Research Article |
Corresponding author: Zhu L. Yang ( fungi@mail.kib.ac.cn ) Academic editor: María P. Martín
© 2021 Zheng-Mi He, Zhu L. Yang.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
He Z-M, Yang ZL (2021) A new clitocyboid genus Spodocybe and a new subfamily Cuphophylloideae in the family Hygrophoraceae (Agaricales). MycoKeys 79: 129-148. https://doi.org/10.3897/mycokeys.79.66302
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Phylogenetically, the genera Cuphophyllus, Ampulloclitocybe and Cantharocybe are treated as basal in the family Hygrophoraceae, despite weak support. However, the exact phylogenetic positions of the three genera have remained unresolved, and taxa related to these genera are poorly known. In this study, a new clitocyboid genus Spodocybe was proposed based on multigenic phylogenetic inference datasets and morphological evidence. The analyses of ITS as well as two combined datasets ITS-nrLSU-rpb2 and ITS-nrLSU-rpb1-rpb2-tef1-α-atp6 supported that (1) Spodocybe formed a well-supported monophyletic clade; and (2) sisters Spodocybe and Ampulloclitocybe, along with Cantharocybe and Cuphophyllus also formed a monophyletic lineage, as sister to the rest of the Hygrophoraceae. Meanwhile, two new species, namely S. rugosiceps and S. bispora, from southwestern China, were documented and illustrated. These results support the new proposed genus Spodocybe, and that Spodocybe, Ampulloclitocybe, Cantharocybe and Cuphophyllus should be retained in the Hygrophoraceae as a new subfamily Cuphophylloideae.
Ampulloclitocybe, Cantharocybe, Cuphophyllus, morphological characters, phylogenetic analysis, taxonomy
The widespread genus Clitocybe (Fr.) Staude currently encompasses large numbers of species with clitocyboid habit, sharing the features of saprophytic nutrition, funnel-shaped pileus, decurrent lamellae, a usually white, cream or pale colored spore-deposit and smooth and inamyloid spores (
Based on phylogenetic analyses over the past 20 years, (i) many new genera within the Tricholomatoid clade were proposed to accommodate previous Clitocybe species deviating from the core Clitocybeae clade (
The molecular phylogenetic relationships among members of the Hygrophoraceae Lotsy were well studied by
Recently, some collections were shown to be closely related to Clitocybe trulliformis (Fr.) P. Karst. based on ITS-BLAST searches while at the same time they were surprisingly related to taxa of the genus Cuphophyllus based on nrLSU-BLAST searches. As far as we know, C. trulliformis and allied species were lacking taxonomic revision, especially regarding their molecular phylogenetic status. Furthermore, the phylogenetic delimitation of the Hygrophoraceae was ambiguous due to the uncertain positions of Cuphophyllus, Ampulloclitocybe and Cantharocybe. Hence, the aims of this study were (a) to propose and describe a new genus of the Hygrophoraceae for species related to C. trulliformis based on morphological and molecular analyses and (b) to reconstruct the phylogeny of the Hygrophoraceae for determining the exact phylogenetic placements of Cuphophyllus, Ampulloclitocybe and Cantharocybe with multi-gene data.
Twenty-three specimens of species similar to C. trulliformis and related species were collected from southwestern and northeastern China and western Germany, during 2007 to 2020. The fresh fruitbodies were dried using heat or silica gel. Voucher specimens were deposited in the Herbarium of Kunming Institute of Botany, Chinese Academy of Sciences (
Specimens used in phylogenetic analysis and their GenBank accession numbers. The newly generated sequences are shown in bold.
Species | Voucher | Locality | GenBank accession number | |||||
---|---|---|---|---|---|---|---|---|
ITS | nrLSU | rpb2 | rpb1 | tef1-α | atp6 | |||
Acantholichen pannarioides | MDF352 | Costa Rica | KT429795 | KT429807 | KT429817 | |||
Acantholichen campestris | DIC595b | Brazil | KT429798 | KT429810 | KT429818 | |||
Acantholichen galapagoensis | MDF058 | Ecuador | KT429785 | KT429800 | KT429812 | |||
Ampulloclitocybe clavipes |
|
China: Jilin | MW616462 | MW600481 | MW656471 | MW656467 | MW656461 | MW656478 |
AFTOL-ID 542 | AY789080 | AY639881 | AY780937 | AY788848 | AY881022 | |||
DJL06TN40 | USA | FJ596912 | KF381542 | KF407938 | ||||
Arrhenia auriscalpium | TUB 011588 | DQ071732 | ||||||
Arrhenia acerosa | Lueck2 | Germany | KP965766 | KP965784 | ||||
Cantharellula umbonata | CBS 398.79 | France | MH861222 | MH872990 | ||||
Cantharocybe gruberi | AFTOL-ID 1017 | USA | DQ200927 | DQ234540 | DQ385879 | DQ435808 | DQ059045 | |
AH24539 | Spain | JN006422 | JN006420 | |||||
Cantharocybe brunneovelutina | DJL-BZ-1883 | Belize | NR160458 | NG068731 | ||||
Cantharocybe virosa | TENN63483 | India | KX452405 | JX101471 | ||||
Iqbal-568 | Bangladesh | KX452403 | KF303143 | |||||
Chromosera cyanophylla | AFTOL-ID 1684 | USA | DQ486688 | DQ457655 | KF381509 | |||
Chromosera ambigua | GE18008-1 | France | MK645573 | MK645587 | MK645593 | |||
Chromosera lilacina | GE18035 | Canada | MK645577 | MK645591 | MK645597 | |||
Chromosera xanthochroa | GE18033 | Canada | MK645576 | MK645590 | MK645596 | |||
Chrysomphalina chrysophylla | AFTOL-ID 1523 | USA | DQ192180 | DQ457656 | ||||
Chrysomphalina grossula | OSC 113683 | EU644704 | EU652373 | |||||
Clitocybe aff. costata | DJL06TN80 | USA | FJ596913 | |||||
Clitocybe herbarum | G0171 | Hungary | MK277719 | |||||
Clitocybe trulliformis | 14562 | Italy | JF907809 | |||||
4804 | Russia | MH930178 | ||||||
Clitocybe cf. trulliformis | G0460 | Hungary | MK277728 | |||||
Clitocybe sp. | NAMA 2015-206 | USA | MH910535 | |||||
Clitocybe sp. | NAMA 2015-318 | USA | MH910563 | |||||
Clitocybe sp. | Mushroom Observer 302917 | USA | MK607556 | |||||
Cora pavonia | DIC215 | Ecuador | KF443238 | KF443261 | KF443275 | |||
Cora aspera | DIC110 | Bolivia | KF443230 | KF443257 | KF443267 | |||
Cora reticulifera | DIC119 | Ecuador | KF443239 | KF443262 | KF443269 | |||
Cora squamiformis | DIC146 | Bolivia | KF443240 | KF443263 | KF443273 | |||
Corella brasiliensis | MDF017 | Bolivia | KF443229 | KF443255 | KF443276 | |||
Corella aff. Melvinii | MDF200 | Brazil | KJ780569 | KY861725 | ||||
Cuphophyllus pratensis | Lueck7 | Germany | KP965771 | KP965789 | ||||
DJL-Scot-8 | UK | KF291057 | KF291058 | |||||
Cuphophyllus aurantius | CFMR PR-6601 | Puerto Rico | KF291099 | KF291100 | KF291102 | |||
Cuphophyllus aff. pratensis | AFTOL-ID 1682 | USA | DQ486683 | DQ457650 | DQ435804 | |||
Cuphophyllus sp. |
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China: Tibet | MW762875 | MW763000 | MW789179 | MW789163 | ||
Cyphellostereum galapagoense | CDS 41163 | Ecuador | NR158415 | NG068806 | ||||
Cyphellostereum imperfectum | DIC115 | Guatemala | KF443218 | KF443243 | KF443277 | |||
Dictyonema interruptum | Ertz 10475 | Portugal | EU825967 | KF443282 | ||||
Dictyonema schenckianum | DIC113 | Brazil | KF443225 | KF443251 | KF443285 | |||
Eonema pyriforme | G1063 | Poland | MK278075 | |||||
Gliophorus psittacinus | CFMR DEN-25 | Denmark | KF291075 | KF291076 | KF291078 | |||
Gliophorus graminicolor | TJB-10048 (CORT) | Australia | KF381520 | KF381545 | KF407936 | |||
Gliophorus aff. laetus | CFMR PR-5408 | Puerto Rico | KF291069 | KF291070 | ||||
Gloioxanthomyces nitidus | GDGM41710 | China: Jilin | MG712283 | MG712282 | MG711911 | |||
Haasiella splendidissima | Herb. Roux n. 4044 | France | JN944400 | JN944401 | ||||
Herb. Roux n. 3666 | Moldova | JN944398 | JN944399 | |||||
Haasiella venustissima | A. Gminder 971488 | Italy | KF291092 | KF291093 | ||||
E. C. 08191 | Italy | JN944393 | JN944394 | |||||
Humidicutis marginata | JM96/33 | AF042580 | ||||||
Humidicutis auratocephalus | AFTOL-ID 1727 | USA | DQ490624 | DQ457672 | DQ472720 | DQ447906 | ||
Humidicutis dictiocephala | QCAM6000 | Ecuador | KY689661 | KY780120 | ||||
Humidicutis sp. | CFMR BZ-3923 | Belize | KF291110 | KF291111 | ||||
Hygroaster nodulisporus | AFTOL-ID 2020 | USA | EF561625 | |||||
Hygroaster albellus | AFTOL-ID 1997 | Puerto Rico | KF381521 | EF551314 | ||||
Hygrocybe conica | FO 46714 | DQ071739 | ||||||
Hygrocybe cf. acutoconica | CFMR NC-256 | USA | KF291117 | KF291118 | KF291120 | |||
Hygrocybe coccinea | AFTOL-ID 1715 | USA | DQ490629 | DQ457676 | DQ472723 | DQ447910 | GU187705 | |
Hygrocybe aff. conica | AFTOL-ID 729 | AY854074 | AY684167 | AY803747 | ||||
Hygrophorus eburneus | US97/138 | Germany | AF430279 | |||||
GDGM70059 | USA | MT093608 | ||||||
Hygrophorus chrysodon |
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China: Tibet | MW616463 | MW600482 | MW656472 | MW656462 | MW656479 | |
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China: Tibet | MW762876 | MW763001 | MW789180 | MW789164 | MW773440 | MW789195 | |
Hygrophorus flavodiscus |
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China: Yunnan | MW616464 | MW600483 | MW656473 | MW656468 | MW656463 | MW656480 |
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China: Yunnan | MW616465 | MW600484 | MW656474 | MW656469 | MW656464 | MW656481 | |
Hygrophorus gliocyclus |
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China: Tibet | MW616466 | MW600485 | MW656475 | MW656465 | MW656482 | |
Hygrophorus hypothejus |
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Germany | MW616467 | MW600486 | MW656476 | MW656470 | MW656483 | |
Hygrophorus pudorinus | AFTOL-ID 1723 | USA | DQ490631 | DQ457678 | DQ472725 | DQ447912 | GU187710 | |
Hygrophorus sp. 1 |
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China: Yunnan | MW762877 | MW763002 | MW789181 | MW789165 | MW773441 | MW789196 |
Hygrophorus sp. 2 |
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China: Jilin | MW616468 | MW600487 | MW656477 | MW656466 | MW656484 | |
Hygrophorus sp. 3 |
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China: Tibet | MW762878 | MW763003 | MW789182 | MW789166 | MW773442 | MW789197 |
Hygrophorus sp. 4 |
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China: Tibet | MW762879 | MW763004 | MW789183 | MW789167 | MW773443 | MW789198 |
Lichenomphalia hudsoniana | GAL18249 | USA | JQ065873 | JQ065875 | ||||
Lichenomphalia meridionalis | S-270-FB1 | Japan | LC428308 | LC428307 | ||||
Neohygrocybe ovina | GWG H. ovina Rhosisaf (ABS) | UK | KF291233 | KF291234 | KF291236 | |||
Neohygrocybe griseonigra | GDGM 44492 | China | MG779451 | MG786565 | ||||
Neohygrocybe ingrata | DJL05TN62 (TENN) | USA | KF381525 | KF381558 | KF381516 | |||
Neohygrocybe subovina | GRSM 77065 | USA | KF291140 | KF291141 | ||||
Spodocybe bispora |
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China: Yunnan | MW762880 | MW763005 | MW789184 | MW789168 | MW773444 | MW789199 |
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China: Yunnan | MW762881 | MW763006 | MW789185 | MW789169 | MW773445 | MW789200 | |
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China: Yunnan | MW762882 | MW763007 | MW789186 | MW789170 | MW773446 | MW789201 | |
Spodocybe rugosiceps |
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China: Yunnan | MW762883 | MW763008 | MW789187 | MW789171 | MW773447 | MW789202 |
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China: Yunnan | MW762884 | MW763009 | MW789188 | MW789172 | MW789203 | ||
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China: Yunnan | MW762885 | MW763010 | MW789189 | MW789173 | MW773448 | MW789204 | |
Spodocybe rugosiceps |
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China: Yunnan | MW762886 | MW763011 | MW789190 | MW789174 | MW773449 | MW789205 |
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China: Yunnan | MW762887 | MW763012 | MW789191 | MW789175 | MW789159 | MW789206 | |
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China: Yunnan | MW762888 | MW763013 | MW789192 | MW789176 | MW789160 | MW789207 | |
Spodocybe sp. 1 |
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China: Jilin | MW762889 | MW763014 | MW789193 | MW789177 | MW789161 | MW789208 |
Spodocybe sp. 2 |
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China: Yunnan | MW762890 | MW763015 | MW789194 | MW789178 | MW789162 | MW789209 |
Porpolomopsis calyptriformis | CFMR ENG-3 | UK | KF291242 | KF291243 | KF291245 | |||
Porpolomopsis aff. calyptriformis | DJL05TN80 (TENN) | USA | KF291246 | KF291247 | KF291249 | |||
Porpolomopsis lewelliniae | TJB-10034 (CORT) | Thailand | KF291238 | KF291239 | KF291241 | |||
Pseudoarmillariella ectypoides | AFTOL-ID 1557 | USA | DQ192175 | DQ154111 | DQ474127 | DQ516076 | GU187733 | |
Pseudoarmillariella bacillaris |
|
China | KC222315 | KC222316 | ||||
Sinohygrocybe tomentosipes | GDGM 50075 | China: Hunan | MG685873 | MG696902 | MG696906 | |||
GDGM 43351 | China: Sichuan | MG685872 | MG696901 | MG696905 | ||||
Amylocorticium cebennense | CFMR HHB-2808 | USA | GU187505 | GU187561 | GU187770 | GU187439 | GU187675 | |
Aphroditeola olida | DAOM 226047 | Canada | KF381518 | KF381541 | ||||
Macrotyphula fistulosa | IO. 14. 214 | Spain | MT232352 | KY224088 | MT242317 | MT242354 | ||
Macrotyphula juncea | IO. 14. 177 | Sweden | MT232353 | MT232306 | MT242337 | MT242355 | ||
Macrotyphula phacorrhiza | IO. 14. 167 | Sweden | MT232364 | MT232315 | MT242348 | MT242326 | MT242367 | |
IO.14. 200 | France | MT232363 | MT232314 | MT242347 | MT242366 | |||
Phyllotopsis nidulans | IO. 14. 196 | Spain | MT232308 | MT242338 | MT242319 | MT242357 | ||
Phyllotopsis sp. | AFTOL-ID 773 | DQ404382 | AY684161 | AY786061 | DQ447933 | DQ059047 | ||
Pleurocybella porrigens | UPS F-611822 | Sweden | MT232355 | MT232309 | MT242339 | |||
Plicaturopsis crispa | AFTOL-ID 1924 | USA | DQ494686 | DQ470820 | GU187816 | |||
Pterulicium echo | ZRL20151311 | LT716065 | KY418881 | KY419026 | KY418979 | KY419076 | ||
Pterulicium gracilis | IO. 14. 142 | Sweden | MT232356 | MT232310 | MT242358 | |||
Sarcomyxa serotina | AFTOL-ID 536 | USA | DQ494695 | AY691887 | DQ859892 | DQ447938 | GU187754 | |
Serpulomyces borealis | CFMR L-8014 | USA | GU187512 | GU187570 | GU187782 | GU187686 | ||
Tricholomopsis decora | AFTOL-ID 537 | DQ404384 | AY691888 | DQ408112 | DQ447943 | DQ029195 | ||
Tricholomopsis osiliensis | ZRL20151760 | LT716068 | KY418884 | KY419029 | KY419079 | |||
Typhula capitata | IO. 15. 122 | Spain | MT232357 | MT232312 | MT242341 | MT242321 | MT242360 | |
Typhula incarnata | IO. 14. 92 | Sweden | MT232362 | MT232313 | MT242346 | MT242325 | ||
Typhula micans | IO. 14. 165 | Sweden | MT232361 | KY224102 | MT242345 | MT242324 | MT242364 |
Macroscopic characters of species were described based on the raw field record data and photographs. Colors used in description referred to
Total genomic DNA was extracted using the Ezup Column Fungi Genomic DNA Purificaton Kit (Sangon Biotech, Shanghai, China) according to the manual. For the PCR amplification, (1) Primers ITS5 and ITS4 (
The PCR mixtures contained 1× PCR buffer, 1.5mM MgCl2, 0.2mM dNTPs, each primer at 0.4 μM, 1.25U of Taq polymerase (Sangon Biotech, Shanghai, China), and 1 μL of DNA template in a total volume of 25 μL. Reactions were performed with the following program: initial denaturation at 94 °C for 5 min, 35 cycles at 94 °C for 30 s, 50 °C (atp6), 52 °C (nrLSU, tef1-α, rpb1 and rpb2) or 54 °C (ITS) for 30 s, and 72 °C for 30 s (ITS and atp6), 50 s (nrLSU and rpb2) or 75 s (tef1-α and rpb1), and for terminal elongation, the reaction batches were incubated at 72 °C for 10 min. All PCR products were detected by 2% agarose gel electrophoresis and then sent to the Kunming branch of Tsingke Biological Technology Co., Ltd. (Beijing, China) for sequencing.
Sequences used for phylogenetic analysis (presented in Table
As shown in Table
The combined dataset ITS-nrLSU-rpb2 comprised 221 sequences from 88 samples with a total of 3135 positions. In the three-gene tree (Fig.
ML analysis of Hygrophoraceae combined ITS, nrLSU and rpb2 sequence data, with Macrotyphula juncea, Macrotyphula phacorrhiza and Phyllotopsis sp. as outgroups. Bootstrap values (BP) ≥ 50% from ML analysis and Bayesian posterior probabilities (PP) ≥ 0.90 are shown at nodes. The newly generated sequences are shown in bold.
In order to accurately determine the position of Spodocybe in the family Hygrophoraceae and better clarify the phylogenetic relationships of Spodocybe, Ampulloclitocybe, Cantharocybe and Cuphophyllus, a further six-gene matrix ITS-nrLSU-rpb1-rpb2-tef1-α-atp6 composed of 179 sequences from 54 samples with 5405 positions was used to rebuild the Hygrophoraceae tree. As revealed by the six-gene phylogenetic analysis (Fig.
ML analysis of Hygrophoraceae combined ITS, nrLSU, rpb1, rpb2, tef1-α and atp6 sequence data, with representatives of Amylocorticiaceae, Pterulaceae and the Hygrophoroid clade (Aphroditeola, Macrotyphula, Phyllotopsis, Pleurocybella, Sarcomyxa, Tricholomopsis and Typhula) as outgroups. Bootstrap values (BP) ≥ 50% from ML analysis and Bayesian posterior probabilities (PP) ≥ 0.90 are shown at nodes. Branches with BP ≥ 75% and PP ≥ 0.95 are bolded. The newly generated sequences are shown in bold. Lamellar trama type B for bidirectional, D for divergent, I for interwoven, P for pachypodial, R for regular, S for subregular, T for tri-directional. Lamellar trama types of specimens collected in this study were identified by ourselves and others referred to
In addition, an ITS dataset (23 sequences, 1053 positions) was applied to phylogenetic analysis for displaying the relationships among Spodocybe species from this study and species of Clitocybe treated from GenBank. In the ITS tree (Fig.
Phylogram showing the phylogenetic relationships among Spodocybe species and species of Clitocybe treated from Genbank based on ITS sequence data, with representatives of Ampulloclitocybe, Cuphophyllus and Hygrophorus as outgroups (rooted with Hygrophorus eburneus). Bootstrap values (BP) ≥ 50% from ML analysis and Bayesian posterior probabilities (PP) ≥ 0.90 are shown at nodes. The newly generated sequences are shown in bold. EA, NA and SE refer to East Asia, North America and South Europe, respectively.
Characterized generally by clitocyboid basidiomes, convex to funnel-shaped pileus, decurrent lamellae, absence of veils, inamyloid basidiospores and presence of clamps.
From the type genus Cuphophyllus.
Cuphophyllus (Donk) Bon.
Basidiomes small, medium-sized to large, mostly clitocyboid, rarely omphalinoid or mycenoid; veils absent. Pileus convex, applanate to funnel-shaped; surface usually dry, smooth, lubricous or rarely viscid. Lamellae decurrent to deeply decurrent. Basidiospores ellipsoid, oblong or subglobose, thin-walled and inamyloid. Pileipellis usually a cutis, sometimes ixocutis or trichoderm. Lamellar trama regular, subregular, interwoven or bidirectional. Clamp connections present.
Usually gregarious or caespitose on ground, rarely on wood; widespread in temperate and tropical regions.
The genera Ampulloclitocybe, Cantharocybe, Cuphophyllus and Spodocybe are included in the subfamily Cuphophylloideae, which is in correspondence with Cuphophylloid grade of
Differs from Ampulloclitocybe by its small basidiomes and subregular lamellar trama rather than medium-sized basidiomes and bidirectional lamellar trama. Differs from Cuphophyllus in the ratio of basidia to basidiospore length less than 5, and lamellar trama subregular rather than interwoven. Differs from Cantharocybe in its absence of cheilo- and caulocystidia, having small basidiomes rather than large ones and having subregular lamellar trama rather than regular one.
Spodo- refers to grey; -cybe refers to head; that is a Clitocybe-like genus with gray pileus.
Spodocybe rugosiceps Z. M. He & Zhu L. Yang.
Basidiomes small, clitocyboid. Pileus convex, applanate to infundibuliform; surface dry, greyish (2B1), grey-brown (5C4) to dark grey-brown (5E4); center depressed with age. Lamellae decurrent to deeply decurrent, white (1A1) to cream (1A2), thin, moderately crowded, sometimes furcate and interveined. Stipe central, subcylindrical, concolorous with pileus. Basidiospores ellipsoid, oblong to cylindrical, colourless, hyaline, smooth, thin-walled, inamyloid; ratio of basidia to basidiospore length less than 5. Pileipellis and stipitipellis a cutis. Lamellar trama subregular. Clamp connections abundant, present in all parts of basidiome.
Saprophytic, usually gregarious or caespitose on the ground of coniferous or coniferous and broad-leaved mixed forest; distributed in the temperate and subtropical zones from June to November.
Differs from S. bispora in having a rugose pileus, smaller basidiospores and 4-spored rather than 2-spored basidia. Differs from C. trulliformis in having smaller basidiospores and a rugose rather than felty-squamulose pileus.
rugosiceps refers to the rugose pileus.
China. Yunnan Province: Kunming City, near Yeya Lake, at 25.136658°N, 102.873027°E, alt. 2000 m, 11 Aug 2020, Z. M. He 72 (
Basidiomes small, clitocyboid. Pileus 0.5–2 cm in diam, at first nearly applanate, then concave; surface dry and rugose, gray-brown (5E2-4) to gray-black (4F2-4) in the center and gray-brown (5C2-4) or gray (5B1-2) towards margin; center often slightly umbonate; margin straight and undulating; context thin and white (1A1) to cream (1A2). Lamellae deeply decurrent, white (1A1) to cream (1A2), thin (up to 2 mm high), crowded, sometimes forked and intervenose. Stipe 2.5–6 × 0.2–0.4 cm, central, narrowly cylindrical to subcylindrical, sometimes flexuous, hollow; surface dry and nearly smooth, concolorous with pileus; context white (1A1).
Basidiospores [60/3/3] 5–6 (6.5) × (2.5)3–3.5(4) μm, Q = (1.38)1.55–1.95(2), Qm = 1.73 ± 0.14, elongate, colorless, hyaline, smooth, thin-walled, inamyloid. Basidia 20–24 × 5–6 μm, clavate, 4-spored, colorless, hyaline, thin-walled; sterigmata up to 4 μm long; ratio of basidia to basidiospore length values about 3–5. Cystidia absent. Lamellar trama subregular; hyphae colorless, hyaline, cylindrical, thin-walled, 3–10 µm wide. Pileipellis a cutis, but in places upright or trichodermial in appearance, made up with thin-walled cylindrical hyphae 3–9 µm wide. Stipitipellis a cutis, composed of thin-walled cylindrical hyphae 3–10 μm wide. Clamp connections present in all parts of basidiome.
Gregarious or caespitose, growing saprotrophically in forest litter, often under conifers, on the ground, known from subtropical zone of Yunnan, China; from July to October.
China. Yunnan Province: Dali Bai Autonomous Prefecture, Yunlong Country, Tianchi National Nature Reserve, at 25.850365°N, 99.274236°E, alt. 2509 m, 28 Sep 2019, X. H. Wang 7471 (
Differs from S. rugosiceps in having a nearly smooth pileus, larger basidiospores and 2-spored rather than 4-spored basidia. Differs from C. trulliformis in having a nearly smooth rather than felty-squamulose pileus.
Bispora refers to 2-spored.
China. Yunnan Province: Baoshan City, Longyang District, Shuizhai Village, at 25.273967°N, 99.306216°E, alt. 2400 m, 12 Aug 2011, J. Qin 324 (
Basidiomes small, clitocyboid. Pileus 1.5–3 cm in diam, plano-convex to funnel-shaped; surface dry and nearly smooth, greyish-brown (4B2-3) to grey-brown (4E3-5); center depressed, usually with a low umbo, somewhat darker; margin generally straight and undulating, incurved when old; context thin and white (1A1). Lamellae deeply decurrent, white (1A1) to cream (1A2), thin, 1–2 mm high, relatively crowded, sometimes forked and intervenose. Stipe 1–3 × 0.2–0.4 cm, central, subcylindrical, hollow; surface dry and nearly smooth, concolorous with pileus; context white (1A1).
Basidiospores [60/3/3] (7)7.5–10.5(11.5) × 3–4 μm, Q = (2.05)2.11–3(3.33), Qm = 2.56 ± 0.3, cylindrical, colorless, hyaline, smooth, thin-walled, inamyloid. Basidia 20–30 × 4–5.5 μm, clavate, 2-spored, colorless, hyaline, thin-walled; sterigmata up to 10 μm long; ratio of basidia to basidiospore length less than 5 (about 2–4). Cystidia absent. Lamellar trama subregular, colorless, hyaline, made up of thin-walled cylindrical hyphae with 3–10 µm wide. Pileipellis a cutis, composed of thin-walled cylindrical hyphae 3–11 µm wide. Stipitipellis a cutis, composed of thin-walled cylindrical hyphae 3–10 μm wide. Clamp connections in all parts of basidiomes.
Saprophytic, usually gregarious on the ground of coniferous or coniferous and broad-leaved mixed forest, known from Yunnan, China; July to September.
China. Yunnan Province: Kunming City, Qipan Mountain, at 26.060020°N, 102.576823°E, alt. 1900 m, 25 Jul 2020, Z. M. He 35 (
In our current study, the new clitocyboid species were clustered into a monophyletic lineage (BP = 100%, PP = 1.00) in the Hygrophoraceae according to the multi-gene phylogenetic analysis (Figs
Besides the molecular analyses, morphological data also support its separation from the relative genera. Spodocybe shares clitocyboid basidiomes, decurrent lamellae, inamyloid basidiospores and the presence of clamps with the other genera Ampulloclitocybe, Cuphophyllus and Cantharocybe. However, the genus Ampulloclitocybe, typified by A. clavipes, differs from Spodocybe in having medium-sized basidiomes and bidirectional lamellar trama (
For a long time, C. trulliformis has been placed in the genus Clitocybe based on the clitocyboid feature and habit since 1879 (
In previous studies, Cuphophyllus, Ampulloclitocybe and Cantharocybe were treated as basal in Hygrophoraceae (
The authors are very grateful to their colleagues at Kunming Institute of Botany, Chinese Academy of Sciences, including Drs. Xiang-Hua Wang, Jiao Qin, Bang Feng, Qi Zhao and Master students Hua Qu and Si-Peng Jian for collecting and providing specimens; and Drs. Gang Wu, Yang-Yang Cui, Qing Cai for providing help on morphological observation and phylogenetic analysis. This study was financed by Yunnan Ten-Thousand-Talents Plan – Yunling Scholar Project and Postdoctoral Directional Training Foundation of Yunnan Province.
Alignment of ITS-LSU-RPB2 dataset used in the three-gene phylogenetic analysis
Data type: fasta file
Explanation note: ITS: 1-1380, LSU: 1381–2356, RPB2: 2357–3135.
Alignment of ITS-LSU-RPB1-RPB2-TEF1-ATP6 dataset used in the six-gene phylogenetic analysis
Data type: fasta file
Explanation note: ITS: 1–1217, LSU: 1218–2158, RPB1: 2159–3358, RPB2: 3359–4089, TEF1: 4090–4967, ATP6: 4968–5405
Alignment of ITS dataset used in the single-gene phylogenetic analysis
Data type: fasta file