Abstracts of papers published in other scientiﬁc journals during 2007-2015

The genetic constitution of B incompatibility factors
in Lentinula edodes was estimated by a recombinationt e s t . We i s o l a t e d B f a c t o r r e c o m b i n a n t s i nmonosporous isolates from a cultivated strain fromJapan and a wild strain from Papua New Guinea. By
an intrastrain mating test between B factor
recombinants, these recombinants were classiﬁed into
two groups in a cultivated strain from Japan and at
least four groups in a wild strain from Papua New
Guinea. These results suggest that the third matingtype locus, Bγ, is linked to Bα or Bβ in a wild strain
from Papua New Guinea and imply that a putative Bγ
locus is the same in allelic speciﬁcity or absent in thetwo component homokaryons in a cultivated strain
from Japan.

Papers with TMI contribution numbers
No. 391
Intersterility between populations of Lentinula
edodes from Papua New Guinea
Norihiro Shimomura, Kazuhisa Terashima, andKozaburo Hasebe
Mycosciense 50: 240-243, 2009.
Mating tests among strains of Lentinula edodes
distributed in Asia-Australasia were conducted. As aresult, 26 strains were classified into three groups: 2
strains from Mt. Wilhelm in Papua New Guinea（PN1group）showed intersterility with 7 strains from Mt.
Albert Edward and Mt. Kaisenik in Papua New Guinea（PN2 group）and semicompatibility（clamp formation
restricted to contact zone between paired monokaryons）with 17 strains from Asia-Australasia（AAgroup）, whereas the strains of the PN2 group showedcompatibility with the AA group. These results suggestthat the shiitake populations distributed in AsiaAustralasia including Papua New Guinea are in theprocess of speciation.

Key words: B incompatibility factor, Lentinula
edodes, mating test, recombination.
シイタケの B 不和合性因子における 3 遺伝子座の可能性
霜村典宏・下廣慶宜・会見忠則・長谷部公三郎
日本きのこ学会誌 19: 22-24, 2011.
シイタケの B 不和合性因子組換え型菌株間の交配試験により，
本因子の構成について解析した．
日本産の栽培品種 1 菌株およびパプアニューギニ
ア産野生菌株 1 菌株より B 不和合性因子組換え型菌株を分離した．B 不和合性因子組換え型菌株
間の系統内交配をした結果，今回用いた日本産栽
培品種では 2 群に，そしてパプアニューギニア産野生菌株では 4 群に分類できた．これらの結果か
ら，パプアニューギニア産野生菌株の B 不和合性因子においては，Bαまたは Bβに第 3 の座位
Bγが連鎖している可能性が考えられた．また，日本産栽培品種では Bγがホモであるか，あるいは存在しない可能性があると思われた．

Key words: geographic lineage, intersterility,
Lentinula edodes, mating compatibility.
Contribution No.391 of the Tottori Mycological Institute. 菌
蕈研究所研究業績 第 391 号．

No. 392
Possibility of three loci for B incompatibility
factor in Lentinula edodes
Norihiro Shimomura, Yoshitaka Shimohiro, Tadanori
Aimi and Kozaburo HasebeMushroom Science and Biotechnology 19: 22-24,
2011.

Contribution No. 392 of the Tottori Mycological Institute. 菌
蕈研究所研究業績第 392 号．

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No. 393
PCR-based gene marker with a high efficiency
genome scanning（HEGS）system:
application for distinguishing among cultivars
in Lentinula edodes
Kazuhisa Terashima, Chisato Funato, Teruyuki
Matsumoto, Asa Maeda, and Kozaburo Hasebe
World J Microbiol Biotechnol 28: 1315-1319, 2012.
To establish a high throughput and cost-efficiency
procedure for distinguishing among cultivars in
Lentinula edodes, the polymorphism in the genes
reported previously in this fungus was examined using
PCR or PCR–RFLP techniques with a high-efﬁciency
genome scanning（HEGS）system. As a result, PCRbased markers derived from eight genes（tyr, cap, ppa,
IGS-RFLP, pri B-RFLP, mfb C-RFLP, gla-RFLP, xyRFLP）showed polymorphisms among cultivars in thisfungus and consequently, enabled to distinguishseventy-nine cultivars used in this study.
Key words: the shiitake mushroom, HEGS genemarker, breederʼs right, premium production.
Contribution No.393 of the Tottori Mycological Institute. 菌
蕈研究所研究業績 第 393 号．

No. 394
Intraspecific comparison of three complete
mitochondrial genome sequences from theoyster mushroom, Pleurotus ostreatus
Mai Morinaga, Yasuhito Okuda, Rika Takagi, Teruyuki
Matsumoto and Yukitaka Fukumasa-Nakai
Mushroom Science 18: 293-299, 2012. Proceedings ofthe 18th Congress of the International Society for
Mushroom Science.

presumably important; however, these effects have notbeen eluciadated. Intraspeciﬁc variation of mt genome
structures, which is indispensable information forassessing the effects of mt genome structure, has beenassessed mainly by using restriction fragment length
polymorphism（RFLP）patterns. Here, we usedcomplete mt genome sequences of Pleurotus ostreatus
isolates to examine intraspecific variation among mt
genome structure among three genome types, I（accession number: AB573642）, II（accession
number: AB573692）, and III（NC_009905）, andcompared this variation with that determined using
RFLP patterns.
The mt genome of each P. ostreatus type was acircular DNA molecule ranging in size from 71 947 bp（type I）to 73 242 bp（type III）
. Each of the genomes
contained 14 common mt genes, one ribosomal small
subunit protein 3 gene, one RNA polymerase gene
（only in type III）, two DNA polymerase genes, andtwo rRNA genes. Twenty-four tRNA genes were
determined to form a common set among the three
types of mt genomes. The type I mt genome had two
additional tRNA genes. The gene orders in each mt
genome were almost identical, but each had different
open reading frames（ORFs）and introns. Comparisonof nucleotide sequences showed that there were many
substitutions and indels in the intergenic regions.
Futhermore, nucleotide sequence differences wereobserved in gene-coding regions, the majority ofwhich were related to polymorphisms in amino-acidcoding sequences. These results indicated bothconservation of mt genome primary structure in P.
ostreatus species and sequence polymorphisms that
corresponded to the mtDNA RFLPs.
Key words: comparative genomics, genome
organization, mitochondrial genome（mtDNA）,
Pleurotus ostreatus.

Mitochondria, ATP-producing organelles in cells,
have their own independent genomes called themitochondrial（mt）genome. The effects of the mt
genome structure on cellular phenotype during the
fruiting body formation of basidiomycetes areContribution No. 394 of the Tottori Mycological Institute. 菌
蕈研究所研究業績第 394 号．

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No. 395
Characterization of vitamin B12 compounds inthe fruiting bodies of shiitake mushroom（Lentinula edodes）and bed logs after fruitingof the mushroom
Tomohiro Bito, Fei Teng, Noriharu Ohishi, Shigeo
Takenaka, Emi Miyamoto, Emi Sakuno, Kazuhisa
Terashima, Yukinori Yabuta, Fumio Watanabe
Mycoscience 55: 462-468, 2014. DOI: 10.1016/j.
myc.2014.01.008
This study determined the vitamin B12 content incommercially available dried fruiting bodies ofshiitake mushroom, Lentinula edodes. The vitamin B12contents in dried donko-type fruiting bodies withclosed caps（5.61 ± 3.90 μg/100 g dry weight）, did
not significantly differ from those of dried koushintype fruiting bodies with open caps（4.23 ± 2.42 μg/
100 g dry weight）. The bed logs after fruiting of themushroom also contained the vitamin B 12 levels
similar to that in the dried shiitake fruiting bodies. To
determine whether the dried shiitake fruiting bodies
and their bed logs contained vitamin B 12 or othercorrinoid compounds that are inactive in humans, we
puriﬁed corrinoid compounds using an immunoafﬁnity
column and identiﬁed vitamin B12 using vitamin B12dependent Escherichia coli 215 bioautograms andliquid chromatography-electrospray ionization tandem
mass spectrometry（LC/ESI-MS/MS）chromatograms.
Dried shiitake fruiting bodies rarely contained an
unnatural corrinoid vitamin B 12[c-lactone] that isinactive in humans. Given that shiitake mushroom
lacks the ability to synthesize vitamin B12 de novo, thevitamin B12 found in dried shiitake fruiting bodies
must have been derived from the bed logs.

No. 396
Development of the specific rDNA-ITS andSSR primers to detect the ectomycorrhizal
fungus Rhizopogon roseolus（In Japanese）
Yasuhito Okuda, Chisato Funato, Norihiro Shimomura
and Teruyuki Matsumoto
Japanese Journal of Mycology 55: 29-34, 2014.
By the comparison of sequences of rDNA-ITS（internal transcribed spacer）region and SSR（simple
sequence repeat）loci from 45 isolates of Rhizopogonroseolus, the ﬁve speciﬁc primer sets, one rDNA-ITS
primer and four SSR primers, were designed for
detection of R. roseolus. All these primer sets ampliﬁed
the specific PCR products for the total DNAs frommycelia of 45 R. roseolus isolates, respectively, but noamplified product was detected for the DNAs fromhost plant（Pinus thunbergii）and 16 ectomycorrhizal
isolates consist of 13 species including two
Rhizopogon species, R. luteolus and R. superiorensis.
Furthermore, target regions of these primer sets werealso specifically amplified using by DNA templates
from ectomycorrhizas of this fungus. Thus, the fivesets of primer pair represent a potent tool for themonitoring of introduced isolates of R. roseolus andfor molecular ecology applications.
Key words: ectomycorrhiza, molecular marker,
multiplex PCR, Shoro.
外生菌根菌ショウロの検出用特異的 rDNA-ITS お
よび SSR プライマーの開発
奥田康仁・舩戸知聖・霜村典宏・松本晃幸
日本菌学会会報 55: 29-34, 2014.
日本産ショウロ 45 株の rDNA-ITS および SSR
座位領域の配列比較を行い，ショウロに特異的な
rDNA-ITS および SSR 座位増幅用プライマーセッ
トを作製した．各プライマーにより，ショウロ
45 株の菌糸から抽出した全 DNA すべてから特異的な PCR 増幅が認められたが，宿主クロマツや本菌と同属の 2 種を含む 13 種 16 株の外生菌根菌の全 DNA では認められなかった．ショウロの菌

K e y w o r d s : c h l o r a m i n e - T, c o b a l a m i n ,pseudovitamin B12, vegetarians, vitamin B12[c-lactone].
Contribution No.395 of the Tottori Mycological Institute. 菌
蕈研究所研究業績 第 395 号．

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High-resolution melting（HRM）解 析 を 用 い た
シイタケ品種「菌興 115 号」の迅速 DNA 品種識別法の開発（和文）前田亜紗・寺島和寿・長谷部公三郎
日本きのこ学会誌 23: 114-119, 2015.

根から抽出した DNA からも特異的な増幅が認められた．このことはこれらのプライマーが菌根からのショウロの検出および分子生態学的調査に利用できることを示している．
Contribution No.396 of the Tottori Mycological Institute. 菌
蕈研究所研究業績 第 396 号．

No. 397
Development of a method for rapid straintyping of a shiitake cultivar, Kinko 115, by
high-resolution melting（HRM）analysis（InJapanese）
Asa Maeda, Kazuhisa Terashima and Kozaburo
HasebeMushroom Science and Biotechnology 23: 114-119,
2015.
High-resolution melting（HRM）analysis is a
closed-tube method for detecting nucleotide
polymorphisms, enabling rapid genotyping. In thisstudy, we have developed a rapid strain-typing method
for Kinko 115, a premium shiitake cultivar, by HRM
analysis. When HRM analyses of tyrosinase（tyr）
genes of 64 shiitake cultivars were carried out, 3
cultivars including Kinko 115 could be distinguishedfrom another 61 cultivars. After that, HRM analyses of
the ribosomal intergenic spacer（IGS）region in these3 cultivars revealed that Kinko 115 was distinguishedfrom the other 2 cultivars. Consequently, Kinko 115
could be distinguished from other cultivars by HRM
analyses of the tyr gene and the IGS region.
Furthermore, Kinko 115 could be distinguished fromthe others by HRM analyses using not only purified
DNA, but also crude DNA extracts from mycelia orgills of dried fruiting bodies. Thus, the HRM analyses
developed in this study would be useful for rapidly
and easily identifying Kinko 115.
Key words: high-resolution melting（HRM）
analysis, Lentinula edodes, premium shiitake cultivar,
strain-typing.

S4

本研究では，High-resolution melting（HRM）解析を用い，各地でブランド化が進行中である，極
厚肉で食味に優れた原木シイタケ用品種「菌興115 号」の，迅速・簡便な識別法を開発した．まず，
チロシナーゼ遺伝子座の 3' 末端領域を標的として，64 品種のシイタケを解析した結果，菌興 115
号を他 61 品種と識別することができた．さらに，
IGS 領域を標的として残る 3 品種の解析を行った
結果，菌興 115 号を他の 2 品種と識別することができた．これらの結果から，上記 2 箇所の遺伝子
領域の HRM 解析を行うことにより，菌興 115 号
を他の 63 品種と区別できることが明らかになった．また，培養菌糸体および乾シイタケのヒダから粗抽出した DNA を用いても菌興 ll5 号を識別できることを明らかにした．以上の結果から，
HRM 解析法は，菌興 115 号の迅速，簡便な検出に有効であることが示された．
Contribution No.397 of the Tottori Mycological Institute. 菌
蕈研究所研究業績 第 397 号．

Papers without TMI contribution numbers
Luminescent Mycena: new and noteworthy
speciesDennis E. Desjardin, Brian A. Perry, D. Jean Lodge,
Cassius V. Stevani and Eiji Nagasawa
Mycologia 102: 459-477, 2010. DOI: 10.3852/09197.
Seven species of Mycena are reported as luminescent,representing specimens collected in Belize, Brazil,
Dominican Republic, Jamaica, Japan（Bonin Islands）,
Malaysia（Borneo）and Puerto Rico. Four of them
represent new species（Mycena luxaeterna, M.
luxarboricola, M. luxperpetua, M. silvaelucens）andthree represent new reports of luminescence inpreviously described species（M. aff. abieticola, M.
aspratilis, M. margarita）. Mycena subepipterygia issynonymized with M. margarita, and M. chlorinosma

78 sequences from the fast-evolving nuclear internaltranscribed spacers and are able to recognize 18
reciprocally monophyletic species. To address whether
or not porcini form a monophyletic group, wecompiled a broadly sampled dataset of 41 taxa,
including other members of the Boletineae, and used
separate and combined phylogenetic analysis ofsequences from the nuclear large subunit ribosomal
DNA, the largest subunit of RNA polymerase II, andthe mitochondrial ATPase subunit six gene. Contrary
to previous studies, our separate and combined
phylogenetic analyses support the monophyly of
porcini. We also report the discovery of two taxa that
expand the known distribution of porcini to Australia
and Thailand and have ancient phylogenetic
connections to the rest of the group. A relaxedmolecular clock analysis with these new taxa dates theorigin of porcini to between 42 and 54 million years
ago, coinciding with the initial diversification ofangiosperms, during the Eocene epoch when the
climate was warm and humid. These results reveal an
unexpected diversity, distribution, and ancient originof a group of commercially valuable mushrooms thatmay provide an economic incentive for conservationand support the hypothesis of a tropical origin of theectomycorrhizal symbiosis.

is proposed as a possible synonym. Comprehensive
descriptions, illustrations, photographs andcomparisons with phenetically similar species areprovided. A redescription of M. chlorophos, based on
analyses of type specimens and recently collected
topotypical material, is provided. The addition of theseseven new or newly reported luminescent species ofMycena brings the total to 71 known bioluminescentspecies of fungi.
Key words: Agaricales, bioluminescence,
Mycenaceae, mycenoid fungi, taxonomy.

Molecular phylogenetics of porcini
mushrooms（Boletus section Boletus）
Bryn T.M. Dentinger, Joseph F. Ammirati, Ernst E.
Both, Dennis E. Desjardin, Roy E. Halling, Terry W.
Henkel, Pierre-Arthur Moreau, Eiji Nagasawa, Kasem
Soytong, Andy F. Taylor, Roy Watling, Jean-Marc
Moncalvo and David J. McLaughlin
Molecular Phylogenetics and Evolution 57: 1276–
1292, 2010.
Porcini（Boletus section Boletus: Boletaceae:
Boletineae: Boletales）are a conspicuous group ofwild, edible mushrooms characterized by fleshy
fruiting bodies with a poroid hymenophore that is“stuffed”with white hyphae when young. Theirreported distribution is with ectomycorrhizal plants
throughout the Northern Hemisphere. Little progresshas been made on the systematics of this group using
modern molecular phylogenetic tools because
sampling has been limited primarily to European
species and the genes employed were insufficient toresolve the phylogeny. We examined the evolutionary
history of porcini by using a global geographic
sampling of most known species, new discoveries
from little explored areas, and multiple genes. We used

Key words: molecular systematics, molecular clock,biogeography, Boletales, evolution, synapomorphy,
partial veil, ITS, RPB1, ATP6, stuffed pores,
sustainable non-timber forest product, conservation.

Taxonomic reevaluation of a fungus describedas “Mucidula mucida” in Japan
Shuji Ushijima, Norihiro Shimomura, Eiji Nagasawa
and Nitaro Maekawa
Mushroom Science and Biotechnology 20: 22-30,
2012.
In Japan, a fungus described as Mucidula mucida（“Japanese M. mucida”, Japanese name: “Numeritsubatake”）was identiﬁed as M. mucida var. asiatica based

S5

捻性な子実体を形成した．さらに，ITS 領域に基づく分子系統樹において，両菌は 1 つのクレード
内 に 混 在 し た． こ れ ら の 結 果 は， 両 菌 を M.
mucida の種内分類群として分割すべきでないこ

on morphological features and phylogenetic analysisof the internal transcribed spacer（ITS）region of
nuclear ribosomal DNA. This variety differs
macromorphologically from M. mucida var. venosolamellata（Japanese name: “Numeritsubatakemodoki”）mainly inとを強く示唆する．また，日本産ヌメリツバタケ
that the latter possesses reticulate, anastomosed veins （ヌメリツバタケモドキを含む）は，担子胞子のon the lamellae. However, mating compatibility tests
大きさ，傘表皮組織の構造および ITS 領域を用いM.
mucidaasiatica
M.
mucida”）
betweenvar.
（= “Japaneseた 分 子 系 統 解 析 結 果 に お い て，M. mucida var.
mucida とは明らかに種レベルで異なった．
and var. venosolamellata appeared compatible in all
pairings. Furthermore, the basidiomata derived fromthe mating-compatible mycelia possessed intermediate
macro- and micromorphological features between M.
The genus Ponticulomyces（Physalacriaceae,
mucida var. asiatica and var. venosolamellata. In ourAgaricales）from Japan
molecular phylogenetic tree, the two fungi formed a
Shuji Ushijima, Eiji Nagasawa, Hiroto Suhara, Nitaro
clade of “Japanese M. mucida”/M. mucida var.
Maekawa
asiatica/M. mucida var. venosolamellata, andMycoscience 53: 156-160, 2012. DOI:10.1007/
specimens of the two fungi were randomly scattered
S10267-011-0147-Y
within this clade. These results strongly suggest thatfungi belonging to the clade should not be divided into
Two species of the genus Ponticulomyces collected
i n t r a s p e c i f i c t a x a . M u c i d u l a m u c i d a v a r.
from Japan for the first time are described andvenosolamellata（= var. asiatica）differed from M. mucida
illustrated. Ponticulomyces kedrovayae is characterized
var. mucida in their pileipellis structure, basidiospore
by its lamellae staining yellow when bruised and insize and ITS-based phylogenetic analysis. Theseage, stipe lacking an annulus and a pseudorhiza,
differences indicate that M. mucida var. venosolamellata
scattered pileal hairs, and large amygdaliform
mucidaand var.
are different at the speciﬁc level.
basidiospores. It mainly occurs on dead wood of
Fagus crenata. Ponticulomyces orientalis is
Key words: mating compatibility, molecularcharacterized by its scattered pileal hairs and broadly
phylogenetic analysis, morphology, Mucidula
ellipsoid to ellipsoid basidiospores. It was collectedvenosolamellata, taxonomy.
mostly on dead wood of Cameria japonica.
日本産ヌメリツバタケ（Mucidula mucida）の分
類学的再検討
牛島秀爾・霜村典宏・長澤栄史・前川二太郎
日本きのこ学会誌 20: 22-30, 2012.

Key words: Agaricomycetidae, new geographic
distribution, Oudemansiella, taxonomy, Xerula.

Productivity of the toxic substance, fasciculol
日 本 産 ヌ メ リ ツ バ タ ケ（Japanese M. mucida） E, in fruiting bodies of Hypholoma fasciculareとして知られている菌は，形態的特徴および核and some other members of Strophariaceae
M.
rDNA の ITS 領域を用いた分子系統解析の結果，（In Japanese）
mucida var. asiatica と同定された．本変種は，ひKumiko Oka, Marina Nishida, Eiji Nagasawa, Shuji
だに皺を持つ日本固有種のヌメリツバタケモドキ
Ushijima, Atsushi Ishihara and Nitaro Maekawa（M. mucida var. venosolamellata）と は， 肉 眼 的 特Mushroom Science and Biotechnology 22: 147-152,
徴に基づいて区別されてきた．しかし，両菌は交2015.
配可能であり，両菌の中間的な形態的特徴を示す

S6

The poisonous mushroom Hypholoma fasciculare
属種のいくつかの子実体中においても極めて少量（Japanese name: Nigakuritake）, belonging to theであるが検出された．検出された子実体には食用
family Strophariaceae（Basidiomycota, Agaricales）,
きのこ種も含まれていた．さらに，分子系統解析
produces fasciculol E as a major toxic agent.
の結果，ファシキュロール E 生産性はモエギタ
Fasciculol E has also been reported from a few related
ケ科内の Hypholoma 属との系統的類縁性と密接
species, but the range of species producing it is notに関係していることが示唆された．
clear. In the present study, the content of fasciculol Ein fruiting bodies of H. fasciculare and related species
in the family Strophariaceae was analyzed by UPLC/
A new species of Dactylosporina
MS/MS. The results revealed that all species of （Physalacriaceae, Agaricales）from Japan
Hypholoma examined in this study stably produce
Shuji Ushijima, Eiji Nagasawa, Shiro Kigawa, Nitaro
fasciculol E. Moreover, a very small amount of theMaekawa
toxin was detected from several species belonging toMycoscience 56: 10-13, 2015. DOI: 10.1016/j.
the genera Pholiota and Stropharia. Of these, somemyc.2014.01.012
species are known as edible mushrooms, but the
Dactylosporina brunneomarginata sp. nov.,
amount contained in their fruiting bodies was muchl o w e r t h a n t h e L D 50 v a l u e s o f f a s c i c u l o l E .
belonging to the family Physalacriaceae（Agaricales）
Phylogenetic analysis using the internal transcribedis described and illustrated. This fungus differs fromspacer（ITS）region sequences suggests that thepreviously described species of Dactylosporina
productivity of fasciculol E is correlated withmacroscopically by its viscid pileus and stipe surfacephylogenetic distance from H. fasciculare within thewhen wet, the lamellae with dark brown edges, andfamily Strophariaceae.
microscopically by its gloeocystidioid pileo-, cheilo-,and caulocystidia with olive to yellow-brown oily
Key words: fasciculol E, Hypholoma fasciculare,content. This is the ﬁrst report on Dactylosporina fromphylogenetic analysis, Strophariaceae, UPLC/MS/MS.
Asia.
ニガクリタケおよびモエギタケ科属種子実体における毒成分ファシキュロール E の生産性（和文）岡久美子・西田麻理奈・長澤栄史・牛島秀爾・石原 亨・前川二太郎
日本きのこ学会誌 22: 147-152, 2015

Key words: Agaricomycetidae, echinulatebasidiospores, taxonomy, Xerula/Oudemansiellacomplex.

Relationship between Panus lecomtei and P.
strigellus inferred from their morphological,
molecular and biological characteristics
Ruby Vargas-Isla, Marina Capelari, Nelson MenolliJr., Eiji Nagasawa, Keisuke Tokimoto, Noemia Kazue
Ishikawa
Mycoscience 56: 561-571, 2015. DOI: 10.1016/j.
myc.2015.05.004

モエギタケ科に所属する Hypholoma fasciculare（ニガクリタケ）は毒きのこであり，主要な毒成分としてファシキュロール E を生産する．これまでに本種以外のきのこ種子実体にもファシキュ
ロール E が含まれていることが報告されているが，詳細な調査は行われていなかった．そこで本
研究では，H. fasciculare およびモエギタケ科属種
子実体におけるファシキュロール E の含有量を調査した．その結果，供試した Hypholoma 属種の子実体すべてよりファシキュロール E が検出され，本属の近縁属である Pholiota 属や Stropharia

Panus strigellus was first recorded in Amazonas
State of Brazil. This edible mushroom has macroscopic
characteristics similar to those of P. lecomtei. In thisS7

s t u d y, w e u s e d e v i d e n c e o b t a i n e d f r o m amorphological comparison, molecular analyses andmating tests to clarify the taxonomic status of P.
strigellus and show differences compared to thesympatric species of Panus. In addition, this paper

Identification and symbiotic ability ofPsathyrellaceae fungi isolated from a
photosynthetic orchid, Cremastra
appendiculata（Orchidaceae）

bodies of the one isolate, SI1-1 were induced. In
addition, two fungal isolates, SI1-1 and KI1-1, weregrown in symbiotic cultures with C. appendiculata toverify their status as mycobionts.
• Key results: In phylogenetic analyses, all isolates
clustered with fungi belonging to Coprinellus in
Psathyrellaceae of Agaricales. Phylogenetic analysesof these DNA sequences showed that five fungal
isolates from C. appendiculata, including SI1-1 andtwo mycobionts isolated from the mycoheterotrophic
orchid Epipogium roseum, have very similar ITS
sequences. Isolate SI1-1 was identiﬁed as Coprinellus
domesticus based on the morphological characteristicsof the fruiting body. Isolates SI1-1 and KI1-1 induced
seed germination of C. appendiculata as mycobionts.
• Conclusions: This report is the ﬁrst of a mycorrhizal
symbiosis between a fungus in Psathyrellaceae and aphotosynthetic orchid, revealing a new pathway to fullmycoheterotrophy and contributing to ourunderstanding of the evolution of mycoheterotrophy.

Takahiro Yagame, Eriko Funabiki, Eiji Nagasawa,
Toshimitsu Fukiharu and Koji Iwase
Am. J. Bot. 100（9）: 1823-1830, 2013. DOI: 10.3732/
ajb.1300099

Key words: Coprinellus domesticus, mycorrhizal
symbiosis, Orchidaceae, photosynthetic orchids,
saprobic fungi.

discusses and reports some morphologicalcharacteristics of P. strigellus. The tetrapolar mating
system of P. strigellus and the incompatibility between
P. lecomtei and P. strigellus was confirmed. The
geographical distributions of both species in theAmericas are presented.
Key words: edible mushroom, Lentinus strigellus,
Lentinus strigosus, Panus rudis.

• Premise of the study: Photosynthetic orchids foundin highly shaded forests are often mixotrophic,receiving part of their carbon energy via
ectomycorrhizal fungi that had originally received
carbohydrate from trees. A photosynthetic orchid,Cremastra appendiculata, is also found under highly
shaded forest, but our preliminary data suggested thatits associated fungi were not ectomycorrhizal. We
tested whether their relation is an unusual example ofa mixotrophic orchid associating with saprotrophic
fungi by direct detection of fungal DNAs in
conjunction with isolation of the fungus in pure culture
and experimental inoculation of orchid seeds with thefungus.
• Methods: For isolated mycobionts of C.
appendiculata plants, two regions of nuclear ribosomal
DNA, the internal transcribed spacer（ITS）and thelarge subunit（LSU）, were sequenced, and fruiting

A comparison of dried shiitake mushroom inlog cultivation and mycelial cultivation fromdifferent geographical origins using stablecarbon and nitrogen isotope analysis（InJapanese）
Yaeko Suzuki, Rumiko Nakashita, Noemia Kazue
Ishikawa, Akiko Tabuchi, Emi Sakuno and Keisuke
Tokimoto
Bunseki Kagaku 64（12）: 859-866, 2015.
We determined carbon and nitrogen isotopic
compositions（δ13C and δ15N）of dried shiitake mushroom（Lentinula edodes）samples from Japan, China, South
Korea and Brazil in order to discriminate theirgeographical origins. In log cultivation, the δ13C valuesof Japanese dried shiitake samples were lower thanthose of Chinese samples, depending on the δ13C values

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of log and their growth conditions. In mycelial
cultivation, the δ13C and δ15N values of Japanese dried
shiitake samples were higher than those of Chinese
samples. By using the δ13C and δ15N values, 87.4% ofJapanese dried shiitake samples（n = 95）and 87.9%
of Chinese dried shiitake samples（n = 66）in log
cultivation, 90.0% of the Japanese dried shiitake
samples（n = 50）and 93.9% of Chinese dried shiitake
samples（n = 114）in mycelial cultivation, werecorrectly classified according to the production site.
These results suggested that the δ13C and δ15N values
will be potentially useful for tracing their geographicalorigin of dried shiitake samples.
Key words: geographical origin, stable isotope
ratio, Lentinula edodes, Mushroom.
炭素・窒素安定同位体比分析による原木栽培及び菌床栽培乾シイタケの産地間比較（和文）鈴木彌生子・中下留美子・Noemia Kazue Ishikawa・
田淵諒子・作野えみ・時本景亮
分析化学 64: 859-866, 2015

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原木栽培及び菌床栽培の乾シイタケについて，
炭素・窒素安定同位体比（δ13C，δ15N）を用いて，栽培方法及び産地判別の可能性を検証した． 原
木栽培乾シイタケは，国産は中国産よりもδ13C は低く，δ15N は高い傾向が得られ，韓国産は国産と中国産の中間的な値を示した． 菌床栽培乾シイ
タケは，国産は中国産よりもδ13C，δ15N が高い傾向を示した．ブラジル産については，原木栽培品・菌床栽培品ともにほかの地域よりもδ15N が高い
特徴が得られた． 国産・中国産の乾シイタケについて，炭素・窒素安定同位体比分析を行い，産地判別の可能性を検証した結果，原木栽培の正答率は，国産 87.4%，中国産 87.9%，菌床栽培の判別率は，国産 90.0%，中国産 93.9% となった． 以上より，原木栽培及び菌床栽培の乾シイタケのδ13C，
δ15N は，栽培方法及び産地によって特徴的な分布を示し，中国産と国産については産地判別の可能性が示唆された．