MycoKeys 90: | 19-1 45 (2022) er-reviewed open-access journal doi: 10.3897/mycokeys.90.78880 < MycoKkeys https://mycokeys.pensoft. net Launched to accelerate biodiversity research Taxonomic studies of bluish Mycena (Mycenaceae, Agaricales) with two new species from northern China Qin Na', Zewei Liu', Hui Zeng”?, Binrong Ke??, Zhizhong Song', Xianhao Cheng', Yupeng Ge' I Shandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, China 2 Institute of Edible Mushroom, Fujian Academy of Agricultural Sciences, Fuzhou 350011, China 3 National and Local Joint Engineering Research Center for Breeding and Cultivation of Featured Ed- ible Mushroom, Fuzhou 350011, China Corresponding author: Yupeng Ge (gaiyupeng@126.com) Academic editor: Thorsten Lumbsch | Received 5 December 2021 | Accepted 2 June 2022 | Published 17 June 2022 Citation: Na Q, Liu Z, Zeng H, Ke B, Song Z, Cheng X, Ge Y (2022) Taxonomic studies of bluish Mycena (Mycenaceae, Agaricales) with two new species from northern China. MycoKeys 90: 119-145. https://doi.org/10.3897/ mycokeys.90.78880 Abstract Bluish Mycena are rare, but constitute a taxonomically complex group. A total of eight bluish species in four sections have previously been reported from North America, Europe, Oceania and Asia. Two species with a blue pileus, collected in China during our taxonomic study of Mycena s.|., are described here as new to science: Mycena caeruleogrisea sp. nov. and M. caeruleomarginata sp. nov. Detailed descriptions, line drawings and a morphological comparison with closely-related species, especially herbarium specimens of M. subcaerulea from the USA, are provided. The results of Bayesian Inference and Maximum Likelihood phylogenetic analyses of a dataset of 96 nuclear rDNA ITS and 20 nLSU sequences of 43 Mycena species are also presented. The morphological data and the results of the phylogenetic analyses support the intro- duction of M. caeruleogrisea and M. caeruleomarginata as new species. A taxonomic key to bluish Mycena species of sections Amictae, Cyanocephalae, Sacchariferae and Viscipelles is provided. Keywords Mycenoid fungi, phylogeny, taxonomy, two new taxa Copyright Qin Na et al. 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. 120 Qin Na et al. / MycoKeys 90: 119-145 (2022) Introduction Mycena (Pers.) Roussel, with almost 600 species distributed worldwide, is one of the largest genera in Agaricales (He et al. 2019). Maas Geesteranus (1980, 1992a, 1992b) proposed an infrageneric classification of Mycena, based on a combination of mac- roscopic and microscopic features. In this classification, the species are defined mac- roscopically based on basidiomata colour (pileus, stipe and lamellae face and edge). Within Mycena, species of sect. Adonideae (Fr.) Quél., now treated as Atheniella Red- head, Moncalvo, Vilgalys, Desjardin & B.A. Perry, sect. Aciculae Kiithner ex Singer and sect. Oregonenses Maas Geest., are well characterised by their bright colours, such as pink, red, white or yellow (Maas Geesteranus 1980). Members of sect. Calodontes (Fr. ex Berk.) Quél. are prominently violet and dark colours can also be observed in sect. Rubromarginatae Singer ex Maas Geest. (Robich 2003, 2016; Aravindakshan and Manimohan 2015; Aronsen and Lessge 2016). In addition, the microscopic charac- ters are also considered to be very important in the infrageneric division of Mycena, containing basidiospores, cheilocystidia, pileipellis and stipitipellis (Maas Geesteranus 1992a, 1992b; Robich 2003, 2016; Aravindakshan and Manimohan 2015; Aronsen and Lessge 2016). No current published framework exists for the genus as a whole, however and the morphologically based classification of Maas Geesteranus (1992a, 1992b) has not been fully tested and validated. Some recent studies indicate that several Mycena sections, for example, sects. Amparoina T. Bau & Q. Na, Calodontes (Fr. Ex Berk.) Quél and Sacchariferae Kithner ex Singer, are apparently monophyletic, whereas others are not (Harder et al. 2010; Na and Bau 2019b). Several taxa, tradi- tionally assigned to Mycena, such as the Atheniella group, have been removed from the genus and others may need to be incorporated into genera, such as Cruentomycena R.H. Petersen, Kovalenko & O.V. Morozova, Favolaschia (Pat.) Pat., Hemimycena Singer, Panellus P. Karst., Resinomycena Redhead & Singer and Roridomyces Rexer (Redhead and Singer 1981; Rexer 1994; Antonin and Noordeloos 2004; Petersen et al. 2008; Redhead et al. 2012). Eight bluish Mycena in four sections have been documented so far. Amongst these species, five have been reported from the Northern Hemisphere: M. subcaerulea Sacc. in North America, M. amicta (Fries) Quél. and M. cyanorhiza Quél. in Europe, M. indigotica Wei & Kirschner and M. lazulina Har. Takah., Taneyama and Terashi- ma & Oba in Asia (Smith 1947; Maas Geesteranus 1980, 1992a, 1992b; Perry 2002; Robich 2003; Aronsen and Lzssge 2016; Terashima et al. 2016; Wei and Kirschner 2019; Perry et al. 2020). A bluish tint is often present on the pileus or stipe of these five species, four of them being classified into three sections: sect. Amictae Alexander H. Smith ex Maas Geesteranus, sect. Sacchariferae and sect. Viscipelles Kiihner, but M. indigotica has tubes confused with members of Favolaschia (Pat.) Pat. and not as- signed any section (Smith 1947; Maas Geesteranus 1980, 1992a, 1992b; Perry 2002; Robich 2003; Aronsen and Lzssge 2016; Terashima et al. 2016; Wei and Kirschner 2019; Perry et al. 2020). The three known bluish Mycena species from the Southern Two novo bluish Mycena species from northern China 121 Hemisphere are M. caesiocana Singer, M. cyanosyringea Singer and ™. interrupta (Berkeley) Sacc. (Singer 1969; Singer and Gomez 1982; Grgurinovic 2003). These species are distributed in Oceania and South America, Australia, Chile, Costa Rica, New Caledonia and New Zealand, where they usually grow on dead woods, decaying logs or tree stumps in deciduous forests of trees, such as Eucalyptus robusta Smith and Persea lingue (Ruiz & Pav.) Nees and develop basidiomata under high temperatures (Singer 1969; Singer and Gomez 1982; Grgurinovic 2003). The three allied species can be easily recognised: M. caesiocana and M. cyanosyringea are well characterised by the presence of a storm-grey pileus and extremely small basidiomata (pileus diameter and stipe length all less than 3 mm) and M. interrupta has a blue stipe base (Singer 1969; Singer and Gomez 1982; Grgurinovic 2003). In addition, M. cyanocephala Singer described from Chile, is considered to be synonymous with M. interrupta (Grgurinovic 2003). Although MZ. cyanorhiza, from the Northern Hemisphere, also has a blue stipe base similar to M. interrupta, but differs in pale brown to pale grey pi- leus and smaller basidiospores and cheilocystidia (Robich 2003; Aronsen and Lessge 2016; Perry et al. 2020). To date, fewer than 100 species of Mycena have been documented from China; amongst them, 14 new species have been described in recent years (He and Fang 1994; Guo et al. 1999; Shih et al. 2014; Li et al. 2015; Na and Bau 2018, 2019a, 2019b; Liu et al. 2021). During our investigations of mycenoid fungi in north-western China, we discovered two putative new taxa possessing a blue pileus with a greyish or brownish tint and a gelatinous pileipellis, clearly distinct from other species of Mycena, in the Liupan and Changbai Mountains. The results of our morphological observations and phylogenetic analyses support the introduction of these two new taxa. Materials and methods Morphology Macromorphological observations were made on fresh specimens in the field and from photographs, with colour terms and notation following Kornerup and Wan- scher (1978). Specimen pieces were mounted in 5% potassium hydroxide (KOH) and stained with Congo red when necessary. The prepared specimens were observed under a Lab Al microscope (Carl Zeiss AG, Jena, Germany) and photographed and recorded using the supplied ZEN 2.3 (blue edition) software (Carl Zeiss AG). Melzer’s Reagent was used to test whether spores and tissues were amyloid and dextrinoid (Horak 2005). The dimensions of basidiospores were recorded according to Ge et al. (2021), Liu et al. (2021) and Na et al. (2021, 2022). The examined collections have been deposited in the Fungarium of the Fujian Academy of Agricultural Sciences (FFAAS), China. In the subsequent taxonomic description, author abbreviations follow Index Fungorum (http://www.indexfungorum.org). 122 Qin Na et al. / MycoKeys 90: 119-145 (2022) Phylogenetic analysis Genomic DNAs of the putative new species were extracted from dried materials using a NuClean PlantGen DNA kit (Kangwei Century Biotechnology, Beijing, China). The internal transcribed spacer (ITS) region and the nuclear large subunit (nLSU) of nuclear ribosomal DNA were amplified using the PCR cycling protocol detailed in Ge et al. (2021) with primers ITS1/ITS4 and LROR/LR7, respectively (White et al. 1990; Hopple and Vilgalys 1999). In addition, no sequence information has been published for M. subcaerulea and only a few ITS sequences of M. cyanorhiza and M. amicta, which were found to be phylogenetically closely related to the new species, are avail- able in GenBank. For three M/. subcaerulea specimens, we tried to obtain our target sequences by using next-generation sequencing (NGS) technology and whole-genome sequencing of the specimens was performed on the Illumina sequencing platform (HiSeg PE150) with standard procedures. The 150 bp paired-end libraries were pre- pared to generate approximately 3G raw data. ITS (GenBank accessions KT900146, NR_154169) and nLSU (GenBank accessions MK629349 and NG_070530) were randomly selected for using as custom seed and custom label databases according to the instructions (https://github.com/Kinggerm/GetOrganelle/wiki/FAQ: How to as- semble custom loci?) of the software programme GetOrganelle (Jin et al. 2020). Fi- nally, two ITS sequences (GenBank accessions OL711671 and OL711672) and three nLSU sequences (OL711666, OL711667 and OL711668) were captured from next- generation sequencing data of three specimens (TENN-F-051121, TENN-F-057919 and CUP-A-015335) of M. subcaerulea and used for subsequent analysis. Thirteen sequences (six ITS and seven nLSU) newly generated in this study were deposited in GenBank. Additionally, a total of 103 ITS and nLSU sequences (including Xerom- phalina campanella |Batsch] Kithner & Maire, which is often chosen as an outgroup for Mycena) were retrieved from GenBank for use in the phylogenetic analysis. In- formation on all analysed sequences (116) is given in Table 1. Generated sequences and those retrieved from GenBank were aligned and manually checked using BioEdit 7.0.4.1 and Clustal X 1.81 (Thompson et al. 1997; Hall 1999), with gaps in the align- ment treated as missing data. The ITS and nLSU datasets were aligned separately. After estimating the optimal model of nucleotide evolution for the two partitions indepen- dently using Modeltest 3.7 (Posada and Crandall 1998), the two datasets were con- catenated. The combined aligned dataset, which was deposited in TreeBase (submis- sion ID 29069; study accession URL: http://purl.org/phylo/treebase/phylows/study/ TB2:S29069), was subjected to Bayesian Inference (BI) and Maximum Likelihood (ML) phylogenetic analyses. The BI analysis was performed in MrBayes 3.2.6 (Ron- quist and Huelsenbeck 2003). For the BI analysis, Markov Chain Monte Carlo chains were run for two million generations, with sampling carried out every 100" genera- tion until the critical value for the topological convergence diagnostic was less than 0.01 (Ronquist and Huelsenbeck 2003). The ML analysis, with a rapid bootstrapping algorithm involving 1,000 replicates, was performed in raxmlGUI 1.5b1 (Stamatakis et al. 2005). Two novo bluish Mycena species from northern China 125 Table |. Specimens along with GenBank accession numbers used in the phylogenetic analysis. Sequences newly generated in this study are indicated in bold. No. Species Voucher Origin ITS ID LSU ID References 1. = Mycena abramsii 231a Venice JF908400 — Unpublished 2. M. abramsii HMJAU 43282 China MH396626 MK629348 Unpublished 3. M. abramsii HMJAU 43468 China MH396627 — Unpublished 4. M. abramsii KA12-0434 Korea KR673481 = Kim et al. (2015) 5. M. adscendens Aronsen 120803 Norway KT900140 — Aronsen and Larsson (2015) 6. M. adscendens Orstadius329-05 Norway KT900141 — Aronsen and Larsson (2015) 7. M. adscendens Aronsen061119 Norway KT900142 — Aronsen and Larsson (2015) 8. M. adscendens Aronsen 120826 Norway KT900143 — Aronsen and Larsson (2015) 9. M. albiceps MGW 1504 USA KY744173, =MF797661 Unpublished 10. M. albiceps SAT 1518708 USA KY777372 |= MF797659 Unpublished 11. M. alnetorum CM14-RG2 USA KU295552 — Unpublished 12. M. amicta 189f Italy JF908394 — Osmundson et al. (2013) 13. M. amicta 4745-HRL 1312 Canada KJ705188 — Unpublished 14. M. amicta CBS 352.50 France MH856655 — Vu et al. (2019) 15. M. amicta CBS 254.53 France MH857183 — Vu et al. (2019) 16. M. amicta CBS 257.53 France MH857184 MH868722 Vu et al. (2019) 17. M. amicta H6036851 Finland = MW/540687 — Unpublished 18. M. arcangeliana 252b Italy JF908401 — Osmundson et al. (2013) 19. M. arcangeliana 252f Italy JF908402 — Osmundson et al. (2013) 20. MM. caeruleogrisea FFAAS 0001 Holotype China MW051896 OL711662 This study 21. M. caeruleogrisea FFAAS 0002 China MW051897 OL711663 This study 22. M. caeruleomarginata FFAAS 0357 Holotype China OL711669 OL711664 This study 23. M. caeruleomarginata FFAAS 0358 China OL711670 OL711665 This study 24. M. chlorophos ACL257 Malaysia KJ206983 — Chew et al. (2015) 25. M. chlorophos ACL271 Malaysia KJ206986 — Chew et al. (2015) 26. W&M. cinerella Aronsen051014 Norway KT900146 — Aronsen and Larsson (2015) 27. M. cinerella 173 Russia MF926553 — Malysheva et al. (2017) 28. M. citrinomarginata 317h Italy JF908416 — Osmundson et al. (2013) 29. M. citrinomarginata AD4TN Tunisia KU973883 — Unpublished 30. M. clavicularis 615i Italy JF908466 — Osmundson et al. (2013) 31. M. clavicularis 615b Italy JF908467 — Osmundson et al. (2013) 32. M. cyanorhiza 120b Italy JF908385 — Osmundson et al. (2013) 33. M. deeptha DM334g India JX481737 — Aravindakshan et al. (2012) 34. M. diosma KA13-1230 Korea KR673698 — Kim et al. (2015) 35. M. diosma 320f Italy JF908417 — Osmundson et al. (2013) 36. M. entolomoides HMJAU 43048 China MG654736 — Na and Bau (2018) 37. M. entolomoides HMJAU 43052 China MG654737. =MK722348 Na and Bau (2018) 38. M. entolomoides HMJAU 43126 China MG654738 MK722349 Na and Bau (2018) 39. M. filopes 3782 Canada KJ705175 = Unpublished 40. M. filopes KA12-1699 Korea KR673631 — Kim et al. (2015) 41. M. filopes 287£ Italy JF908410 — Osmundson et al. (2013) 42. M. galericulata DM136-40516 USA OM212953 — Unpublished 43. WM. galericulata LXL71 China MZ669083 = Unpublished 44. M. galericulata F26441 USA MZ317346 — Unpublished 45. M. galericulata EP.19-A1625 Greece MT458520 — Unpublished 46. M. galericulata 50 Norway =MW576935 — Unpublished 47. M. galericulata TFB14649 USA MN088382 — Unpublished 48. M. illuminans ACL161 Malaysia KJ206975 — Chew et al. (2015) 49. M. illuminans ACL175 Malaysia KJ206976 — Chew et al. (2015) 50. MM. illuminans ACL212 Malaysia KJ206980 — Chew et al. (2015) 51. MM. leaiana 1028 Italy JF908376 — Osmundson et al. (2013) 52. M. leaiana CNH03 (TENN) USA MF686520 — Unpublished 53. M. meliigena 39 Italy JF908423 — Osmundson et al. (2013) 54. M. meliigena 39d Italy JF908429 — Osmundson et al. (2013) 55. M. metata 313b Italy JF908412 — Osmundson et al. (2013) 56. M. olivaceomarginata GG436-86 Svalbard = GU234119 — Geml et al. (2015) 57. M. olivaceomarginata CBS 228.47 France MH856228 MH867756 Vu et al. (2019) 124 Qin Na et al. / MycoKeys 90: 119-145 (2022) No. Species Voucher Origin ITS ID LSU ID References 58. MM. olivaceomarginata CBS 229.47 France MH856229 MH867757 Vu et al. (2019) 59. M. olivaceomarginata HK47-15 Norway MT153141 = Thoen et al. (2020) 60. M. pachyderma 979a Italy JF908491 — Osmundson et al. (2013) 61. M. pearsoniana FCME25817 USA JN182198 — Harder et al. (2012) 62. M. pearsoniana TENN61544 USA JN182199 — Harder et al. (2012) 63. M. pearsoniana TENNG1384 USA JN182200 — Harder et al. (2012) 64. M. pelianthina CBH164 Denmark = FN394548 — Unpublished 65. M. pelianthina 108b Italy JF908379 — Osmundson et al. (2013) 66. MM. pelianthina 108f Italy JF908380 — Osmundson et al. (2013) 67. M. plumbea JN198391 China JN198391 — Wu et al. (2013) 68. M. plumbea 420526MF0010 China MG719769 a Unpublished 69. M. polygramma 439b Italy JF908433 — Osmundson et al. (2013) 70. M. polygramma 439f Italy JF908434 — Osmundson et al. (2013) 71. M. pura TENNG5043 USA JN182202 — Harder et al. (2012) 72. WM. pura f alba CBH410 USA FN394595 _ Unpublished 73. M. purpureofusca SL09-06 Canada HQ604766 — Unpublished 74. M. purpureofusca G. Alfredsen Norway JQ358809 — Unpublished 75. M. rosea 938a Italy JF908488 — Osmundson et al. (2013) 76. &M. rosea Champ-21 Spain KX449424 — Pérez-Izquierdo et al. (2017) 77. M. rubromarginata 407q Italy JF908430 _- Osmundson et al. (2013) 78. M. rubromarginata TL-12780 USA KX513845 = KX513849 Perry and Desjardin (2016) 79. M. seminau ACL136 Malaysia KP537250 KJ206952 Chew et al. (2015) 80. M. seminau ACL308 Malaysia KP537252 KJ206964 Chew et al. (2015) 81. M. seynesii 711 Italy JF908469 — Osmundson et al. (2013) 82. M. seynesii 71h Italy JF908470 — Osmundson et al. (2013) 83. M. silvae-nigrae 515 Italy JF908452 — Osmundson et al. (2013) 84. M. silvae-nigrae CC 13-12 USA KF359604 — Baird et al. (2014) 85. M. stylobates 455 Italy JF908439 — Osmundson et al. (2013) 86. M. subcaerulea TENN-F-051121 USA OL711671 OL711666 This study 87. M. subcaerulea TENN-F-057919 USA OL711672 OL711667 This study 88. M. subcaerulea CUP-A-015335 USA — OL711668 This study 89. M. supina 128a Italy JF908388 — Osmundson et al. (2013) 90. MM. tenax p187i USA EU669224 — Unpublished 91. M. tenax OSC 113746 USA EU846251 — Unpublished 92. M. viridimarginata 104h Italy JF908378 — Osmundson et al. (2013) 93. M. vulgaris 447h Italy JF908435 — Osmundson et al. (2013) 94. M. vulgaris 3781 Canada KJ705177 — Unpublished 95. M. zephirus KA13-1265 Korea KR673722 — Kim et al. (2015) 96. Xeromphalina campanella TFB14487 USA KP835678 KM011910 Aldrovandi et al. (2015) 97. X. campanella TFB7283A USA KM024575 KM024671 Aldrovandi et al. (2015) Results Phylogenetic analysis BI and ML reconstructions, based on the optimal evolutionary model selected for the ITS and nLSU partitions (GTR + I + G), recovered similar topologies. The BI tree was selected as a representative phylogeny (Fig. 1). In the tree shown in Fig. 1, which is based on 116 concatenated ITS+nLSU se- quences of 43 Mycena species and the new taxa, the two samples of M. caeruleogrisea and the two samples of MM. caeruleomarginata each form monophyletic lineages with high statistical support (MZ. caeruleogrisea, ML bootstrap support [BS] = 100, Bayes- ian posterior probability [BPP] = 1.00; MZ. caeruleomarginata, BS = 100, BPP = 1.00). According to the tree topology, MZ. subcaerulea is the species most closely related to M. caeruleogrisea and M. caeruleomarginata, consistent with morphology and clusters Two novo bluish Mycena species from northern China 125 --/-- — Mycena diosma KA 13-1230 aus se diosma 320 eee pura TENN65043 96/T.00 ena pene f. a na [08D --/-4 100/1.00 cena pel lanthiut --/).98 x reens peltanthina CBH164 a ( a Aap pearsoniana F ene 17 LO0/1.00} | cena pearsoniana CBH16 saith Sa TENN61384 cena rosea }O0/1. fycena rosea © ‘Mycena sen Cae 4A Oe Mycena masta dur Gr dati adscendens Aronsen1]120826 100/1.00f Mycena adscendens Aronsen061119 --/0.96) cena adscendens Orstadius329-05 : i eng 4 adsc lens Aronsen120803 aaa Mycona F pe ae ACL161 ao I 87/1.00 100/1.00) cena HJuminans ACL 212 sii uns cena Ms ATS —/1,00 ycena chloro pphros ACT Arba 79/0.97 Mycena coqqorhici of be g = naaicae cena pachyderma 979a at sitar a ieta Wie HRL 1312 a cee amicta CBS 352.50 83/1.00 100/1.00] cone palin oe CBS ah 23 ha Meare ame SR eRst sso, LUDA Mycena cacruleogrisea FFAAS0001@ Holotype "Mycena caepu leogrisea FAA 31933 e 100/1,00 cena sup caerulea 5@ 83/1.00 Bercena SU peaerutea AEN F- S112, jr : ce Supca 100/1.00 -/0.95L¢ Mycena eure en LENNsE ie i : Holotype 100/1.00 gruleomars inata FFAA 100/1.00 Mycena albigeps cv 808 & cena a 96/1.00 Mes na cinereffa je eae cinereli 7 nsen051014 Wycon eaiana | ~-/-s 100/1 ‘60 28 18 Mycena leaiana CNH03 (TENN) cena tenax = lopt.o0 cena tenax Bsc 113746 OOD Mpcena val vulgaris 447h td |p Mycerta entolontt lp aS SRL TAU 43048 tL ycena entol. omar es J a3 146 a entolomoides , 430 100/1.00— Mycena clayicularis 1 1 laris 6|5b 100/1 Mie pagcer ie sz 252b 96/1.00 Mycena arcangeligna 252f erat Gicead is I, cena filopes 49/T.00 st ma el ena BE 99/1.00 ry cena Pe rare, 439b --/0.99 Seen e pO ya cena Gt stvccenlerne 515 00/1.00 cena sivas a fava CC 13- 213 é uifl ae cena aler| cul fata = a1 uae rea alertcu Hiern 1934 --/- ; fycena’ galeric 100/1.00 cena gilericu tas fycena BeleriCulae ee eiG cen. iculata 190/1.00- Mycena pabromase peena gle --/. [ “Mycena rub gd ed TL-12780 100/1,00 Ld nid ii aah A tandiclags citri omargi 100/1.00}--— “Mycena olivaceo inanimate tevez 6-86 | Mycéna o fivaceomarginata --/=| Mycena olivaceomarginata EBS. 356° 47 ~-/1. 09} lycena olivaceoma eginata | CBS 228.47 98/0,99 Mycena purp pureofusca eens Puy ureofusca G., A a --/1,00 --/0.99 p 2? Cend Sé eyes lige Be afa.d ie he vi ae bea SE 09 0 ena A -/- Cc ra aN a eae a rams AU 43282 a cena ra hee HMJAU 43486 Mycena alnetorum CM14-RG2 100/1.00-— Xeromphalina Campanella TFB14487 ‘Xeromphalina campanella TFB7283A on Figure |. Phylogenetic tree inferred from partial ITS+nLSU sequence data by Bayesian inference and maxi- mum likelihood. The tree is rooted with Xeromphalina campanella. Maximum likelihood support values (BS) > 75 and Bayesian posterior probabilities (BPP) > 0.95 are indicated above or below branches (BS/BPP). Red dots indicate two new species, while green dots indicate Mycena subcaerulea specimens from’ TENN and CUP. 126 Qin Na et al. / MycoKeys 90: 119-145 (2022) with the latter two species with high statistical support (BS = 100, BPP = 1.00). The M. subcaerulea clade comprises three samples: CUP-A-015335 (originally identified as M. cyanothrix G.F. Atk.), TENN-F-051121 and TENN-F-057919 (BS = 100, BPP = 1.00). By its morphological features and phylogenetic placement, sample CUP- A-015335 should be re-assigned to M. subcaerulea. The clade comprising M. subcaeru- lea and the two new taxa are sister to M. amicta, with the clade constituted by these four species in turn sister to VM. cyanorhiza. Despite the close relationships, the two new species are strongly supported as distinct from M. amicta and M. cyanorhiza (Fig. 1). It is noteworthy that the six samples of M/. amicta from Europe and North America cluster together with strong support (BS = 100, BPP = 1.00), but the Canadian materi- al (voucher no. 189f) seems to be closer to the Italian sample (voucher no. 4745-HRL 1312) than to the specimens from France and Finland. In addition, MM. pachyderma Kiihner, a non-bluish species in sect. Viscipelles, is a sister taxon (BS = 79, BPP = 0.97) to M. cyanorhiza in the same section. Taxonomy In addition to morphological studies of the new taxa collected in China, morphologi- cal observations were made on 17 bluish specimens of Mycena loaned from fungal her- baria in the USA, namely, four specimens from the University of Tennessee (TENN) and 13 specimens from University of Cornell (CUP). Our morphological observations using a light microscope confirmed the identity of 12 specimens as M. subcaerulea: TENN-F-014183, TENN-F-051121, TENN-F-052683, TENN-F-057919, CUP-A-002382, CUP-A-009686, CUP-A-014679, CUP-A-015138, CUP-A-015335, CUP-A-022677, CUP-A-023037 and CUP-A-023304. Another specimen, CUP-A-021234, previously identified as M. iris, was well characterised as M. amicta, based on its elongated ellipsoid basidiospores and clavate cheilocystidia with a rounded apex. As already noted by Smith (1947), the basidiomata of CUP-A-018443, CUP-A-022667, CUP-A-051322 and CUP-A-051323 were too small to be examined. Mycena caeruleogrisea Q. Na, Y.P. Ge & H. Zeng, sp. nov. MycoBank No: MB837656 Figs 2, 3, 4 Diagnosis. This species is characterised by blue pileus, turning bluish-grey with age, pileus covered by a separable, gelatinous pellicle, stipe pruinose and with a blue base and stipe basal disc and acanathocysts of pileipellis absent. Mycena subcaerulea differs from M. caeruleogrisea by a greenish-blue to greyish-brown pileus that turns yellow and remains blue at the centre and margin with age, a greenish-blue to brownish-blue stipe and smaller, globose to subglobose basidiospores. Two novo bluish Mycena species from northern China lens J te _ ; Ag > > “i Figure 2. Fresh basidiomata of Mycena caeruleogrisea a—d M. caeruleogrisea (FFAAS 0001, holotype) e, f M. caeruleogrisea (FFAAS 0002) g-i entirely pruinose stipe j, k bluish base. Scale bars: 10 mm (a-=i); 2 mm (j-k). Photographs by Yupeng Ge (a, b) and Qin Na (c=k). 128 Qin Na et al. / MycoKeys 90: 119-145 (2022) : a a 4 * ot 2 @ >. ty whe AR Sy PEAS Se eT = eee" pe COS ae SLES 2IGE ‘ Figure 3. Microscopic features of Mycena caeruleogrisea (FFAAS 0001, holotype) a-d basidiospores e-g cheilocystidia h pileipellis i stipitipellis and caulocystidia. Scale bars: 10 um (a=i). Structures were stained with Congo Red medium before photographing. Two novo bluish Mycena species from northern China 129 Figure 4. Morphological features of Mycena caeruleogrisea (FFAAS 0001, holotype) a basidiomata b basidia c basidiospores d cheilocystidia e stipitipellis and caulocystidia f pileipellis. Scale bars: 10 mm (a); 10 um (b-f). Drawings by Qin Na and Yupeng Ge. 130 Qin Na et al. / MycoKeys 90: 119-145 (2022) Holotype. Cutna. Ningxia Hui Autonomous Region: Liangdianxia, Liupan Mountains National Forest Park, Jingyuan County, Guyuan City, 35°21'74"N, 106°18'37"E, 19 July 2020, Qin Na, Yupeng Ge, Hui Zeng, Junqing Yan and Zewei Liu, FEAAS 0001 (collection number MY0164). Etymology. Refers to the pileus colour: blue when young, becoming bluish-grey with age. Description. Pileus 12—25 mm in diameter, hemispherical when young, conical, obtusely conical, campanulate with age, shallowly sulcate, translucently striate, almost smooth when young, becoming slightly brownish scaly at the centre, pruinose, with a glabrescent margin, dull blue (23D5) at the centre, margin pallescent to pastel blue (23A4), turning bluish-grey (23D2-—23D3), a bit sticky, covered by a separable, gelati- nous pellicle. Context white, thin, fragile. Lamellae 16-28 reaching the stem, adnate to slightly adnexed with a short tooth, narrowly spaced, white, with intervenose veins, edges concolorous with the face. Stipe 48-76 x 1.5—2.0 mm, equal or slightly broad- ened below, hollow, fragile, entirely pruinose (Fig. 2g—i), white, base greyish-blue (23B5) (Fig. 2j, k), covered with white fibrils, a basal disc absent. Odour and taste indistinctive. Basidiospores [60/3/2] (8.8) 9.3-10.4-11.3 (11.8) x (5.5) 5.7-6.5-6.9 (7.3) um [Q = 1.57-1.68, Q = 1.60 + 0.072] [holotype [40/2/1] (9.1) 9.4-10.3-11.3 (11.6) x (5.6) 6.0-6.5-6.9 (7.2) um, Q = 1.55-1.63, Q = 1.59 = 0.049], ellipsoid, hyaline in 5% KOH, smooth, guttulate, thin-walled, amyloid. Basidia 22-29 x 7-9 um, 4- or 2-spored, clavate. Cheilocystidia 40-62 x 4—6 um, clustered, abundant, elongated clavate or cylindrical, apically broadly rounded, thin-walled, hyaline, forming a sterile lamellae edge. Pleurocystidia absent. Pileipellis an ixocutis with 1-4 um wide hyphae, smooth or sparsely coated with simple cylindrical excrescences or inflated cells, 3-11 x 1—2 um, embedded in gelatinous matter; acanathocysts absent. Hypodermium undif- ferentiated. Hyphae of the stipitipellis 3-8 um in diameter, smooth, hyaline; caulocys- tidia 38-69 x 6-8 um, long cylindrical, smooth, transparent. All tissues dextrinoid. Clamps present in all tissues. Habit and habitat. Scattered on humus and fallen leaves in mixed forests of Acer, Populus, Pinus and Quercus. Known distribution. Ningxia Hui Autonomous Region, China. Additional material examined. Ningxia Hui Autonomous Region: Xiaonan- chuan, Jingyuan County, Guyuan City, 20 July 2020, Qin Na, Yupeng Ge, Hui Zeng, Junging Yan and Zewei Liu, FFAAS 0002 (collection number MY0169). Remarks. ‘The original description of M. subcaerulea Sacc. was as follows: “Pileo ten- uissimo, campanulato v. convexo, striato, glabro, pallide ceruleo-viridi; stipite tenui, equali, roseo-albo, subtiliter pruinoso; lamellis angustis, confertis, antice attenuatis, candidis; sporis subglobosis. 4 yw. d. Hab. In trunco fagineo in montibus Adirondack Amer. bor. — Caspitosa, 5 cm. alta; pileus 8-13 mm. latus. Discus margine saturatius coloratus atque pileus cuticula secernibili obtectus.” (Saccardo 1887). This North American species, which also has bluish basidiomata, is the taxon most closely resembling MM. caeruleogrisea in both macro- and microscopic features; however, M/. subcaerulea differs by a greenish-blue to greyish-brown pileus that turns yellow and remains blue at the centre and margin with age, a greenish- Two novo bluish Mycena species from northern China 1 Boa blue to brownish-blue stipe and smaller, globose to subglobose, basidiospores [6-8 x 6—7(8) um] (Saccardo 1887; Smith 1947). In addition, M. subcaerulea was found solitary, scattered or gregarious on debris, decaying wood or bark around the bases of living trees, especially of oak, but also occurring quite frequently on decaying wood of basswood, elm, beech and other hardwoods (Smith 1947). The following microscopic characteris- tics of M. subcaerulea were also observed on the 11 CUP-A and TENN-F specimens in our study: basidiospores 5.6—-8.3 x 5.3—-7.9 um, globose to subglobose; basidia 19-24 x 6-8 um, clavate, 4-spored; cheilocystidia 36-55 x 3—6 um; pileipellis hyphae 2-4 um wide, coated with cylindrical excrescences or inflated cells, 1.1-14.9 x 0.7—1.4 um, em- bedded in gelatinous matter; hyphae of the stipitipellis 4-10 um in diameter; caulocyst- idia 42-70 x 4-10 um, fusiform or cylindrical, smooth; clamps present (Figs 5, 6). In M. cyanorhiza, the base of the stipe can be strikingly sky blue, but it has a pale brown, grey to almost white pileus, a stipe base arising from a patch of fine fibrils, clavate to obpyriform cheilocystidia with finger-like excrescences and basidiospores that are elongated ellipsoid (Q = 1.6-2.2); these features all contrast with those of the new species (Aronsen and Lessoe 2016; Perry et al. 2020) (Table 2). In addition, MZ. amicta can be easily mistaken for M. caeruleogrisea, as it sometimes also has a bluish pileus when mature and similarly- shaped basidiospores, cheilocystidia and caulocystidia, but /Z. amicta can be distinguished from the latter species in having a pileus generally more brownish with a bluish tinge more or less present, an indistinct to raphanoid odour, a greyish-brown stipe that has a blue to blue-green base and is covered with a dense, fairly coarse, white pubescence and smaller cheilocystidia (16-45 x 3.5—7 pm); in addition, M. amicta is restricted to growth on wood and woody debris (Robich 2003; Aronsen and Lessoe 2016) (Table 2). Mycena interrupta, which is well characterised by its acid blue to dull blue pileus and translu- cent stipe, is easily distinguished from M. caeruleogrisea by having smaller basidiomata, free lamellae, a white hirsute basal disc with blue margins on the stipe, broadly ellipsoid to subglobose spores and cheilocystidia covered with coarse excrescences (Grgurinovic 2003) (Table 2). Mycena lazulina, a new taxon reported from south-western Japan, pos- sesses a blue stipe and cheilocystidia with numerous excrescences, which can be used to differentiate it from M. caeruleogrisea (Terashima et al. 2016). Another recently-described species of Mycena from Taiwan, M. indigotica, has blue basidiomata; however, the cap has tubes similar to Boletus and possesses globose basidiospores (Wei and Kirschner 2019). Mycena caeruleomarginata Q. Na & Y.P. Ge, sp. nov. MycoBank No: MB842100 Figs 7, 8, 9 Diagnosis. This species is characterised by dark brown pileus with a blue margin and the stipe densely pruinose, entirely covered with puberulous hairs and stipe basal disc and acanathocysts of pileipellis absent. Mycena subcaerulea differs from M. caeruleogrisea in having a pileus that is distinctly greyish-brown with a blue centre and margin, turning yellow with age, a stipe tinged greenish-blue and globose to subglobose basidiospores. Qin Na et al. / MycoKeys 90: 119-145 (2022) 4 b S RE e f | h | | Eres, J Figure 5. Microscopic features of Mycena subcaeruleaa,b basidiospores (TENN-F-0579 19) € basidiospores (CUP-A-002382) d basidiospores (CUP-A-015138) e=g cheilocystidia (TENN-F-057919) h, i pileipel- lis (TENN-F-057919) j stipitipellis and caulocystidia (TENN-F-057919). Scale bars: 5 um (a=d); 10 um (e-j). Structures were stained with Congo Red medium before photographing. Two novo bluish Mycena species from northern China 1:33 Aas Figure 6. Morphological features of Mycena subcaerulea a basidiospores b basidia € stipitipellis and caulocystidia d cheilocystidia e pileipellis. Scale bars: 10 um (a—e). Drawings by Qin Na and Yupeng Ge. 134 Qin Na etal. / MycoKeys 90: 119-145 (2022) Holotype. Cuina. Jilin Province: Chixi Protection Station, Erdaobaihe Town, Antu County, Yanbian Korean Autonomous Prefecture, 42°46'35"N, 128°15'04"E, 3 July 2021, Qin Na, Yupeng Ge, Binrong Ke and Chi Yang, FFAAS 0357 (collection number MY0337). Etymology. Refers to the pileus, which is blue at the margin. Description. Pileus 3.5-13 mm in diameter, parabolic, obtusely conical when young, hemispherical, campanulate with age, with an umbo at the centre, shallowly sulcate, translucently striate, smooth, slightly gelatinous, the margin infrequently out of flatness, dark brown (6F5—GF7), disc brown (6E6—GE7), becoming greyish-blue (23B5) to blue (23B7) towards the margin (Fig. 7c, d, i), margin grey (23B1) (Fig. 7c, d, i), cov- ered by a separable, viscid pellicle. Context white, fragile, thin. Lamellae 14—25 reaching the stem, adnate to slightly adnexed with a short tooth, white, inconspicuously inter- venose, edges concolorous with the face. Stipe 32-46 x 1.0—2.0 mm, equal, base some- times slightly broadened, fragile, hollow, pruinose, entirely puberulous when young (Fig. 7h), becoming sparingly so, especially in the middle part, when old (Fig. 7e), egreyish-brown (5E3) to brown (5E4), base with an greyish-blue (23B5) tinge (Fig. 7a, f), sparsely covered with white fibrils, a basal disc absent. Odour and taste indistinctive. Basidiospores [60/3/2] (6.2) 6.4-7.1-7.7 (7.9) x (4.4) 4.7-5.2-5.8 (6.0) um [Q = 1.23-1.54, Q = 1.36 = 0.071] [holotype [40/2/1] (6.4) 6.6-7.2-7.7 (7.8) x (4.7) 4,9-5.2—5.3 (5.7) um, Q= 1.26-1.53, Q = 1.39 + 0.070], broadly ellipsoid to ellipsoid, hyaline in 5% KOH, guttulate, smooth, thin-walled, amyloid. Basidia 26-35 x 6-12 um, 4- or 2-spored, clavate. Cheilocystidia 32-48 x 4-6 um, abundant, clustered, cylindri- cal or elongated clavate, apically broadly rounded, thin-walled, hyaline, forming a sterile lamellae edge. Pleurocystidia absent. Pileipellis an ixocutis with 2-4 um wide hyphae, simple, cylindrical excrescences, 2—6 x 1-2 um, embedded in gelatinous matter; acana- thocysts absent. Hypodermium undifferentiated. Hyphae of the stipitipellis 3-6 um in diameter, smooth, hyaline; caulocystidia smooth, transparent, of two shapes: (1) fusiform or cylindrical, 19-40 x 4-8 pm; (2) extremely long cylindrical, sometimes with a narrow apex, 115-178 x 5—9 um. All tissues dextrinoid. Clamps present in all tissues. Habit and habitat. Scattered on rotten wood in Picea, Pinus, Quercus, Robinia and Tilia mixed forests. Known distribution. Jilin Province, China. Additional material examined. Jilin Province: Hancongling, Erdaobaihe Town, Antu County, Yanbian Korean Autonomous Prefecture, 42°46'36"N, 128°15'04"E, 4 July 2021, Qin Na, Yupeng Ge, Binrong Ke and Chi Yang, FFAAS 0358 (collection number MY0343). Remarks. ‘The diagnostic features of M. caeruleomarginata can be used to distin- guish this new taxon from the closely-related bluish species M. subcaerulea, M. cyanorhi- za, M. amicta and M. interrupta (Table 2). Mycena subcaerulea, the species most similar to M. caeruleomarginata, differs in having a pileus that is distinctly greyish-brown with a blue centre and margin, turning yellow with age, a stipe tinged greenish-blue and glo- bose to subglobose basidiospores (Q = 1.01—1.14) according to the original description Two novo bluish Mycena species from northern China WZ YZ x A ‘eis QO, [— "Cia 7 type) g-j Z. caeruleomarginata (FFAAS 0358) a, f stipe with a bluish base ¢, d, i pileus with blue mar- gin e, h densely white, pruinose to pubescent stipe. Scale bars: 10 mm (a, b, e, f, g, h); 5 mm (c, d); 2 mm (i,j). Photographs by Qin Na (a=f) and Yupeng Ge (g-j). 136 Qin Na et al. / MycoKeys 90: 119-145 (2022) b basidia c basidiospores d cheilocystidia e stipitipellis and caulocystidia f pileipellis. Scale bars: 10 mm (a); 10 pm (b-f). Drawings by Qin Na and Yupeng Ge. Two novo bluish Mycena species from northern China Figure 9. Microscopic features of Mycena caeruleomarginata (FFAAS 0357, holotype) ad basidiospores e-g cheilocystidia h-j pileipellis j stipitipellis and caulocystidia. Scale bars: 5 um (a=d); 10 um (e-j). Structures were stained with Congo Red medium before photographing. 119-145 (2022) Qin Na et al. / MycoKeys 90 138 *propAure ‘snqnorde onbryqo azo0ys quoururord ym ‘asogoysqns Ajarer peprosdyys Afpeosq HT = © “url (90 $= CS OL = ¥) 8'8-L'S x (LO + = CS ‘66 = X) 9 TI-¥'8 “(E/¥S) “SATIOUNISIP JOU INOPE, “OSEQ eng Tp pousney ® WO DUO SsIp asournsd ayy PIA UIMENSqNs 07 poypene Soy usaonysuen ‘aseq spremoy ATTeDadso asoutnid uaiyo ‘Arp 0} ystour qeoupuréo Buoy wu 7z 01 dq *propAure ‘padeys-did ‘g°] =~ ae?) TIT =O ‘UM cH x 6-69 ‘peprooar ou aise} ‘snonru Apurey se povsodas Jo suou Tous ‘spqy arya ‘Sunerpes ‘ouy jo ypred & wos 8uIsutds ‘(ysay ay} UT osfe) anyq Ays ‘oansiry aseq Sourym-ourpedy 01 Aaus aed aed oypprur oy) ur yososarqeys ‘snopniaqnd Ajemmua ‘qestrpuryéd “uur [—G'0 x (0Z-) 0€-¢ “propdure ‘podeys-did ‘g"] = aedy GI-GT = 0 ‘wm 9-Cy x L:01-G'Z | ‘esoqo]8qns 10 asoqoys “um (g) /-9 x g-9 ‘prouryder 02 OUNsSIpUT ‘anyq poureis aq Aeur ayersqns oy} "YstuMOIq PIpros aad ‘ong Ajamua Jo sureys ude13 Ayjeuy Jo ystderd 01 Surpry uoos sry ye -oN]q SUTOS TIT IO snoIO;OOUOD aaoge anyq Ystusers 07 Ysinyq uy 01 ‘BUNOO TYMUTOS sou) Ie aseq | SUTpey UOOs Inq IsIy ye aNTq UNTAOAUT ay} UN snosdxjOIA JO snossepy] WYSIYs & | ‘proraodur aseq “Waosarqe|s WeYyMIUTOS ym AyTeuorsvoo0 “xode oy ye Joped “eiprsAoojnes Jo SUNLOS asap & WHO} yeymouros ATTensn ‘umorq ystéoI3 [Ie 2240 quaosaqnd Ajaynurur so asournsd ‘souadsaqnd arya ‘asteoo AprTey Ajpsuap asiy ye ‘Onseya ‘snoursepnseo TepNgn) ‘iNs Jo snonxopF 939191 ‘genba Spry “urur (¢°Z) Z| “Suo] wo g—¢ pue esuop eB YA PetoAod Ajommua ‘TeorupuyfAd “ur 7—G'0) x 0/-OF *propAure ‘prosdiyja 01 prosdiype Appeosq “7G [-€7"T = 0 ‘wm (09) S-LY (VY) x (6'L) LL-¥'9 (79) *propAure ‘prosdyyp “Z-T-G T =O ‘unl (/°Z) €°Z-7'9 (0°9) x (STL) 9 TI-€'6 (0°6) SATOUNISIPUT dANOUNSIPUT “sTQy outa 1g ® YM paraaoo O8uN anyq ploe YIM aseq “UMOI 1ST] 01 UMOIq Ystmoyad 1.6) and lacks smooth cheilocystidia and caulocystidia (Aronsen and Lessge 2016; Perry et al. 2020). In addition, M/. amicta resembles M. caeruleomarginata in its bluish pileus, pruinose stipe and pileipellis embedded in a layer of gelatinous matter, but the former differs in having a pale grey-brown pileus that is sometimes ochraceous yellow and greenish when young and bluish when old, a rapha- noid odour and elongated ellipsoid basidiospores (7.5—10.7 x 4.5-6.0 um) (Robich 2003; Aronsen and Lzssge 2016). The Southern Hemisphere species M. interrupta is well characterised by its blue pileus at maturity, a translucent stipe with a basal disc and cheilocystidia with excrescences (Grgurinovic 2003). Furthermore, two new species with bluish basidiomata reported from East Asia, M. lazulina and M. indigotica, can be easily distinguished from the new species in their whitish pileus or tubes similar to Boletus; M. lazulina having cheilocystidia with numerous excrescences and M. indig- otica possesses globose basidiospores (Terashima et al. 2016; Wei and Kirschner 2019). Mycena caeruleogrisea and M. caeruleomarginata share the same bluish pileus and stipe base, smooth and cylindrical cheilocystidia and pileipellis embedded in a layer of ge- latinous matter. Mycena caeruleomarginata can be readily distinguished, however, based on the dark brown colour of the pileus with a blue margin, yellowish-brown to light brown stipe, broadly ellipsoid to ellipsoid spores and caulocystidia of two shapes. Key to seven bluish Mycena species of sections Amictae, Cyanocephalae, Sacchariferae, and Viscipelles 1 @heilocysviciastiomstmOOn bh, 1! 08k. tele leans ate sce e eeeke ashes seated etdegn eeu aden’ 2 ~ Cheilocystidiasimoath\ sect: 4A 7:76fte)* vel wnat needa vncev eee. eeeeeedeceedten ears 4 2 Acanthocysts préesent(sect.. Sacchariferde) tscccassccteestscueescnsvccntanes M. lazulina — A@ALIEMO CYSES-AD SCI 2.0. wasasea crtowsderunedey eatees peamteseeonats seat ha ponte sbemeatagebeweneuaee 3 3 Stipe with basal disc (sect. Cyanocephalae) ..icsescesesesseeseneee M. interrupta = Stipe without basal disc (sect. Viscipelles)....icicecessseeseeseerees M. cyanorhiza = Basidiospores Sub glODOSe ./-\.scta.pseosthosusennPetnurcasonnsan thas os M. subcaerulea - Basidiospores broadly ellipsoid to ellipsoid... ee eeeeeeeeeeseeeseeeeeeneeeeeeens > 5 Caulocystidia of two types: (1) fusiform or cylindrical, 19-40 x 4-8 um; (2) extremely long, cylindrical (length > 100 pm)........... M. caeruleomarginata - Caulocystidia of one type, fusiform, subcylindrical to cylindrical (length < OO eet, hes, 8 Be oa del tere en et aon elec, pee, error coe 6 6 Pileus pale grey-brown or pale sepia brown, sometimes with an olivaceous, greenish or bluish-green shade; margin often bluish-green or rarely dingy cit- RIT eMO. OCMLACEO Use Ve LOWE n, he enc erovevanre exeobeeede Gr ocvterse cect aewerm every s M. amicta — Pileus sky blue, greyish-blue with age; margin blue when young, turning blu- Iga Chey Wi CMEOLCS. as54s, Pirate oe. es sentences at aenns acackt M. caeruleogrisea Two novo bluish Mycena species from northern China 141 Discussion With their blue pileus and gelatinous pileipellis, the new taxa M. caeruleogrisea and M. caeruleomarginata are unique in China. Similar species described from North America and Europe, namely, M. subcaerulea, M. cyanorhiza and M. amicta, have blu- ish basidiomata as well, but with age, these species often change colours—to green, brown or yellow and the sizes and shapes of their basidiospores and cheilocystidia are also different (Saccardo 1887; Smith 1947; Maas Geesteranus 1980, 1992a, 1992b; Grgurinovic 2003; Robich 2003; Aronsen and Lzssge 2016) (Table 2). Mycena inter- rupta, described from the Southern Hemisphere, can be distinguished from the two newly-described species, based on both habitat and morphology (Grgurinovic 2003). Mycena lazulina (sect. Sacchariferae), which has a white pileus, blue stipe base, acantho- cysts and a non-gelatinised pileipellis, seems to be the most distinct bluish species and is not included in Table 2 (Terashima et al. 2016). According to taxonomic research based on morphology and phylogeny, our newly-described species are more similar to M. subcaerulea and M. amicta and should, thus, be classified into sect. Amictae. Although pileus colour has been used as a basis for sectional division in Mycena, this character does not seem to be satisfactory for species identification, especially within the same section (Smith 1947; Maas Geesteranus 1980, 1992a, 1992b; Grgurinovic 2003; Robich 2003; Aronsen and Lzssge 2016). In sect. Viscipelles, for example, M. cy- anorhiza can be distinctly characterised by the presence of a sky blue stipe, but M. ulmi B.A. Perry & H.W. Keller, M. pachyderma and M. pseudocyanorrhiza Robich do not exhibit any bluish tint (Robich 2003; Aronsen and Lessge 2016; Perry et al. 2020). A combination of macroscopic and microscopic features, such as the colour of basidi- omata and the shapes and sizes of spores, cheilocystidia, pileipellis, caulocystidia and dextrinoid tissues, is, thus, generally regarded as more important for the identification of Mycena taxa. Acknowledgements This study was supported by the Natural Science Foundation of Shandong Province (grant No. ZR2020QC001), the National Natural Science Foundation of China (grant No. 3190012), the Natural Science Foundation of Shandong Province (grant No. ZR- 2019PC028) and the Innovation Team of Shandong Agricultural Industry Technology System (grant No. 26, SDAIT-07-03). We are extremely grateful for the assistance of the Herbaria of Cornell University and the University of Tennessee and especially appreciate the kind help of Collections Manager Margaret Oliver, Dr P. Brandon Ma- theny and Curator Teresa Iturriaga with specimen loan requests. We sincerely thank Dr Jianwei Liu for help with phylogenetic analyses and Dr Xiaojuan Deng, Mr Bai Wang, Mr Chi Yang and colleagues of the Guyuan Branch Institute of Ningxia Academy of Agriculture and Forestry Sciences for help with fieldwork. We also thank the editors and reviewers for their corrections and suggestions to improve our work. 142 Qin Na et al. / MycoKeys 90: 119-145 (2022) References Aldrovandi MS, Johnson JE, O’ Meara BC, Petersen RH, Hughes KW (2015) The Xeromphalina campanellalkauffmanii complex: Species delineation and biogeographical patterns of speciation. Mycologia 107(6): 1270-1284. https://doi.org/10.3852/15-087 Antonin V, Noordeloos ME (2004) A monograph of the genera Hemimycena, Delicatula, Fayodia, Gamundia, Myxomphalia, Resinomycena, Rickenella, and Xeromphalina (Tribus Mycenae sensu Singer, Mycena excluded) in Europe. IHW-Verlag, 280 pp. Aravindakshan DM, Manimohan P (2015) Mycenas of Kerala. SporePrint Books, Calicut, India, 213 pp. Aravindakshan DM, Kumar TK, Manimohan P (2012) A new bioluminescent species of Mycena sect. Exornatae from Kerala State, India. Mycosphere: Journal of Fungal Biology 3(5): 556-561. https://doi.org/10.5943/mycosphere/3/5/4 Aronsen A, Lessge T (2016) The Genus Mycena s.l. Fungi of Northern Europe Vol. 5. Narayana Press, Gylling, Denmark, 373 pp. Aronsen A, Larsson E (2015) Studier i slaktet Mycena (hattor). Svensk Mykologisk Tidskrift 36(3): 23-29. Baird R, Stokes CE, Wood-Jones A, Watson C, Alexander M, Taylor G, Johnson K, Threadgill P, Diehl S (2014) A molecular clone and culture inventory of the root fungal community associated with Eastern Hemlock in Great Smoky Mountains National Park. Southeastern Naturalist (Steuben, ME) 13(6): 219-237. https://doi.org/10.1656/058.013.s601 Chew A, Desjardin DE, Tan YS, Musa MY, Sabaratnam V (2015) Bioluminescent fungi from peninsular Malaysia—A taxonomic and phylogenetic overview. Fungal Diversity 70(1): 149-187. https://doi.org/10.1007/s13225-014-0302-9 Ge YP, Liu ZW, Zeng H, Cheng XH, Na Q (2021) Updated description of Atheniella (Mycenaceae, Agaricales), including three new species with brightly coloured pilei from Yunnan Province, southwest China. MycoKeys 81: 139-164. https://doi.org/10.3897/ mycokeys.8 1.67773 Geml J, Timling I, Robinson CH, Lennon N, Nusbaum HC, Brochmann C, Noordeloos MC, Taylor DL (2015) An arctic community of symbiotic fungi assembled by long-distance dispersers: Phylogenetic diversity of ectomycorrhizal basidiomycetes in Svalbard based on soil and sporocarp DNA. Journal of Biogeography 39(1): 74-88. https://doi.org/10.1111/ j.1365-2699.2011.02588.x Grgurinovic CA (2003) The genus Mycena in south-eastern Australia. Fungal Diversity Press, 327 pp: Guo SX, Fan L, Cao WQ, Chen XM (1999) Mycena dendrobii, a new mycorrhizal fungus. Junwu Xuebao 18: 141-144. https://doi.org/10.13346/j.mycosystema.1999.02.007 [in Chinese] Hall TA (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98. https://doi. org/10.1021/bk-1999-0734.ch008 Harder CB, Lessoe T, Kjoller R, Froslev TG (2010) A comparison between ITS phylogenetic relationships and morphological species recognition within Mycena sect. Calodontes in Northern Europe. Mycological Progress 9(3): 395-405. https://doi.org/10.1007/s11557- 009-0648-7 Two novo bluish Mycena species from northern China 143 Harder CB, Lodge DJ, Petersen RH, Hughes KW, Blanco JC, Froslev TG, Lessoe T (2012) Amyloidity is not diagnostic for species in the Mycena pearsoniana complex (Mycena section Calodontes). Mycological Progress 2012(11): 725-732. https://doi.org/10.1007/ s11557-011-0782-x He X, Fang X (1994) Three new species of the genus Mycena. Junwu Xuebao 13(2): 92-98. https://doi.org/10.13346/j.mycosystema.1994.02.003 [in Chinese] He MQ, Zhao RL, Hyde KD, Begerow D, Kemler M, Yurkov A, McKenzie EHC, Raspé O, Kakishima M, Sanchez-Ramirez S, Vellinga EC, Halling R, Papp V, Zmitrovich IV, Buyck B, Ertz D, Wijayawardene NN, Cui B-K, Schoutteten N, Liu X-Z, Li T-H, Yao Y-J, Zhu X-Y, Liu A-Q, Li G-J, Zhang M-Z, Ling Z-L, Cao B, Antonin V, Boekhout T, da Silva BDB, De Crop E, Decock C, Dima B, Dutta AK, Fell JW, Geml J, Ghobad-Nejhad M, Giachini AJ, Gibertoni TB, Gorjon SP, Haelewaters D, He S-H, Hodkinson BP, Horak E, Hoshino T, Justo A, Lim YW, Menolli Jr N, Mesi¢ A, Moncalvo J-M, Mueller GM, Nagy LG, Nilsson RH, Noordeloos M, Nuytinck J, Orihara T, Ratchadawan C, Rajchenberg M, Silva-Filho AGS, Sulzbacher MA, Tkalcéec Z, Valenzuela R, Verbeken A, Vizzini A, Wartchow F, Wei T-Z, Weifs M, Zhao C-L, Kirk PM (2019) Notes, outline and divergence times of Basidi- omycota. Fungal Diversity 99(1): 105-367. https://doi.org/10.1007/s13225-019-00435-4 Hopple Jr JS, Vilgalys R (1999) Phylogenetic relationships in the mushroom genus Coprinus and dark-spored allies based on sequence data from the nuclear gene coding for the large ribosomal subunit RNA: Divergent domains, outgroups, and monophyly. Molecular Phy- logenetics and Evolution 13(1): 1-19. https://doi.org/10.1006/mpev.1999.0634 Horak E (2005) Rohrlinge und Blatterpilze in Europa. Elsevier, Spektrum Akad. Verlag, 555 pp. Jin JJ, Yu WB, Yang JB, Song Y, dePamphilis CW, Yi T-S, Li D-Z (2020) GetOrganelle: A fast and versatile toolkit for accurate de novo assembly of organelle genomes. Genome Biology 21(1): e241. https://doi.org/10.1186/s13059-020-02154-5 Kim CS, Jo JW, Kwag YN, Sung GH, Lee SG, Kim SY, Shin CH, Han SK (2015) Mushroom Flora of Ulleung-gun and a Newly Recorded Bovista Species in the Republic of Korea. Mycobiology 43(3): 239-257. https://doi.org/10.5941/MYCO.2015.43.3.239 Kornerup A, Wanscher JH (1978) Methuen handbook of colour. Eyre Methuen, London. Li Y, LiTH, Yang ZL, Bau T, Dai YC (2015) Atlas of Chinese macrofungal resources. Central Chinese Farmer Press, Zhengzhou, China, 1351 pp. Liu ZW, Na Q, Cheng XH, Wu XM, Ge YP (2021) Mycena yuezhuoi sp. nov. (Mycenaceae, Ag- aricales), a purple species from the peninsula areas of China. Phytotaxa 511(2): 148-162. https://doi.org/10.11646/phytotaxa.511.2.3 Maas Geesteranus RA (1980) Studies in Mycenas-15, A tentative subdivision of the genus Mycena in the Northern Hemisphere. Persoonia 11: 93-120. Maas Geesteranus RA (1992a) Mycenas of the Northern Hemisphere I. Studies in Mycenas and other papers. Koninklijke Nederlandse Akademie van Wetenschappen, Amsterdam. Maas Geesteranus RA (1992b) Mycenas of the Northern Hemisphere II. Conspectus of the Mycenas of the Northern Hemisphere. Koninklijke Nederlandse Akademie van Weten- schappen, Amsterdam. Malysheva V, Malysheva E, Voronina EY, Fedosova A, Bibikov N, Kiseleva DS (2017) Mycorrhiza of Pyroloids (Pyrola rotundifolia, P media and Orthilia secunda): Species composition of sym- bionts and trophic status of plants. Mikologia i Fitopatologia 51(6): 350-364. [in Russian] 144 Qin Na et al. / MycoKeys 90: 119-145 (2022) Na Q, Bau T (2018) New species of Mycena (Mycenaceae, Agaricales) with colored lamellae and three new species records from China. Phytotaxa 361(3): 266-278. https://doi. org/10.11646/phytotaxa.361.3.2 Na Q, Bau T (2019a) Mycena section Sacchariferae: Three new species with basal discs from Chi- na. Mycological Progress 18(3): 483-493. https://doi.org/10.1007/s11557-018-1456-8 Na QC, Bau T (2019b) Recognition of Mycena sect. Amparoina sect. nov. (Mycenaceae, Agaricales), including four new species and revision of the limits of sect. Sacchariferae. MycoKeys 52: 103-124. https://doi.org/10.3897/mycokeys.52.34647 Na Q, Hu YP, Liu ZW, Zeng H, Qi LL, Ding H, Cheng XH, Ge YP (2021) The first report- ed occurrence of Leucoinocybe (Porotheleaceae, Agaricales) in China: Leucoinocybe lishu- iensis sp. nov. from Zhejiang Province. Nova Hedwigia 113(3—4): 453-469. https://doi. org/10.1127/nova_hedwigia/2021/0661 Na Q, Hu YP, Zeng H, Song ZZ, Ding H, Cheng XH, Ge YP (2022) Updated taxonomy on Gerronema (Porotheleaceae, Agaricales) with three new taxa and one new record from China. MycoKeys 89: 87-120. https://doi.org/10.3897/mycokeys.89.79864 Osmundson TW, Robert VA, Schoch CL, Baker LJ, Smith A, Robich G, Mizzan L, Garbelotto MM (2013) Filling Gaps in Biodiversity Knowledge for Macrofungi: Contributions and Assessment of an Herbarium Collection DNA Barcode Sequencing Project. PLoS ONE 8(4): e62419. https://doi.org/10.1371/journal. pone.0062419 Pérez-Izquierdo L, Morin E, Maurice JP, Martin F, Rincén A, Buée M (2017) A new promising phylogenetic marker to study the diversity of fungal communities: The Glycoside Hydro- lase 63 gene. Molecular Ecology Resources 2017(6): 1-17. https://doi.org/10.1111/1755- 0998.12678 Perry BA (2002) A taxonomic investigation of Mycena in California. Doctoral dissertation, San Francisco State University, San Francisco, the United States. Perry BA, Desjardin DE (2016) New species of Mycena (Basidiomycota, Agaricales) from California. Phytotaxa 269(1): 33-40. https://doi.org/10.11646/phytotaxa.269.1.4 Perry BA, Keller HW, Forrester ED, Stone BG (2020) A new corticolous species of Mycena sect. Viscipelles (Basidiomycota: Agaricales) from the bark of a Living American elm tree in Texas, U.S.A. Journal of the Botanical Research Institute of Texas 14(2): 167-185. https:// doi.org/10.17348/jbrit.v14.i2.1000 Petersen RH, Hughes KW, Lickey EB, Kovalenko AE, Psurtseva V (2008) A new genus, Cruentomycena, with Mycena viscidocruenta as type species. Mycotaxon 105(4): 119-136. Posada D, Crandall KA (1998) Modeltest: Testing the model of DNA substitution. Bioin- formatics (Oxford, England) 14(9): 817-818. https://doi.org/10.1093/bioinformat- ics/14.9.817 Redhead SA, Singer R (1981) Resinomycena gen. nov. (Agaricales), an ally of Hydropus, Mycena and Baeospora. Mycotaxon 8(1): 150-170. [Fungi] Redhead SA, Moncalvo JM, Vilgalys R, Desjardin DE, Perry BA (2012) Index Fungorum: Published Numbers 14: 1-1. Rexer KH (1994) Die Gattung Mycena s.l., Studien zu ihrer Anatomie, Morphologie und Systematik. Eberhard-Karls-Universitat Tiibingen, Tubingen, Germany, 132 pp. Two novo bluish Mycena species from northern China 145 Robich G (2003) Mycena d’Europa. Associazione Micologica Bresadola, Trento, 728 pp. Robich G (2016) Mycena @ Europa Volume 2. Associazione Micologica Bresadola, Trento, 796 pp. Ronquist FE, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics (Oxford, England) 19(12): 1572-1574. https://doi.org/10.1093/ bioinformatics/btg180 Saccardo PA (1887) Sylloge Hymenomycetum, Vol. I. Agaricineae (in Latin). Sylloge Fungo- rum, Berlin, 1146 pp. Shih YS, Chen CY, Lin WW, Kao HW (2014) Mycena kentingensis, a new species of luminous mushroom in Taiwan, with reference to its culture method. Mycological Progress 13(2): 429-435. https://doi.org/10.1007/s11557-013-0939-x Singer R (1969) Mycoflora Australis. Beihefte zur Nova Hedwigia 29: 110. Singer R, Gomez LD (1982) Basidiomycetes of Costa Rica. Brenesia 19-20: 31-47. Smith AH (1947) North American species of Mycena. University of Michigan Press, Ann Ar- bor, Michigan, 521 pp. Stamatakis A, Ludwig T, Meier H (2005) RAxML-IIE: A fast program for maximum likelihood- based inference of large phylogenetic trees. Bioinformatics maximum likelihood-based in- ference of large phylogenetic trees. Bioinformatics (Oxford, England) 21(4): 456-463. https://doi.org/10.1093/bioinformatics/btil91 Terashima Y, Takahashi H, Taneyama Y (2016) The fungal flora in southwestern Japan, Agarics and boletes. Tokai University Press, Kanagawa, Tokyo, 349 pp. Thoen E, Harder CB, Kauserud H, Botnen SS, Vik U, Taylor AF, Menkis A, Skrede I (2020) In vitro evidence of root colonization suggests ecological versatility in the genus Mycena. The New Phytologist 227(2): 601-612. https://doi.org/10.1111/nph.16545 Thompson JD, Gibson TJ, Plewniak F Jeanmougin E Higgins DG (1997) The Clustal-X win- dows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 63: 215-228. https://doi.org/10.1093/nar/25.24.4876 Vu D, Groenewald M, de Vries M, Gehrmann T; Stielow B, Eberhardt U, AlHatmi A, Groe- newald JZ, Cardinali G, Houbraken J, Boekhout T, Crous PW, Robert V, Verkley GJM (2019) Large-scale generation and analysis of filamentous fungal DNA barcodes boosts cov- erage for kingdom Fungi and reveals thresholds for fungal species and higher taxon delimita- tion. Studies in Mycology 92(1): 135-154. https://doi.org/10.1016/j.simyco.2018.05.001 Wei CL, Kirschner R (2019) A new Mycena species with blue basidiomata and porioid hymeno- phore from Taiwan. Mycoscience 60(1): 10-13. https://doi.org/10.1016/j.myc.2018.06.001 White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribo- somal RNA genes for phylogenetics. In: Innis, Seliger H, Gelfand DH, Sinsky JJ, White TJ (Eds) PCR Protocols: a Guide to Methods and Applications, 315-322. https://doi. org/10.1016/B978-0-12-372180-8.50042-1 Wu LS, Han T, Li WC, Jia M, Xue LM, Rahman K, Qin LP (2013) Geographic and tissue influences on endophytic fungal communities of Taxus chinensis var. mairei in China. Cur- rent Microbiology 66(1): 40-48. https://doi.org/10.1007/s00284-012-0235-z