全 文 ：
菌物学报
jwxt@im.ac.cn 15 July 2015, 34(4): 581‐588
Http://journals.im.ac.cn Mycosystema ISSN1672‐6472 CN11‐5180/Q © 2015 IMCAS, all rights reserved.
研究论文 Research paper DOI: 10.13346/j.mycosystema.150051
Supported by the National Basic Research Program of China (973 Program, 2014CB138305)
Corresponding author. E‐mail: fungi@mail.kib.ac.cn
Received: 10‐02‐2015, accepted: 10‐05‐2015
A new edible mushroom resource, Pleurotus abieticola, in southwestern
China
LIU Xiao‐Bin1, 2 LIU Jian‐Wei1, 2 YANG Zhu‐Liang1*
1Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences,
Kunming, Yunnan 650201, China
2University of Chinese Academy of Sciences, Beijing 100039, China
Abstract: Species of the genus Pleurotus are very important edible mushrooms and many of them can be cultivated in
commercial scale. Although P. abieticola was originally described from Russian Far East, and then reported from northeastern
China and northwestern Russia, its distribution range is still largely unknown. Our morphological and molecular phylogenetic
evidence indicated that this species is also distributed in subalpine mountains of southwestern China. This paper documented
the taxon based on morphological and ecological features, and DNA sequences generated from materials collected from Sichuan
Province and the Tibet Autonomous Region.
Key words: Basidiomycetes, new distribution, edible mushroom, taxonomy
冷杉侧耳——中国西南一种新的食用菌资源
刘晓斌 1, 2 刘建伟 1, 2 杨祝良 1*
1中国科学院昆明植物研究所东亚植物多样性与生物地理学院重点实验室 云南 昆明 650201
2中国科学院大学 北京 100039
摘 要：侧耳属 Pleurotus 真菌具有重要经济价值，该属不少种类可以商业化人工栽培。冷杉侧耳 P. abieticola 原初报道于
俄罗斯远东地区，后来在我国东北和俄罗斯西北也有记载，但因为文献中记载的标本有限，我国研究人员对该种并不十
分了解。在开展侧耳属的研究中，作者发现该种在我国西南亚高山地区也有分布。基于采自四川和西藏的标本，利用形
态、生态特征及 DNA 序列证据，作者对该种进行了描述，以期为该种的资源开发利用提供科学依据。
关键词：担子菌，新分布，食用菌，分类
ISSN1672‐6472 CN11‐5180/Q Mycosystema July 15, 2015 Vol. 34 No. 4
http://journals‐myco.im.ac.cn
582
INTRODUCTION
Many species of the genus Pleurotus (Fr.) P.
Kumm. are very important edible mushrooms (Dai et
al. 2010). Several of them are cultivated in large
commercial scales (Sánchez 2010). Due to their
importance, very rich studies on the genus were
carried out in the past (Corner 1981; Hilber 1982;
Singer 1986; Vilgalys et al. 1993; Vilgalys & Sun 1994;
Segedin et al. 1995; Petersen & Krisai‐Greilhuber
1996; Li & Yao 2004, 2005; Huang et al. 2010; Li et al.
2014).
During the study of edible mushroom resources
in Pleurotus, we encountered an interesting species,
P. abieticola R.H. Petersen & K.W. Hughes, which was
originally described from Far East Russia, and then
reported from northeastern China with a limited
number of collections (Petersen & Hughes 1997;
Albertó et al. 2002; Li et al. 2014). Our
morphological and molecular phylogenetic evidence
indicated that this species is also distributed in
subalpine regions in southwestern China. This paper
documented the taxon based on materials collected
from Sichuan and Tibet.
1 MATERIALS AND METHODS
1.1 Specimens and morphological descriptions
Basidiomata were photographed and collected
with field‐notes. Specimens were dried and then
kept in the Herbarium of Cryptogams of Kunming
Institute of Botany, Chinese Academy of Sciences
(HKAS). Morphological descriptions of the
basidiomata are based on field notes. Method for
microscopic observation and explanations of
basidiospore data followed Yang & Feng (2013).
1.2 DNA extraction, PCR and sequencing
Total DNA was isolated from silica‐gel dried
materials using the CTAB method (Doyle & Doyle
1987). The internal transcribed spacer (ITS) region
was amplified with primer pair ITS1/ITS4
(http://www.biology.duke.edu/fungi/mycolab/prime
rs.htm) in an ABI 2720 thermal cycler (Applied
Biosystems, Foster City, CA, USA). The PCR program
was as follows: pre‐denaturation at 94°C for 4min;
then followed by 35 cycles of denaturation at 94°C
for 40s, annealing at 50°C for 50s, elongation at 72°C
for 90s; afterwards, a final elongation at 72°C for
8min was included. PCR products were depurated
with the Gel Extraction & PCR Purification Combo Kit
(Spin‐column, Bioteke, Beijing, China), and then
sequenced on an ABI‐3730‐XL sequence analyzer
(Applied Biosystems, Foster City, CA, USA) using the
same primer combinations used for the PCR.
Forward and reverse sequences were assembled and
edited with SeqMan (DNA STAR package; DNAStar
Inc., Madison, WI, USA). Sequences used in this
study and additional sequences obtained by us were
listed in Table 1.
1.3 Phylogenetic analyses
ITS sequences of genus Pleurotus were
retrieved from GenBank and were combined with
the ITS sequences generated in this study to form a
dataset. Pleurotus purpureo‐olivaceus were chosen
as outgroups (Moncalvo et al. 2002). The dataset
was then aligned using MAFFT v7.130b (Katoh &
Standley 2013) and manually optimized on Bioedit
v7.0.9 (Hall 1999).
Maximum l ikelihood (ML) and Bayesian
inference (BI) analyses were applied using RaxML
(Stamatakis 2008) and MrBayes (Ronquist &
LIU Xiao‐Bin et al. / A new edible mushroom resource, Pleurotus abieticola, in southwestern China
菌物学报
583
Table 1 Specimens used in molecular phylogenetic studies and their GenBank accession numbers
Species Collections GenBank accession #
ITS tef1‐α rpb1 rpb2 LSU
Pleurotus abieticola HKAS45720 KP771696 KP867895 KP867886 KP867879 KP867907
Pleurotus abieticola TENN52359 AY450348 ‐ ‐ ‐ ‐
Pleurotus abieticola HKAS45507 KP771697 KP867896 KP867887 KP867880 KP867908
Pleurotus abieticola HKAS46100 KP771695 KP867897 KP867888 KP867881 KP867909
Pleurotus abieticola TENN58284 AF345656 ‐ ‐ ‐ ‐
Pleurotus albidus Duke327 AF345658 ‐ ‐ ‐ ‐
Pleurotus albidus BAFC 50.261 AF345659 ‐ ‐ ‐ ‐
Pleurotus australis CBS100127 EU424276 ‐ ‐ ‐ ‐
Pleurotus australis PDD87/021XP AY315764 ‐ ‐ ‐ ‐
Pleurotus citrinopileatus HKAS85956 KP867919 KP867898 ‐ ‐ KP867910
Pleurotus citrinopileatus HMAS63344 AY696301 ‐ ‐ ‐ ‐
Pleurotus citrinopileatus TFM‐M‐E793 AB115043
Pleurotus cornucopiae TENN55191 AY450341
Pleurotus cornucopiae H‐14 JQ837484
Pleurotus cystidiosus subsp. abalonus CBS80391 AY315806 ‐ ‐ ‐ ‐‐
Pleurotus cystidiosus subsp. abalonus VT2476 AY315802 ‐ ‐ ‐ ‐
Pleurotus cystidiosus IFO30607 AY315778 ‐ ‐ ‐ ‐
Pleurotus cystidiosus AG55 FJ608592 ‐ ‐ ‐ ‐
Pleurotus eryngii HIK135 HM998833 ‐ ‐ ‐ ‐
Pleurotus eryngii HIK154 HM998841 ‐ ‐ ‐ ‐
Pleurotus eryngii HIK139 HM998837 ‐ ‐ ‐ ‐
“Pleurotus cf. eryngii” C24 FJ514570 ‐ ‐ ‐ ‐
Pleurotus eryngii var. tuoliensis HIK152 HM998839 ‐ ‐ ‐ ‐
Pleurotus eryngii var. tuoliensis HIK138 HM998836 ‐ ‐ ‐ ‐
Pleurotus eryngii var. tuoliensis CCMSSC01433 KP867912 ‐ ‐ KP867873 KP867900
Pleurotus fossulatus HIK127 HM998828 ‐ ‐ ‐ ‐
Pleurotus fossulatus ATCC 52666 AY265833 ‐ ‐ ‐ ‐
Pleurotus levis TENN58298 AF345662
Pleurotus nebrodensis HIK125 HM998826 ‐ ‐ ‐ ‐
Pleurotus nebrodensis UPA6 HM998816 ‐ ‐ ‐ ‐
Pleurotus nebrodensis HIK137 HM998835 ‐ ‐ ‐ ‐
Pleurotus ostreatus HKAS84903 KP867913 KP867889 ‐ KP867874 KP867901
Table 1 continued
ISSN1672‐6472 CN11‐5180/Q Mycosystema July 15, 2015 Vol. 34 No. 4
http://journals‐myco.im.ac.cn
584
Table 1 continued
Pleurotus ostreatus HKAS53480 KP867914 KP867890 ‐ KP867875 KP867902
Pleurotus ostreatus CCMSSC06141 KP867915 KP867891 ‐ ‐ KP867903
Pleurotus ostreatus TENN 53662 AY854077 ‐ ‐ ‐ ‐
Pleurotus populinus TENN56749 AY450346 ‐ ‐ ‐ ‐
Pleurotus populinus ATCC 90083 AY368667 ‐ ‐ ‐ ‐
Pleurotus pulmonarius HMAS76672 AY696299
Pleurotus pulmonarius HKAS76382 KP867916 KP867892 KP867883 KP867876 KP867904
Pleurotus pulmonarius ECS‐0158 GU722283 ‐ ‐ ‐ ‐
Pleurotus purpureo‐ olivaceus ICMP17077 GQ411512
Pleurotus purpureo‐ olivaceus PDD91632 GQ411523
Pleurotus tuber‐regium DMC172 EU908193
Pleurotus tuber‐regium RV95/947.1 AF109966
Note: KP771695‐KP771697, KP867873‐KP867920 are sequences generated in this study.
Huelsenbeck 2003), respectively. For phylogenetic
analysis, GTR+G was chosen as the best fit model for
the dataset by using Mrmodeltest 2.3 (Nylander
2004). As GTR is the only model available in RAxML,
we thus used GTRGAMMA with the default setting in
ML analysis. Statistic supports were calculated using
nonparametric bootstrapping with 1 000 replicates.
For BI analysis, GTR+G model was used with the
default setting. We set the generations to 2 million
and used the stoprul command with the value of
stopval set to 0.01. Trees were sampled every 100
generation. Statistic supports were obtained by the
using sumt command implemented in MrBayes by
discarding the first 25% of generations as burn‐ins.
2 RESULTS
2.1 Molecular phylogeny
Our target species, P. abieticola, was related to
the P. ostreatus ‐ P. pulmonarius species complex,
including P. nebrodensis (Inzenga) Quél., P. eryngii
(DC.) Quél., P. fossulatus Cooke, P. eryngii var.
tuoliensis C.J. Mou, P. ostreatus (Jacq.) P. Kumm., P.
populinus O. Hilber & O.K. Mill., P. albidus (Berk.)
Pegler, and P. pulmonarius (Fr.) Quél. with high
statistical supports (Bootstrap values 100%, and
Bayesian posterior probabilities 1) (Fig. 1). Three
sequences generated from different collections of P.
abieticola collected from southwestern China were
clustered to the sequences generated from the same
species collected from Russia (Bootstrap values 88%,
and Bayesian posterior probabilities <0.90).
2.2 Taxonomy
Pleurotus abieticola R.H. Petersen & K.W. Hughes,
Mycologia 89: 175, 1997. Figs. 2–3
Basidiomata small to medium‐sized. Pileus
flabelliform, 3–10cm from attachment to margin,
3–8cm in width; surface greyish, grey to brownish
grey, becoming paler when mature, glabrous,
smooth, finely innately streaked toward margin,
appearing hygrophanous or slightly viscid when
LIU Xiao‐Bin et al. / A new edible mushroom resource, Pleurotus abieticola, in southwestern China
菌物学报
585
Fig. 1 Phylogenetic analysis of Pleurotus species inferred from Maximum Likelihood (ML) analysis of ITS sequences. Bootstrap
values (ML, >50%) are shown above or beneath individual branches, Bayesian posterior probabilities (BI, ≥0.95) are indicated
with thick branches. Sequences of the target species obtained in this study are in bold face.
ISSN1672‐6472 CN11‐5180/Q Mycosystema July 15, 2015 Vol. 34 No. 4
http://journals‐myco.im.ac.cn
586
Fig. 2 Basidiomata of Pleurotus abieticola. A: HKAS 45720; B: HKAS 46100.
Fig. 3 Microscopic characters of Pleurotus abieticola (HKAS
45720). A: Basidiospores; B: Basidia; C: Cheilocystidia. Bars:
A= 10µm, B and C= 20µm.
wet; margin inrolled when young, becoming straight
by maturity; context white, unchanging, relatively
thin (up to 8mm in thickness near attachment).
Lamellae strongly decurrent, dense to subdistant,
white; lamellar edge entire, concolorous with
lamella surface; lamellulae 2–3 tiers. Stipe lateral to
strongly eccentric, 0.5–2×0.5–1.5cm, subcylindric,
whitish to white, longitudinally striate as extensions
of lamellae but forming a reticulum on surface of
stipe. Odour none; taste mild. Spore print white to
cream‐colored.
Basidiospores [70/3/3] (8–) 8.5–13 (–14)×4–5
(–5.5)µm, Q= (1.7–) 1.82–2.75 (–2.89) (Q= 2.26 ±
0.28), elongate to nearly cylindrical, colorless and
hyaline, thin‐walled, smooth, non‐amyloid,
non‐dextrinoid. Basidia 27–40×5.5–8.5µm, narrowly
clavate, hyaline, thin‐walled, 4‐spored, sometimes
2‐spored; sterigmata 3–5µm long. Cheilocystidia
abundant, broadly clavate, clavate, narrowly clavate
to nearly cylindrical, 15–40×5–14µm, colorless and
hyaline, thin‐walled. Pleurocystidia absent. Lamellar
trama monomitic, composed of ± irregularly arranged
thin‐ to thick‐walled (up to 2µm thick), colorless and
hyaline, filamentous hyphae 3–10µm wide. Pileipellis
a 40–60µm thick cutis composed of repent, radially
arranged, yellowish to brownish filamentous hyphae
2–5µm wide. Trama of pileus monomitc, composed
of radially to irregularly arranged thin‐ to slightly
LIU Xiao‐Bin et al. / A new edible mushroom resource, Pleurotus abieticola, in southwestern China
菌物学报
587
thick‐walled (up to 1µm thick), colorless and hyaline,
filamentous hyphae 3–15µm wide. Trama of stipe
monomitc, composed of vertically to irregularly
arranged thin‐ to thick‐walled (up to 2µm thick),
colorless and hyaline, filamentous hyphae 3–10µm
wide. Stipitipellis composed of vertically arranged,
branching and sometimes anastomosing hyphae
3–5µm wide. Clamp connections abundant in all
tissues.
Habitat and known distribution: growing on
rotten wood of Picea in subalpine forests dominated
by Picea; in summer at elev. 3 600–4 100m in
southwestern China. Also known from northeastern
China and Russia.
Specimens examined: CHINA, Sichuan Province,
Xiangcheng County, Reda, alt. 3 600m, 14 July 2004,
Z.L. Yang 4122 (HKAS 45507). Tibet Autonomous
Region, Leiwuqi County, Mengda, alt. 4 100m, 9
August 2004, Z.L. Yang 4341 (HKAS 45720); Leiwuqi
County, Haola, alt. 3 900m, 10 August 2004, Z.W. Ge
320 (HKAS 46100).
3 DISCUSSION
Although the genus Pleurotus harbors many
economically important species due to their well
known usage as vegetable or food (Dai et al. 2010),
species of Pleurotus are morphologically not easily
separated from each other. Both morphological and
molecular phylogenetic data should be employed in
the characterization of the species of the genus. In
addition, cultural characters and mating tests can
also provide useful evidence for species delimitation
(Petersen & Hughes 1993, 1997).
Morphologically, Pleurotus abieticola is very
similar to the other species of the P. ostreatus ‐ P.
pulmonarius complex, including P. pulmonarius, P.
ostreatus, P. populinus, P. albidus, P. nebrodensis, P.
eryngii, P. fossulatus, and P. eryngii var. tuoliensis,
which produce monomitic basidiomata. However, it
differs from the other taxa by its occurrence mainly
on coniferous rotten wood and the common
presence of cheilocystidia (Petersen & Hughes 1997;
Albertó et al. 2002; our observations).
Phylogenetically, Pleurotus abieticola, was
related to the P. ostreatus ‐ P. pulmonarius species
complex (Fig. 1). Pleurotus abieticola was basal to all
the other species mentioned above, which is
consistent with the results of Albertó et al. (2002).
Geographically, P. abieticola was originally
described from far‐eastern Russia (Sichote Alin
Biosphere Preserve), and then reported from
northeastern China (Songjianghe and Baihe in Jilin
Prov.) and northwestern Russia (north of St.
Petersburg) based on all the five collections available
then (Petersen & Hughes 1997; Albertó et al. 2002;
Li et al. 2014). Our collections made in southwestern
China largely extend the known distribution range of
the species.
Although the epithet “abieticola” may indicate
that the species has a preference for substrates of
Abies, P. abieticola can also grow on rotten wood of
Picea (Albertó et al. 2002; our observations in the field).
In addition, according to Albertó et al. (2002), this
taxon was on Alnus or Salix in northwestern Russia.
Acknowledgements: The authors are very grateful to Dr. Z.W.
Ge (Kunming Institute of Botany of the Chinese Academy of
Sciences) for providing a collection and an image of P.
abieticola, and Prof. Dr. T. Bau (Jilin Agriculture University,
China) for providing literature.
[REFERENCES]
Albertó EO, Petersen RH, Hughes KW, Lechner B, 2002.
Miscellaneous notes on Pleurotus. Persoonia, 18: 55‐69
ISSN1672‐6472 CN11‐5180/Q Mycosystema July 15, 2015 Vol. 34 No. 4
http://journals‐myco.im.ac.cn
588
Corner EJH, 1981. The agaric genera Lentinus, Panus and
Pleurotus with particular reference to Malaysian species.
Beihefte zur Nova Hedwigia, 69: 1‐169
Dai YC, Zhou LW, Yang ZL, Wen HA, Bau T, Li TH, 2010. A
revised checklist of edible fungi in China. Mycosystema,
29: 1‐21 (in Chinese)
Doyle JJ, Doyle JL, 1987. A rapid DNA isolation procedure for
small quantities of fresh leaf material. Phytochemical
Bulletin, 19: 11‐15
Hall TA, 1999. BioEdit: a user‐friendly biological sequence
alignment editor and analyses program for Windows
95/98/NT. Nucleic Acids Symposium Series, 41: 95‐98
Hilber O, 1982. Die Gattung Pleurotus. Bibliotheca
Mycologica, 87: 1‐448
Huang CY, Chen G, Gao S, Gao W, Zhang JX, 2010. Analysis of
internal transcribed spacer regions of main species of the
genus Pleurotus. Mycosystema, 29: 365‐372 (in Chinese)
Katoh K, Standley DM, 2013. MAFFT multiple sequence
alignment software version 7: improvements in
performance and usability. Molecular Biology and
Evolution, 30: 772‐780
Li XL, Yao YJ, 2004. Assembling phylogenetic tree of Pleurotus
based on 28S rDNA sequences. Mycosystema, 23:
345‐350 (in Chinese)
Li XL, Yao YJ, 2005. Revision of the taxonomic position of the
Phoenix Mushroom. Mycotaxon, 91: 61‐73
Li Y, Bau T, Liu Y, Li TH, 2014. Flora fungorum sinicorum. Vol.
45. Pleurotoid‐lentinoid fungi. Science Press, Beijing. 1‐
206 (in Chinese)
Moncalvo JM, Vilgalys R, Redhead SA, Johnson JE, James TY,
Aime MC, Hofstetter V, Verduin SJW, Larsson E, Baroni TJ,
Thorn RG, Jacobsson S, Clémençon H, Miller OKJr, 2002.
One hundred and seventeen clades of euagarics.
Molecular Phylogenetics and Evolution, 23: 357‐400
Nylander J, 2004. MrModeltest 2.2. Computer software
distributed by the University of Uppsala.
Petersen RH, Hughes KW, 1993. Intercontinental
interbreeding collection of Pleurotus pulmonarius with
notes on P. ostreatus and other species. Sydowia, 45:
139‐152
Petersen RH, Hughes KW, 1997. A new species of Pleurotus.
Mycologia, 89: 173‐180
Petersen RH, Krisai‐Greilhuber I, 1996. An epitype specimen
for Pleurotus ostreatus. Mycological Research, 100:
229‐235
Ronquist F, Huelsenbeck JP, 2003. MrBayes 3: Bayesian
phylogenetic inference under mixed models.
Bioinformatics, 19: 1572‐1574
Sánchez C, 2010. Cultivation of Pleurotus ostreatus and other
edible mushrooms. Applied Microbiology and
Biotechnology, 85: 1321‐1337
Segedin BP, Buchanan PK, Wilkie JP, 1995. Studies in the
Agaricales of New Zealand 4: new species, new records
and renamed species of Pleurotus (Pleurotaceae).
Australian Systematic Botany, 8: 453‐482
Singer R, 1986. The Agaricales in modern taxonomy. 4th
edition. Koeltz Scientific Books, Koenigstein. 1‐981
Stamatakis A, Hoover P, Rougemont J, 2008. A rapid
bootstrap algorithm for the RAxML Web‐Servers.
Systematic Biology, 75: 758‐771
Vilgalys R, Smith A, Sun BL, 1993. Intersterility groups in the
Pleurotus ostreatus complex from the continental United
States and Canada. Canadian Journal of Botany, 71:
113‐128
Vilgalys R, Sun BL, 1994. Ancient and recent patterns of
geographic speciation in the oyster mushroom Pleurotus
revealed by phylogenetic analysis of ribosomal DNA
sequences. Proceedings of the National Academy of
Sciences of the United States of America, 91: 4599‐4603
Yang ZL, Feng B, 2013. The genus Omphalotus
(Omphalotaceae) in China. Mycosystema, 32: 545‐556
Zhang JX, 2014. Powerful supports of scientific research are
indispensable to the development of mushroom industry.
Mycosystema, 33: 175‐182 (in Chinese)