Biodiversity Data Journal 11: 99027 OO) doi: 10.3897/BDJ.11.e99027 open access Taxonomy & Inventories Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from Yunnan Province, China Zheng-Quan Zhang}, Chao-Hai Li*+$, Lin Lit, Hong-Wei Shen*!1, Jun He*, Xi-Jun Suf, Zong-Long Luot + College of Agriculture and Biological Science, Dali University, Dali, China § College of Pharmacy, Dali University, Dali, China | Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, Thailand | School of Science, Mae Fah Luang University, Chiang Rai, Thailand # Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming, China Corresponding author: Zong-Long Luo (luozonglongfungi@163.com) Academic editor: Alfredo Vizzini Received: 20 Dec 2022 | Accepted: 01 Mar 2023 | Published: 15 Mar 2023 Citation: Zhang Z-Q, Li C-H, Li L, Shen H-W, He J, Su X-J, Luo Z-L (2023) Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from Yunnan Province, China. Biodiversity Data Journal 11: e99027. https://doi.org/10.3897/BDJ.11.e99027 Abstract Background Geastrum is the largest genus of Geastraceae and is widely distributed all over the world. Four specimens which belong to Geastrum were collected during our scientific expedition to Cangshan Mountain, Yunnan, China. Based on morphological characteristics and phylogenetic analysis, a new species was introduced. New information Geastrum suae is characterised by its large basidiomata (height 35-70 mm, diameter 18-— 37 mm) with long stipe (height 10-45 mm), smooth pink exoperidium and sessile globose endoperidial body. Phylogenetic analysis has been carried out, based on the internal © Zhang Z 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. 2 Zhang Z et al transcribed spacer (ITS) and large subunit ribosomal ribonucleic acid (nrLSU) sequence data. The illustration and description for the new taxa are provided. Keywords Geastraceae, ITS, nrLSU, taxonomy, phylogeny Introduction Geastrum Pers. is the largest genus of Geastraceae and was established by Persoon (1794). Geastrum is commonly known as the earthstars with worldwide distribution and the most species-diverse in the family Geastraceae. Up to now, there are 140 valid species in this genus (Wijayawardene et al. 2022, Zhou et al. 2022, Cabral et al. 2022, Wang and Bau 2023). Geastrum clearly differs from Myriostoma by a single endodermal stoma (Sousa et al. 2014). Due to the non-splitting ectoderm and the poorly-developed endoperidium being different from Geastrum, researchers thought that Radiigera is one of the genera closely related to Geastrum (Sunhede 1989, de Toledo and Castellano 1996). Later, some studies have found that specimens in Radiligera are nested in Geastrum (Hosaka et al. 2006, Hosaka and Castellano 2008, da Silva et al. 2013), but the relationship between these two genera has not been studied in depth until Jeppson et al. (2013) classified the species of Radiigera into the genus Geastrum. Species of this genus are distributed globally, especially in temperate and tropical regions, such as Brazil- Amazon and Europe (de Leén 1968, da Silva et al. 2013, Jeppson et al. 2013, Zamora et al. 2014, Cabral et al. 2014a, Cabral et al. 2014b, Sousa et al. 2015, Crous et al. 2016, Cabral et al. 2017, Crous et al. 2017, Sousa et al. 2017, Crous et al. 2018a, Crous et al. 2018b, Assis et al. 2019, Finy et al. 2021, Rodrigues et al. 2021). However, the taxonomic relationship under the genus was chaotic (Zamora et al. 2013) until Zamora et al. (2014) divided it into 14 Sections using polygenic analysis, viz. Sect. Campestria, Corollina, Elegantly, Exareolata, Fimbriata, Fornicata, Geastrum, Hariotia, Hieronymia, Myceliostroma, Papillata, Pseudoilmbata, Schmidelia and Trichaster. In China, the early systematic report of Geastrum can be found in "Fungi in China" (Deng 1963) and "The Confluence of Chinese Fungi" (Dai 1979). Zhou et al. (2007) detailed descriptions of 16 species of Geastrum in China in "Flora Fungorum Sinicorum- Geastraceae and Nidulariaceae". Later, three new records and nine new species were reported (Han and Bau 2016, Zhou et al. 2022, Wang and Bau 2023). Four specimens which belong to Geastrum were collected during our scientific expedition to Cangshan Mountain, Yunnan, China. Morphological and phylogenetic analysis revealed that these specimens are the same species and are different from other species in Geastrum. Therefore, we introduced it as a new species and provided the detailed description and illustration. Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from ... 3 Materials and methods Morphological description Macro-morphological descriptions were based on fresh specimens, which were photographed in the field with notes and laboratory supplemental measurements. The colour is compared with the standard colours in the colorhexa website (https:// www.colorhexa.com). Micro-morphological data were obtained from the fresh specimens and observed by using a light microscope, following Accioly et al. (2019). Sections were studied at magnification of up to 1000 using a Nikon eclipse Ni microscope and phase contrast illumination and scanning electron microscope (SEM) analysis was done under a Shimadzu SSX-—550. Preparation of the material examined under SEM followed da Silva et al. (2011). Microscopic features and measurements were made from slide preparations stained with 5% potassium hydroxide (KOH). Basidiospore features, hyphal system, colour, sizes and shapes were recorded and photographed. Measurements were made using the Image Framework v.0.9.7 to represent variation in the size of basidiospores, 5% of measurements were excluded from each end of the range and extreme values are given in parentheses. ih u it 0 The abbreviation for spore measurements (n/m/p) denote “n” spores measured from “m ih 0 basidiocarps of “p” specimens. Basidiospore dimensions (and “Q” values) are given as (a) b—av—c (d), where “a” represents the minimum, “d’ the largest, “av” the average “b” and “c’” covers a minimum of 90% of the values. “Q” is the length/width ratio of a spore inside view and “Qm” for the average of all spores + standard deviation. Voucher specimens are deposited in the Herbarium of Cryptogams, Kunming Institute of Botany Academia Sinica (KUN-HKAS). DNA extraction, PCR amplification and sequencing The DNA extractions were performed from a small piece of the dried basidioma by using Trelief ™ Plant Genomic DNA Kit from Tsingke Biotechnology Co., Ltd (Beijing, China). Two DNA regions were amplified: the internal transcribed spacer nuclear ribosomal DNA (ITS), nuclear ribosomal large subunit (nrLSU) with the primer pairs ITS1F/ITS4 and LROR/ LR5, respectively (Table 1). Table 1. Amplification primers information used in this study. Gene Primer Primer sequence (5'-3') References ITS ITS1F CTTGGTCATT TAGAGGAAGTAA Gardes and Bruns (1993) ITS4 TCCTCCGCTTATTGATATGC White et al. (1990) nrLSU LROR ACCCGCTGAACTTAAGC Vilgalys and Hester (1990) LR5 ATCCTGAGGGAAACTTC Vilgalys and Hester (1990) Zhang Z et al PCR reactions (25 ul) contained mixture: 12.5 yl 2X SanTaq PCR Master Mix (including MgClo, dNTP, Taq DNA Polymerase, PCR buffer, loading etc.), 1 ul each of primer, 2 ul DNA solution and 9.5 ul sterilised distilled HzO. The PCR cycling for ITS and nrLSU was as follows: initial denaturation at 94°C for 5 min, followed by 35 cycles at 94°C for 30 sec, 53°C for 30 sec and 72°C for 50 sec and a final extension of 72°C for 10 min. The PCR products were visualised via UV light after electrophoresis on 1% agarose gels stained with ethidium bromide. Successful PCR products were sent to Sangon Biotech Limited Company (Shanghai, China), using forward PCR primers. When sequences have heterozygous INDELS or ambiguous sites, samples were sequenced bidirectionally to make contigs of the amplified regions or verify the ambiguous sites. Raw DNA sequences were assembled and edited in Sequencher 4.1.4 and the assembled DNA sequences were deposited in GenBank (Table 2). Table 2. Species, specimens, Collection locality and GenBank accession numbers of sequences used in this study (newly-generated sequences are indicated in bold). Species Geastrum mirabile Geastrum javanicum Geastrum mirabile Geastrum parvistriatum Geastrum parvistriatum Geastrum striatum Geastrum campestre Geastrum aff. arenarium Geastrum lageniforme Geastrum cf. calceum Geastrum cf. calceum Geastrum aff. hariotii Geastrum cf. saccatum Geastrum hieronymi Geastrum cf. stipitatum Geastrum albonigrum Geastrum aff. arenarium Geastrum cf. arenarium Geastrum argentinum Geastrum argentinum Geastrum berkeleyi Strain/Voucher strain: 228-394 TNS:TKG-GE-90902 TNS:KH-JPN10-714 MA-Fungi 69583 Herb. Zamora 272 Herb. Zamora 257 Herb. Zamora 283 Herb. Zamora 76 Herb. Zamora 316 UFRN-Fungos 723 MA-Fungi 83761 Bérge Petterson 2070 Herb. Sunhede 7749 MA-Fungi 83767 Herb. Zamora 528 MA-Fungi 36140-2 MA-Fungi 68191 MA-Fungi 83760 LPS 48446 MA-Fungi 82605 MA-Fungi 74668 Collection locality GenBank Accession No. Japan Japan Japan Spain Spain Spain Spain Spain Spain Brazil Argentina Mozambique Australia Argentina Brazil Panama Spain Argentina Argentina Argentina Spain ITS AB509736 JN845100 JN845109 JN943160 JN943162 JN943164 JN943167 KF988338 KF988339 KF988340 KF988341 KF988342 KF988343 KF 988344 KF988345 KF 988349 KF 988350 KF988351 KF988352 KF988353 KF988354 nrLSU JN845218 JN845227 JN939560 JN939572 JN939557 JN939575 KF988470 KF 988514 KF 988477 KF988478 KF988507 KF988556 KF988509 KF988576 KF988468 KF988469 KF988471 KF988472 KF988473 KF988474 Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from ... Species Geastrum berkeleyi Geastrum berkeleyi Geastrum campestre Geastrum campestre Geastrum corollinum Geastrum corollinum Geastrum coronatum Geastrum coronatum Geastrum coronatum Geastrum aff. coronatum Geastrum aff. coronatum Geastrum elegans Geastrum elegans Geastrum entomophilum Geastrum fimbriatum Geastrum fimbriatum Geastrum flexuosum Geastrum floriforme Geastrum floriforme Geastrum fornicatum Geastrum fornicatum Geastrum fuscogleba Geastrum fuscogleba Geastrum glaucescens Geastrumglaucescens Geastrum aff. glaucescens Geastrum hariotii Geastrum aff. hariotii Geastrum aff. hariotii Geastrum hieronymi Geastrum kotlabae Geastrum kotlabae Geastrum aff. kotlabae Geastrum lageniforme Geastrum aff. lageniforme Strain/Voucher Herb. Sunhede 7724 Herb. Zamora 504 Herb. Sunhede 7575 MICH 28566 MA-Fungi 5746 Herb. Sunhede 7744 Herb. Zamora 266 Herb. Zamora 522 MICH 28567 MICH 72012 MICH 72014 Herb. Zamora 189 UPS F-560810 MA-Fungi 70785 Herb. Zamora 234 Herb. Sunhede 7739 UPS F-119844 MA-Fungi 69173 Herb. Zamora 453 Herb. Zamora 255 MA-Fungi 30749 NY Trappe 1071 NY Trappe 9500 MA-Fungi 83762 MA-Fungi 83763 MA-Fungi 83764 MA-Fungi 83765 MA-Fungi 78296 MA-Fungi 78289 MA-Fungi 83766 MA-Fungi 39563 Herb. Zamora 440 MA-Fungi 33300 Herb. Zamora 207 MA-Fungi 83768 Collection locality GenBank Accession No. Sweden Sweden Sweden USA Spain Sweden Spain Sweden USA USA USA Spain Sweden Brazil Spain Sweden Sweden Spain Spain Spain Spain USA USA Argentina Argentina Argentina Argentina Brazil Brazil Argentina Spain Spain Tanzania Spain Argentina ITS KF988355 KF988356 KF988357 KF 988358 KF988359 KF988360 KF 988361 KF988362 KF988363 KF988364 KF988365 KF 988366 KF 988367 KF988368 KF988369 KF988370 KF 988371 KF 988372 KF 988373 KF988374 KF988375 KF 988376 KF 988377 KF988378 KF988379 KF988380 KF988381 KF988382 KF988383 KF988384 KF988385 KF988386 KF 988387 KF 988388 KF 988389 nrLSU KF988475 KF988476 KF988479 KF988480 KF988481 KF988482 KF988483 KF988484 KF988485 KF988486 KF988487 KF988488 KF988489 KF988490 KF988491 KF988492 KF988493 KF988494 KF988495 KF988496 KF988497 KF988498 KF988499 KF988500 KF988501 KF988502 KF988504 KF988505 KF988506 KF988508 KF988510 KF988511 KF988512 KF988513 KF988516 Species Geastrum aff. lageniforme Geastrum aff. lageniforme Geastrum aff. lageniforme Geastrum aff. lageniforme Geastrum aff. lageniforme Geastrum melanocephalum Geastrum melanocephalum Geastrum michelianum Geastrum michelianum Geastrum aff. michelianum Geastrum minimum Geastrum minimum Geastrum minimum Geastrum minimum Geastrum minimum Geastrum minimum Geastrum morganii Geastrum aff. morganii Geastrum aff. morganii Geastrum aff. morganii Geastrum aff. morganii Geastrum ovalisporum Geastrum pectinatum Geastrum pectinatum Geastrum pectinatum Geastrum pectinatum Geastrum pleosporum Geastrum pouzarii Geastrum pouzarii Geastrum pseudolimbatum Geastrum pseudolimbatum Geastrum quadrifidum Geastrum quadrifidum Geastrum quadrifidum Geastrum rufescens Zhang Z et al Strain/Voucher COFC Hama 327 MA-Fungi 83770 MA-Fungi 83769) MA-Fungi 78398 Herb. Ribes 221210-01 Herb. Zamora 34 Herb. Sunhede 7737 Herb. Sunhede 7738 Herb. Zamora 227 MA-Fungi 83771 Herb. Zamora 191 Herb. Sunhede 7746 MICH 72010 MICH 28119 MA-Fungi 31530 MA-Fungi 86669 Herb. Lebeuf HRLO177 Herb. Zamora 367 Herb. Zamora 525 MA-Fungi 83772 MA-Fungi 83773 MA-Fungi 47184 Herb. Zamora 252 UPS F-560803 UPS F-09935 (161483) MA-Fungi 83774 MA-Fungi 56971 MA-Fungi 2944 Herb. Sunhede 7494 Herb. Zamora 231 UPS F-560804 Herb. Zamora 170 MA-Fungi 86671 MICH 72512 Herb. Zamora 253 Collection locality Niger Argentina Argentina Portugal Spain Spain Sweden Sweden Spain Argentina Spain Sweden USA Spain USA Sweden Canada Spain Spain Argentina Argentina Bolivia Spain Sweden Tanzania Argentina Cameroon Czechoslovakia Czechoslovakia Spain Sweden Spain Sweden USA Spain GenBank Accession No. ITS KF988390 KF988391 KF988392 KF988393 KF988394 KF988395 KF988396 KF 988397 KF988398 KF988399 KF988400 KF988401 KF 988402 KF988403 KF 988404 KF988405 KF 988406 KF 988407 KF 988408 KF988409 KF988410 KF988411 KF988412 KF988413 KF988414 KF988415 KF988416 KF988417 KF988418 KF988419 KF988420 KF988421 KF988422 KF988423 KF 988424 nrLSU KF988517 KF988518 KF988519 KF988520 KF988521 KF988522 KF988523 KF988524 KF988525 KF988527 KF988528 KF988529 KF988530 KF988531 KF988532 KF988533 KF988534 KF988535 KF988536 KF988537 KF988538 KF988539 KF988540 KF988541 KF988542 KF988543 KF988544 KF988545 KF988546 KF988547 KF988548 KF988549 KF988550 KF988551 KF988552 Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from ... Species Geastrum rufescens Geastrum cf. saccatum Geastrum cf. saccatum Geastrum cf. saccatum Geastrum cf. saccatum Geastrum cf. saccatum Geastrum cf. saccatum Geastrum cf. saccatum Geastrum cf. saccatum Geastrum schmidelii Geastrum schmidelii Geastrum cf. schweinitzii Geastrum cf. schweinitzii Geastrum cf. schweinitzii Geastrum cf. schweinitzii Geastrum smardae Geastrum smardae Geastrum smithii Geastrum striatum Geastrum “triplex” Geastrum “triplex” Geastrum cf. velutinum Geastrum cf. velutinum Geastrum cf. velutinum Geastrum cf. velutinum Geastrum violaceum Geastrum violaceum Geastrum sp.1 Geastrum sp.1 Geastrum sp.2 Geastrum sp.2 Geastrum sp.3 Geastrum sp.4 Geastrum sp.5 Geastrum sp.5 Strain/Voucher Herb. Zamora 274 MA-Fungi 47185-2 MA-Fungi 83775 UPS F-530056 MA-Fungi 83777 Herb. Zamora 260 Herb. Zamora 461 COFC Hama 343 MA-Fungi 83778 Herb. Zamora 279 UPS F-560805 S Henrik Kylin 1983 30.X MA-Fungi 83779 MA-Fungi 36141 MA-Fungi 83780 Herb. Lebeuf HRL 0160 Herb. Zamora 527 MA-Fungi 83783 MA-Fungi 86672 UPS F-014630 (213863) MA-Fungi 83784 MA-Fungi 83785 MA-Fungi 83786 Herb. Ribes 311207-62 MA-Fungi 83787 BAFC 51671 MA-Fungi 82487 MA-Fungi 83788 MA-Fungi 83789 MA-Fungi 31143 MA-Fungi 37546 MA-Fungi 83790 MA-Fungi 83791 Herb. Zamora 145 Herb. Zamora 450 Collection locality GenBank Accession No. Spain Bolivia Argentina Japan Argentina Spain Spain Niger Argentina Spain Sweden Papua New Guinea Argentina Panama Argentina Canada Spain Argentina Sweden Madagascar Argentina Argentina Argentina Spain Peru Argentina Argentina Argentina Argentina Spain Spain Argentina Peru Spain Spain ITS KF988425 KF988426 KF988427 KF988428 KF988429 KF988430 KF988431 KF988432 KF 988433 KF 988434 KF988435 KF 988436 KF 988437 KF 988438 KF988439 KF 988440 KF988441 KF988442 KF988443 KF988444 KF988445 KF988446 KF988447 KF 988448 KF988449 KF988450 KF988451 KF988452 KF988453 KF988454 KF988455 KF 988456 KF 988457 KF 988458 KF 988459 nrLSU KF988553 KF988554 KF988555 KF988558 KF988559 KF988560 KF988561 KF988562 KF988563 KF988564 KF988565 KF988566 KF988567 KF988568 KF988569 KF988573 KF988574 KF988575 KF988577 KF988578 KF988579 KF988581 KF988582 KF988583 KF988584 KF988585 KF988586 KF988587 KF988588 KF988589 KF988590 KF988591 KF988592 KF988593 KF988594 Species Geastrum sp.6 Geastrum sp.7 Geastrum sp.7 Geastrum sp.8 Geastrum hirsutum Geastrum javanicum Geastrum minutisporum Geastrum minutisporum Geastrum pusillipilosum Geastrum pusillipilosum Geastrum piquiriunense Geastrum hirsutum Geastrum rubropusillum Geastrum baculicrystallum Geastrum brunneocapillatum Geastrum rubellum Geastrum neoamericanum Geastrum courtecuissei Geastrum rubellum Geastrum rubellum Geastrum neoamericanum Geastrum suae Geastrum suae Geastrum suae Geastrum suae Geastrum hariotii Geastrum aff. lageniforme Geastrum cf. saccatum Geastrum cf. schweinitzii Geastrum cf. velutinum Geastrum michelianum Geastrum setiferum Geastrum setiferum Geastrum velutinum Geastrum velutinum Zhang Z et al Strain/Voucher MA-Fungi 83792 MA-Fungi 83793 MA-Fungi 83794 MA-Fungi 83795 UFRN-Fungos 1214 UFRN-Fungos 1215 CORD14 CORD15 UFRN:Fungos 2315 UFRN:Fungos 2759 UFRN:Fungos:2892 INPA:259950 UFRN:Fungos:2308 UFRN:Fungos:2835 UFRN:Fungos:2286 UFRN:Fungos:2844 UFRN:Fungos:2302 LIP:FH 2004090503 LIP:CL/MART 8067B LIP:PAM/MART 12.100 LIP:JLC 12030103 HKAS 123795 (Holotype) HKAS 123794 HKAS 123793 HKAS 123796 (Paratype ) MA-Fungi 80070 MA-Fungi 79056 MA-Fungi 83776 S Henrik Kylin 842 MA-Fungi 73247 Herb. Ribes 231208-31 MA-Fungi 83781 MA-Fungi 83782 BJTC 221 BJTC 598 Collection locality Argentina Argentina Argentina Argentina Brazil Brazil Argentina Argentina Brazil Brazil Brazil Brazil Brazil Brazil Brazil Brazil Brazil Guadeloupe Martinique Martinique French China China China China Dominican Republic Brazil Argentina Fiji India Spain Argentina Argentina China China GenBank Accession No. ITS KF988460 KF988461 KF 988462 KF 988463 KJ127029 KJ127031 KM260664 KM260665 KX761175 KX761177 MH260269 MH634993 MH634994 MH634995 MH634996 MH634999 MH635001 MH635003 MH635009 MH635010 MH635014 ON529511 ON529512 ON529513 ON529514 MZ508877 nrLSU KF988595 KF988596 KF988597 KF988598 KX761176 KX761178 MH260270 MH635026 MH635027 MH635028 MH635029 MH635031 MH635040 MH635033 MH635037 MH635038 ON529515 ON529516 ONS29517 ON529518 KF988503 KF988515 KF988557 KF988570 KF988580 KF988526 KF988571 KF988572 MZ509382 Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from ... 9 Species Strain/Voucher Collection locality GenBank Accession No. ITS nrLSU Geastrum yanshanense BJTC 381 China MZ508878 MZ509383 Geastrum yanshanense BJTC 057 China MZ508879 MZ509384 Geastrum yanshanense BJTC 255 China MZ508880 Schenella pityophila Herb. Zamora 530 Spain KF 988346 KF988464 Schenella pityophila Herb. Zamora 531 Spain KF 988347 KF988465 Myriostoma coliforme MA-Fungi 83759 Argentina KF 988348 KF988467 Sequence alignment Sequence data of two partial loci, internal transcribed spacer region (ITS) and the large subunit ribosomal RNA gene (nrLSU) were analysed. All the sequences, except those which were obtained from this study, were selected from GenBank for phylogenetic analyses (Table 2). Sequences were aligned using the online version of MAFFT v.7 (http:// mafft.cbrc.jp/alignment/server/) (Katoh and Standley 2013) and adjusted using BioEdit v. 7.0.9 (Hall 1999) by hand to allow maximum alignment and minimise gaps. Ambiguous regions were excluded from the analyses and gaps were treated as missing data. AliView 1.19-beta was used to convert the alignment fasta file to Phylip and Nexus format for phylogenetic analysis. Phylogenetic analyses were obtained from Maximum Likelihood (ML) and Bayesian Inference (BI). Molecular phylogenetic analyses The Maximum Likelihood (ML) and Bayesian Inference (BI) methods were used to analyse the combined dataset of ITS and nrLSU sequences. ML analysis was conducted with RAXML-HPC2 on the CIPRES Science Gateway (Miller et al. 2010), involving 100 ML searches; all model parameters were estimated by the programme. The ML bootstrap values (ML-BS) were obtained with 1000 rapid bootstrapping replicates. Bayesian analysis was performed with MrBayes v.3.2 (Ronquist et al. 2012), with the best- fit model of sequence evolution estimated with MrModelTest 2.3 (Nylander et al. 2008) to evaluate posterior probabilities (PP) (Rannala and Yang 1996, Zhaxybayeva and Gogarten 2002) by Markov Chain Monte Carlo (MCMC) sampling. Six simultaneous Markov chains were run for 100,000,000 generations, trees were sampled every 500" generation and 200,000 trees were obtained. The first 50,000 trees, representing the burn-in phase of the analyses, were discarded, while the remaining 150,000 trees were used for calculating posterior probabilities in the majority rule consensus tree (the critical value for the topological convergence diagnostic is 0.01). The phylogenetic tree was visualised with Figlree version 1.4.0 (Rambaut 2012) and made in Adobe Illustrator CS5 (Adobe Systems Inc., USA). Sequences derived in this study were deposited in GenBank (http://www.ncbi.nim.nih.gov). 10 Zhang Z et al Taxon treatment Geastrum suae Z.Q. Zhang C.H. Li & Z.L. Luo, sp. nov. MycoBank IMB845193 Materials Holotype: a. scientificName: Geastrum suae; kingdom: Fungi; phylum: Basidiomycota; class: Agaricomycetes; order: Geastrales; family: Geastraceae; genus: Geastrum; verbatimElevation: 2160 m; locationRemarks: China, Yunnan Province, Dali City, Cangshan Mountain; verbatimLatitude: 25°43'36.97"N; verbatimLongitude: 100°07'16.46"E; year: 2020; month: September; day: 4; habitat: Terrestrial; fieldNotes: grows in groups on the ground in mixed coniferous and broad-leaved forests, with thick humus; recordNumber: SJ582; recordedBy: Zheng-Quan Zhang; type: KUN-HKAS 123795; occurrence!D: 6D676216-9572-5A8D-A7C3-4C445C671395 Paratype: a. scientificName: Geastrum suae; kingdom: Fungi; phylum: Basidiomycota; class: Agaricomycetes; order: Geastrales; family: Geastraceae; genus: Geastrum; verbatimElevation: 2221 m; locationRemarks: China, Yunnan Province, Dali City, Cangshan Mountain; verbatimLatitude: 25°40'16.38"N; verbatimLongitude: 100°09'08.42"E; year: 2020; month: October; day: 14; habitat: Terrestrial; fieldNotes: grows in groups on the ground in mixed coniferous and broad-leaved forests, with thick humus; recordNumber: MB015; recordedBy: Chao-Hai Li; type: KUN-HKAS 123796; occurrence!D: F58D4D0C-33BB-541B-A92B-7AF436F12F49 Other materials: a. scientificName: Geastrum suae; kingdom: Fungi; phylum: Basidiomycota; class: Agaricomycetes; order: Geastrales; family: Geastraceae; genus: Geastrum; verbatimElevation: 2208 m; locationRemarks: China, Yunnan Province, Dali City, Cangshan Mountain; verbatimLatitude: 25°40'28"N; verbatimLongitude: 100°08'59"E; year: 2021; month: September; day: 3; habitat: Terrestrial; fieldNotes: grows in groups on the ground in mixed coniferous and broad-leaved forests, with thick humus; recordNumber: SJ2501; recordedBy: K. Wang; type: KUN-HKAS 123793; occurrencelD: 9213A508-19C3-5A9C-8E34-8DCDD9D4C170 scientificName: Geastrum suae; kingdom: Fungi; phylum: Basidiomycota; class: Agaricomycetes; order: Geastrales; family: Geastraceae; genus: Geastrum; verbatimElevation: 2350 m; locationRemarks: China, Yunnan Province, Yangbi County, Cangshan Mountain; verbatimLatitude: 25°41'59"N; verbatimLongitude: 100°02'00"; year: 2021; month: October; day: 1; habitat: Terrestrial; fieldNotes: grows in groups on the ground in mixed coniferous and broad-leaved forests, with thick humus; recordNumber: $J2500; recordedBy: G. H. Yang; type: KUN-HKAS 123794; occurrencelD: CE65C858-1CD7-53B3-B545-F 1750391FB8E Description Unexpanded basidiomata 13-28 mm, cylindrical to ellipsoidal, very light grey (#fdfdfd) to very pale red (#ffe6e6) with a slight protrusion, rough. Expanded basidiomata height Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from ... 11 35-70 mm, diameter 18-37 mm, deep saccate, Exoperidium splitting into 6, arched, not hygrometric, prosthecae length 23-35 mm, diameter 5-13 mm, exoperidium attached to the rhizomorphs. Rhizomorphs with 0.1—-5.4 um hyphae, fibrous and transparent, white (#ffffff). Mycelial layer 49.5-59.0 um, consisting of transparent hyphae (1.0—3.5 um) with thin walls and no septum, curved. Fibrous layer 6.5—16.5 um, transparent, curved, thick-walled hyphae (1.1-5.0 um) smooth, transparent to cream (#fffddO), pure red (#e60000) to dark red (#9a0000) when stained with Congo red. Pseudoparenchymatous layer 2.5-19.3 = 2.7-30.4 wm, irregular shape, mycelium is transparent when fresh, pure orange (#ffa500) to moderate pink (#cc6691) when stained with Congo red, the thickness of the pseudoparenchyma layer is about 1.0-1.3 mm, very soft pink (#d98ca0). Endoperidial body 11-23 mm, globose, sessile, very light grey (#dfdfdf) to dark grey (#a0a0a0), with lighter reticulation. Endoperidial surface with some protruding hyphae, endoperidium is interwoven by transparent hyphae, fibrous. Peristome fibrillose, unpleated, wide conical, with obvious oral margin ring. Columella obvious very light grey (#f4f4f4 to #e0e0e0). Eucapillitium hyphae 1.0—5.5 um, thick-walled, with distinct cavities, smooth, the ends tapering and are bluntly rounded (Fig. 1). Figure 1. Geastrum suae (KUN-HKAS 123795, holotype). a fresh unexpanded fruiting bodies; b, c fresh mature fruiting bodies; d mycelial layer, fibrous layer and pseudoparenchymatous layer; e hyphae of mycelial layer; f pseudoparenchymatous layer (cells in the stack); g, h eucapillitium hyphae; i-k basidiospores (LM); I-n basidiospores (SEM). Scale bars: a = 10 mm; b, c, e = 20 mm; d = 80 um; f, g, i-k = 10 um; h = 70 um; 1 = 1 um; m, n = 500 nm. 12 Zhang Z et al Basidiospores globose: Holotype (40/2/1) 4.5-5.3-6.0 x (4.5)5.0-5.4-6.0 um, Q = (0.80)0.83—1.12(1.14), Qm= 0.98 + 0.08, n = 40, including spines truncated at the apex ornamentation, with 0.2—0.5 ym high warts, ornamentation isolated or coalescing crest- like warts. Basidia not observed. Diagnosis Geastrum suae is characterised by long stipes and larger basidiomata; Pseudoparenchymatous layer is pink, smooth; globose endoperidial body, grey; the ends of eucapillitium hyphae taper and are bluntly rounded; and they live in groups. Etymology The species is named suae (Lat.), in memory of the Chinese mycologist Prof. Hong- Yan Su, who kindly helped the authors in many ways and sadly passed away on 3 May 2022 during the preparation of the current paper. Habit It grows in groups on the ground in mixed coniferous and broad-leaved forests where there are Alnus nepalensis and Pinus yunnanensis, with thick humus. Currently, it is known only from Cangshan Mountain. Analysis Phylogenetic analysis Firstly, we constructed the ML tree of Geastrum genus, based on ITS (1-540 bp) and nrLSU (541-1498 bp) genes and found that G. suae is in Sect. Mycelioatroma. The Maximum Likelihood bootstrap values (ML) equal to or greater than 70% are given above each node (Fig. 2), with the Final ML Optimisation Likelihood: -24127.230142. The aligned matrix had 856 distinct alignment patterns, with 6.78% completely undetermined characters or gaps. The base frequency and rate are as follows: A = 0.274187, C = 0.208839, G = 0.265219, T = 0.251755; rate AC = 1.202699, AG = 3.054698, AT = 1.472914, CG = 0.671195, CT = 5.726232, GT = 1.000000; gamma distribution shape: a = 0.269052. Therefore, we constructed the ML tree and Bayesian tree of Sect. Mycelioatroma, based on ITS and nrLSU genes and clarified the position of G. suae in this Section. The dataset is composed of ITS and nrLSU genes, comprising a total of 1478 characters including gaps, ITS (1-591 bp) and nrLSU (592-1478 bp), including 35 taxa with Myriostoma coliforme (MA-Fungi 83759) as the outgroup taxon (Fig. 3). The best fit model for the combined 2-gene dataset estimated and applied in the Bayesian analysis was GTR+I+G, Iset nst = 6, rates = invgamma; prset statefreqpr = dirichlet (1,1,1,1). The phylogenetic analysis of ML and BI produces similar topology. The combined dataset analysis of RAxML generates a best-scoring tree (Fig. 3), with the Final ML Optimisation Likelihood value of -7513.207751. The aligned matrix had 584 distinct alignment patterns, Geastrum suae sp. nov. (Geastraceae, Basidiomycota) a new species from 13 with 21.33% completely undetermined characters or gaps. The base frequency and rate are as follows: A = 0.272494, C 1.093594, AG = 2.765430, AT 0.207593, G 0.257821, T = 0.262093; rate AC 1.755140, CG = 0.441983, CT = 5.721217, GT 1.000000; gamma distribution shape: a = 0.243957. Bootstrap support values with ML greater than 70% and Bayesian posterior probabilities (PP) greater than 0.95 are given above the nodes (Fig. 3). 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EES] Phylogenetic tree of Geastrum species data. Cement mee nacre AY JLCH wee ‘angers 2 Geastram ef scheeiiten 'S Herik Kylin #42 UPS of spate Vers Zartore $28 PWAIPOIPIAPY “POS and related taxa, based on ITS and nrLSU sequence 14 Zhang Z et al Geastrum velutinum Herb. Ribes. 1120 17-62 Geastrum javanicum TNS: TKG-GE-90902 Geastrum javanicum UFRN: Fungos: 1215 Geastrum velutinum MA-Fungi_ 83787 Geastrum velutinum BJTC S' Geastrum velutinum MA-Fungi Geastrum velutinum MA-Fungi 83785 Geastrum yanshanense BJTC 255 100/1 ‘Geastrum yanshanense BITC 381 Geastrum yanshanense BJTC 057 880,99 Geastrum suae KUN. 0 Geastrum suae Geastrum suae Geastrum suae