Shiga Forest Research Center

Yasu, Japan

Shiga Forest Research Center

Yasu, Japan
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Yamaguchi M.,Mushroom | Narimatsu M.,Iwate Prefectural Forestry Technology Center | Fujita T.,Kyoto Prefectural Forestry Technology Center | Kawai M.,Nara Forest Research Institute | And 7 more authors.
Mycorrhiza | Year: 2016

Tricholoma matsutake is an ectomycorrhizal basidiomycete that produces prized, yet uncultivable, “matsutake” mushrooms along densely developed mycelia, called “shiro,” in the rhizosphere of coniferous forests. Pinus densiflora is a major host of this fungus in Japan. Measuring T. matsutake biomass in soil allows us to determine the kinetics of fungal growth before and after fruiting, which is useful for analyzing the conditions of the shiro and its surrounding mycorrhizosphere, predicting fruiting timing, and managing forests to obtain better crop yields. Here, we document a novel method to quantify T. matsutake mycelia in soil by quantifying a single-copy DNA element that is uniquely conserved within T. matsutake but is absent from other fungal species, including close relatives and a wide range of ectomycorrhizal associates of P. densiflora. The targeted DNA region was amplified quantitatively in cultured mycelia that were mixed with other fungal species and soil, as well as in an in vitro co-culture system with P. densiflora seedlings. Using this method, we quantified T. matsutake mycelia not only from shiro in natural environments but also from the surrounding soil in which T. matsutake mycelia could not be observed by visual examination or distinguished by other means. It was demonstrated that the core of the shiro and its underlying area in the B horizon are predominantly composed of fungal mycelia. The fungal mass in the A or A0 horizon was much lower, although many white mycelia were observed at the A horizon. Additionally, the rhizospheric fungal biomass peaked during the fruiting season. © 2016 Springer-Verlag Berlin Heidelberg


Ota Y.,Japan Forestry and Forest Products Research Institute | Yamanaka T.,Japan Forestry and Forest Products Research Institute | Murata H.,Mushroom | Neda H.,Mushroom | And 5 more authors.
Mycologia | Year: 2012

Tricholoma matsutake (S. Ito & S. Imai) Singer and its allied species are referred to as matsutake worldwide and are the most economically important edible mushrooms in Japan. They are widely distributed in the northern hemisphere and established an ectomycorrhizal relationship with conifer and broadleaf trees. To clarify relationships among T. matsutake and its allies, and to delimit phylogenetic species, we analyzed multilocus datasets (ITS, megB1, tef, gpd) with samples that were correctly identified based on morphological characteristics. Phylogenetic analyses clearly identified four major groups: matsutake, T. bakamatsutake, T. fulvocastaneum and T. caligatum; the latter three species were outside the matsutake group. The haplotype analyses and median-joining haplotype network analyses showed that the matsutake group included four closely related but clearly distinct taxa (T. matsutake, T. anatolicum, Tricholoma sp. from Mexico and T. magnivelare) from different geographical regions; these were considered to be distinct phylogenetic species. © 2012 by The Mycological Society of America.


Kojima N.,Shiga Forest Research Center | Hosoda I.,Japan Forestry and Forest Products Research Institute | Nakamura S.,Oji Paper Co.
Hydrological Processes | Year: 2012

Understanding the effects of severe human induced forest disturbances with soil loss on rainfall-runoff responses is important for future forest management. However, few studies have addressed this issue, which is methodologically difficult compared with the hydrological assessments of the effects of logging. In this study, several small catchments in Japan with different soil and geological conditions were compared using the runoff model HYCYMODEL to reveal their runoff characteristics. The results were then examined on the basis of runoff mechanisms to demonstrate the possible ranges of the effects derived from human disturbances for each geological type. For granite mountains, bare land can be considered the severest case of disturbances leading to high stormflow peaks, although a large baseflow remains because of the water storage fluctuation in weathered bedrock. For sedimentary rock mountains, the severest case may be a forest on the clayey soil without brown forest soil producing flashy runoff characteristics including a large stormflow volume with a sensitive response to the antecedent dryness and a low baseflow rate. © 2011 John Wiley & Sons, Ltd.


Hatoh K.,Kyoto University | Izumitsu K.,Kyoto University | Morita A.,Kyoto University | Shimizu K.,Chiba University | And 6 more authors.
Mycoscience | Year: 2013

Agrobacterium tumefaciens-mediated transformation (AMT) was successfully applied to mycelia of the 3 economically important mushrooms Hypsizigus marmoreus, Flammulina velutipes, and Grifola frondosa. We used the hygromycin B resistance gene (hph) under the control of the Cryptococcus neoformans actin promoter. Eighty-six resistant strains of H. marmoreus, 4 of F. velutipes, and 2 of G. frondosa were obtained. All transformants were highly resistant to hygromycin B, suggesting that the C. neoformans actin promoter has a potential universal promoter activity in basidiomycetes. Southern analysis revealed random but single integration of the hph gene. © 2012 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.


Izumitsu K.,Kyoto University | Hatoh K.,Kyoto University | Sumita T.,Kyoto University | Kitade Y.,Kyoto University | And 8 more authors.
Mycoscience | Year: 2012

We have optimized a simple and rapid preparation procedure for mushroom DNA extraction from colonies on media or from fruiting bodies for PCR amplification. The protocol combines microwaving twice for 1 min, cooling for 10 min, and centrifuging for 5 min. By using this procedure, more than 100 samples of mushroom DNA can be prepared within 1 h. The DNA obtained can be used for (1) identifying mushroom species by PCR and subsequent sequencing, (2) amplifying low copy number genes (at least 2,000 bp), and (3) screening genetic transformants. This technique will contribute to the mycology of mushroom species. © 2012 The Mycological Society of Japan and Springer.


Yamanaka T.,Japan Forestry and Forest Products Research Institute | Ota Y.,Japan Forestry and Forest Products Research Institute | Konno M.,Miyagi Prefectural Forestry Technology Institute | Kawai M.,Nara Forest Research Institute | And 4 more authors.
Mycologia | Year: 2014

Tricholoma matsutake is the most commercially important edible mushroom in pine forests in Japan. Tricholoma bakamatsutake and T. fulvocastaneum, species closely related to T. matsutake, occur in Fagaceae forests. We examined ectomycorrhizal (EM) formation by these Tricholoma species by in vitro synthesis among seven strains (two of T. matsutake, four of T. bakamatsutake, one of T. fulvocastaneum) and axenic plants of pine (Pinus densiflora) and oak (Quercus serrata, Q. phillyraeoides). All strains, except for one of T. matsutake, formed EM associations with both pine and oak. Plant growth and mycelial development were differently affected by EM formation depending on the plant-fungus combination. © 2014 by The Mycological Society of America.


Murata H.,Mushroom | Ota Y.,Japan Forestry and Forest Products Research Institute | Yamada A.,Shinshu University | Ohta A.,Shiga Forest Research Center | And 2 more authors.
Mycoscience | Year: 2013

Tricholoma dulciolens is an ectomycorrhizal basidiomycete that produces "matsutake"-like mushrooms in association with Picea abies in Fennoscandia. The phylogenetic position of T. dulciolens relative to matsutake species is, however, unknown. In the present study, we demonstrated that T. dulciolens is phylogenetically independent from T. matsutake and its allied species. In addition, herbarium specimens identified as T. caligatum from Abies alba forests in Spain were phylogenetically related to but distinct from T. dulciolens. Based on ribosomal internal transcribed spacer DNA sequence data, T. dulciolens appears to exist in North America, as well. This is the first phylogeny of T. dulciolens in Fennoscandia and its related species in southern Europe. © 2013 The Mycological Society of Japan.


Yamada A.,Shinshu University | Endo N.,Shinshu University | Murata H.,Japan Forestry and Forest Products Research Institute | Ohta A.,Shiga Forest Research Center | Fukuda M.,Shinshu University
Mycoscience | Year: 2013

Tricholoma matsutake produces commercially valuable yet uncultivable matsutake mushrooms during an ectomycorrhizal association with coniferous trees. In the Far East, most matsutake are harvested in managed Pinus densiflora forests. To determine whether T. matsutake has host plant specificity, we synthesized mycorrhiza in vitro between T. matsutake Y1 that originated from a P. densiflora forest and various Pinaceae and oak hosts. The strain Y1 formed a continuous Hartig net, a sign of ectomycorrhization, in the lateral roots of Pinus sylvestris, Pinus koraiensis, Pinus parviflora var. pentaphylla, Picea glehnii, Picea abies, and Tsuga diversifolia seedlings in vitro, which resembled those formed with the natural host Pinus densiflora. The strain conferred a discontinuous Hartig net with Pinus thunbergii, Picea yezoensis, Abies veitchii, and Larix kaempferi. However, no such development by this strain was observed on the roots of Quercus serrata, unlike T. bakamatsutake B1, a false matsutake that is symbiotic with oak trees. The data suggest that T. matsutake can be associated with diverse conifers but may establish ectomycorrhizal relationships only with specific host plant species. © 2013 The Mycological Society of Japan.


Murata H.,Mushroom | Ohta A.,Shiga Forest Research Center | Yamada A.,Shinshu University | Horimai Y.,Shinshu University | And 3 more authors.
Mycoscience | Year: 2015

We document here that monosporous isolates of the ectomycorrhizal basidiomycete Tricholoma matsutake were initially uni- or bi-nucleate. During pure culture, however, many uninucleate isolates became multinucleate. While the parent strain had two patterns of single nucleotide polymorphisms (SNPs) within its single-copy DNA, 19 of 20 monosporous isolates exhibited one of the two parent SNPs, and an isolate that was binucleate upon germination had both, indicating the former were monokaryotic and the latter dikaryotic. Of those, two isolates carrying SNPs different from one another have been predominantly uninucleate for 9 years. These isolates may be useful in genetics/breeding of "matsutake" mushrooms. © 2014 The Mycological Society of Japan.

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