Front. Plant Sci. Frontiers in Plant Science Front. Plant Sci. 1664-462X Frontiers Media S.A. 10.3389/fpls.2023.1226794 Plant Science Original Research Diversity and taxonomy of the genus Amanita (Amanitaceae, Agaricales) in the Yanshan Mountains, Northern China Zhou Hao † Guo MeiJun † Zhuo Lan Yan HuiFang Sui XiaoNan Gao Yue Hou ChengLin * College of Life Science, Capital Normal University, Beijing, China

Edited by: Daniel Pinero, National Autonomous University of Mexico, Mexico

Reviewed by: Jing Si, Beijing Forestry University, China; Tolgor Bau, Jilin Agriculture University, China

*Correspondence: ChengLin Hou, chenglin-hou@cnu.edu.cn

†These authors have contributed equally to this work

 14 09 2023 2023 14 1226794 22 05 2023 23 08 2023 Copyright © 2023 Zhou, Guo, Zhuo, Yan, Sui, Gao and Hou 2023 Zhou, Guo, Zhuo, Yan, Sui, Gao and Hou

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Globally, the species of Amanita are key components of ectomycorrhizal ecosystems. Some of them are widely known as poisonous or edible fungi. Although many new Amanita species from China have been described, the species diversity of Yanshan Mountains remains unknown. We here describe three new species, namely, A. borealis sp. nov. (Sect. Amanita), A. brunneola sp. nov. (Sect. Caesareae), and A. yanshanensis sp. nov. (Sect. Validae), based on morphological observations and molecular phylogenetic analyses. In addition, nine known species, namely, A. caesareoides (Sect. Caesareae), A. chiui (Sect. Vaginatae), A. muscaria (Sect. Amanita), A. oberwinklerana (Sect. Roanokenses), A. ovalispora (Sect. Vaginatae), A. subglobosa (Sect. Amanita), A. subjunquillea (Sect. phalloideae), A. vaginata var. vaginata (Sect. Vaginatae), and A. virosa (Sect. phalloideae), were reported from Yanshan Mountains for the first time. Our results emphasize that China has a high diversity of Amanita species and that additional studies are required to understand the exact species number. These findings play a crucial role in Amanita toxin research and ecological conservation. This study investigated the areas where Amanita species-related research is lacking. The study also attempted to better understand Amanita distribution and thus contribute to related research. This study enriches the species diversity of Amanita in Yanshan Mountains and offers additional data supporting the macrofungal systematics, toxin research, and diversity and ecological studies of Amanita in future studies.

 new taxa poisonous fungi ectomycorrhizal species diversity molecular systematics 32270012 National Natural Science Foundation of China10.13039/501100001809 section-in-acceptance Plant Systematics and Evolution Introduction

Amanita Pers., the largest genus of the family Amanitaceae E.-J. Gilbert, was established by Persoon (Persoon, 1797). It is an almost cosmopolitan genus comprising approximately 650 accepted species (Yang et al., 2004; Tulloss, 2005; Yang, 2005; Menolli et al., 2009; Yang, 2015; Kim et al., 2013a, b; Cai et al., 2014; Ariyawansa et al., 2015; Cho et al., 2015; Cai et al., 2016; Cui et al., 2018; Yang et al., 2018; Fraiture et al., 2019; Mighell et al., 2019; Mighell et al., 2021; Su et al., 2022).

Most Amanita species are ectomycorrhizal fungi of ecological importance, and more than 10 plant families are known to be symbiotically associated with Amanita (Beeli et al., 1935; Reid, 1980; Pegler and Shah-Smith, 1997; Wood, 1997; Yang, 1997; Yang, 2005; Davison et al., 2017; Cui et al., 2018). However, some Amanita species may be saprotrophic (e.g., A. pruittii A. H. Sm. ex Tulloss) (Cui et al., 2018).

Some species are commonly known as edible fungi, such as A. caesarea (Scop.) Pers., A. sinensis Zhu L. Yang, and A. yuaniana Zhu L. Yang et al. In addition, some Amanita species are poisonous, including A. subjunquillea S. Imai, A. virosa Bertill., and A. tenuifolia (Murrill) Murrill et al. (Yang, 2005). In China, deaths caused by consuming poisonous Amanita species are common (Li et al., 2020; Li et al., 2021a; Li et al., 2021b).

Based on the traditional morphological and anatomical characteristics, and the molecular phylogeny evidence, the classification of Amanita has also undergone many changes. Corner and Bas (1962) and Bas (1969) considered observing the natural characteristics of Amanita species in the wild important. They applied microscopic characteristics to taxonomy and split the species into two subgenera and six sections. Many mycologists have accepted this taxonomic method as a great historical advance in the Amanita classification (Jenkins, 1977; Hongo, 1982; Jenkins, 1986; Mao, 1990; Pegler and Shah-Smith, 1997). However, the classification of its subgenera remains disputed (Moser, 1967; Garcin, 1984; Singer, 1986). Subsequently, the systematic research on Amanita is gradually deepening with the development and advancement of molecular systematics. A recent comprehensive phylogenetic study introduced the latest classification system for Amanita. According to this system, Amanita was divided into 3 subgenera and 11 sections (Cui et al., 2018). This system is followed by other mycologists (Kumar et al., 2021; Suwannarach et al., 2022; Huang et al., 2023).

The Yanshan Mountains (115°–119°47′E, 39°40′–41°20′N) is located in northern China and has a high plant diversity ( Figure 1 ). The main forest types on these mountains are deciduous broad-leaved forests and mixed coniferous and broad-leaved forests. The original dominant ectomycorrhizal plants included Quercus mongolica Fisch. ex Ledeb., Betula platyphylla Suk., Abies nephrolepis (Trautv.) Maxim., Populus tomentosa Carrière, and Pinus tabuliformis Carr. (Wang et al., 2021). The Yanshan Mountains region has a warm–temperate continental monsoon climate with an annual precipitation of 350–700 mm. The peak of precipitation occurs in June–August. The altitude of these mountains ranges from 200 to 2,200 m (Zhou et al., 2022a; Zhou et al., 2022b; Zhou et al., 2022c). Some past records of Amanita species in this area are available (Yang, 2004; Chen et al., 2006; Zhang et al., 2017; Cui et al., 2018; Wu et al., 2020). However, information available on Amanita species on these mountains is incomplete.

Geographical location and topography of the study area.

Taxonomic research has never been considered a popular study, but it can be considered the basis for understanding the world and research in related professional fields and can only be studied and applied if we figure out what the species really is. Especially think of Amanita, which are a very attractive taxa of macrofungi. It plays an important role in toxin research and ecological conservation. In the present study, 36 fresh Amanita specimens were collected from Yanshan Mountains. There were 20 herbarium specimens were loaned from the Herbarium Mycologicum Academiae Sinicae (HMAS, Institute of Microbiology, Chinese Academy of Sciences) for further research. On the basis of morphological examination and inference of phylogeny, three new species and nine known species were reported herein. The study aimed to determine the taxonomic status and phylogenetic position of Amanita species represented by these specimens so as to establish a comprehensive database of macrofungal diversity in Yanshan Mountains, especially Amanita species, and to use this database as a basis for future studies on macrofungal systematics, fungal toxin, diversity, and ecology in this region.

 Materials and methods Sample collection and morphological analyses

The specimen collection area is shown in Figure 1 (Figure and data provided by the Chinese Academy of Environmental Sciences). The specimens were collected during 2019–2022 from Yanshan Mountains and photographed in the field. Macroscopic features of fresh specimens such as colors and odors were noted. Color codes and designations were assigned after referring to the website ColorHexa (https://www.colourhexa.com). The specimens were dried using a Dorrex dryer at 50°C for approximately 12 h and deposited in the Herbarium of the College of Life Science, Capital Normal University, Beijing, China (BJTC). Other 20 herbarium specimens from the research area were obtained from the HMAS.

To observe microscopic characters, thin sections of the dried material were mounted in 3% KOH or sterilized water. Then, the materials were stained with 1% Congo red to increase the visibility of the structures. Microscopic features (e.g., basidiospores, pileipellis, and volval remnants) were observed and measured under a light microscope (Olympus DP71, Tokyo, Japan). Basidiospore measurements of new species are presented as (a)b − c(d). Among them, b − c, a, and d represent a minimum of 90% of the measured values, minimum extreme values, and maximum extreme values, respectively. Q represents the length/width ratio of the basidiospores, and Qm is the average Q values of all basidiospores measured (Bas, 1969). Qm values ± sample standard deviations are provided. The descriptive terms are in accordance with Cui et al. (2018).

 DNA extraction, and PCR amplification and sequencing

DNA was extracted using the M5 Plant Genomic DNA Kit (Mei5 Biotechnology, Co., Ltd., Beijing, China). The extracted DNA was solubilized in 1× TE buffer/sterile water and stored at −20°C for further use. The following primer sets were used for amplification: nrITS1f/nrITS4 (White et al., 1990; Gardes and Bruns, 1993) for the nuclear ribosomal DNA internal transcribed spacer (nrITS rDNA) region, LR5/LR0R (Vilgalys and Hester, 1990) for the large subunit nuclear rDNA (nrLSU rDNA) region, rpb2-6f/rpb2-7r (Cai et al., 2014) for the second largest subunit of the RNA polymerase II (rpb 2) region, Am-b-tubulin F/Am-b-tubulin R (Cai et al., 2014) for the beta-tubulin (β-tubulin) region, and EF1-983F/EF1-1567R (Rehner and Buckley, 2005) for the translation elongation factor 1-α (tef1-α) region. PCRs were performed in a reaction volume of 25 μL. The obtained DNA was subjected to Sanger dideoxy sequencing (Sangon Biotechnology, Co., Ltd, Shanghai, China). The new sequences obtained in this study were deposited in GenBank (https://www.ncbi.nlm.nih.gov). Table 1 lists the accession numbers of the sequences used for phylogenetic analysis.

Information of sequences used in the nrITS-nrLSU-rpb2-tef1-α phylogenetic analysis in this study.

Taxa Voucher Location GenBank accession numbers
nrITSi nrLSU rpb2 tef1-α
Amanita cf. angustilamellata HKAS89451 Mount Taishan, Shandong, China MH508292 MH486431 MH485910 MH508716
Amanita cf. angustilamellata HKAS83453 Puer, Yunnan, China — MH486430 — —
Amanita albidostipes HKAS57358(T) Baoshan, Yunnan, China MH508500 MH486756 — —
Amanita alboradicata MHHNU10531 (T) Jilin, China MW016759 MW016757 MW546614 MW546620
Amanita altipes HKAS91125 Ganzi, Sichuan, China MH508254 MH486367 MH485862 MH508669
Amanita aporema FRI62674 Malaysia KU714575 KU714551 KU714593 KU714538
Amanita ballerina OR1014 Thailand KY747466 — KY656883 —
Amanita ballerina OR1026(T) Thailand KY747467 — KY656884 —
Amanita battarrae HKAS92090 Mount Changbai, Jilin, China MH508266 MH486388 MH485880 MH508689
Amanita bisporigera RET 377-9 Tennessee, USA KJ466374 KJ466434 — KJ481936
Amanita borealis BJTC Z110 Changping, Beijing, China OR058499 OR042383 OR051502 OR051531
Amanita borealis BJTC L169 (T) Pinggu, Beijing, China OR058500 OR042384 OR051503 OR051532
Amanita breckonii NY00066695 (T) USA — KJ535440 — —
Amanita brunneitoxicaria BZ2015-01 Thailand NR_151655 — KY656879 —
Amanita brunneitoxicaria BZ2015-02 Thailand KY747463 — KY656880. —
Amanita aff. brunneofuliginea MB-000633 (duplicate HKAS84873) Germany MH508252 MH486364 MH485861 MH508667
Amanita brunneofuliginea HKAS89226 Daofu, Sichuan, China MH508269 MH486393 MH485885 MH508693
Amanita brunneola BJTC Z087 Changping, Beijing, China OR058501 OR042385 — OR051533
Amanita brunneola BJTC C650 (T) Miyun, Beijing, China OR058502 OR042386 OR051504 OR051534
Amanita brunneomaculata HKAS68393 Lijiang, Yunnan, China MH508278 MH486410 MH485892 MH508698
Amanita brunneostrobilipes HKAS60291 (T) Hainan, China MH508281 MH486415 – MH508703
Amanita caesarea HKAS96166 Italy MH508283 MH486418 MH485898 MH508705
Amanita caesareoides HKAS71021 Japan MH508284 MH486419 MH485899 MH508706
Amanita caesareoides HKAS92009 Benxi, Liaoning, China MH508285 MH486421 MH485901 MH508708
Amanita caesareoides BJTC 630 Yanqing, Beijing, China OR058503 OR042387 OR051505 OR051535
Amanita caesareoides BJTC C654 Xinglong, Hebei, China OR058504 OR042388 OR051506 OR051536
Amanita caojizong HKAS79673 (T) Yunnan, China MH508291 MH486429 MH485908 MH508714
Amanita castanea MFLU 15-1424 Thailand KU904823 KU877539 — —
Amanita changtuia HKAS92100 Mudanjiang, Heilongjiang, China MH508299 MH486442 MH485919 MH508724
Amanita chepangiana HKAS56718 Yunnan, China KU714569 KU714545 KU714588 KU714534
Amanita chiui HKAS76328 Yanyuan, Sichuan, China MH508303 MH486447 MH485930 MH508727
Amanita chiui BJTC L130 Pinggu, Beijing, China OR058505 OR042389 — OR051537
Amanita cinctipes HKAS78465 Guangdong, China MH508305 MH486449 – –
Amanita cinctipes HKAS101388 Heishiding, Guangdong, China MH508304 MH486448 — —
Amanita cingulata HKAS75600 (T) Yanling, Zhuzhou, China MH508310 NG_058602 — —
Amanita citrina HKAS101397 France MH508311 MH486456 MH485936 MH508732
Amanita citrinoannulata HKAS83459 (T) Chongqing, China MH508318 MH486464 MH485944 MH508740
Amanita citrinoindusiata HKAS100522 (T) Lijiang, Yunnan, China MH508320 MH486468 MH485947 MH508744
Amanita collariata MHHNU31095 Hunan, China OM955206 OM955204 OM949814 OM949813
Amanita concentrica HKAS87061 Yunnan, China MH508327 KR824785 KR824794 KR824827
Amanita cruzii BARONI8998 (T) Dominican Republic KC855222 KC855222 — MH508750
Amanita detersa HKAS71476 (T) Lushui, Yunnan, China MH508328 MH486475 MH485954 MH508752
Amanita eijii HKAS70229 Yunnan, China MH508333 MH486484 MH485963 MH508761
Amanita elata HKAS83449 Xishuangbanna, Yunnan, China MH508334 MH486486 MH485965 MH508763
Amanita elliptica HKAS79602 Hainan. China MH508335 MH486487 – MH508764
Amanita elongata RET 384-5 Canada MH508337 MH486489 MH485967 MH508766
Amanita esculenta HKAS89035 Xishuangbanna, Yunnan, China MH508338 MH486490 MH485968 MH508767
Amanita exitialis HKAS75774 Mount Baiyun, Guangdong, China JX998027 JX998052 KJ466591 JX998001
Amanita farinosa HKAS67958 Tengchong, Yunnan, China MH508341 MH486498 MH485973 MH508773
Amanita flavipes HKAS57650 Dali, Yunnan, China MH508343 MH486501 MH485975 MH508776
Amanita flavoconia RET 485-6 New York, USA MH508351 MH486514 MH485985 MH508790
Amanita flavopantherina HKAS82613 Shangri-La, Yunnan, China MH508355 MH486519 MH485989 MH508795
Amanita fritillaria HKAS91952 Xishuangbanna, Yunnan, China MH508367 MH486548 MH486015 MH508820
Amanita frostiana RET7-25-92-E USA AF024453
Amanita fuliginea HKAS75780 Heishiding, Guangdong, China JX998023 JX998048 KJ466595 JX997995
Amanita fuligineoides HKAS52727 Hunan, China JX998024 JX998047 KJ466599
Amanita fulva HKAS96168 Austria MH508371 MH486555 MH486022 MH508826
Amanita fulva HKAS96168 Australia MH508371 MH486555 MH486022 MH508826
Amanita fulvopyramidalis MHHNU10581 Zhejiang, China – MW471097 MW546619 MW546625
Amanita fulvopyramidalis MHHNU8814 (T) Hunan, China MT878220 MT878535 MW546617 MW546623
Amanita fuscoflava HKAS59800 (T) Limushan, Hainan, China MH508372 MH486557 MH486023 MH508827
Amanita fuscosquamosa PDD92862 Australia MH508373 MH486558 — MH508828
Amanita gemmata C. Bas8942 L Unknown AF024457
Amanita griseofarinosa HKAS80926 Hubei, China MH508375 MH486559 MH486025 MH508830
Amanita griseofolia HKAS38159 (T) Yunnan, China AY436448 AY436488 — —
Amanita griseopantheirna HKAS83560 (T) Basu, Tibet, China MH508385 MH486573 — MH508842
Amanita griseorosea HKAS77332 (T) Mount Limu, Hainan, China KJ466411 KJ466474 — KJ481992
Amanita griseoumbonata HKAS92103 (T) Mudanjiang, Heilongjiang, China MH508389 MH486578 MH486040 MH508847
Amanita gymnopus HKAS71618 Yunnan, China MH508393 MH486582 MH486044 MH508851
Amanita hamadae HKAS79076 Heishiding, Guangdong, China MH508394 MH486584 MH486046 —
Amanita heishidingensis HKAS76122 Guangdong, China NR_151651 KC429045 – –
Amanita heishidingensis HKAS81484 Guangdong, China KJ922999 KJ922993 MH486049 MH508854
Amanita hemibapha HMAS 54784 Ninger, Yunnan, China — MH486587 — —
Amanita hunanensis HKAS100632 Jinzhai, Anhui, China MH508396 MH486588 MH486050 MH508856
Amanita ibotengutake HKAS56045 Xifeng, Jilin, China MH486589 MH508857
Amanita imazekii HKAS92011 Benxi, Liaoning, China MH508398 MH486592 MH486053 MH508860
Amanita incarnatifolia HKAS100593 Anhui, China MH508400 MH486594 MH486056 MH508862
Amanita japonica TMI26147 Japan KJ922994 KJ922990 – –
Amanita javanica FRIM:61503 Malaysia KU714572 KU714548 — KU714536
Amanita kitamagotake EN-4 Japan AB721450 AB721450 — —
Amanita kotohiraensis HKAS100577 Anhui, China MH508415 MH486614 MH486068 MH508874
Amanita lippiae RET418-2 Brazil NR_154692 KP258992 — —
Amanita liquii HKAS36611 Yunnan, China AY436462 AY436493 — —
Amanita longistriata HKAS68331 Lanping, Yunnan, China MH508428 MH486631 MH486081 MH508888
Amanita longistriata HKAS54578 Lijiang, Yunnan, China — MH486629 MH486079 —
Amanita melleialba HKAS83446 (T) Puer, Yunnan, China MH508430 KR824767 KR824792 KR824813
Amanita melleiceps HKAS80145 Macheng, Hubei, China MH508432 MH486640 MH486089 MH508897
Amanita minutisquama HKAS100504 (T) Hainan, China MH508435 MH486644 MH486095 MH508902
Amanita mira HKAS91953 Mengla, Yunnan, China MH508437 MH486646 MH486097 —
Amanita molliuscula HKAS75555 Shennongjia, Hubei, China KJ466408 KJ466471 KJ466638 KJ481973
Amanita molliuscula HKAS77324 (T) Zhouzhi, Shaanxi, China KJ466409 KJ466472 KJ466639 KJ481974
Amanita molliuscula HMJAU20469 Mount Changbai, Jilin, China KJ466410 KJ466473 KJ466640 KJ481975
Amanita muscaria HKAS61888 Huzhong, Heilongjiang, China MH508439 MH486651 MH486100 MH508908
Amanita muscaria HMJAU4549 Kyrov, Russia MH508440 KR824787 — KR824829
Amanita muscaria BJTC L491 Chicheng, Hebei, China OR058506 OR042390 — OR051538
Amanita neoovoidea HKAS89025 Yunnan, China MH508445 MH486656 MH486106 MH508913
Amanita oberwinkleriana HKAS77330 Hainan, China KJ466380 KJ466441 KJ466606 KJ481946
Amanita oberwinkleriana BJTC C659 Pinggu, Beijing, China — OR042391 OR051507 OR051539
Amanita oberwinkleriana BJTC Z328 Jizhou, Tianjin, China — OR042392 — OR051540
Amanita oberwinkleriana BJTC Z318 Jizhou, Tianjin, China — OR042393 OR051508 OR051541
Amanita oberwinkleriana BJTC Z862 Jizhou, Tianjin, China — OR042394 — OR051542
Amanita oberwinkleriana BJTC C320 Xinglong, Hebei, China — OR042395 — OR051543
Amanita oberwinkleriana HMAS 263406 Yanqing, Beijing, China OR058507 — — —
Amanita oberwinkleriana HMAS 253800 Mentougou, Beijing, China OR058508 OR042396 OR051509 OR051544
Amanita oberwinkleriana HMAS 253801 Mentougou, Beijing, China — OR042397 OR051510 OR051545
Amanita oberwinkleriana HMAS 253802 Mentougou, Beijing, China — — OR051511 —
Amanita oberwinkleriana HMAS 253796 Mentougou, Beijing, China OR058509 OR042398 OR051512 OR051546
Amanita ochracea HKAS87986 Shangri-La, Yunnan, China MH508454 MH486686 MH486123 —
Amanita ocreata HKAS79686 California, USA KJ466381 MH486688 KJ466607 KJ481947
Amanita olivaceofusca HKAS97581 (T) Lancang, Yunnan, China MH508457 MH486691 MH486127 —
Amanita orienticrocea HKAS90455 (T) Jingdong, Yunnan, China MH508465 MH486701 MH486133 —
Amanita orientifulva HKAS87937 Shangri-La, Yunnan, China MH508468 MH486704 MH486136 —
Amanita orientigemmata HKAS80978 Lanping, Yunnan, China MH508469 MH486708 MH486140
Amanita orsonii HKAS52264 Kunming, Yunnan, China MH508474 MH486714 MH486145 —
Amanita ovalispora HKAS79625 Heishiding, Guangdong, China MH508479 MH486722 MH486150 —
Amanita ovalispora HKAS101406 Hainan, China MH508478 MH486720 MH486148 —
Amanita ovalispora BJTC Z057 Changping, Beijing, China — OR042399 OR051513 OR051547
Amanita ovalispora BJTC Z311 Jizhou, Tianjin, China OR058510 OR042400 OR051514 OR051548
Amanita pachycolea HKAS101422 USA MH508480 MH486724 MH486152 —
Amanita pallidocarnea HKSA97678 Lancang, Yunnan, China MH508482 MH486728 MH486156 —
Amanita pallidorosea HKAS82350 Mount Taishan, Shandong, China MH508485 MH486737 MH486163 —
Amanita pallidorosea HKAS61937 Meixian, Shaanxi, China KJ466382 KJ466443 KJ466609 KJ481949
Amanita pallidorosea HKAS82350 Shandong, China MH508485 MH486737 MH486163 MH508971
Amanita pallidorosea HKAS82350 Shandong, China MH508485 MH486737 MH486163 MH508971
Amanita pantherina MB-102863 (duplicate HKAS84852) Germany MH508488 MH486743 MH486167 —
Amanita parvifritillaria HKAS83737 (T) Ailaoshan, Yunnan, China MH508494 MH486749 MH486173 —
Amanita parvipantherina HKAS67828 Yunnan, China MH508496 MH486750 MH486174 MH508979
Amanita persicina RET151-4 USA NR_154668 EU071969 EU071862
Amanita phalloides HKAS75773 California, USA JX998031 JX998060 KJ466612 JX998000
Amanita porphyria HKAS92088 Changbai Mountain, Jilin, China MH508506 MH486761 MH486180 —
Amanita aff. princeps TRTC-150309 Thailand JX844734 KF877274 — KF877160
Amanita princeps FRI:62849 Malaysia KU714576 KU714552 KU714594 KU714539
Amanita pseudopantheirna HKAS80007 (T) Binchuan, Yunnan, China MH508514 MH486777 MH486191 —
Amanita pseudoporphyria HKAS89074 Yunnan, China MH508524 MH486785 MH486199 MH509012
Amanita pseudoprinceps HKAS97523 (T) Heishiding, Guangdong, China MH508527 MH486788 MH486202 —
Amanita pseudosychnopyramis HKAS87999 (T) Lijiang, Yunnan, China MH508530 KR824778 KR824790 KR824824
Amanita pseudovaginata HKAS70138 Mount Wuliang, Yunnan, China MH508531 MH486791 MH486205 —
Amanita pyramidata HKAS87943 (T) Yunnan, China MH508535 MH486795 MH486209 MH509021
Amanita regalis HKAS56699 Czech Republic MH508537 MH486797
Amanita retenta HKAS70020 (T) Lijiang, Yunnan, China MH508543 MH486802 MH486215 —
Amanita rimosa HKAS75777 Baisha, Hainan, China JX998018 JX998044 KJ466615 JX998005
Amanita roseitincta RET 284-10 Texas, USA MH508550 KC855224 — —
Amanita roseolifolia HKAS101403 (T) Hainan, China MH508548 MH486807 MH486219 MH509032
Amanita rubescens HKAS100631 Anhui, China MH508554 MH486810 – MH509037
Amanita rubescens HKAS92034 Anshan, Liaoning, China MH508558 MH486815 MH486226 —
Amanita rubiginosa HKAS52216 (T) Yunnan, China MH508561 MH486817 MH486229 MH509045
Amanita rubroflava HKAS83089 (T) Lingbaoshan, Yunnan, China MH508568 MH486827 MH486238 —
Amanita rubrovolvata BZ2015-68 Thailand KY747465 KY747477 KY656882
Amanita rufoferruginea HKAS79616 Guangdong, China MH508579 MH486842 MH486252 MH509068
Amanita sepiacea HKAS56799 Baoshan, Yunnan, China MH508584 MH486847 MH486256 —
Amanita shennongjiana HKAS75553(T) Shennongjia, Hubei, China MH508590 MH486862 MH486270 —
Amanita siamensis HKAS83680 Lingbaoshan, Yunnan, China — MH486865 MH486272 —
Amanita similis FRI:3740 Malaysia KU714566 JF710796 — KU714531
Amanita sinensis HKAS74388 Xundian, Kunming, Yunnan MH508596 MH486870 MH486277 —
Amanita sinocitrina HKAS100530 Suiyang, Guizhou, China MH508598 MH486873 MH486279 —
Amanita sp. HKAS101390 Heishiding, Guangdong, China MH508603 MH486878 MH486284 MH509100
Amanita sp. BJTC S233 Yanqing, Beijing, China OR058511 OR042401 OR051515 OR051549
Amanita sp. HMAS 26491 Mentougou, Beijing, China OR058512 OR042402 — OR051550
Amanita spissa HKAS92089 Changbai mountain, Jilin, China MH508612 MH486893 MH486296 MH509115
Amanita spissacea HKAS57649 Dali, Yunnan, China KJ466373 KJ466480 KJ466645 KJ481980
Amanita spissacea HKAS57754 Yulong, Yunnan, China MH508609 MH486886 MH486290 MH509108
Amanita squarrosipes HKAS76359 (T) Muli, Sichuan, China MH508613 MH486894 MH486297 MH509116
Amanita suballiacea RET 490-1 Connecticut, USA KJ466420 KJ466485 KJ466601 KJ481941
Amanita subfrostiana HKAS58750 Sichuan, China – MH486897 MH486299 MH509119
Amanita subfrostiana HKAS57042 Yunnan, China JN943173 JN941162 JQ031118 KJ482003
Amanita subfuliginea HKAS77326 Lechang, Guangdong, China KJ466404 KJ466467 KJ466636 KJ481971
Amanita subglobosa HMAS 253798 Miyun, Beijing, China OR058513 OR042403 OR051516 OR051551
Amanita subglobosa HKAS67914 Baoshan, Yunnan, China MH508619 MH486902 MH486303 MH509123
Amanita subjunquillea HKAS75770 Meixian, Shaanxi, China JX998034 JX998062 KJ466653 JX997999
Amanita subjunquillea HKAS75771 Shennongjia, Hubei, China JX998032 JX998063 KJ466654 JX997997
Amanita subjunquillea BJTC 217 Yanqing, Beijing, China OR058514 OR042404 OR051517 OR051552
Amanita subjunquillea BJTC 085 Yanqing, Beijing, China OR058515 OR042405 — —
Amanita subjunquillea BJTC C558 Huairou, Beijing, China OR058516 OR042406 OR051518 OR051553
Amanita subjunquillea BJTC 033 Yanqing, Beijing, China OR058517 OR042407 OR051519 OR051554
Amanita subjunquillea BJTC 704 Yanqing, Beijing, China OR058518 OR042408 — —
Amanita subjunquillea BJTC Z276 Xinglong, Hebei, China OR058519 OR042409 OR051520 OR051555
Amanita subjunquillea BJTC 112 Yanqing, Beijing, China OR058520 OR042410 — OR051556
Amanita subjunquillea BJTC Z172 Shunyi, Beijing, China OR058521 OR042411 OR051521 OR051557
Amanita subjunquillea HMAS 253775 Mentougou, Beijing, China OR058522 OR042412 OR051522 OR051558
Amanita submembranacea MB-001174 (duplicate HKAS84857) Germany MH508626 MH486916 — MH509135
Amanita subpallidorosea HKAS77350 Taichung, Taiwan, China KJ466400 KJ466462 KJ466631 KJ481966
Amanita subpallidorosea LHJ140923-41 (T) Guizhou, China KP691683 KP691692 KP691701 KP691670
Amanita subparcivolvata MHHNU32907 (T) Hunan, China OM955207 OM955205 OM949819 —
Amanita subparcivolvata MHHNU33169 Hunan, China OM955215 OM955217 OM949820 —
Amanita subparvipantherina HKAS56986 (T) Yongping, Yunnan, China KR824776 KR824820
Amanita sychnopyramis f. subannulata HKAS26144 Yunnan, China — AF024480 — —
Amanita sychnopyramis f. sychnopyramis HKAS83454 Wenshan, Yunnan, China — MH486927 — MH509144
Amanita timida HKAS83228 Guangdong, China MH508636 MH486930 MH486323 MH509147
Amanita torrendii LOU Fungi 17408 Spain GQ925386 GQ925369 — —
Amanita umbrinolutea HKAS89201 Luhuo, Sichuan, China MH508637 MH486933 MH486326 MH509150
Amanita vaginata var. vaginata HMAS 253281 Mentougou, Beijing, China OR058523 OR042413 OR051523 —
Amanita vaginata var. vaginata BJTC Z521 Huairou, Beijing, China OR058524 OR042414 — —
Amanita vaginata var. vaginata BJTC 682 Yanqing, Beijing, China OR058525 OR042415 — —
Amanita vaginata var. vaginata BJTC 677 Yanqing, Beijing, China OR058526 OR042416 OR051524 —
Amanita vaginata var. vaginata H.A.v.d.Aa s.n Unknow — AF024482 — —
Amanita velatipes RET 489-2 Michigan, USA MH508643 MH486938 MH486331 MH509155
Amanita vestita HKAS79687 Hainan, China MH508647 KJ466494 KJ466662 KJ481995
Amanita virgineoides HKAS100518 Henan, China MH508648 MH486944 MH486339 MH509165
Amanita virosa HKAS56694 Juva, Finland JX998030 JX998058 KJ466664 JX998007
Amanita cf. xanthogala HKAS84707 Yunnan, China MH508298 MH486441 MH485918 MH508723
Amanita yanshanensis BJTC Z083 Changping, Beijing, China OR058527 OR042417 OR051525 OR051559
Amanita yanshanensis BJTC Z760 Changping, Beijing, China OR058528 OR042418 OR051526 OR051560
Amanita yanshanensis BJTC Z049 (T) Changping, Beijing, China OR058529 OR042419 OR051527 OR051561
Amanita yanshanensis BJTC Z824 Changping, Beijing, China OR058530 OR042420 — OR051562
Amanita yanshanensis BJTC C182 Pinggu, Beijing, China OR058531 OR042421 — OR051563
Amanita yanshanensis BJTC Z815 Changping, Beijing, China OR058532 OR042422 OR051528 OR051564
Amanita yanshanensis BJTC Z819 Changping, Beijing, China OR058533 OR042423 OR051529 OR051565
Amanita yanshanensis BJTC Z820 Changping, Beijing, China OR058534 OR042424 OR051530 OR051566
Amanita yenii HKAS87047 Yunnan, China MH508652 MH486951 MH486344 MH509171
Amanita yuaniana HKAS58807 Lijiang, Yunnan, China MH508653 MH486954 MH486347 MH509174
Limacella glioderma HKAS90169 Heilongjiang, China MH508658 KT833808 KT833823 KT833836
Limacellopsis Asiatica HKAS76497 Gansu, China — KT833811 KT833826 KT833839
Limacellopsis Asiatica HKAS82561 (T) Aba, Sichuan, China — KT833812 KT833827 KT833840
Myxoderma ochraceoluteum MEL2305332 Australia MH508660 MH486965 MH486358 MH509185
Myxoderma ochraceoluteum MEL2341329 Australia MH508661 MH486966 MH486359 MH509186

The new generated sequences are emphasized in bold; “–” show no sequence.

The new species is in bold. “—” means no sequence.

 Molecular data analyses and species delimitation

The nrITS-nrLSU-rpb2-tef1-α multi-locus dataset included 216 ingroup samples. These samples were used to infer the phylogenetic status of our Amanita specimens at the phylloclade level. The nrLSU dataset included 204 ingroup samples. These samples were used to analyze the subsection in which the species were located. Furthermore, nrITS was used to infer phylogenetic relationships between new and known Amanita species, as GenBank contains a large amount of nrITS sequence data for this genus. These nrITS sequences of new species were divided into different datasets, given that these sequences are too variable to obtain reliable genus-wide comparisons. Based on previous study findings and the GenBank database of the National Center for Biotechnology Information, reference sequences of all Amanita species in the dataset were selected for phylogenetic analysis (Cui et al., 2018; Su et al., 2022, Table 1 ). Limacella glioderma (Fr.) Maire (HKAS 90169), Limacellopsis asiatica Zhu L. Yang, Q. Cai & Y.Y. Cui (HKAS 76497 and 82561), Myxoderma ochraceoluteum (P.D. Orton) Zhu L. Yang, Q. Cai & Y.Y. Cui (MEL 2305332), and M. ochraceoluteum (MEL2341329) as outgroup refers to Cui et al. (2018).

All sequences were compared using MAFFT v.6 (Katoh and Toh, 2010) and trimmed automatically using Gblocks 0.91b (http://phylogeny.lirmm.fr/phylo_cgi/one_task.cgi?task_type=gblocks) (Dereeper et al., 2008). Bayesian inference (BI) analysis was performed using MrBayes v.3.1.2 (Ronquist and Huelsenbeck, 2003), and maximum likelihood (MI) analysis gene trees were estimated using RAxML 7.4.2 Black Box (Stamatakis, 2006; Stamatakis et al., 2008).

The BI analysis was performed using a Markov chain Monte Carlo (MCMC) algorithm (Rannala and Yang, 1996) and MrBayes 3.1.2 (Ronquist and Huelsenbeck, 2003) based on the best substitution model determined by MrModeltest 2.3 (Nylander, 2004), GTR + I + G for nrITS, nrLSU, rpb2, and tef-1α. Two MCMC chains were run from random trees for 10,000,000 generations, stopping when the average standard deviations of split frequencies were less than 0.01. Trees were stored for each 1,000 generations. The first 25% of the trees were excluded as the burn-in stage for each analysis. Branches with significant Bayesian posterior probability (BPP) values were then estimated in the resulting trees (Posada and Crandall, 1998). The ML analysis was performed using a GTR + GAMMA + I locus replacement model (Guindon et al., 2010). The branch support was obtained using the bootstrapping (BS) method of 1,000 replications (Hillis and Bull, 1993). Branches with a bootstrap (BS) support of ≥50% and BPP of ≥0.95 were considered significant (Hillis and Bull, 1993).

 Results Phylogenetic analyses

The nrLSU dataset contained 209 sequences, including 42 newly obtained sequences. The length of the aligned dataset was 790 bp long. The nrLSU phylogenetic analysis results revealed that our specimens belonged to six sections under Amanita, namely, sections Amanita Pers., Caesareae Singer, Roanokenses Singer, Phalloideae Quél., Validae Quél, and Vaginatae Quél ( Figure S1 ), and were divided into 12 clades. Subsequently, the nrITS-nrLSU-rpb2-tef1-α multi-locus phylogenetic analysis was performed to infer the phylogenetic status of our Amanita specimens at the phylloclade level ( Figure 2 ). The combined nrITS-nrLSU-rpb2-tef1-α dataset had 726 sequences, including 143 newly obtained sequences in this study. The aligned dataset was 2,150 bp long including alignment gaps (245 bp for nrITS, 790 bp for nrLSU, 660 bp for rpb2, and 365 bp for tef1-α). Results of the nrITS-nrLSU-rpb2-tef1-α and nrLSU phylogenetic analyses revealed that the subgenera and sections proposed by Cui et al. (2018) were strongly supported with significant BPP values and ML bootstrap (MLB).

The nrITS-nrLSU-rpb2-tef1-α multi-locus phylogenetic tree obtained from the Bayesian analysis.

The nrITS-nrLSU-rpb2-tef1-α multi-locus phylogenetic analysis showed that our specimens were clustered into 14 clades. Moreover, they formed three distinct and strongly supported new branches, which were nested in sections Amanita, Caesareae, and Validae, respectively. These three new lineages were as follows: two specimens (BJTC Z110 and BJTC L169) formed one clade (BPP = 1.00, MLB = 100%) and were closely related to A. griseopantherina Yang-Yang Cui, Qing Cai & Zhu L. Yang, A. pantherina (DC.) Krombh., and A. subglobosa Zhu L. Yang on Figure 2 . The two specimens (BJTC Z087 and BJTC C650) clustered into a branch with a high support (BPP = 1.00, MLB = 100%), which further clustered into a clade containing A. longistriata S. Imai., with moderate support. Eight specimens (BJTC Z049, BJTC Z820, BJTC Z083, BJTC Z824, BJTC Z760, BJTC Z819, BJTC C182, and BJTC Z815) were clustered together, forming a completely supported clade (BPP = 0.99, MLB = 99%). The new branches clustered with A. spissacea S. Imai and formed a sister clade in the phylogenetic tree. The nrLSU phylogenetic analysis revealed topologies similar to those of the multi-locus phylogenetic tree, and the specimens also formed three new lineages. Therefore, based on the results of phylogenetic and morphological analyses, these new clades were identified as three new species herein.

In addition, through morphological and phylogenetic analyses, we identified nine known Amanita species collected from Yanshan Mountains, northern China, including A. caesareoides Lj. N. Vassiljeva, A. chiui Yang-Yang Cui, Qing Cai & Zhu L. Yang, A. muscaria (L.: Fr.) Lam., A. oberwinkleriana Zhu L. Yang & Yoshim. Doi, A. ovalispora Boedijn, A. virosa Bertillon, A. subglobosa Zhu L. Yang, A. subjunquillea S. Imai, and A. vaginata var. vaginata (Bull.) Lam. Because we could not obtain the sequence information of the herbarium specimens (HMAS 40501 and HMAS 40503) of A. virosa, we could only identify them through morphological observation.

Although the specimen BJTC S233 was clustered with A. cf. angustilamellata (HKAS 89451 and HKAS 83453) in the nrITS-nrLSU-rpb2-tef1-α and nrLSU phylogenetic analyses, additional specimens are required to elucidate its phylogenetic position and morphological characters. The specimens BJTC C654 and HMAS 26491 probably represented undescribed species. However, they could not be described in the present study because of the poor condition of their basidiomata, inadequate number of specimens, and uncertain phylogenetic position in the nrITS-nrLSU-rpb2-tef1-α phylogenetic analysis. Therefore, we only temporarily termed them as Amanita sp.

The numbers above the branches represent strong support (BPP ≥0.95 and/or MLB ≥50%). Red font represents t the location of the newly acquired sequences. Table 1 presents the accession numbers of sequence information used.

 Taxonomy

Based on our phylogenetic and morphological data, three new species and nine known species of Amanita from Yanshan Mountains are described below.

<italic>Amanita borealis</italic> H. Zhou & C. L. Hou, sp. nov.

Figures 3A, B , 4

Fresh basidiomata of new species of Amanita in this study. (A, B) Amanita borealis sp. nov. (type, BJTC L169); (C) Amanita brunneola sp. nov. (type, BJTC C650); (D) Amanita brunneola sp. nov. (BJTC Z087); (E, F) Amanita yanshanensis sp. nov. (type, BJTC Z049). Bars: (A–E) = 2 cm.

Microscopic characters of Amanita borealis (type, BJTC L169). (A). Hymenium and subhymenium; (B). Basidiospores; (C). Longitudinal section of volval remnants on pileus. Bars: (A, B) = 10 μm, (C) = 40 μm.

MycoBank: MB 847659

Etymology: The specific epithet “borealis” means “northern,” referring to the native occurrence of this species in the Northern China.

Type: CHINA. Beijing, Pinggu district, Dongniujiaoyu village, 40.323599 N, 117.156964 E, alt. 477 m, 19 Aug., 2020, coll. G.Q.C., C.L.H. and Y.T.Z. (L169/BJTC L169).

Basidiomata small- to medium-sized. Pileus 2–6 cm in diam., plano-convex to applanate, lacking an obvious depression or umbo at the center; surface brownish (#915b25), brown (#7d4e20) to dark brown (#402810), darker at the center, pyramidal, subverrucose to subconical, dirty white (#f2f2f2) to white (#ffffff) volval remnants on the pileus (ca. 2–8-mm diam.), densely arranged over the disk; margin slightly striate (ca. 0.05–0.1 R), non-appendiculate; trama white (#ffffff), unchanging. Free, crowded, white (#ffffff) lamellae; truncated, plentiful lamellulae. Stipe 5–10 cm long, 0.4–1.1 cm wide at the apex, subcylindric or slightly attenuate upward, surface white (#ffffff), with silk luster, glabrous or covered with concolorous, floccose squamules; white (#ffffff) to yellowish (#ffffe7) context; basal bulb subglobose to fusiform (1.5–2.5-cm diam.), white (#ffffff) to yellowish (#ffffe7); floccose volval remnants on the stipe base, arranged in belts on the lower part of the stipe, and often forms a collar-like or shortly limbate volva on limit between the stipe and basal bulb, white (#ffffff) to yellowish (#ffffe7). Annulus apical, subapical to fugacious, thick, with white (#ffffff) and silk luster on the upper surface. Spore print not observed. Odor indistinct.

Bilateral lamellar trama, 20–60-μm-wide mediostratum, composed of abundant subfusiform, ellipsoid to clavate inflated cells (20–80 × 12–50 μm); abundant, 2–9-μm-wide filamentous hyphae; scarce vascular hyphae. Lateral stratum is composed of abundant subfusiform to ellipsoid inflated cells (20–45 × 6–25 μm), diverging at an angle of ca. 30°–45° to the mediostratum; abundant, 3–8-μm-wide filamentous hyphae. A 30–60-μm-thick subhymenium, with two to three layers of ovoid, subglobose, fusiform, ellipsoid, or irregular cells (10–30 × 8–20 μm). Basidia (30–50 × 7.5–14 μm), slenderly clavate, four-spored, with clamps, hyaline; 3–5-μm-long sterigmata; basidiospores [60/2/2] (8.0–)8.7–9.6(–11.5) × (6.0–)6.5–8.2(–8.5) μm, Q = (1.10–)1.22–1.45(–1.51), Qm = 1.31 ± 0.10, broadly ellipsoid to ellipsoid, thin-walled, hyaline, pale yellow, smooth, small apiculus, inamyloid; sterile lamellar edge, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (15–40 × 10–30 μm), single and terminal or in chains of 2–3, thin-walled, hyaline; abundant, 3–9-μm-wide filamentous hyphae, irregularly arranged or ± running parallel to the lamellar edge. Pileipellis 50–200 μm thick, gelatinized upper layer (30–75 μm thick), composed of radially, thin-walled, colorless, 2–8-μm-wide filamentous hyphae; lower layer (40–100 μm thick) composed of radially and compactly arranged, colorless, 2–8-μm-wide filamentous hyphae; scarce vascular hyphae. Volval remnants on the pileus are composed of somewhat vertically to irregularly arranged elements: 3–7-μm-wide scarce to scattered, subcolorless, thin-walled, branching, anastomosing filamentous hyphae; globose, subglobose, fusiform to ellipsoid, sometimes irregular, inflated cells (30–70 × 10–55 μm) that are nearly colorless, slightly thick-walled, and terminal or in chains of 2–3; scarce vascular hyphae. Volval remnants on the stipe base similar to that on the pileus, but with more abundant filamentous hyphae and fairly abundant vascular hyphae. Longitudinally acrophysalidic stipe trama; acrophysalides (50–300 × 15–50 μm); scattered to abundant, 3–15-μm-wide filamentous hyphae. Annulus is composed of radially arranged elements: abundant, subglobose, fusiform to ellipsoid inflated cells (20–40 × 10–30 μm) that were hyaline and thin-walled; abundant, 2–7-μm-wide filamentous hyphae that were hyaline and thin-walled. Clamps present in all parts of basidioma.

Habitat and distribution: This species is scattered in the broad-leaved forests of Q. mongolica Fisch. Ex Ledeb. Basidioma occurs in summer and autumn.

Additional specimens examined: CHINA. Beijing, Changping district, Beitaizi, 40.272906 N, 116.420298 E, alt. 149 m, 15 Aug., 2019, coll. H.Z., X.Y.S. and Y.T.Z. (ZH110/BJTC Z110).

Commentary: A. borealis is well circumscribed by its brownish to dark-brown pileus densely covered with pyramidal, subverrucose to subconical, floccose volval remnants on the stipe base arranged in incomplete belts, and broadly ellipsoid to ellipsoid basidiospores (8.0–11.5 × 6.0–8.5 μm). Furthermore, it is found in association with Fagaceae (Quercus mongolica) trees.

A. borealis belongs to section Amanita and is closely related to A. griseopantherina, A. pantherine, and A. subglobosa on the multi-locus and nrITS phylogenetic trees ( Figures 2 , S2 ). In addition, A. borealis, A. subglobosa, and A. pantherine have similar morphologies. A. pantherine is a species originally described from Europe but is not found in China. It has a relatively lower annulus, narrower basidiospores, and no clamps (Cui et al., 2018). From the morphological viewpoint, A. subglobosa can be separated from A. borealis based on its relatively darker-colored pileus, usually with the presence of clamps and wider basidiospores (8.5–12.0 × 7.0–9.5 μm) (Yang, 1997; Yang, 2005; Yang, 2015; Cui et al., 2018).

 <italic>Amanita brunneola</italic> H. Zhou & C. L. Hou, sp. nov.

Figures 3C, D , 5

Microscopic characters of Amanita brunneola (type, BJTC C650). (A). Hymenium and subhymenium; (B). Basidiospores; (C). Longitudinal section of outer part of volval remnants on stipe base. Bars: (A, B) = 10 μm, (C) = 40 μm.

MycoBank: MB 847660

Etymology: The epithet “brunneola” refers to the brown tone of pileus.

Type: CHINA. Beijing, Miyun district, Heilongtan, 40.560617 N, 116.782229 E, alt. 260 m, 27 Aug., 2020, coll. G.Q.C., C.L.H. and Y.T.Z. (C650/BJTC C650).

Basidiomata small- to medium-sized. Pileus 3–7 cm in diam., convex to applanate, with an umbo at the center, brown tone (#808080) to dark orange (#a5682a) over the disk; verrucose to subconical, dirty white (#f2f2f2) to white (#ffffff) volval remnants on the pileus; margin slightly striate (ca. 0.25–0.3 R), non-appendiculate; trama white (#ffffff), unchanging. Free, crowded, white (#ffffff) to cream (#fffdd0) lamellae; truncated, plentiful lamellulae. Stipe 8–15 cm long, 0.5–1.5 cm wide at the apex, subcylindric or slightly tapering upward, with the apex slightly expanded, white (#ffffff) to pale grayish (#a6a6a6), glabrous above the annulus, and densely covered with gray (#a1a1a1) to dark gray (#888888) squamules under the annulus; white (#ffffff) context, hollow in the center; absence of basal bulb; volva saccate (ca. 2 × 2.5 cm) membranous, both surfaces white (#ffffff) to dirty white (#f2f2f2). Annulus subapical, pendant from attachment ca. 2 cm below the apex of the stipe, membranous, pale grayish upper surface (#808080), brownish gray lower surface (#a5682a). Spore print not observed. Odor indistinct.

Bilateral lamellar trama, 20–60-μm-wide mediostratum, composed of abundant subfusiform, ellipsoid to clavate inflated cells (35–80 × 10–40 μm); abundant, 4–12-μm-wide filamentous hyphae; scarce vascular hyphae. Lateral stratum is composed of abundant subfusiform to ellipsoid inflated cells (25–55 × 9–15 μm), diverging at an angle of ca. 30–45° to the mediostratum; abundant, 2–6-μm-wide filamentous hyphae. A 30–55-μm-thick subhymenium, with two to three layers of subglobose to ellipsoid or irregular cells (10–20 × 8–20 μm). Basidia ( Figure 5A ) 35–50 × 8.5–15 μm, slenderly clavate, four-spored, with clamps, hyaline; 2–5-μm-long sterigmata; basidiospores [60/2/2] (9.5–)9.9–11(–12) × (7.5–)8.5–9.2(–9.5) μm, Q = (1.12–)1.22–1.42(–1.53), Qm = 1.34 ± 0.12, mostly ellipsoid, occasionally broadly ellipsoid, thin-walled, hyaline, smooth, occasionally with a small apiculus, inamyloid; sterile lamellar edge, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (10–40 × 10–30 μm), single and terminal or in chains of 2–3, thin-walled, hyaline; abundant, 2–5-μm-wide filamentous hyphae, irregularly arranged or ± running parallel to the lamellar edge. Pileipellis 80–150 μm thick, slightly gelatinized upper layer (20–50-μm thick), composed of radially arranged, thin-walled, colorless, 2–5-μm-wide filamentous hyphae; lower layer (40–90-μm thick) composed of radially arranged, colorless to brownish (#a53f2a), 2–8-μm-wide filamentous hyphae; scarce vascular hyphae. Volval remnants on the pileus are composed of inflated cells up to 20–40 × 15–30 μm and abundant filamentous hyphae, subglobose, ovoid to ellipsoid, clavate or sphaeropedunculate, with hyaline to yellowish (#ffff9a) vacuolar pigments; septa with clamps; inner part of volval remnants on the pileus often with conspicuous vascular hyphae. Volval remnants on the stipe base are similar to those on the pileus, but with more abundant filamentous hyphae and fairly abundant vascular hyphae. Longitudinally acrophysalidic stipe trama; acrophysalides (35–280 × 15–50 μm); scattered to abundant, 3–13-μm-wide filamentous hyphae. Annulus is composed of radially arranged elements: scarce, ellipsoid to cylindrical, colorless, thin-walled inflated cells (30–80 × 10–20 μm); very abundant to dominant, 2–8 μm wide, colorless, thin-walled filamentous hyphae; scarce vascular hyphae. Clamps are present in all parts of basidioma.

Habitat and distribution: This species is scattered in the broad-leaved forests of Castanea mollissima Blume and Carpinus turczaninowii Hance. Basidioma occurs in summer and autumn.

Additional specimens examined: CHINA. Beijing, Changping district, Dayangshan Mountains National Forest Park, 40.30799 N, 116.424929 E, alt. 260 m, 14 Aug., 2019, coll. H.Z., X.Y.S.and Y.T.Z. (ZH087/BJTC Z087).

Commentary: A. brunneola is well circumscribed by its brownish to dark orange pileus, with an umbo at the center, covered with verrucose to subconical, densely covered with gray to dark gray squamules under the annulus, and mostly ellipsoid, occasionally broadly ellipsoid basidiospores (9.5–12 × 7.5–9.5 μm). Furthermore, it is found in association with Fagaceae and Betulaceae trees.

A. brunneola belongs to section Caesareae and is closely related to A. longistriata, A. fense M. Mu & L.P. Tang, and A. incarnatifolia Zhu L. Yang on the multi-locus and nrITS phylogenetic trees ( Figures 2 , S3 ). A. brunneola can be distinguished from A. fense and A. longistriata on the basis of its longer striations (0.3–0.5 R) on the pileal margin, white to cream lamellae, and a white stipe. Basidiospores of A. brunneola (10.0–13.0 × 8.0–11.0 μm) are longer than those of A. longistriata, whereas they are shorter than those of A. fense (Imai, 1938; Gilbert, 1940; Gilbert, 1941; Hongo, 1959; Yang and Doi, 1999; Yang, 2005; Yang, 2015; Cui et al., 2018; Wu et al., 2021).

 <italic>Amanita caesareoides</italic> Lj. N. Vassiljeva, Notul. syst. Sect. cryptog. Inst. bot. Acad. Sci. U. S. S. R. 6: 199 (1950)

Figures 6A, B

Fresh basidiomata of known species of Amanita in this study. (A, B) A caesareoides (BJTC C630); (C, D) A chiui (BJTC L130); (E, F) A muscaria (BJTC L491); (G) oberwinkleriana (BJTC C320); (H, I) A oberwinkleriana (BJTC C659); (J, K) A ovalispora (BJTC Z311); (L) A subjunquillea (BJTC 085); (M) A subjunquillea (BJTC 704); (N) A vaginata var. vaginata (BJTC 677); (O) A vaginata var. vaginata (BJTC 682). Bars: (A–O) = 2 cm.

Basidiomata small- to medium-sized. Pileus 5–10 cm in diam., applanate, often umbonate at the center, orange-red (#ffa500) to orange (#ffb733); absence of volval remnants on the pileus; margin striate (0.3–0.5 R), non-appendiculate. Cream (#fffdd0) to yellowish (#ffff4d) lamellae; truncated lamellulae. Stipe 8–18 cm long, 0.7–2 cm wide, yellowish (#ffff4d) to orange-red (#ffa500), with its surface covered with snakeskin-shaped, orange (#ffb733) squamules; absence of basal bulb; volval remnants on the stipe base saccate. Annulus subapical, orange-red (#ffa500) to orange (#ffb733). Spore print not observed. Odor indistinct.

Basidia (33–45 × 8–12 μm), clavate, four-spored. Basidiospores [60/4/2] (7.0–)7.5–9.5(–10.5) × (6.0–)6.5–8.2(–8.5) μm, Q = (1.06–)1.12–1.34(–1.40), Qm = 1.23 ± 0.06, broadly ellipsoid, rarely ellipsoid or subglobose, inamyloid. Clamps are present in all parts of basidioma.

Distribution: This species is known to be found in northeastern China (Yang, 2015; Cui et al., 2018), India (Bhatt et al., 2003; Bhatt et al., 2017), Japan (Imazeki and Hongo, 1987; Endo et al., 2016), Republic of Korea (Cho et al., 2015), and Russian Far East (Vassiljeva, 1950).

Habitat and distribution: It is present individually or is scattered in the broad-leaved forests of C. viminea Lindley and P. davidiana Dode. Basidioma occurs in summer and autumn.

Specimens examined: CHINA. Beijing, Yanqing district, Yudushan Mountains, 40.550099 N, 115.875254 E, alt. 963 m, 20 Aug., 2018, coll. C.L. Hou, H. Zhou and J.Q. Li (630/BJTC 630).

Commentary: A. caesareoides was first described from the Russian Far East by Vassiljeva (1950). It was subsequently reported from China, India, Japan, and Republic of Korea (Imazeki and Hongo, 1987; Bhatt et al., 2003; Cho et al., 2015; Yang, 2015; Endo et al., 2016; Bhatt et al., 2017). The morphological description of our specimen is consistent with that provided by Vassiljeva (1950). In our multi-locus phylogenetic analysis, the specimen BJTC 630 clustered with A. caesareoides (HKAS 92009 and HKAS 71021), forming a completely supported clade (BPP = 1.00, MLB = 88%) ( Figure 2 ). Based on these characters, we described our specimen BJTC 630 as A. caesareoides. Detailed descriptions, line drawings, and images of A. caesareoides can be found in Yang (2015).

 <italic>Amanita chiui</italic> Yang-Yang Cui, Qing Cai & Zhu L. Yang, Fungal Diversity 91: 77 (2018)

Figures 6C, D

Basidiomata small- to medium-sized. Pileus 3–8 cm in diam., often dark gray (#a9a9a9), brown (#a5682a) to brownish (#d18e4a); volval remnants on the pileus mostly absent or occasionally retained as small, white (#ffffff) patches; margin striate (0.25–0.3 R), non-appendiculate; trama white (#ffffff), unchanging. Free, white (#ffffff) to cream (#fffdd0) lamellae. Stipe 6–10 cm long, 0.5–1.5 cm wide at the apex, white (#ffffff), dirty white (#fcfcfc), brownish (#a5682a) to brown (#cc8236). Annulus absent. Spore print not observed. Odor indistinct.

Basidia (45–65 × 14–17 μm), clavate, four-spored; 4–6-μm-long sterigmata; basal septa lacking clamps. Basidiospores [30/2/1] (9.0–)9.5–12.0(–12.5) × (8.0–)9.0–11.0(–11.5) μm, Q = (1.00–)1.05–1.25(–1.30), Qm = 1.15 ± 0.06, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; small apiculus. Volval remnants on the stipe base are composed of longitudinally arranged elements: abundant to very abundant, branching, anastomosing filamentous hyphae. Clamps are absent in all parts of basidioma.

Distribution: This species is known to be found in northwestern and southwestern China (Cui et al., 2018).

Habitat and distribution: It is present individually or scattered in the broad-leaved forests of Castanea mollissima Blume, with basidioma occurring in summer and autumn.

Specimens examined: CHINA. Beijing, Pinggu district, Sizuolou, 40.272187 N, 117.135187 E, elev. 223 m, 16 August 2019, coll. C.L. Hou, G.Q. Cheng and R.T. Zhang (L130/BJTC L130).

Commentary: Generally, A. chiui is characterized by a dark gray, brown to brownish pileus; a white to dirty white stipe that is often densely covered with brownish squamules; and subglobose to broadly ellipsoid basidiospores (10.0–12.5 × 9.0–11.0 μm) (Cui et al., 2018). Our multi-locus phylogenetic analysis revealed that the specimen BJTC L130 clustered together with A. chiui (HKAS 77330, type), forming a completely supported clade (pp = 1.00, MLB = 100%). The nrLSU phylogenetic analysis exhibited topologies similar to those of the multi-locus phylogenetic tree ( Figure S1 ). Based on these characters and phylogenetic analysis results, the specimen BJTC L130 was described as A. chiui. Detailed descriptions, line drawings, and images of A. caesareoides can be found in Cui et al. (2018).

 <italic>Amanita muscaria</italic> (L.: Fr.) Lam., Encycl. Méth. Bot. (Paris) 1(1): 111 (1783)

Figures 6E, F

Basidiomata small- to large-sized. Pileus 5–15 cm in diam., orange-red (#ffa500); pyramidal, conical, verrucose to felted, white (#fffff), sometimes yellowish (#ffffd8), removable volval remnants on the pileus; margin striate (up to 0.2 R), non-appendiculate; trama white (#ffffff), unchanging. White lamellae (#fffff), truncated lamellulae. Stipe 7–16 cm long, 0.5–2.0 cm wide at the apex., white (#fffff), covered with white (#fffff) fibrils; white (#fffff) context, unchanging; ovoid, fusiform to subglobose basal bulb (1–4-cm diam.); verrucose to conical, white (#fffff) to yellowish (#ffffd8) volval remnants on the stipe base, sometimes arranged in incomplete rings. Subapical to submedian annulus. Spore print not observed. Odor indistinct.

Basidia (40–60 × 12–15 μm), clavate, four-spored. Basidiospores [30/1/1] (9–)9.5–10.8(–12.5) × (7–)7.3–8.5(–9.0) μm, Q = (1.2–)1.24–1.45(–1.47), Qm = 1.32 ± 0.06, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on the pileus are composed of vertically arranged elements: scattered filamentous hyphae; very abundant inflated cells. The structure of volval remnants on the stipe base is similar to that of volval remnants on the pileus, but with irregularly arranged elements. Clamps are present in all parts of basidioma.

Distribution: This species is known to be found in Asia (Imai, 1933; Imai, 1938; Kumar et al., 1990; Yang, 2005; Geml et al., 2006; Geml et al., 2008; Yang, 2015), Europe (Neville and Poumarat, 2004), North America (Geml et al., 2006; Geml et al., 2008), and northeastern and northwestern areas of China (Cui et al., 2018).

Habitat and distribution: It is present individually or is scattered in the broad-leaved forests of B. platyphylla Suk. Basidioma occurs in summer and autumn.

Specimens examined: CHINA. Hebei Province, Chicheng County, Dahaituo Mountains National Nature Reserve, 40.563828 N, 115.798698 E, elev. 1640 m, 22 August 2018, coll. C.L. Hou, J.Q. Li and G.Q. Cheng (L491/BJTC L491).

Commentary: A. muscaria was described from Europe and represents the type species of Amanita (Gilbert, 1940; Gilbert, 1941; Jenkins and Petersen, 1976; Galli, 2001; Neville and Poumarat, 2004; Yang, 2005; Yang, 2015; Cui et al., 2018). Yang and Oberwinkler (1999) conducted a detailed study on A. muscaria basidiomal development and anatomy. The morphological description of our specimen is consistent with that given by Cui et al. (2018). In our multi-locus analysis ( Figure 2 ), A. muscaria was found to be closely related to A. persicina (D.T. Jenkins) Tulloss & Gem. These results are consistent with those of Cui et al. (2018). Based on these characters, our specimen BJTC L491 was described as A. muscaria. Detailed descriptions of A. muscaria can be found in Cui et al. (2018).

 <italic>Amanita oberwinkleriana</italic> Zhu L. Yang & Yoshim. Doi, Bull. Natn. Sci. Mus., Tokyo, Ser. B 25 (3): 120 (1999)

Figures 6G–I , 7

Microscopic characters of Amanita oberwinkleriana (BJTC C320). (A). Hymenium and subhymenium; (B). Basidiospores; (C). Longitudinal section of outer part of volval remnants on stipe base. Bars: (A, B) = 10 μm, (C) = 40 μm.

Basidiomata small- to medium-sized. Pileus 3–8 cm in diam., applanate, no slightly depressed at the center, surface gray-white (#e6e6e6) to white (#ffffff), large, gray-white (#e6e6e6) volval remnants on the pileus (ca. 2–4 (–6)-mm diam.), densely arranged over the disk; margin slightly striate (ca. 0.05–0.15 R), non-appendiculate. Free, white (#ffffff), unchanging, somewhat crowded lamellae; attenuate, plentiful lamellulae. Stipe 8–15 cm long, 0.5–2.0 cm wide at the apex, subcylindric or slightly tapering upward, with the apex slightly expanded, gray-white (#e6e6e6) to white (#ffffff), with minute concolorous squamules; grayish (#e6e6e6), unchanging context; subglobose basal bulb (1.0–2.0-cm diam.), dirty white (#f2f2f2), with upper part covered with verrucose, dirty grayish (#e6e6e6) to dirty white (#f2f2f2) volval remnants arranged in irregular concentric rings. Annulus absent. Spore print not observed. Odor indistinct.

Basidia (28–45 × 8–13 μm), slenderly clavate, four-spored, with clamps, hyaline to pale yellow (#ffffd8); 5–6-μm-long sterigmata, clamped basal septa; basidiospores [80/4/3] (6–)7.0–10(–10.5) × (5–)5.5–6.8(7.5) μm, Q = (1.25–)1.35–1.58(–1.62), Qm = 1.41 ± 0.11, mostly ellipsoid, occasionally broadly ellipsoid, thin-walled, hyaline, pale yellow, smooth, with a small to medium-large apiculus, amyloid. Volval remnants on the stipe base are similar to those on the pileus, but with more abundant filamentous hyphae and fairly abundant vascular hyphae. Clamps are present in all parts of basidioma.

Distribution: This species is known to be present in central, eastern, southern, and southwestern China (Yang and Li, 2001; Yang, 2005; Yang, 2015; Cui et al., 2018); India (Bhatt et al., 2003); Japan (Yang and Doi, 1999); and Republic of Korea (Kim et al., 2013a).

Habitat and distribution: It is present individually or is scattered in the coniferous forests and mixed coniferous and broad-leaved forests of Pinus tabuliformis Carr., Q. mongolica Fisch. ex Ledeb., and P. davidiana Dode. Basidioma occurs in summer and autumn.

Specimens examined: CHINA. Beijing, Hebei Province, Xinglong country, Babaziling village, 40.310326 N, 117.585117 E, alt. 878 m, 22 Aug., 2020, coll. G.Q. Cheng., C.L. Hou.and Y.T. Zhang (C320/BJTC C320); CHINA. Tianjin, Jizhou district, JiulongShan Mountains National Forest Park, 40.152701 N, 117.508008 E, alt. 300 m, 18 Aug., 2019, coll. H. Zhou, X.Y. Shen and Y.T. Zhou (ZH328/BJTC Z328); CHINA. Beijing, Pinggu district, Dongxinzhuang village, 40.560714 N, 116.779865 E, alt. 286 m, 27 Aug., 2020, coll. G.Q. Cheng., C.L. Hou and Y.T. Zhang (C659/BJTC C659); CHINA. Tianjin, Jizhou district, JiulongShan Mountains National Forest Park, 40.151969 N, 117.509612 E, alt. 226 m, 18 Aug., 2019, coll. H. Zhou, X.Y. Shen and Y.T. Zhou (ZH318/BJTC Z318); CHINA. Beijing, Huairou district, Baiquanshan, 40.496637 N, 116.649129 E, alt. 268 m, 17 Aug., 2020, coll. H. Zhou, X.Y. Shen and Y.T. Zhou (ZH862/BJTC Z862); CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.27 N, 115.57 E, elev. unknown, 11 July 2011, coll. unknown (HMAS 263406); CHINA. Beijing, Mentougou district, Donglingshan Mountains, 40.22 N, 116.50 E, elev. unknown, 29 July., 2013, coll. W.L. Lu (HMAS 253800); CHINA. Beijing, Mentougou district, Donglingshan Mountains, 40.22 N, 116.50 E, elev. unknown, 29 July., 2013, coll. W.L. Lu (HMAS 253801); CHINA. Beijing, Mentougou district, Donglingshan Mountains, 40.22 N, 116.50 E, elev. unknown, 29 July., 2013, coll. W.L. Lu (HMAS 253802); CHINA. Beijing, Mentougou district, Yunmengshan Mountains, 40.22 N, 116.50 E, elev. unknown, 29 July., 2013, coll. T.Z Wei (HMAS 253796).

Commentary: Yang and Doi (1999) described A. oberwinkleriana from Japan, and it was subsequently reported from China, India, and Republic of Korea (Yang and Li, 2001; Bhatt et al., 2003; Yang, 2005; Kim et al., 2013a; Yang, 2015; Cui et al., 2018). The morphological description of our specimen is consistent with that provided by Yang and Doi (1999). In our multi-locus phylogenetic analysis, 10 specimens clustered together with A. oberwinkleriana (HKAS 77330), forming a completely supported clade (BPP = 1.00, MLB = 96%). Based on these characters, our specimens were described as A. oberwinkleriana. Of note, the samples from the herbarium specimens (HMAS) exhibited poor sequence quality because of a long time. The multi-locus and nrLSU phylogenetic trees revealed that the branches of these herbarium specimens were longer ( Figures 2 , S1 ).

 <italic>Amanita ovalispora</italic> Boedijn, Sydowia 5(3-6): 320 (1951)

Figures 6J, K , 8

Microscopic characters of Amanita ovalispora. (BJTC Z311). (A). Hymenium and subhymenium; (B). Basidiospores; (C). Longitudinal section of outer part of volval remnants on stipe base. Bars: (A, B) = 10 μm, (C) = 40 μm.

Basidiomata small- to medium-sized. Pileus 3–6 cm in diam., plano-convex to applanate, distinctly umbonate at the center, gray-brown (#75615a) to gray (#808080), absence of volval remnants on the pileus, margin striate (0.2–0.5 R), non-appendiculate; trama white (#ffffff), unchanging. Free, crowded, white (#ffffff) lamellae; white (#ffffff) lamellar edge; truncated, plentiful lamellulae. Stipe 8–16 cm long, 0.5–1.5 cm wide at the apex, subcylindric or slightly tapering upward, with the apex slightly expanded, white (#ffffff) to dirty white (#f2f2f2), glabrous or covered with minute, concolorous fibrils; white (#ffffff), hollow in the center context; absence of basal bulb; volva saccate, membranous, both white (#ffffff) surfaces. Annulus absent. Spore print not observed. Odor indistinct.

Basidia (30–55 × 8–14 μm), clavate, four-spored, basal septa lacking clamps, hyaline; 4–6-μm-long sterigmata; basidiospores [60/2/2] (8–)9.5–11.8(–12.5) × (7–)7.3–9.1(–10.2) μm, Q = (1.15–)1.23–1.49(–1.52), Qm = 1.34 ± 0.10, globose to subglobose, thin-walled, hyaline, smooth, with a small apiculus, inamyloid. Volval remnants on the stipe are composed of longitudinally arranged elements: very abundant filamentous hyphae. Outer surface of the volval remnants on the stipe base is similar to the structure of the outer part, but with more abundant filamentous hyphae; gelatinized inner surface with structure similar to that of the inner part. Clamps are absent in all parts of basidioma.

Distribution: This species is known to be present in southern and southwestern China (Yang, 1997; Yang, 2005; Yang, 2015; Cui et al., 2018), and Indonesia (Boedijn, 1951).

Habitat and distribution: It is present individually or scattered in the coniferous forests and mixed coniferous and broad-leaved forests of Pinus tabuliformis Carr. and Juglans mandshurica Maxim., with basidioma occurring in summer and autumn.

Specimens examined: CHINA. Tianjin, Jizhou district, JiulongShan National Forest Park, 40.147837 N, 117.510365 E, alt. 174 m, 18 Aug., 2019, coll. H. Zhou, X.Y. Shen and Y.T. Zhang (ZH311/BJTC Z311); CHINA. Beijing, Changping district, Tibiyinshan, 40.317323 N, 116.321293 E, alt. 353 m, 14 Aug., 2019, coll. H H. Zhou, X.Y. Shen and Y.T. Zhang (ZH057/BJTC Z057).

Commentary: Boedijn (1951) was first described A. ovalispora from Indonesia. Subsequently, Yang (1997) examined the holotype, and described their basidiospores. According to Cui et al. (2018), no sequence of A. ovalispora is available from its type locality to delimit this species accurately. The morphological description of our specimens is consistent with that provided by Boedijn (1951). In our multi-locus phylogenetic analysis, two specimens clustered together with A. oberwinklerana (HKAS 79625 and HKAS 101406), forming a completely supported clade (BPP = 1.00, MLB = 96%) ( Figure 2 ). The nrLSU phylogenetic analysis exhibited topologies similar to those of the multi-locus phylogenetic tree ( Figure S1 ). According to these characters, our specimens were described as A. ovalispora.

 <italic>Amanita subglobosa</italic> Zhu L. Yang, Bibl. Mycol. 170: 18 (1997)

Basidiomata small- to medium-sized. Pileus 4–8 cm in diam., brownish (#a5682a) to dark brown (#68421a); pyramidal to verrucose, white (#ffffff) to yellowish (#ffff9a), removable volval remnants on the pileus; margin striate (0.1–0.4 R), non-appendiculate; trama white (#ffffff), unchanging. White (#ffffff) to cream (#fffeea) lamellae, truncated lamellulae. Stipe 5–15 cm long, 0.5–2 cm wide, white (#ffffff) to dirty white (#f2f2f2); white (#ffffff), unchanging context; subglobose basal bulb (1.5–3.5-cm diam.), with its upper part covered with conical to pulverulent, yellowish (#ffff9a) to brownish (#a5682a) volval remnants, often forming a collar between the limit of the stipe and basal bulb. Subapical to submedian, white (#ffffff), persistent annulus. Spore print not observed. Odor indistinct.

Basidia (35–55 × 10–15 μm), clavate, four-spored. Basidiospores [60/2/2] (7.0–)8.3–11.0 (–13.0) × (6.0–)6.5–9.5(–11.5) μm, Q = (1.05–) 1.13–1.40(–1.60), Qm = 1.29 ± 0.08, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on the pileus are composed of vertically arranged elements: fairly abundant filamentous hyphae; abundant inflated cells. Clamps are present in all parts of basidioma.

Distribution: This species is known to be present in central, northeastern, and southwestern China (Yang, 1997; Yang, 2005; Yang, 2015; Cui et al., 2018); India (Semwal et al., 2007); Republic of Korea (Kim et al., 2013b); and Thailand (Sanmee et al., 2008).

Habitat and distribution: It is present solitary or is scattered in the pine, broad-leaved, or mixed forests of Fagaceae and Pinaceae trees. Basidioma occurs in summer and autumn (Cui et al., 2018).

Specimens examined: CHINA. Beijing, Mentougou district, Baihuashan Mountains, 39.52 N, 115.36 E, elev. unknown, 22 Aug., 1964, coll. Y.C. Zong and Q.T. Tao (HMAS 34658); CHINA. Beijing, Mentougou district, Tanzhe Temple, 39.54 N, 116.01 E, elev. unknown, 22 Aug., 1965, coll. Q.M. Ma (HMAS 253798).

Commentary: Yang (1997) first described A. subglobosa from China. Later, it was reported from India, Republic of Korea, and Thailand (Semwal et al., 2007; Sanmee et al., 2008; Kim et al., 2013b). The morphological description of our specimen is consistent with that provided by Yang (1997). Our multi-locus phylogenetic analysis revealed that a specimen HMAS 253798 clustered together with A. subglobosa (HKAS 67914), forming a completely supported clade (BPP = 1.00, MLB = 100%) ( Figure 2 ). The nrLSU phylogenetic analysis revealed topologies similar to those of the multi-locus phylogenetic tree ( Figure S1 ). Based on these characters, we described our specimen as A. subglobosa. In addition, DNA sequences from another herbarium specimens (HMAS 34658) could not be generated. However, we also examined the morphology of these specimens, and the results proved that these specimens were A. subglobosa.

 <italic>Amanita subjunquillea</italic> S. Imai, Bot Mag (Tokyo) 47: 424 (1933)

Figures 6L, M , 9

Microscopic characters of Amanita subjunquillea (BJTC 217). (A). Hymenium and subhymenium; (B). Basidiospores. Bars: (A, B) = 10 μm.

Synonym: Amanita subjunquillea var. alba Zhu L. Yang, Bibl. Mycol. 170: 174 (1997)

Basidiomata small- to medium-sized. Pileus 3–6 cm in diam., globose when young, hemispherical when expanding, later plano-convex to applanate, surface pale yellow (#ffff00) to light yellow (#ffffa1), becoming paler toward the margin; absence of volval remnants on the pileus. Free, white (#ffffff) to cream (#fffdd0), plentiful lamellae. Stipe 4–10 cm long, 0.8–2 cm wide at the apex, subcylindric or slightly tapering upward, with the apex slightly expanded, surface pale yellow (#ffff00) to light yellow (#ffffa1), with silk luster, upper part often covered with pale yellow (#ffff00) floccose to farinose squamules, lower part often covered with white (#ffffff) to pale yellow (#ffff00) floccose squamules; white (#ffffff) to pale yellow (#ffff00) context; marginate, dirty white (#fcfcfc) to grayish (#cccccc) basal bulb (1–2-cm diam.), with upper edge shortly limbate, dirty white (#fcfcfc) to grayish (#cccccc) volval remnants. Annulus absent. Spore print not observed. Odor indistinct.

Basidia (25–55 × 6.5–12 μm), slenderly clavate, four-spored, with clamps, hyaline; 3–5-μm-long sterigmata; basidiospores [60/4/4] (6–)6.5–8.4(–9) × (4–)4.1–6.6(–7.5) μm, Q = (1.02)1.18–1.36(1.51), Qm = 1.34 ± 0.20, mostly ellipsoid, occasionally spherical, thin-walled, hyaline, smooth, with a small apiculus, weakly amyloid. Volval remnants on the pileus are composed of irregularly to subvertically arranged elements: fairly abundant, branching, anastomosing filamentous hyphae; very abundant, globose, subglobose to ellipsoid inflated cells. Clamps are present in all parts of basidioma.

Distribution: This species is known to be from central, northern, northeastern, northwestern, and southwestern China (Yang, 1997; Yang, 2005; Yang, 2015; Cai et al., 2016; Cui et al., 2018); India (Bhatt et al., 2003; Bhatt et al., 2007); Japan (Imai, 1933; Imai, 1938; Yang and Doi, 1999; Imazeki et al., 2011); and Republic of Korea (Kim et al., 2013a; Cho et al., 2015).

Habitat and distribution: It is present individually or is scattered in coniferous forests and mixed coniferous and broad-leaved forests of Pinus tabuliformis Carr., J. mandshurica Maxim, and P. davidiana Dode., with basidioma occurring in summer and autumn.

Specimens examined: CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.596637 N, 115.849129 E, alt. 648 m, 17 Sep., 2017, coll. C.L. Hou, H. Zhou and J.Q. Li (217/BJTC 217); CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.316637 N, 115.459129 E, alt. 1102 m, 17 Sep., 2017, coll. C.L. Hou, H. Zhou and J.Q. Li (085/BJTC 085); CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.517899 N, 115.820799 E, alt. 906 m, 20 Aug., 2018, coll. C.L. Hou, H. Zhou and J.Q. Li (704/BJTC 704); CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.30024 N, 115.499129 E, alt. 860 m, 17 Sep., 2017, coll. C.L. Hou, H. Zhou and J.Q. Li (033/BJTC 033); CHINA. Beijing, Mentougou district, Donglingshan Mountains, 40.22 N, 116.50 E, elev. unknown, coll. unknown (HMAS 253775); CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.31 N, 115.45 E, alt. 1216 m, 16 Sep., 2017, coll. C.L. Hou, H. Zhou and J.Q. Li (112/BJTC 112); CHINA. Beijing, Hebei Province, Xinglong country, Changgou, 40.205334 N, 117.625724 E, alt. 878 m, 18 Aug., 2019, coll. G.Q. Cheng., C.L. Hou.and H. Zhou (ZH276/BJTC Z276); CHINA. Beijing, Huairou district, Sunzhazi Village 40.943482 N, 116.507391 E, alt. 789 m, 25 Aug., 2020, coll. C.L. Hou, G.Q. Cheng and Y.T. Zhang (C558/BJTC C558); CHINA. Beijing, Shunyi district, Mulin Town, 40.235317 N, 116.819856 E, alt. 45 m, 16 Aug., 2019, coll. G.Q. Cheng., C.L. Hou.and H. Zhou (ZH172/BJTC Z172).

Commentary: Imai (1933) first described A. subjunquillea from Japan. Later, it was reported from China, India, and Republic of Korea (Yang, 1997; Bhatt et al., 2003; Yang, 2005; Bhatt et al., 2007; Kim et al., 2013a; Cho et al., 2015; Yang, 2015; Cai et al., 2016; Cui et al., 2018). This species is deadly poisonous (Imazeki et al., 1988; Kawase et al., 1992; Chen et al., 2016; Tang et al., 2016). The morphological description of our specimen is consistent with that provided by Imai (1933). In our multi-locus phylogenetic analysis, nine specimens clustered together with A. oberwinklerana (HKAS 75771, HKAS 75770, and HKAS 75773), forming a completely supported clade (BPP = 1.00, MLB = 99%) ( Figure 2 ). The nrLSU phylogenetic analysis showed topologies similar to those of the multi-locus phylogenetic tree ( Figure S1 ). Based on these characters, our specimens were described as A. subjunquillea.

 <italic>Amanita vaginata</italic> var. <italic>vaginata</italic> (Bull.) Lam., Encycl. Méth. Bot. (Paris) 1(1): 109 (1783)

Figures 6N, O

Basionym: Agaricus vaginatus Bull., Herb. Fr. (Paris) 3: tab. 98 (1783) [1782-83].

Basidiomata small- to medium-sized. Pileus 3–7 cm in diam., somewhat umbonate, grayish (#a6a6a6) to gray (#b3b3b3); volval remnants on the pileus absent or retained as white (#ffffff) patches; margin striate (0.1–0.3 R), non-appendiculate. White (#ffffff) lamellae; truncated lamellulae. Stipe 5–10 cm long, 0.5–2.0 cm wide at the apex., white (#ffffff) to dirty white (#fcfcfc), glabrous or covered with fibrous, grayish (#a6a6a6) to gray (#b3b3b3) fibrils; absence of basal bulb; volval remnants on the stipe base saccate, outer surface white (#ffffff) to dirty white (#fcfcfc), inner surface white (#ffffff). Annulus absent. Spore print not observed. Odor indistinct.

Basidia (48–60 × 13–18 μm), clavate, four-spored. Basidiospores [60/3/3] (9.2–) 9.5–11.2 (–13.0) × (8.0–) 9.0–10.9 (–13.0) μm, Q = (1.0–)1.02–1.08 (–1.10), Qm = 1.01 ± 0.03, globose to subglobose, inamyloid. Clamps are absent in all parts of basidioma.

Distribution: This species is known to be from Asia (Imai, 1933; Imai, 1938; Tulloss et al., 2001; Yang, 2005; Imazeki et al., 2011; Kim et al., 2013b; Cui et al., 2018), North America (Coker, 1917; Thiers, 1982; Jenkins, 1986; Tulloss et al., 1995), and Europe (Horak, 1968; Consiglio, 2000).

Habitat and distribution: It is present individually or is scattered in broad-leaved forests of Fagaceae. Basidioma occurs in summer and autumn.

Specimens examined: CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.512712 N, 115.817818 E, elev. 555 m, 26 August 2018, coll. C.L. Hou, J.Q. Li and H. Zhou (682/BJTC 682); CHINA. Beijing, Yanqing district, Songshan Mountains National Nature Reserve, 40.512154 N, 115.817489 E, elev. 901 m, 26 August 2018, coll. C.L. Hou, J.Q. Li and H. Zhou (677/BJTC 677); CHINA. Beijing, Huairou district, Shangdian, 40.927121 N, 116.697837 E, elev. 681 m, 19 August 2019, coll. H. Zhou, X.Y. Shen and R.T. Zhang (ZH521/BJTC Z521); CHINA. Beijing, Mentougou district, Donglingshan Mountains, 39.55 N, 116.24 E, elev. unknown, coll. unknown (HMAS 253281); CHINA. Beijing, Miyun district, 40.22 N, 116.5 E, elev. unknown, August 1998, coll. X.L. Mao (HMAS 78411); CHINA. Beijing, Mentougou district, Donglingshan Mountains, 39.55 N, 116.24 E, elev. unknown, 19 August 1998, coll. H.A. Wen and S.X. Sun (HMAS 75237).

Commentary: A. vaginata var. vaginata is characterized by a gray pileus with striations at the margin, a white stipe lacking an annulus, and globose to subglobose basidiospores (9.0–13.0 (–14.0) μm) (Lange, 1935; Huijsman, 1959; Bas, 1967; Horak, 1968; Jenkins, 1986; Breitenbach and Kränzlin, 1995; Tulloss et al., 1995). In our multi-locus phylogenetic analysis, four specimens clustered together with A. vaginata var. vaginata (LAF 024482, type), forming a completely supported clade (BPP = 1.00, MLB = 99%) ( Figure 2 ). The nrLSU phylogenetic analysis displayed topologies similar to those of the multi-locus phylogenetic tree ( Figure S1 ). Based on these characters and results of phylogenetic analyses, our specimens were described as A. vaginata var. vaginata. Detailed descriptions, line drawings, and images of A. vaginata var. vaginata are available in Yang (2015). Unfortunately, DNA sequences could not be generated from two herbarium specimens (HMAS 78411 and HMAS 75273). However, on examining the morphology of these specimens, we confirmed that these specimens were A. vaginata var. vaginata.

<italic>Amanita virosa</italic> Bertillon, Dict. Encycl. Sci. Médic.: 497 (1866)

Replaced synonym: Agaricus virosus Fr., Epicr. syst. mycol. (Upsaliae): 3 (1838) [1836-1838]; Agaricus virosus Sowerby, Col. fig. Engl. Fung. Mushr. (London) 3: tab. 407 (1809).

Basidiomata small- to medium-sized. Pileus 5–10 cm in diam., umbonate at the center, white (#ffffff), often cream (#fffdd0) at the center; absence of volval remnants on the pileus; margin non-striate, non-appendiculate; white (#ffffff), unchanging trama. White (#ffffff) lamellae; attenuate lamellulae. Stipe 8–13 cm long, 0.7–2.0 cm wide, white (#ffffff), covered with concolorous squamules; white (#ffffff), unchanging context; Subapical, white (#ffffff), persistent annulus. Spore print not observed. Odor not observed.

Basidia (30–50 × 10–12 μm), clavate, four-spored. Basidiospores [60/2/2] (7.5–)8.0–11.0(–12.0) × (7.0–)8.0–10.0(–11.0) μm, Q = (1.00–)1.05–1.15 (–1.21), Qm = 1.05 ± 0.05, globose to subglobose, amyloid. Volval remnants on the stipe base are composed of longitudinally to irregularly arranged elements: very abundant to nearly dominant filamentous hyphae; scarce to scattered inflated cells. Clamps are absent in all parts of basidioma.

Distribution: This species is known to be present in Europe (Konrad and Maublanc, 1924; Chiusa, 2000; Contu, 2000; Neville and Poumarat, 2004) and East Asia (Imai, 1933; Imai, 1938; Zhang et al., 2010; Imazeki et al., 2011; Li et al., 2015; Yang, 2015; Cai et al., 2016; Cui et al., 2018).

Habitat and distribution: It is present solitary or is scattered on soil in the subtropical to temperate forests of Fagaceae and Pinaceae. Basidioma occurs in summer and autumn (Cui et al., 2018).

Specimens examined: CHINA. Beijing, Miyun district, 40.22 N, 116.50 E, elev. unknown, 4 August 1987, coll. Y.C. Zong (HMAS 40501); CHINA. Beijing, Miyun district, 40.22 N, 116.50 E, elev. unknown, 29 August 1987, coll. Y.C. Zong (HMAS 40530).

Commentary: A. virosa is widely distributed across Europe and temperate to subtropical Asia (Contu 2000; Chiusa, 2000; Neville and Poumarat, 2004; Zhang et al., 2010; Li et al., 2015; Yang, 2015; Cai et al., 2016; Cui et al., 2018). The morphological description of our specimen is consistent with that provided by Cui et al. (2018). Unfortunately, we could not obtain sequence data from any of the herbarium specimens (HMAS 40501 and HMAS 40503). We also examined the morphology of these specimens, and the results proved that these specimens were A. virosa.

<italic>Amanita yanshanensis</italic> H. Zhou & C. L. Hou, sp. nov.

Figures 3E, F , 10

Microscopic characters of Amanita yanshanensis (type, BJTC Z049). (A). Hymenium and subhymenium; (B). Basidiospores; (C). Longitudinal section of outer part of volval remnants on stipe base; (D). longitudinal section of inner part of volval remnants on stipe base. Bars: (A, B) = 10 μm, (C, D) = 40 μm.

MycoBank: MB 847661

Etymology: The epithet “yanshanensis” refers to the locality where the type specimen was collected.

Type: CHINA. Beijing, Changping district, Yanshou temple, 40.373152 N, 116.322892 E, alt. 268 m, 14 Aug., 2019, coll. H.Z., X.Y.S.and Y.T.Z. (ZH049/BJTC Z049).

Basidiomata small- to medium-sized. Pileus 4–8 cm diam., sub-hemisphere when young, hemispherical when expanding, later convex, plano-convex to applanate, with an umbo at the center; surface gray-white (#e6e6e6) when young, then gray-white (#e6e6e6) to dark grayish red (#755a5a), large, dark grayish red (#755a5a) to very dark (mostly black) red (#1f1818), or very dark grayish red (#4a3939) volval remnants on the pileus (ca. 2–6 mm diam.), densely arranged over the disk; margin slightly striate (ca. 0.05–0.2 R), non-appendiculate; trama white (#ffffff), unchanging. Free, white (#ffffff), unchanging, somewhat crowded lamellae; attenuate, plentiful lamellulae. Stipe 7–12 cm long, 0.5–1.5 cm wide at the apex, subcylindric or slightly attenuate upward, surface white (#ffffff) to gray-white (#e6e6e6), with silk luster, upper part often covered with gray-white (#e6e6e6) to gray-brown (#755a5a) floccose to farinose squamules, lower part often covered with gray-brown (#755a5a) to dark-brown (#1f1818) verrucose, floccose squamules; white (#ffffff) context; subglobose to clubbed, dirty white (#f2f2f2) to white (#ffffff) basal bulb (1–2 cm diam.); floccose to felted, dark grayish red (#755a5a) to dark (mostly black) red (#1f1818) volval remnants, arranged irregularly or in incomplete belts or rings on the stipe base. Apical, subapical to fugacious, thin annulus, with cream color (#fffeea) and silk luster on the upper surface, a matte cream-colored (#fffeea) lower surface with floccose to farinose warts. Spore print not observed. Odor indistinct.

Bilateral lamellar trama. Mediostratum 20–50 μm wide, composed of abundant ellipsoid to clavate inflated cells (45–60 × 20–40 μm); abundant, 2–6-μm-wide filamentous hyphae; scarce vascular hyphae. Lateral stratum is composed of abundant subfusiform to ellipsoid inflated cells (20–40 × 5–15 μm), diverging at an angle of ca. 30–45° to the mediostratum; abundant, 2–5-μm-wide filamentous hyphae. A 30–55-μm-thick subhymenium, with two to three layers of subglobose to ellipsoid or irregular cells (8–15 × 5–10 μm). Basidia (25–40 × 6–12 μm), slenderly clavate, four-spored, with clamps, hyaline to pale yellow (#ffffd8); 5–6-μm-long sterigmata; basidiospores [120/5/4] 6.0–8.0(–9.0) × 4.0–5.5(–6.0) μm, Q = 1.24–1.45(–1.57), Qm = 1.40 ± 0.12, mostly ellipsoid, occasionally broadly ellipsoid, thin-walled, hyaline, pale yellow, smooth, occasionally with a small to medium-large apiculus, amyloid; sterile lamellar edge, composed of subglobose to ellipsoid inflated cells (20–40 × 10–30 μm), single and terminal or in chains of 2–3, thin-walled, hyaline; abundant, 2–5-μm-wide filamentous hyphae, irregularly arranged or ± running parallel to the lamellar edge. Pileipellis 100–200-μm thick, gelatinized upper layer (25–100-μm thick), composed of subradially to somewhat interwoven, thin-walled, colorless, 2–7-μm-wide filamentous hyphae; lower layer (70–100-μm thick) composed of radially and compactly arranged, pale yellow (#ffffd8), 4–8 (–10)-μm-wide filamentous hyphae; scarce vascular hyphae. Volval remnants on the pileus are composed of inflated cells and filamentous hyphae, more or less arranged in erect chains; abundant to dominant, subglobose, ovoid to ellipsoid, clavate or sphaeropedunculate inflated cells (up to 30 × 25 μm), with hyaline to pale yellow (#ffffd8) vacuolar pigments; septa with clamps; inner part of volval remnants on the pileus often with conspicuous vascular hyphae. Volval remnants on the stipe base are similar to those on the pileus, but with more abundant inflated cells. Stipe trama composed of longitudinally arranged, long clavate terminal cells (50–300 × 15–40 μm); scattered to abundant, 5–10-μm-wide filamentous hyphae; scarce vascular hyphae. Annulus is composed of radially arranged elements: abundant, subglobose, fusiform to ellipsoid, hyaline, thin-walled inflated cells (15–25 × 8–15 μm); abundant, hyaline, thin-walled, 2–5-μm-wide filamentous hyphae. Clamps are present in all parts of basidioma.

Habitat and distribution: This species is scattered in the broad-leaved forests of Castanea mollissima Blume., with basidioma occurring in summer and autumn.

Additional specimens examined: CHINA. Beijing, Changping district, Yanshou Temple, 40.368272 N, 116.321008 E, alt. 234 m, 17 Aug., 2020, coll. H. Zhou, X.Y. Shen and X.B. Huang (ZH820/BJTC Z820); CHINA. Beijing, Changping district, Yanshou temple, 40.368309 N, 116.321011 E, alt. 227 m, 17 Aug., 2020, coll. H. Zhou, X.Y. Shen and X.B. Huang (ZH819/BJTC Z819); CHINA. Beijing, Changping district, Yanshou temple, 40.368654 N, 116.322574 E, alt. 216 m, 17 Aug., 2020, coll. H. Zhou, X.Y. Shen and X.B. Huang (ZH815/BJTC Z815); CHINA. Beijing, Changping district, Yanshou temple, 40.371961 N, 116.321044 E, alt. 245 m, 17 Aug., 2020, coll. H. Zhou, X.Y. Shen and X.B. Huang (ZH824/BJTC Z824); CHINA. Beijing, Changping district, DaYangShan Mountains National Forest Park, 40.308138 N, 116.42437 E, alt. 245 m, 25 July, 2020, coll. H. Zhou, Y.T. Zhang.and X.B. Huang (ZH760/BJTC Z760); CHINA. Beijing, Changping district, DaYangShan Mountains National Forest Park, 40.308003 N, 116.425251 E, alt. 265 m, 14 Aug., 2019, coll. H. Zhou, X.Y. Shen and Y.T. Zhang (ZH083/BJTC Z083); CHINA. Beijing, Pinggu district, Dongxinzhuang village, 40.294008 N, 117.051816 E, alt. 198 m, 19 Aug., 2020, coll. G.Q. Cheng, C.L. Hou.and Y.T. Zhang (C182/BJTC C182); CHINA. Beijing, Changping district, Yanshou temple, 40.373152 N, 116.322892 E, alt. 268 m, 14 Aug., 2019, coll. H. Zhou, X.Y. Shen and X.B. Huang (ZH049/BJTC Z049).

Commentary: A. yanshanensis is well circumscribed by its gray-white to dark grayish red pileus densely arranged with pyramidal, subverrucose to subconical, floccose volval remnants on the stipe base arranged irregularly or in incomplete belts or rings, and mostly ellipsoid, occasionally broadly ellipsoid basidiospores (6.0–9.0 × 4.0–6.0 μm). Furthermore, it is associated with the trees of Fagaceae (Castanea mollissima).

This species belongs to section Validae and is closely related to A. spissacea on the multi-locus phylogenetic tree ( Figure 2 ). In the single loci phylogenetic trees of both nrITS and nrLSU, A. yanshanensis clustered into an independent clade ( Figures S1 , S4 ). Moreover, basidiomata of A. yanshanensis with a gray-brown to gray pileus are also comparable with those of A. spissacea and A. fritillaria (Sacc.) Sacc. However, A. fritillaria has a basal bulb covered with conical, blackish, dark-gray to gray-brown volval remnants and a dirty white to gray annulus (Corner and Bas, 1962; Kumar et al., 1990; Yang, 2005; Yang, 2015; Cui et al., 2018). A. spissacea has a stipe covered with grayish to brownish squamules, pulverulent to floccose, a grayish annulus, and wider basidiospores (7.0–9.5× 6.0–7.5 μm) than A. yanshanensis (Imai, 1933; Imai, 1938; Hongo, 1959; Imazeki and Hongo, 1987; Imazeki et al., 1988; Imazeki et al., 2011; Cui et al., 2018).

 Discussion

In our study, the molecular phylogenetic analyses further supported the delineation of Amanita into two subgenera, namely, subgen. Amanita Pers. and Amanitana (E.-J. Gilbert) E.-J. Gilbert, as suggested by Cui et al. (2018) ( Figures 2 , S1 ). Based on the macroscopic morphology and the preliminary comparison of the original sequence we obtained, the sections where the specimens were located were initially known. Therefore, the sequence information of subgen. Lepidella Beauseigneur and sections Amarrendiae (Bougher & T. Lebel) Zhu L. Yang, Y.Y. Cui, Q. Cai & Ling Ping Tang, Arenariae Zhu L. Yang, Y.Y. Cui & Q. Cai, Amidella (J. E. Gilbert) Konrad & Maubl., and Strobiliformes Singer ex Q. Cai, Zhu L. Yang & Y.Y. Cui were not included in the final phylogenetic analyses. Of them, species in sections Amarrendiae and Arenariae have not been found in China. Cui et al. (2018) mentioned that, for a better understanding of the range of variation in characters, new species should be described based on several specimens, but the technology of molecular systematics used in the present study improved the accuracy of our description of species. As specified in the results, the quality of sequences of these old herbarium specimens (HMAS 283800, HMAS 253802, HMAS 263406, and HMAS 253796) identified as A. oberwinkleriana was not good, but its systematic position and combined morphology could still be somewhat helpful in specimen identification. In addition, we could only depend on morphological observations for identifying these specimens as their sequences could not be obtained because of the poor condition of their basidiomata.

In this study, three new species belonged to three sections under the subgenera Amanita and Amanitana, namely, sections Amanita (A. boreqalis), Caesareae (A. brunneola), and Validae (A. yanshanensis). Section Amanita is distinguished by basidioma with agarose; a pileus with persistent volval remnants, a pileal margin striate; truncated lamellae; presence of a basal bulb; and inamyloid basidiospores (Cui et al., 2018). Many species in this section produced neuropsychotoxins (Wieland, 1973; Yang, 2005; Chen et al., 2016). Studies have identified 27 taxa, comprising 23 species, 2 varieties, and 2 forms, in China (Cui et al., 2018; Su et al., 2022). Previous studies have reported 21 species of section Caesareae (Cui et al., 2018; Mu et al., 2021). The species belonging to section Validae were characterized by a pileal margin non-striae and non-appendiculate; annulus membranous, dominance of filamentous hyphae; volval remnants often as verrucae, warts, flocci, patches, or occasionally as short limb; amyloid basidiospores; and absence of clamps. To date, 18 taxa have been identified from this section (Cui et al., 2018).

Studies have found and recorded 169 Amanita species in China, with most of them being concentrated in the southwest, northwest, and south of China, including Yunnan, Guangdong, Heilongjiang, Liaoning, and Hunan provinces (Yang, 1994; Yang, 1997; Yang et al., 2004; Yang, 2005; Deng et al., 2014; Li and Cai, 2014; Ariyawansa et al., 2015; Li et al., 2015; Yang, 2015; Cai et al., 2016; Deng et al., 2016; Liu et al., 2017; Cui et al., 2018; Su et al., 2022). Records of Amanita species in Yanshan Mountains, northern China, are few. In this study, 12 Amanita species from Yanshan Mountains were recognized. Of them, 10 species or approximately 83% of the species were new or recorded for the first time in this area. Therefore, accelerating the discovery and description of Amanita species by using both morphological and molecular approaches in this area is necessary.

Literature review revealed that 14 taxa of Amanita were identified from Yanshan Mountains. Three species were identified by Yang (2004): A. parvipantherina Zhu L. Yang, M. Weiss & Oberw (HMKS 32350) and A. griseofolia Zhu L. Yang (HMKS 22610) collected from Tanzhe Temple in Beijing and A. subjunquillea var. alba Zhu L. Yang (HMKS 35536) collected from Beishicheng in Beijing. A. vaginata var. vaginata was collected from Donglingshan Mountains, Beijing (Cui et al., 2018). The morphology of HMKS 75237 was consistent with that of A. vaginata var. vaginata, but we could not obtain the DNA sequence from the specimen. The specimen HMAS 26491 was collected from Baihuashan Mountains, Beijing, and was originally identified as A. subglobosa Zhu L. Yang. However, the results of morphological and phylogenetic analyses conducted in the present study were inconsistent with the original identification. We tentatively named this specimen as Amanita sp. because of the poor status of the specimen and await the subsequent collection of additional specimens for further study. In addition, some Amanita species have been recorded in the literature even in the absence of any detailed information about the specimen, and so, the accuracy of the information about these species needs to be further verified by collecting specimens and obtaining molecular data. These species with only distribution records available are as follows. A. caesarea, A. inaurata Gillet, and A. yuaniana are distributed in Wulingshan Mountains, Hebei Province (Wang et al., 2005); A. flavoconia Alk., A. phalloides (Vaill. ex Fr.) Link, and A. subjunquillea are distributed in Dahaituo Mountains National Nature Reserve, Hebei Province (Wu et al., 2017); A. orientigemmata is distributed in Songshan Mountains National Nature Reserve, Beijing (Wu et al., 2020); A. verna Bull ex Lam. is distributed in Badaling Forest Park, Beijing (Zhang et al., 2017); and A. panterina (DC. ex Fr.) Schrmm is distributed in Dayangshan National Forest Park, Beijing (Chen et al., 2006). Therefore, specimens of Amanita spp. must be more extensively collected from the Yanshan Mountains region to improve the study of their biodiversity.

Amanita deserves special attention because of its unique research and popular science education value. However, because some Amanita species are similar in morphology and color, distinguishing them in the field is difficult. Moreover, many casualties have been caused through mistakenly consumed poisonous Amanita species in many places in China (Li et al., 2020; Li et al., 2021a; Li et al., 2021b). The China Center for Disease Control and Prevention reported eight incidents of mushroom poisoning in Beijing in 2020, with 23 people poisoned. Seven incidents of mushroom poisoning were reported from Hebei Province around Beijing, with 33 people poisoned. In these incidents, the main species causing poisoning were A. rimosa, A. subjunquillea, and A. oberwinkleriana (Li et al., 2021a; Li et al., 2021b). The present study is the initial report on Amanita’s biodiversity in the region of Yanshan Mountains, including northern part of Beijing, and Tianjin and Hebei provinces. Given the large area of North China and its diverse forest types, many more Amanita species may be discovered in this region in future.

 Conclusions

In this research, 20 Amanita specimens deposited in Chinese herbaria and 36 newly collected specimens from North China were studied based on the results of morphological and phylogenetic analyses. In total, 12 phylogenetic species were found. Of them, three species were described as new species, namely A. borealis sp. nov., A. brunneola sp. nov., and A. yanshanensis sp. nov. Furthermore, nine known species were identified, namely, A. caesareoides, A. chiui, A. muscaria, A. oberwinklerana, A. ovalispora, A. subglobosa, A. subjunquillea, A. vaginata var. vaginata, and A. virosa. Our results underscore that China has a very high biodiversity of Amanita species and that additional studies are required to completely determine the exact number of species. It plays a crucial role in Amanita toxin research and ecological conservation. This study investigated the areas where Amanita species-related research is lacking. The study attempted to better understand Amanita distribution and thus contribute to related research. This study improves the knowledge regarding the species diversity of Amanita in Yanshan Mountains and provides new data for the macrofungal systematics, toxin research, and diversity and ecological studies of Amanita in subsequent studies.

 Data availability statement

The datasets presented in this study can be found in online repositories (www.treebase.org, study S41229). The NCBI accession number(s) can be found in the article.

 Author contributions

HZ wrote the manuscript, conducted phylogenetic analysis and morphological observations; MG conducted phylogenetic analysis and morphological observations, conducted the experiments; LZ conducted phylogenetic analysis and morphological observations conducted the experiments; HY conducted phylogenetic analysis and morphological observations; XS conducted phylogenetic analysis and morphological observations; YG conducted the experiments; CH conceived and designed the study. All authors contributed to the article and approved the submitted version.

 Funding

This study was financed by the National Natural Science Foundation of China (No. 32270012) and the Biodiversity Survey and Assessment Project of the Ministry of Ecology and Environment, China (2019HJ2096001006).

 Acknowledgments

We are grateful to Prof. Zhu-Liang Yang at Kunming Institute of Botany, Chinese Academy of Sciences, for his improvements to the manuscript. We also thank the two reviewers for their constructive criticism and suggestions to improve our work.

 Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

 Publisher's note

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.

 Supplementary material

The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fpls.2023.1226794/full#supplementary-material

 References Ariyawansa H. A. Hyde K. D. Jayasiri S. C. Buyck B. Chethana K. W. T. Dai D. Q. . (2015). Fungal diversity notes 111–252—taxonomic and phylogenetic contributions to fungal taxa. Fungal Divers. 75 (1), 27–274. doi: 10.1007/s13225-015-0346-5 Bas C. (1967). Amanita argentea Huijsman am Teutoburger Wald gefunden. Westfäl Pilzbr 6 (7), 125–129. Bas C. (1969). Morphology and subdivision of Amanita and a monograph of its section Lepidella . Persoonia 5 (4), 285–579. Beeli M. Goossens-Fontana M. Robyns W. (1935). Flore iconographique des champignons du Congo, Fasc.I (Bruxelles: Office de Publicité). Bhatt R. P. Mehmood T. Uniyal P. Singh U. (2017). Six new records of genus Amanita (Amanitaceae) from uttarakhand, india. Curr. Res. Environ. Appl. Mycol. 7 (3), 161–182. doi: 10.5943/cream/7/3/3 Bhatt R. P. Semwal K. C. Upadhyay R. C. (2007). New records of section Phalloideae of the genus Amanita from Garhwal Himalaya. India. Mushroom Res. 16 (2), 61–67. Bhatt R. P. Tulloss R. E. Semwal K. C. Bhatt V. K. Moncalvo J. M. Stephenson S. L. (2003). Amanitaceae reported from India. A critically annotated checklist. Mycotaxon 88, 249–270. Boedijn K. B. (1951). Notes on Indonesian fungi: the genus Amanita . Sydowia 5, 317–327. Breitenbach J. Kränzlin F. (1995). Pilze der Schweiz. Band 4. Blätterpilze, 2. Teil. Luzern (Paris: Verlag Mykologia, Lucerne). Cai Q. Cui Y. Y. Yang Z. L. (2016). Lethal amanita species in China. Mycologia 108 (5), 993–1009. doi: 10.3852/16-008 Cai Q. Tulloss R. E. Tang L. P. Tolgor B. Zhang P. Chen Z. H. . (2014). Multi-locus phylogeny of lethal Amanitas: implications for species diversity and historical biogeography. BMC Evol. Biol. 14 (1), 143. doi: 10.1186/1471-2148-14-143 Chen J. Q. Cheng J. H. Du Y. Y. Wang Q. B. Yao Y. J. (2006). Preliminary investigation of macrofungi resource in Beijing. J. Beijing Agric. Coll. 21 (2), 39–43. Chen Z. H. Yang Z. L. Bau T. Li T. H. (2016). Poisonous mushrooms: recognition and poisoning treatment (Beijing: Science Press). Chiusa L. L. (2000). Amanite bianche tossiche. Boll Gr Micol G Bres NS 43 (2), 105–116. Cho H. J. Park M. S. Lee H. Oh S. Jang Y. Fong J. J. . (2015). Four new species of Amanita in Inje County, Korea. Mycobiology 43 (4), 408–414. doi: 10.5941/MYCO.2015.43.4.408 Coker W. C. (1917). The amanitas of the eastern united states. J. Elisha Mitchell Sci. Soc. 33, 1–88. Consiglio G. (2000). Contributo alla conoscenza dei macromiceti dell’Emilia-romagna. XXI. Genere amanita. Boll Gr Micol G Bres NS 43 (2), 211–232. Contu M. (2000). Saggio di una chiave per la determinazione delle specie del genere Amanita osservate in Sardegna. Boll Gr Micol G Bres NS 43 (2), 67–86. Corner E. J. H. Bas C. (1962). The genus Amanita in Singapore and Malaya. Persoonia 2 (3), 241–304. Cui Y. Y. Cai Q. Tang L. P. Liu J. W. Yang Z. L. (2018). The family Amanitaceae: molecular phylogeny, higher-rank taxonomy and the species in China. Fungal Divers. 91, 5–230. doi: 10.1007/s11557-019-01506-1 Davison E. M. Giustiniano D. Busetti F. Gates G. M. Syme K. (2017). Death cap mushrooms from southern Australia: additions to Amanita (Amanitaceae, Agaricales) section Phalloideae Clade IX. Aust. Syst. Bot. 30, 371–389. doi: 10.1071/SB17032 Deng W. Q. Li T. H. Hosen M. I. (2016). Amanita rufobrunnescens, a new species of Amanita section Amidella from South China. Phytotaxa 243 (2), 147–154. doi: 10.11646/phytotaxa.243.2.4 Deng W. Q. Li T. H. Li P. Yang Z. L. (2014). A new species of Amanita section Lepidella from South China. Mycol Prog. 13 (2), 211–217. doi: 10.1007/s11557-013-0906-6 Dereeper A. Guignon V. Blanc G. Audic S. Buffet S. Chevenet F. . (2008). Phylogeny.fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Res. 36, W465–W469. doi: 10.1093/nar/gkn180 Endo N. Fangfuk W. Sakuma D. Phosri C. Matsushita N. Fukuda M. . (2016). Taxonomic consideration of the Japanese redcap Caesar’s mushroom based on morphological and phylogenetic analyses. Mycoscience 57 (3), 200–207. doi: 10.1016/j.myc.2016.01.005 Fraiture A. Amalfi M. Raspé O. Kaya E. Akata I. Degreef J. (2019). Two new species of Amanita sect. Phalloideae from Africa, one of which is devoid of amatoxins and phallotoxins. MycoKeys 53, 93–125. doi: 10.3897/mycokeys.53.34560 Galli R. (2001). Le Amanite (Milano: Edinatura). Garcin R. (1984). Les amanites européennes (Marlioz: Fédération mycologique Dauphiné-Savoie). Gardes M. Bruns T. D. (1993). ITS primers with enhanced specificity for basidiomycetes-application to the identification of mycorrhizae and rusts. Mol. Ecol. 2 (2), 113–118. doi: 10.1111/j.1365-294X.1993.tb00005.x Geml J. Laursen G. A. O’neil K. Nusbaum H. C. Taylor D. L. (2006). Beringian origins and cryptic speciation events in the fly agaric (Amanita muscaria). Mol. Ecol. 15, 225–239. doi: 10.1111/j.1365-294X.2005.02799.x Geml J. Tulloss R. E. Laursen G. A. Sazanova N. A. Taylor D. L. (2008). Evidence for strong inter- and intracontinental phylogeographic structure in Amanita muscaria, a wind-dispersed ectomycorrhizal basidiomycete. Mol. Phylogenet Evol. 48, 694–701. doi: 10.1016/j.ympev.2008.04.029 Gilbert E. J. (1940). Amanitaceae. BRESADOLA J. Iconogr Mycol 27 Suppl. 1 (1), 1–200. Gilbert E. J. (1941). Amanitaceae. BRESADOLA J. Iconogr Mycol 27 Suppl. 1 (2–3), 201–427. Guindon S. Dufayard J. F. Lefort V. Anisimova M. Hordijk W. Gascuel O. (2010). New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Sys. Biol. 59, 307–321. doi: 10.1093/sysbio/syq010 Hillis D. M. Bull J. J. (1993). An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Sys. Biol. 42, 182–192. doi: 10.1093/sysbio/42.2.182 Hongo T. (1959). The agaricales of Japan I-(1). Mem Fac Liberal Arts Shiga Univ Nat. Sci. 9, 47–94. Hongo T. (1982). The amanitas of Japan. Acta Phytotaxon Geobot 33, 116–126. Horak E. (1968). Synopsis generum agaricalium. Beitr Krypt Fl Schweiz 13, 1–741. Huang T. Su L. J. Zeng N. K. Lee S. M. L. Lee S. S. Thi B. K. . (2023). Notes on amanita section validae in Hainan Island, China. Front. Microbiol. 13. doi: 10.3389/fmicb.2022.1087756 Huijsman H. S. C. (1959). Deux amanites méconnues. Bull. Soc. Mycol Fr 75, 14–32. Imai S. (1933). Studies on the agaricaceae of Japan. I: volvate agarics in Hokkaido. Bot. Mag (Tokyo) 47, 423–432. doi: 10.15281/jplantres1887.47.423 Imai S. (1938). Studies on the agaricaceae of Hokkaido: I. J. Facul Agric. Hokkaido Imp Univ Sapporo 43 (1), 1–178. Imazeki R. Hongo T. (1987). Colored illustrations of mushrooms of Japan 1 (Osaka: Hoikusha). Imazeki R. Otani Y. Hongo T. (2011). Fungi of Japan (Revisied and enlarged edition) (Tokyo: Yama-kei Publishers Co.). Imazeki R. Otani Y. Hongo T. Izawa M. (1988). Fungi of Japan (Tokyo: Yamakei Publishers). Jenkins D. T. (1977). A taxonomic and nomenclatural study of the genus Amanita section Amanita for North America. Bibl Mycol 57, 3–126. Jenkins D. T. (1986). Amanita of North America (Eureka: Mad River Press). Jenkins D. T. Petersen R. H. (1976). A neotype specimen for Amanita muscaria . Mycologia 68 (3), 463–469. doi: 10.1080/00275514.1976.12019935 Katoh K. Toh H. (2010). Parallelization of the MAFFT multiple sequence alignment program. Bioinformatics 26, 1899–1900. doi: 10.1093/bioinformatics/btq224 Kawase I. Shirakawa H. Watanabe M. (1992). Deaths caused by Amanita subjunquillea poisoning and the distribution of this mushroom in Hokkaido. Trans. Mycol Soc. Jpn 33, 107–110. Kim C. S. Jo J. W. Kwag Y. N. Kim J. H. Shrestha B. Sung G. H. . (2013a). Taxonomic study of Amanita subgenus Lepidella and three unrecorded Amanita species in Korea. Mycobiology 41 (4), 183–190. doi: 10.5941/MYCO.2013.41.4.183 Kim C. S. Jo J. W. Kwag Y. N. Oh J. Shrestha B. Sung G. H. . (2013b). Four newly recorded Amanita species in Korea: Amanita sect. Amanita and sect. Vaginatae . Mycobiology 41 (3), 131–138. doi: 10.5941/MYCO.2013.41.3.131 Konrad P. Maublanc A. (1924). Icones selectae fungorum. Tome 1. (Paris: Paul Lechevalier). Kumar A. Bhatt R. Lakhanpal T. N. (1990). The Amanitaceae of India (Dehra Dun: Bishen Singh Mahendra Pal Singh). Kumar A. Mehmood T. Atri N. S. Sharma Y. P. (2021). Revised and an updated checklist of the Amanitaceae from India with its specific distribution in the Indian States. Nova Hedwigia 112 (1–2), 223–240. doi: 10.1127/nova_hedwigia/2021/0617 Lange J. E. (1935). Flora agaricina danica, vol I. Recato (Copenhagen: printed by Recato A/S Copenhagen). Li F. Cai Q. (2014). Amanita heishidingensis, a new species of Amanita sect. Lepidella from China. Mycol Prog. 13 (4), 1191–1197. doi: 10.1007/s11557-014-1008-9 Li W. W. Pires S. M. Liu Z. T. Liang J. J. Wang Y. F. Chen W. . (2021b). Mushroom poisoning outbreaks China 2010-2020. China CDC Week. 3 (24), 518–522. doi: 10.46234/ccdcw2021.134 Li H. J. Xie J. W. Zhang S. Zhou Y. J. Ma P. B. Zhou J. . (2015). Amanita subpallidorosea, a new lethal fungus from China. Mycol Prog. 14 (6), 43. doi: 10.1007/s11557-015-1055-x Li H. J. Zhang H. S. Zhang Y. Z. Zhang K. P. Zhou J. Yin Y. . (2020). Mushroom poisoning outbreaks China. 2019. China CDC Week. 2 (2), 19–24. doi: 10.46234/ccdcw2020.005 Li H. J. Zhang H. S. Zhang Y. Z. Zhou J. Yin Y. He Q. . (2021a). Mushroom poisoning outbreaks China 2020. China CDC Week. 3 (3), 41–45. doi: 10.46234/ccdcw2021.014 Liu J. W. Cai Q. Cui Y. Y. Yang Z. L. (2017). Amanita cingulata, a new annulate species of Amanita sect. Vaginatae from subtropical China. Phytotaxa 326 (1), 41–53. doi: 10.11646/phytotaxa.326.1.3 Mao X. L. (1990). Taxonomic study on the genus Amanita from Xizang, China. Acta Mycol Sin. 9, 206–217. Menolli N. J. Capelari M. Baseia I. G. (2009). Amanita viscidolutea, a new species from Brazil with a key to Central and South American species of Amanita section Amanita . Mycologia 101 (3), 395–400. doi: 10.3852/07-079 Mighell K. S. Henkel T. W. Koch R. A. Chin M. L. Brann M. A. Aime M. C. (2021). Amanita in the Guineo-Congolian rainforest: epitypes and new species from the Dja Biosphere Reserve. Cameroon. Mycol. 113 (1), 168–190. doi: 10.1080/00275514.2020.1816386 Mighell K. S. Henkel T. W. Koch R. A. Goss A. Alime M. C. (2019). New species of Amanita subgen. Lepidella from Guyana. Fungal Syst. Evol. 3, 1–12. doi: 10.3114/fuse.2019.03.01 Moser M. (1967). “Die Röhrlinge und Blätterpilze (Agaricales),” in Kleine Kryptogamenflora, Band II b/2. Basidiomyceten II. Teil. 3. Ed. Gams H. H. (Stuttgart/New York: Gustav Fischer Verlag). Mu M. Huang H. Y. Zhang W. H. Tang L. P. (2021). A new species with pink lamellae of Amanita section Caesareae from China. Phytotaxa 478 (1), 141–150. doi: 10.11646/phytotaxa.478.1.10 Neville P. Poumarat S. (2004). Amaniteae: Amanita, Limacella & Torrendia (Italy: Edizioni Candusso, Alassio). Nylander J. A. A. (2004). MrModeltest 2.2. Computer software distributed by the University of Uppsala (Uppsala, Sweden: Evolutionary Biology Centre, University of Uppsala). Pegler D. N. Shah-Smith D. (1997). The genus amanita (Amanitaceae, agaricales) in Zambia. Mycotaxon 61, 389–417. Persoon C. H. (1797). Tentamen dispositionis methodicae fungorum (Leipzig: Petrum Philippum Wolf). Posada D. Crandall K. A. (1998). Modeltest: testing the model of DNA substitution. Bioinformatics 14, 817–818. doi: 10.1093/bioinformatics/14.9.817 Rannala B. Yang Z. (1996). Probability distribution of molecular evolutionary trees: a new method of phylogenetic inference. J. Mol. Evol. 43, 304–311. doi: 10.1007/BF02338839 Rehner S. A. Buckley E. (2005). A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97 (1), 84–98. doi: 10.1080/15572536.2006.11832842 Reid D. A. (1980). A monograph of the Australian species of Amanita Pers. ex Hook. (Fungi). Aust. J. Bot. Suppl. Ser. 10 (8), 1–96. Ronquist F. Huelsenbeck J. P. (2003). MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19, 1572–1574. doi: 10.1093/bioinformatics/btg180 Sanmee R. Tulloss R. Lumyong P. Dell B. Lumyong S. (2008). Studies on amanita (Basidiomycetes: amanitaceae) in northern Thailand. Fungal Divers. 32, 97–123. Semwal K. C. Tulloss R. Bhatt R. Stephenson S. Upadhyay R. (2007). New records of Amanita section Amanita from Garwhal Himalaya, India. Mycotaxon 101, 331–348. Singer R. (1986). The Agaricales in modern taxonomy. 4th edn (Koenigstein: Koeltz Scientific Books). Stamatakis A. (2006). RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 22, 2688–2690. doi: 10.1093/bioinformatics/btl446 Stamatakis A. Hoover P. Rougemont J. (2008). A rapid bootstrap algorithm for the RAxML web servers. Sys. Biol. 57, 758–771. doi: 10.1080/10635150802429642 Su Y. T. Cai Q. Qin W. Q. Cui Y. Y. Chen Z. H. Yang Z. L. (2022). Two new species of Amanita section Amanita from Central China. Mycol Prog. 21, 78. doi: 10.1007/s11557-022-01828-7 Suwannarach N. Kumla J. Khuna S. Wannathes N. Thongklang N. Sysouphanthong P. . (2022). History of Thai mycology and resolution of taxonomy for Thai macrofungi confused with Europe and American names. Chiang Mai J. Sci. 49 (3), 654–683. doi: 10.12982/CMJS.2022.052 Tang S. S. Zhou Q. He Z. M. Luo T. Zhang P. Cai Q. . (2016). Cyclopeptide toxins of lethal amanitas: compositions, distribution and phylogenetic implication. Toxicon 120, 78–88. doi: 10.1016/j.toxicon.2016.07.018 Thiers H. D. (1982). The agaricales (Gilled fungi) of california: Amanitaceae (Eurecka: Mad River Press). Tulloss R. E. (2005). Amanita-distribution in the Americas, with comparison to eastern and southern Asia and notes on spore character variation with latitude and ecology. Mycotaxon 93, 189–232. Tulloss R. E. Bhatt R. Stephenson S. Kumar A. (1995). Studies on Amanita (Amanitaceae) in West Virginia and adjacent areas of the Mid-Appalachians. Preliminary results. Mycotaxon 56, 243–293. Tulloss R. E. Iqbal S. H. Khalid A. N. Bhatt R. P. Bhat V. K. (2001). Studies in Amanita (Amanitaceae) from southern asia. i. some species of pakistan’s northwest frontier province. Mycotaxon 77, 455–490. Vassiljeva L. N. (1950). Species novae fungorum. Notul Syst. Sect Cryptog Inst Bot. Acad. Sci. USSR 6, 188–200. Vilgalys R. Hester M. (1990). Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. J. Bacteriol. 172, 4238–4246. doi: 10.1128/jb.172.8.4238-4246.1990 Wang Y. Huang Z. H. Wang J. Zhang T. Cui G. F. (2021). The population structure and dynamic characteristics of Phellodendron amurense in Yanshan Mountains. Acta Ecol. Sin. 47 (7), 2826–2834. doi: 10.5846/stxb202003300743 Wang Q. Liu H. X. Zhang J. G. Lu J. Liu Y. X. Yang L. H. . (2005). Investigation of wild edible and medical fungi of Wuling mountain area. J. Hebei Univ. (Natural Sci. Edition). 25 (5), 523–525. White T. J. Bruns T. Lee S. Taylor J. (1990). Amplifcation and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: Guide to Methods Appl. 18, 315–322. doi: 10.1016/B978-0-12-372180-8.50042-1 Wieland T. H. (1973). Über die Giftstoffe der Gattung Amanita. Zeitschr f Pilzk 39, 103–112. Wood A. (1997). Studies in the genus amanita (Agaricales) in Australia. Aust. Syst. Bot. 10, 723–854. doi: 10.1071/SB95049 Wu M. Huang H. Y. Zhang W. H. Tang L. P. (2021). A new species with pink lamellae of Amanita section Caesareae from China. Phytotaxa 478 (1), 141–150. doi: 10.11646/phytotaxa.478.1.10 Wu Z. J. Song C. Lin T. M. Yu Q. Y. Yu M. F. Xing S. H. . (2017). Resource and ecological distribution of microfungi in Dahaituo national nature reserve of Hebei province. Edible Fungi China 36 (4), 8–14. Wu J. G. Zhou H. Hou C. L. (2020). Diversity of poisonous macrofungi in Beijing Songshan national nature reserve. J. Capital Normal Univ. (Natural Sci. Edition) 41 (4), 52–56. Yang Z. L. (1994). Studies of the genus Amanita from southwestern China (I). Mycotaxon 51, 459–470. Yang Z. L. (1997). Die amanita-arten von SüdwestChina. Bibl Mycol 170, 1–240. Yang Z. L. (2004). Two new species of Amanita (Basidiomycota) from China. Eds. Agerer R. Piepenbring M. Blanz P. (Frontiers), (Eching: IHW Verlag). Yang Z. L. (2005). Flora fungorum sinicorum. Vol. 27. Amanitaceae (Beijing: Science Press). Yang Z. L. (2015). Atlas of the Chinese species of Amanitaceae (Beijing: Science Press). Yang Z. L. Cai Q. Cui Y. Y. (2018). Phylogeny, diversity and morphological evolution of Amanitaceae. Biosyst. Ecol. Ser. 34, 359–380. Yang Z. L. Doi Y. (1999). A contribution to the knowledge of Amanita (Amanitaceae, Agaricales) in Japan. Bull. Natn Sci. Mus Tokyo Ser. B 25 (3), 107–130. Yang Z. L. Li T. H. (2001). Notes on three white Amanitae of section Phalloideae (Amanitaceae) from China. Mycotaxon 78, 439–448. Yang Z. L. Oberwinkler F. (1999). Die Fruchtköperentwicklung von Amanita muscaria (Basidiomycetes). Nova Hedwigia 68, 441–468. doi: 10.1127/nova.hedwigia/68/1999/441 Yang Z. L. Weiß M. Oberwinkler F. (2004). New species of Amanita from the eastern Himalaya and adjacent regions. Mycologia 96 (3), 636–646. doi: 10.1080/15572536.2005.11832959 Zhang P. Chen Z. H. Xiao B. Tolgor B. Bao H. Y. Yang Z. L. (2010). Lethal amanitas of East Asia characterized by morphological and molecular data. Fungal Divers. 42 (1), 119–133. doi: 10.1007/s13225-010-0018-4 Zhang X. R. Zhou X. Huang Z. H. Pu Z. Zhang X. L. Wang Q. C. . (2017). Ecological distribution of marcofungi in Badaling Forest Park, Beijing. J. Arid Land Resour. Environ. 31 (8), 181–186. doi: 10.1016/j.ecocom.2017.02.005 Zhou H. Cheng G. Q. Hou C. L. (2022b). A new species, Russula luteolamellata (Russulaceae, Russulales) from China. Phytotaxa 556 (2), 136–148. doi: 10.11646/phytotaxa.556.2.3 Zhou H. Cheng G. Q. Sun X. M. Cheng R. Y. Zhang H. L. Dong Y. M. . (2022a). Three new species of Candolleomyces (Agaricomycetes, Agaricales, Psathyrellaceae) from the Yanshan Mountains in China. MycoKeys 88, 109–121. doi: 10.3897/mycokeys.88.81437 Zhou H. Cheng G. Q. Wang Q. T. Guo M. J. Zhuo L. Yan H. F. . (2022c). Morphological characteristics and phylogeny reveal six new species in russula subgenus russula (Russulaceae, russulales) from Yanshan mountains, North China. J. Fungi 8 (12), 1283. doi: 10.3390/jof8121283