bulman for his review of the manuscript. This work was funded by New Zealand’s Foundation for Research, Science & Technology through contract C02X0704, by The New Zealand Institute for Plant & Food Research Limited and by First Light Mushroom Co. Ltd., Gisborne, New Zealand.

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Authors and Affiliations

  1. The New Zealand Institute for Plant & Food Research Limited, Private Bag 4704, Christchurch, New Zealand

    Sandra B. Visnovsky, Nicholas Cummings, Alexis Guerin-Laguette, Yun Wang & Andrew R. Pitman

  2. Faculty of Agriculture, Shinshu University, Minami-minowa, Nagano, 399-4598, Japan

    Akiyoshi Yamada

  3. Ibaraki Prefectural Forestry Research Institute, To 4692, Naka, Ibaraki, 311-0122, Japan

    Hisayasu Kobayashi

  4. Nara Forest Research Institute, Kibi 1, Takatori, Nara, 635-0133, Japan

    Masataka Kawai

  5. Bio-Protection Research Centre, Lincoln University, PO Box 84, Lincoln, Canterbury, New Zealand

    Nicholas Cummings & Andrew R. Pitman

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  1. Sandra B. Visnovsky
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Correspondence to Sandra B. Visnovsky.

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Supplementary Fig. 1

Nucleotide sequence alignment of the consensus DNA sequences for the IGS from L. shimeji, L. fumosum and L. decastes. The alignment shows the location of the primers ShIGSF1 (green arrow) and ShIGSR2 (orange arrow) designed for the specific detection of “Honshimeji” using Geneious Pro v5.3.4 (Biomatters Ltd. 2011). Coloured squares show polymorphic nucleotides on the divergent consensus sequence. A dash represents a missing nucleotide in the associated IGS sequence. The primers were designed to amplify a 345-bp fragment from L. shimeji (PPTX 488 kb)

Supplementary Table S1

Collections of Lyophyllum from this study and the GenBank accession numbers for the respective ITS, IGS and EF1-α DNA sequences (DOCX 18 kb)

Supplementary Table S2

DNA sequences of Lyophyllum retrieved from GenBank used in this study (DOCX 25 kb)

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Visnovsky, S.B., Cummings, N., Guerin-Laguette, A. et al. Detection of the edible ectomycorrhizal fungus Lyophyllum shimeji colonising seedlings of cultivated conifer species in New Zealand. Mycorrhiza 24, 453–463 (2014). https://doi.org/10.1007/s00572-013-0552-5

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