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Amylosporus succulentus sp. nov. (Russulales, Basidiomycota) Evidenced by Morphological Characters and Phylogenetic Analysis

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  • Jiangsu Vocational College of Agriculture and Forestry

Abstract and Figures

Three polypore specimens were collected from Hainan, southern China. They are described and illustrated here as a new species, Amylosporus succulentus, based on a combination of morphological characters and phylogenetic (ITS and nLSU sequences) data. It is characterized by poroid basidiocarps, both simple septate and clamped generative hyphae, hymenial hyphae without clamp connections, and finely asperulate and amyloid basidiospores. These characters are typical for Amylosporus. In the phylogenetic perspective, A. succulentus is closely related to A. campbellii, the generic type, and nested within the Wrightoporiaceae clade. A key to accepted species of Amylosporus worldwide is provided.
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crypt ogami e
mycologie
volume 35 n°3 2014
contents
Qing-Tao ZHANG, QuanLU,MingHE,ConyDECOCK & Xing-Yao
ZHANG — Cytospora palm sp. nov. (Diaporthales, Ascomycota), a canker
agent on Cotinus coggygria(Anacardiaceae) in Northern China........ 211-220
Jie SONG, Yuan-Yuan CHEN&Bao-Kai CUI — Phylogeny and taxonomy
ofClimacocystis(Polyporales) in China .......................... 221-231
Marcia Isabel KAFFER, SuzanaMaria de Azevedo MARTINS, Marcela
Eugenia da Silva CÁCERES & André APTROOT—Anew, locally common
Graphis (Graphidaceae) species from southern Brazil ............... 233-237
Rungtiwa PHOOKAMSAK , Jian-Kui LIU,Dimuthu S. MANAMGODA ,
Dhanushk a N. WANASIN G H E , Hira n A R I Y AWA NSA, P eter E .
MORTIMER, Ekachai CHUKEATIROTE, Eric H.C. McKENZI E & Kevin D.
HYDE — Epitypification of two bambusicolous fungi from Thailand ...... 239-256
Wenjing LI, Jiankui LIU, D. Jayarama BHAT, Erio CAMPORESI, Jianchu XU
&KevinD.HYDE — Introducing th e n ovel s p e cies,Dothiorella
symphoricarposicola, from snowberry in Italy ..................... 257-270
Ji a-Jia CHEN & Lu-L u SHEN — Amylo s por u s succulentus sp. nov.
(Russulales, Basidiomycota) evidenced by morphological characters and
phylogenetic analysis ...................................... 271-282
Shi-Cheng SHAO, Bart BUYCK, Valérie HOFSTETTER, Xiao-Fei TIAN, Yan-
Hong GENG, Fu-Qian g YU & Pei-Gui LIUCantharellushygrophorus, a
new species in subgenusAfrocantharellusfrom tropical southwestern China 283-291
SlavomirADAMIK & Bart BUYCK — Type studies inRussulasubsection
Nigricantesfrom the Eastern United States........................ 293-309
Cryptogamie, Mycologie, 2014, 35 (3): 271-282
© 2014 Adac. Tous droits réservés
doi/10.7872/crym.v35.iss3.2014.271
Amylosporus succulentus sp. nov.
(Russulales, Basidiomycota) evidenced
by morphological characters and phylogenetic analysis
Jia-Jia CHEN#* & Lu-Lu SHEN#
Institute of Microbiology, P.O. Box 61, Beijing Forestry University,
Beijing 100083, P. R. China
Abstract – Three polypore specimens were collected from Hainan, southern China. They
are described and illustrated here as a new species, Amylosporus succulentus, based on a
combination of morphological characters and phylogenetic (ITS and nLSU sequences) data.
It is characterized by poroid basidiocarps, both simple septate and clamped generative
hyphae, hymenial hyphae without clamp connections, and finely asperulate and amyloid
basidiospores. These characters are typical for Amylosporus. In the phylogenetic
perspective, A. succulentus is closely related to A. campbellii, the generic type, and nested
within the Wrightoporiaceae clade. A key to accepted species of Amylosporus worldwide
is provided.
Molecular phylogeny / Polypore / Taxonomy / Wood-inhabiting fungi / Wrightoporiaceae
INTRODUCTION
Amylosporus Ryvarden (1973), typified by A. campbellii (Berk.)
Ryvarden, was introduced for an annual growth habit, poroid basidiocarps, both
simple septate and clamped generative hyphae, hymenial hyphae without clamp
connections, and finely asperulate and amyloid basidiospores (David &
Rajchenberg, 1985, 1987; Hattori, 2008). Four species were recorded worldwide in
Amylosporus, namely A. bracei (Murrill) A. David & Rajchenb., A. campbellii,
A. iobapha (Pat.) A. David & Rajchenb., and A. ryvardenii Stalpers.
During studies on the polypores from southern China, three specimens
previously identified as A. campbellii were re-studied, and they respresent in fact
an undescribed species based on morphological characters and phylogenetic
analysis of ITS and nLSU sequences. Its illustrated description is provided along
with an identification key to the five accepted species of Amylosporus.
MATERIAL AND METHODS
Morphological studies
The studied specimens are deposited at the herbaria of the Institute of
Microbiology, Beijing Forestry University (BJFC) and the Institute of Applied
* Correspondence author: jiaj.chen19@yahoo.com
#Jia-Jia CHEN and Lu-Lu SHEN contributed equally to this work
272 J.-J. Chen & L.-L. Shen
Ecology, Chinese Academy of Sciences (IFP). The microscopic routines followed
Li et al. (2014 ). Sections were studied at magnification up to ×1000 using a Nikon
E80i microscope and phase contrast illumination. Drawings were made with the
aid of a drawing tube. Microscopic features, measurements and drawings were
made from slide preparations stained with Cotton Blue and Melzer’s reagent.
Spores were measured from sections cut from the tubes. Presenting the variation
in the size of the spores, 5% of measurements were excluded from each end of the
range, and were given in parentheses. Basidiospore spine lengths were not
included in the measurements. In the text the following abbreviations were used:
IKI = Melzer’s reagent, IKI+ = amyloid, IKI– = non-dextrinoid and non-amyloid,
KOH =5% potassium hydroxide, CB =Cotton Blue, CB+ =cyanophilous,
CB– = acyanophilous, L = mean spore length (arithmetic average of all spores),
W=mean spore width (arithmetic average of all spores), Q=variation in the
L/W ratios between the specimens studied, n = number of spores measured from
given number of specimens. Special color terms followed Petersen (1996).
Molecular procedures and phylogenetic analysis
A CTAB rapid plant genome extraction kit (China) was used to obtain
PCR products from dried specimens, according to the manufacturer’s instructions
with some modifications (Chen & Cui, 2014). The DNA was amplified with the
primers: ITS4 and ITS5 or ITS1 for ITS (White et al., 1990), and LR0R and LR7
or LR5 for nLSU (http://www.biology.duke.edu/fungi/mycolab/primers.htm). The
PCR procedure for ITS was as follows: initial denaturation at 95°C for 3 min,
followed by 35 cycles at 94°C for 40 s, 54°C for 45 s and 72°C for 1 min, and a final
extension of 72°C for 10 min. The PCR procedure for nLSU was as follows: initial
denaturation at 94°C for 1 min, followed by 35 cycles at 94°C for 30 s, 50°C for
1 min and 72°C for 1.5 min, and a final extension of 72°C for 10 min. The PCR
products were purified and sequenced in Beijing Genomics Institute, China, with
the same primers.
The dataset in our phylogenetic analysis was extended from the previous
studies on Russulales (mainly from Larsson & Larsson, 2003, Miller et al.,
2006, Larsson, 2007). Sequences generated for this study were aligned with
additional sequences downloaded from GenBank (Table 1) using BioEdit (Hall,
1999) and ClustalX (Thompson et al., 1997). Prior to phylogenetic analysis,
ambiguous sequences at the start and the end were deleted and gaps were
manually adjusted to optimize the alignment. Sequence alignment was deposited
at TreeBase (http://purl.org/phylo/treebase; submission ID 16140).
Phylogenetic analysis was done as in Li & Cui (2013). Maximum
parsimony (MP) analysis was applied to the combined ITS and nLSU dataset.
The sequences of Sistotrema brinkmannii (Bres.) J. Erikss., S. coronilla (Höhn.)
Donk ex D.P. Rogers, S. muscicola (Pers.) S. Lundell and S. sernanderi (Litsch.)
Donk were used as outgroups following Larsson & Larsson (2003). The tree
construction procedure was performed in PAUP* version 4.0b10 (Swofford,
2002). All characters were equally weighted and gaps were treated as missing
data. Trees were inferred using the heuristic search option with TBR branch
swapping and 1000 random sequence additions. Max-trees were set to 5000,
branches of zero length were collapsed and all parsimonious trees were saved.
Clade robustness was assessed using a bootstrap (BT) analysis with 1000 replicates
(Felsenstein, 1985). Descriptive tree statistics tree length (TL), consistency index
(CI), retention index (RI), rescaled consistency index (RC), and homoplasy index
Amylosporus succulentus sp. nov. 273
Table 1. A list of species, specimens and GenBank accession number of sequences used in this
study. New sequences are shown in bold
Species Sample no. Locality GenBank accessions
ITS nLSU
Albatrellus ovinus (Schaeff.) Kotl. & Pouzar PV 22-89 AF506396 AF506396
Albatrellus subrubescens (Murrill) Pouzar PV 154-95 AF506395 AF506395
Aleurocystidiellum disciforme (DC.) Boidin et al. NH 13003 RussiaAF506402 AF506402
Aleurocystidiellum subcruentatum (Berk. &
M.A. Curtis) P.A. Lemke NH 12874 Germany AF506403 AF506403
Aleurodiscus amorphus (Pers.) J. Schröt. KHL 4240 Sweden AF506397 AF506397
Amylosporus bracei (Murrill) A. David & Rajchenb. 1008/77 USA KM267724 KJ807076
Amylosporus campbellii (Berk.) Ryvarden 0806/20a Jamaica JF692200 KJ807077
A. succulentus Jia J. Chen & L.L. Shen Dai 7802 China KM213669 KM213671
A. succulentus Dai 7803 China KM213668 KM213670
Amylostereum areolatum (Chaillet ex Fr.) Boidin NH 8041 Romania AF506405 AF506405
Amylostereum laevigatum (Fr.) Boidin NH 2863 Sweden AF506407 AF506407
Auriscalpium vulgare Gray EL 33-95 Sweden AF506375 AF506375
Boidinia aculeata (Sheng H. Wu) E. Larss. &
K.H. Larss. Wu 890714-52 China AF506433 AF506433
Boidinia granulata Sheng H. Wu Wu 9209-34 China AY048880 AY048880
Boidinia propinqua (H.S. Jacks. & Dearden) Hjortstam
& Ryvarden KHL 10931 Jamaica AF506379 AF506379
Bondarzewia montana (Quél.) Singer Canada DQ200923 DQ234539
Bondarzewia podocarpi Y.C. Dai Dai 9261 China KJ583207 KJ583221
Byssoporia terrestris (DC.) M.J. Larsen & Zak Hjm 18172 Sweden DQ389664 DQ389664
Dentipellicula taiwaniana (Sheng H. Wu) Y.C. Dai &
L.W. Zhou Dai 10867 China JQ349115 JQ349101
Dentipellis fragilis (Pers.) Donk Dai 12550 China JQ349110 JQ349096
Dentipellis parmastoi (Nikol.) Stalpers Cui 8513 China JQ349113 JQ349099
Dentipellopsis dacrydicola Y.C. Dai & L.W. Zhou Dai 12004 China JQ349104 JQ349089
Dentipratulum bialoviesense Doma_ski GG 1645 France AF506389 AF506389
Echinodontium tinctorium (Ellis & Everh.) Ellis &
Everh NH 6695 Canada AF506430 AF506430
Gloeocystidiellum bisporum Boidin et al. KHL 11135 Norway AY048877 AY048877
Gloeocystidiellum clavuligerum (Höhn. & Litsch.)
Nakasone NH 11185 SpainAF310088 AF310088
Gloeocystidiellum compactum Sheng H. Wu Wu 880615-21China AF506434 AF506434
Gloeocystidiellum formosanum Sheng H. Wu Wu 9404-16 China AF506439 AF506439
Gloeocystidiellum porosum (Berk. & M.A. Curtis)
Donk NH 10434 Denmark AF310094 AF310094
Gloeocystidiopsis cryptacanthus (Pat.) E. Larss. &
K.H. Larss. KHL 10334 Puerto Rico AF506442 AF506442
Gloeodontia discolor (Berk. & M.A. Curtis) Boidin KHL 10099 Puerto Rico AF506445 AF506445
Gloeodontia pyramidata (Berk. & M.A. Curtis)
Hjortstam Ryvarden 15502Colombia AF506446 AF506446
274 J.-J. Chen & L.-L. Shen
Gloeopeniophorella convolvens (P. Karst.) Boidin et al. KHL 10103 Puerto Rico AF506435 AF506435
Gloiodon nigrescens (Petch) Maas Geest. Desjardin 7287 Bali AF506450 AF506450
Gloiodon strigosus (Sw.) P. Karst. JS 26147 Norway AF506449 AF506449
Gloiothele lactescens (Berk.) Hjortstam EL 8-98 Sweden AF506453 AF506453
Hericium alpestre Pers. NH 13240 Russia AF506457 AF506457
Hericium americanum Ginns DAOMF-21467Canada AF506458 AF506458
Hericium erinaceus (Bull.) Pers. NH 12163 Russia AF506460 AF506460
Heterobasidion annosum (Fr.) Bref. 06129/6 Russia KJ583211 KJ583225
Heterobasidion parviporum Niemelä & Korhonen 04121/3 Finland KJ583212 KJ583226
Lactarius leonis Kytöv. SJ 91016 Sweden AF506411 AF506411
Laxitextum bicolor (Pers.) Lentz NH 5166 Sweden AF310102 AF310102
Lentinellus omphalodes (Fr.) P. Karst. JJ 2077 Sweden AF506418 AF506418
Lentinellus ursinus (Fr.) Kühner EL 73-97 USA AF506419 AF506419
Megalocystidium luridum (Bres.) Jülich KHL 8635 Norway AF506422 AF506422
Peniophora pini (Schleich.) Boidin Hjm 18143 Sweden EU118651 EU118651
Polyporoletus sublividus Snell JA 030918 DQ389663 DQ389663
Pseudoxenasma verrucisporumK.H. Larss. &Hjortstam EL 34-95 Sweden AF506426 AF506426
Russula violacea Quél. SJ 93009 Sweden AF506465 AF506465
Scytinostroma ochroleucum (Bres. & Torrend) Donk TAA 159869 Australia AF506468 AF506468
Scytinostroma odoratum (Fr.) Donk KHL 8546 Sweden AF506469 AF506469
Sistotrema brinkmannii (Bres.) J. Erikss. NH 11412 Turkey AF506473 AF506473
Sistotrema coronilla (Höhn. &Litsch.)Donk ex D.P. Rogers NH 7598 Canada AF506475 AF506475
Sistotrema muscicola (Pers.) S. Lundell KHL 8791 Sweden AF506474 AF506474
Sistotrema sernanderi (Litsch.) Donk KHL 8576 Sweden AF506476 AF506476
Stereum hirsutum (Willd.) Pers. NH 7960 Romania AF506479 AF506479
Vararia ochroleuca (Bourdot & Galzin) Donk JS 24400 Norway AF506485 AF506485
Wrightoporia austrosinensis Y.C. Dai Dai 11579 China KJ807065 KJ807073
Wrightoporia avellanea (Bres.) Pouzar E 7088 AJ537507 AJ537507
W. avellanea Ryvarden 41710 Jamaica AF506488 AF506488
Wrightoporia casuarinicola Y.C. Dai & B.K. Cui Dai 6914 China KJ807068
Wrightoporia lenta (Overh. & J. Lowe) Pouzar Cui 7804 China KJ513292 KJ807081
W. lenta Dai 10462 China KJ513291 KJ807082
Wrightoporia rubella Y.C. Dai Dai 9233 China KJ807071 KJ807084
W. subavellanea Jia J. Chen & B.K. Cui Dai 11484 China KJ513295 KJ807085
W. subavellanea Dai 11488 China KJ513296 KJ807086
W. subavellanea Dai 11492 China KJ513297 KJ807087
Wrightoporia tropicalis (Cooke) Ryvarden TFM F-16446 Japan KJ807072 KJ807088
W. tropicalis Ryvarden 45363 Belize KJ513294 KJ807089
Table 1. A list of species, specimens and GenBank accession number of sequences used in this
study. New sequences are shown in bold (continued)
Species Sample no. Locality GenBank accessions
ITS nLSU
Amylosporus succulentus sp. nov. 275
(HI) were calculated for each maximum parsimonious tree (MPT) generated.
Phylogenetic trees were visualized using Treeview (Page, 1996).
MrModeltest2.3 (Nylander, 2004) was used to determine the best-fit
evolution model for the combined dataset for bayesian inference (BI). BI was
calculated with MrBayes3.1.2 (Ronquist & Huelsenbeck, 2003) with a general
time reversible (GTR) model of DNA substitution and an invgamma distribution
rate variation across sites. Four Markov chains were run for 2 runs from random
starting trees for 5 million generations of the combined ITS and nLSU dataset,
and sampled every 100 generations. The burn-in was set to discard the first 25%
of the trees. A majority rule consensus tree of all remaining trees was calculated.
Branches that received bootstrap support for MP and bayesian posterior
probabilities (BPP) greater than or equal to 75% (MP) and 0.95 (BPP)
respectively were considered as significantly supported.
RESULTS
The ITS+nLSU dataset included sequences from 70 fungal specimens
representing 64 taxa. The dataset had an aligned length of 2098 characters in the
dataset, of which 1010 characters are constant, 173 are variable and parsimony-
uninformative, and 915 are parsimony-informative. Maximum parsimony analysis
yielded one equally parsimonious tree (TL = 6291, CI = 0.349, RI = 0.586,
RC = 0.205, HI = 0.651), and the maximum parsimonious tree was shown in Fig. 1.
Best model estimated and applied in the BI was “GTR+I+G” with equal
frequency of nucleotides. Both MP and BI trees resulted in similar topologies.
Only the MP tree was provided. Both bootstrap values (50%) and BPPs (0.95)
were showed at the nodes (Fig. 1).
The newly sequenced specimens from southern China were embedded
in the Wrightoporiaceae calde as a distinct lineage, and had a close relationship
with Amylosporus campbellii (80% MP and 1.00 BPPs). In addition, A. bracei,
A. campbellii,A. succulentus,Wrightoporia casuarinicola Y.C. Dai & B.K. Cui,
and W. rubella Y.C. Dai formed a distinct and well supported clade (80% MP and
1.00 BPPs) that appears weakly related to W. lenta (Overh. & J. Lowe) Pouzar,
the type species of Wrightoporia.
TAXONOMY
Amylosporus succulentus Jia J. Chen & L.L. Shen, sp. nov. Figs 2-3
MycoBank: MB 809943
Original diagnosis: Differs from other Amylospus species by juicy fruiting
body when fresh, pileate basidiocarps with a cream to pinkish violet pore surface,
dextrinoid and hyaline skeletal hyphae, distinctly inflated contextual skeletal
hyphae in KOH, presence of gloeoplerous hyphae and cystidioles, and ellipsoid,
slightly thick-walled, and cyanophilous basidiospores measuring 4.2-5.2 ×3-3.8 µm.
HOLOTYPUS: CHINA, Hainan Province, Haikou, Jinniuling Park, on
lawn, 1 Sep 2006, Y.C Dai, Dai 7802 (BJFC; IFP).
276 J.-J. Chen & L.-L. Shen
Fig. 1. Strict consensus tree illustrating the phylogeny of the new species and related species
generated by Maximum parsimony based on ITS + nLSU sequences. Parsimony bootstrap
proportions (before the slash markers) higher than 50% and Bayesian posterior probabilities
(after the slash markers) more than 0.95 are indicated along branches.
Amylosporus succulentus sp. nov. 277
Etymology. succulentus (Lat.): referring to watery and juice fruiting body
of the species when fresh.
Basidiocarps annual, pileate, centrally to laterally stipitate, solitary or a
few confluent, watery and juice when fresh, without odour or taste, becoming
corky and light in weight upon drying. Pileus more or less circular, projecting up
to 7 cm long, 15 cm wide, 4 cm thick at the base, sometimes lobed, becoming
thinner towards margins; margins undulating, obtuse to acute. Pileal surface
cream to greyish violet when fresh, becoming pinkish buff to clay-buff when dry,
azonate. Pore surface cream to pinkish violet when fresh, buff upon drying; pores
angular, 2-4 per mm; dissepiments thin, lacerate. Context cream and watery when
fresh, pinkish buff and corky when dry, up to 3 cm thick. Tubes buff and brittle
when dry, up to 1cm long. Stipe short and thick, buff and corky when dry, up to
1 cm long. Type of rot. White rot.
Fig. 2. Basidiocarp of Amylosoporus succulentus from the holotype. a. Fresh basidiocarp in the
wild. b-c. Dried basidiocarp. Bars a, b, c = 1 cm.
278 J.-J. Chen & L.-L. Shen
Fig. 3. Microscopic structures of Amylosporus succulentus (drawn from the holotype).
a. Basidiospores. b. Basidia and basidioles. c. Cystidioles. d. Hyphae from trama. e. Hyphae from
subiculum. Bars: a. 5 µm; b-e. 10 µm.
Amylosporus succulentus sp. nov. 279
Hyphal system dimitic; tramal generative hyphae with simple septa only,
contextual generative hyphae with both simple septa and double or multiple
clamp connections; skeletal hyphae dextrinoid, CB+; contextual skeletal hyphae
distinctly inflated in KOH, up to 20 µm in diam. Context Generative hyphae
domidant, hyaline, thin- to slightly thick-walled, frequently branched, 4-10 µm in
diam; skeletal hyphae frequent, thick-walled with a narrow to wide lumen,
freuqently branched, flexuous, loosely interwoven, 3-8 µm in diam; gloeoplerous
hyphae occasionally present, thin-walled with granular to oily contents appearing
refractive in phase contrast illumination, up to 12 µm in diam. Tubes Generative
hyphae common to dominant, hyaline, thin- to slightly thick-walled, rarely
branched, subparallel along the tubes, 3-6 µm in diam; skeletal hyphae common,
thick-walled with a narrow to wide lumen, freuqently branched, flexuous, loosely
interwoven, 3-8 µm in diam; gloeoplerous hyphae frequently present, thin-walled
with granular to oily contents appearing refractive in phase contrast illumination,
up to 9 µm in diam. Cystidia absent, but cystidioles present, thin-walled, fusoid,
tapering, 13-18 ×3-4 µm; basidia clavate, with four sterigmata and a basal simple
septum, 15-20 ×5-8 µm; basidioles in shape similar to basidia, but slightly smaller.
Basidiospores ellipsoid, hyaline, slightly thick-walled, finely asperulate, IKI+,
CB+, (4.0-)4.2-5.2(-5.4) ×(2.8-)3-3.8(-4) µm, L = 4.73 µm, W = 3.14 µm, Q = 1.46-
1.52 (n = 60/2).
Additional specimens examined:Amylosporus succulentus — CHINA,
Hainan Province, Haikou, Jinniuling Park, on lawn, 1 Sep 2006, Y.C Dai, Dai 7803
(BJFC; IFP) & 7808 (IFP).
DISCUSSION
Morphologically, Amylosporus succulentus is characterized by an annual
growth habit, watery and juicy fruiting body when fresh, pileate basidiocarps with
a cream to pinkish violet pore surface, both simple septate and clamped
generative hyphae, hymenial hyphae without clamp connections, dextrinoid and
hyaline skeletal hyphae, distinctly inflated contextual skeletal hyphae in KOH,
presence of gloeoplerous hyphae and cystidioles, and ellipsoid, slightly
thick-walled, finely asperulate, amyloid and cyanophilous basidiospores which are
4.2-5.2 ×3-3.8 µm. Phylogenetically, two samples of A. succulentus formed a
distinct lineage with strong supports (100% MP, 1.00 BPPs) and are distant from
other taxa in the genus or other genera. Amylosporus succulentus was embedded
in the lineage of the Wrightoporiaceae and clustered with A. campbellii. Both
morphology and rDNA sequence data confirmed that the two samples represent
a new species in Amylosporus.
Amylosporus succulentus is closely related to A. campbellii according to
our rDNA phylogeny (Fig. 1). Morphologically, A. succulentus may be confused
with A. campbellii, as they produce pileate basidiocarps, similar sized pores
(2-4 per mm in A. campbellii), and presence of gloeoplerous hyphae. However,
A. campbellii can be readily distinguished from A. succulentus by its non-
dextrinoid and pale golden yellow skeletal hyphae.
Previously, Amylosporus campbellii was reported in China (Dai, 2007,
2012), but the identifications were based only on morphological characters.
According to the combination of morphological features and rDNA sequences
280 J.-J. Chen & L.-L. Shen
data, the Chinese samples turned out to be different from A. campbellii originally
described from India, and they are described here as a new species, A. succulentus.
It should be noted that previously many Chinese polypores were named after
already described species on the basis of morphology only, and in fact many of
them were later proven to be undescribed species using molecular studies, such as
species in Albatrellus, Hymenochaete, Heterobasidion,Perenniporia,Phellinidium,
Phylloporia,Polyporus etc. (Cui et al., 2008; He & Dai, 2012; Zhou & Dai, 2012;
Cui & Dai, 2013; Zhao & Cui, 2013; Zhao et al., 2013; Chen et al., 2014; Dai et al.,
2014; Zhou et al., 2014).
Amylosporus succulentus may be confused with A. bracei in producing a
pinkish violet pore surface and dextrinoid skeletal hyphae. However, A. bracei
differs from A. succulentus in its resupinate to effused-reflexed basidiocarps with
rhizomorphs, smaller pores (5-7 per mm), absence of gloeoplerous hyphae, and
smaller basidiospores (3-3.5 ×2.5 µm, Ryvarden, 2000). Moreover, the two species
are different in the ITS and nLSU rDNA-based phylogenetic analysis (Fig. 1).
Amylosporus is a white-rot fungal genus belonging to the Russulales. The
previous studies on Amylosporus were mainly based on morphological characters
(Ryvarden, 1977; David & Rajchenberg, 1985; Stalpers, 1996). Recently, Chen &
Cui (2014) proved that Amylosporus was a polyphyletic genus and closely related
with Wrightoporia casuarinicola and W. rubella. However, relationships among
species of Amylosporus appear ambiguous (Fig. 1). Species of Amylosporus were
embedded in the Wrightoporiaceae clade. Nevertheless, species of Wrightoporia
with simple septate generative hyphae clustered with species of Amylosporus,
then formed a different group (92% MP and 1.00 BPPs) that appears distant from
the type species, W. lenta. Species in this clade are characterized by hymenial
hyphae without clamp connections, differing from other species of Wrightoporia.
A further study on subdivision and phylogeny of all the species of Wrightoporia
s.l. and Amylosporus is badly needed by wider taxa sampling and more conserved
gene markers.
KEY TO ACCEPTED SPECIES OF AMYLOSPORUS WORLDWIDE
1.Basidiocarpspileate.................................................. 2
1. Basidiocarps resupinate to effused-reflexed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2. Skeletal hyphae dextrinoid .................................A.succulentus
2. Skeletal hyphae non-dextrinoid ..............................A. campbellii
3.Tramalgenerativehyphae>3µmindiam ............................... 4
3. Tramal generative hyphae < 3 µm in diam. .......................A. iobapha
4. Basidiospores > 4 µm long...................................A.ryvardenii
4. Basidiospores < 4 µm long.......................................A. bracei
Acknowledgements. The research was financed by the Fundamental Research
Funds for the Central Universities (No. BLYJ201403).
Amylosporus succulentus sp. nov. 281
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— We studied sequence variation in 16S rDNA in 204 individuals from 37 populations of the land snail Candidula unifasciata (Poiret 1801) across the core species range in France, Switzerland, and Germany. Phylogeographic, nested clade, and coalescence analyses were used to elucidate the species evolutionary history. The study revealed the presence of two major evolutionary lineages that evolved in separate refuges in southeast France as result of previous fragmentation during the Pleistocene. Applying a recent extension of the nested clade analysis (Templeton 2001), we inferred that range expansions along river valleys in independent corridors to the north led eventually to a secondary contact zone of the major clades around the Geneva Basin. There is evidence supporting the idea that the formation of the secondary contact zone and the colonization of Germany might be postglacial events. The phylogeographic history inferred for C. unifasciata differs from general biogeographic patterns of postglacial colonization previously identified for other taxa, and it might represent a common model for species with restricted dispersal.