Abstract

Introduction

Agarics are mushroom-forming fungi also called euagarics and their hymenium is formed on gills. They belong to the subdivision Agaricomycotina, class Agaricomycetes (Moncalvo et al., 2002; Bauer et al., 2006). They produce important natural substances used in agriculture (e.g., strobilurines), medicine (e.g., pleuromutilines), and biotechnology (e.g., polysaccharides; Pointing et al., 2001; Webster and Weber, 2007; Kück et al., 2014; Hyde et al., 2019; Sandargo et al., 2019). Agaricales is the largest fungal order of agaric mushrooms comprising ca. 13,000 known species (Kirk et al., 2008). Thorough investigations of the agarics phylogeny have recently been provided by He et al. (2019) and Varga et al. (2019). Some agarics are important model organisms for research in genetics and basidiome development such as Coprinopsis cinerea and Cyclocybe cylindrica (Herzog et al., 2019). Among agarics, there are some of the most poisonous mushrooms such as Amanita phalloides, Cortinarius Subgen. Orellani, and Inosperma erubescens, frequently mixed up with edible mushrooms during culinary collecting and thus causing severe fatalities. Nevertheless, there is a large number of edible agaric mushrooms highly prized for culinary purposes such as Agaricus campestris, Coprinus comatus, Cyclocybe cylindrica, Macrolepiota procera, and the worldwide cultivated white button mushroom Agaricus bisporus. Several edible agaric species are saprotrophs and possible to cultivate, but there are also many edible species such as Russula spp. and Lactarius spp. which belong to the ectomycorrhizal ecological guild and thus not cultivable in artificial synthetic media. A number of species such as Lentinula edodes and Flammulina velutipes have culminated as functional mushrooms for developing mushroom-based functional foods and other valued mycochemicals (Chang, 1996; Cateni et al., 2022; Rodríguez-Seoane et al., 2022).

Numerous agarics have also been recognized as sources of antioxidant compounds (e.g., Ferreira et al., 2009; Asatiani et al., 2010; Guo et al., 2012; Wang and Xu, 2014; Sánchez, 2017; Islam et al., 2019; Thu et al., 2020). Antioxidant properties, or the ability to defend against and scavenge/reduce excess free radicals in biological systems, is among the important properties of living organisms and crucial for their survival (Xiao et al., 2020). Mushrooms as one of the most diverse natural antioxidant resources, have received attention in recent decades and are advantageous compared to plants because of their high diversity, fast growth, and culture possibilities (Gargano et al., 2017; Buswell, 2018).

A preliminary checklist of Iranian mushrooms appeared by Ghobad-Nejhad et al. (2020) listing 556 agaric and 29 bolete species. However, the species remain largely unexplored in terms of various important properties. Information about the edible, poisonous, and mycorrhizal agarics in Iran is principally lacking and currently, the antioxidant properties of Iranian agarics have remained largely unexplored.

Due to the lack of knowledge about the diversity of edible, poisonous, and mycorrhizal agarics in Iran, as well as their antioxidant properties, our study aimed to: (i) investigate Iranian agarics and reveal their resources of edible and poisonous species, (ii) present their ecological guilds and bioluminescence potential, and to (iii) explore the antioxidant properties of Iranian agarics and combine it with phylogenetic reconstructions. We believe our results would benefit a wide range of researchers involved in the study of agaric mushrooms.

Materials and methods

Sampling and molecular study

Taxon sampling for the molecular study was primarily done based on the list by Ghobad-Nejhad et al. (2020), supplemented by additional data in the present study. Species current names and species authorities follow Index Fungorum¹ and MycoBank.² Microscopy and morphological studies followed Ghobad-Nejhad et al. (2020). Sequences of the 28S rRNA (nLSU) and the ITS region (covering ITS1, 5.8, and ITS2) were carefully selected from GenBank, with special attention to the quality-controlled sequences (Nilsson et al., 2012) as well as to the authentic sequences obtained from Iranian specimens. For DNA extraction, we sampled more than 20 specimens and 12 samples were successfully sequenced and used in this paper. Genomic DNA was extracted from dried basidiomata using the DNA Extraction Mini Kit (FAVORGEN, Taiwan). The primers used for the amplification cycles were ITS1F/ITS4B or ITS1F/ITS4 (White et al., 1990; Gardes and Bruns, 1993) for the ITS region and LR0R/LR7 or LR0R/LR5 (Hopple and Vilgalys, 1999) for partial nLSU region. All sequences used in the phylogenetic analyses are listed in Table 1.

Table 1

Resources of agarics of Iran and their edibility (, edible; , edible based on own observation in Iran; , poisonous; , poisonous based on own observation in Iran; *, edible if well-cooked but poisonous if raw; X, inedible; ○, uncertain or unknown), ecological guild (♠, soil saprotroph; ☼, ectomycorrhizal; ▐, wood-inhabiting; ♣, leaf/litter-inhabiting; , parasitic), luminescence, and antioxidant potential (S, strong; M, moderate; W, weak; ND, not determined; full details provided in the text).

Species	Edibility	Ecological guild	Luminescent	Antioxidant potential	GenBank accession no.
					ITS	nLSU
Agaricus arvensis Schaeff.		♠	–	S	MT535720	MH872779
Agaricus bisporus (J.E. Lange) Imbach		♠	–	SM	ON952490	DQ071710
Agaricus bitorquis (Quél.) Sacc.		♠	–	SM	MT535709	MT554302
Agaricus bresadolanus Bohus		♠	–	MW	DQ185569	MK277477
Agaricus brunneolus (J.E. Lange) Pilát		♠	–	ND	KU975082	KX083997
Agaricus campestris L.		♠	–	SM	NR_151745.1	MH868030
Agaricus depauperatus (F.H. Møller) Pilát		♠	–	ND	DQ182530	–
Agaricus devoniensis P.D. Orton		♠	–	ND	EU363036	AF059225
Agaricus gennadii (Chatin & Boud.) P.D. Orton		♠	–	ND	KT951318	KR006606
Agaricus iodosmus Heinem.		♠	–	S	MT535702	MT554295
Agaricus iranicus Mahdizadeh, Safaie, Goltapeh, L.A. Parra & Callac	○	♠	–	ND	KY474556	KY474559
Agaricus langei (F.H. Møller) F.H. Møller		♠	–	ND	JF797181	–
Agaricus litoralis (Wakef. & A. Pearson) Pilát		♠	–	ND	MT535711	MT554304
Agaricus moelleri Wasser		♠	–	ND	KT824787	–
Agaricus nevoi Wasser	○	♠	–	ND	MH173866	–
Agaricus phaeolepidotus (F.H. Møller) F.H. Møller		♠	–	ND	MH862921	MH874494
Agaricus pseudolutosus (G. Moreno, Esteve-Rav., Illana & Heykoop) G. Moreno, L.A. Parra, Esteve-Rav. & Heykoop		♠	–	ND	KT951329	KT951453
Agaricus pseudopratensis (Bohus) Wasser		♠	–	S	ON952491	MT554325
Agaricus subrufescens Peck		♠	–	M	KT983412	KT951461
Agaricus xanthodermus Genev.		♠	–	ND	KT824789	KR006612
Agrocybe acericola (Peck) Singer	○	♠	–	ND	MN860126	MK277500
Agrocybe dura (Bolton) Singer		♠ ▐ (saprothrophic on soil, but on decaying woody remnants)	–	S	MT535714	MT554306
Agrocybe ochracea Nauta	X	♠ ▐ (saprothrophic on soil, but on decaying woody remnants)	–	ND	–	–
Agrocybe paludosa (J.E. Lange) Kühner & Romagn. ex Bon	X	♠	–	ND	–	–
Agrocybe pediades (Fr.) Fayo	X	♠	–	M	ON952487, ON952488	AY293582
Agrocybe praecox (Pers.) Fayod		♠ ▐ (saprothrophic on soil, on decaying woody remnants)	–	W	MT535701	MT554294
Agrocybe pusiola (Fr.) R. Heim	X	♠	–	ND	DQ389732	MK277505
Agrocybe tabacina (DC.) Konrad & Maubl.	X	♠	–	ND	–	–
Agrocybe vervacti (Fr.) Singer	X	♠	–	ND	MW425942	MK277506
Alnicola escharioides (Fr.) Romagn.	X	☼	–	ND	MW243076	–
Amanita atkinsoniana Coker	○	☼	–	ND	MZ668014	MK277560
Amanita battarrae (Boud.) Bon	*	☼	–	ND	MH508267	MH486389
Amanita caesarea (Scop.) Pers.		☼	–	M	MZ005548	AF024443
Amanita ceciliae (Berk. & Broome) Bas	*	☼	–	ND	OK299150	OK299170
Amanita crocea (Quél.) Singer	*	☼	–	M	KJ638266	–
Amanita eliae Quél.		☼	–	ND	KF780872	–
Amanita excelsa (Fr.) Bertill.		☼	–	ND	MW258873	MW258922
Amanita gemmata (Fr.) Bertill.		☼	–	ND	MK580689	AF024457
Amanita lividopallescens (Secr. ex Boud.) Kühner & Romagn. agg.	*	☼	–	ND	MT535691	MW013165
Amanita pantherina (DC.) Krombh.		☼	–	M	FR852274	MH486743
Amanita phalloides (Fr.) Link		☼	–	ND	KX449212	KX449230
Amanita rubescens Pers.		☼	–	S	FR852273	MH486816
Amanita strobiliformis (Paulet ex Vittad.) Bertill.	X	☼	–	ND	MH508614	MH486895
Amanita umbrinolutea (Secr. ex Gillet) Bataille	*	☼	–	ND	MH508641	MH486937
Amanita vaginata (Bull.) Lam. s.l.	*	☼	–	M	JF907756	–
Amanita verna (Bull.) Lam.		☼	–	ND	EU909448	HQ539755
Ampulloclitocybe clavipes (Pers.) Redhead, Lutzoni, Moncalvo, and Vilgalys	(seems to be toxic after consumption with alcohol)	♠	–	ND	AY789080	AY639881
Armillaria borealis Marxm. & Korhonen	*	♠	Desjardin et al. (2008)	ND	KP960524	FJ618728
Armillaria cepistipes Velen.	*	♠	Mihail (2015)	ND	FJ903313	KY418876
Armillaria gallica Marxm. & Romagn.	*	♠	Kotlobay et al. (2018)	ND	MW418538	AM269818
Armillaria mellea (Vahl) P. Kumm.	*	♠	Kotlobay et al. (2018)	SM	AF163583	AM269819
Arrhenia griseopallida (Desm.) Watling	X	♠	–	ND	–	–
Asterophora lycoperdoides (Bull.) Ditmar	X		–	ND	MZ159455	MK277604
Atheniella flavoalba (Fr.) Redhead, Moncalvo, Vilgalys, Desjardin, and B.A. Perry	X	♠	–	ND	MH857185	MH868723
Baeospora myosura (Fr.) Singer	X	♠ (on conifer cones)	–	ND	MH856301	MH867849
Battarrea stevenii (Libosch.) Fr.	X	♠	–	ND	AF215648	–
Bolbitius reticulatus (Pers.) Ricken	X	▐	–	ND	JX968249	JX968366
Bolbitius titubans (Bull.) Fr.	X	♠	–	ND	KR425522	KR425552
Calocybe carnea (Bull.) Donk		♠	–	ND	AF357028	MK277666
Calocybe chrysenteron (Bull.) Singer	X	♠	–	ND	KP885639	KP885628
Calocybe gambosa (Fr.) Donk		♠	–	W	MZ159691	AM946414
Calocybe ionides (Bull.) Donk		♠	–	ND	JF907780	MK277668
Calocybe persicolor (Fr.) Singer	X (edible in the Czech Republic)	♠	–	ND	KP192564	AF223176
Candolleomyces candolleanus (Fr.) D. Wächt. & A. Melzer		▐	–	S	MT535718	MT554309
Cantharellus alborufescens (Malençon) Papetti & S. Alberti		☼	–	M	MH463257	MH463258
Cantharellus cibarius Fr.		☼	–	SM	KX907204	KX828805
Cantharellus ferruginascens P.D. Orton		☼	–	ND	MH463294	MH463295
Chlorophyllum brunneum (Farl. & Burt) Vellinga		♠	–	ND	MG742013	MG742022
Chlorophyllum rhacodes (Vittad.) Vellinga		♠	–	M	AY081236	AY176345
Clitocybe angustissima (Lasch) P. Kumm.		♠	–	ND	–	–
Clitocybe barbularum (Romagn.) P.D. Orton	X	♠	–	ND	–	–
Clitocybe diatreta (Fr.) P. Kumm.		♠	–	ND	–	–
Clitocybe metachroa (Fr.) P. Kumm.	X	♠	–	ND	JF907806	AY207155
Clitocybe nebularis (Batsch) P. Kumm.		♠	–	S	DQ149727	AY586685
Clitocybe phyllophila (Pers.) P. Kumm.		♠	–	ND	MH856300	MH867847
Clitocybe rufuloalutacea Métrod ex Bon	○	♠	–	ND	–	–
Clitocybe vibecina (Fr.) Quél.	X	♠	–	ND	JF907821	AY207160
Clitopaxillus alexandri (Gillet) G. Moreno, Vizzini, Consiglio & P. Alvarado		♠	–	W	MG321345	MG321393
Clitopilus prunulus (Scop.) P. Kumm.		♠ ☼	–	M	FJ770408	GU384615
Clitopilus scyphoides (Fr.) Singer	X	♠	–	ND	MH856181	MH867707
Collybia tuberosa (Bull.) P. Kumm.	X	♠	Malakauskienė (2018)	ND	AY854072	AY639884
Conocybe albipes (G.H. Otth) Hauskn.	X	♠	–	ND	–	–
Conocybe apala (Fr.) Arnolds		♠	–	ND	MT535728	MT554318
Conocybe dunensis T.J. Wallace	X	♠	–	ND	JX968227	JX968345
Conocybe juniana (Velen.) Hauskn. & Svrček	X	♠	–	ND	JX968191	JX968307
Conocybe leucopus Kühner ex Kühner & Watling	X	♠	–	ND	–	–
Conocybe macrocephala Kühner & Watling	X	♠	–	ND	JX968182	JX968298
Conocybe microspora (Velen.) Dennis	X	♠	–	ND	JX968160	JX968276
Conocybe ochracea Kühner ex Singer	X	♠	–	ND	–	–
Conocybe olivaceopileata E.F. Malyshevaa	X	♠	–	ND	ON952486	–
Conocybe pilosella (Pers.) Kühner	X	♠	–	ND	JX968231	JX968349
Conocybe rickenii (Jul. Schäff.) Kühner	X	♠	–	ND	AY194541	AY293597
Conocybe subovalis Kühner & Watling	X	♠	–	ND	JX968190	JX968306
Conocybe tenera (Schaeff.) Fayod		♠	–	M	MH855754	MH867266
Contumyces rosellus (M.M. Moser) Redhead, Moncalvo, Vilgalys, and Lutzoni	○	♠	–	ND	OL771755	OL771796
Coprinellus angulatus (Peck) Redhead	X	♠ ▐ (on burnt ground/wood)	–	ND	MN121285	MH868315
Coprinellus disseminatus (Pers.) J.E. Lange	X	▐	–	ND	MK050584	AY207180
Coprinellus domesticus (Bolton) Vilgalys, Hopple & Jacq. Johnson	X	▐	–	S	MH856480	MH868019
Coprinellus flocculosus (DC.) Vilgalys, Hopple & Jacq. Johnson	X	♠ ▐	–	ND	FN396138	FN396208
Coprinellus impatiens (Fr.) J.E. Lange	X	♠	–	ND	MH856810	MH868327
Coprinellus micaceus (Bull.) Vilgalys, Hopple & Jacq. Johnson	X	▐	–	S	ON952489	MT554289
Coprinellus radians (Desm.) Vilgalys, Hopple & Jacq. Johnson	X	▐	–	ND	KU375662	KM272009
Coprinellus silvaticus (Peck) Gminder	X	♠ ▐	–	ND	HQ846986	HQ847072
Coprinellus subimpatiens (M. Lange & A.H. Sm.) Redhead, Vilgalys, and Moncalvo	X	♠ ▐	–	ND	MH857001	MH868522
Coprinellus truncorum (Scop.) Redhead, Vilgalys, and Moncalvo	X	▐	–	S	FM878007	FM876263
Coprinellus xanthothrix (Romagn.) Vilgalys, Hopple & Jacq. Johnson	X	▐	–	ND	JN943112	JN159595
Coprinopsis atramentaria (Bull.) Redhead, Vilgalys, and Moncalvo	(toxic after consumption with alcohol)	▐	–	SM	MH259864	FN396172
Coprinopsis brunneofibrillosa (Dennis) Redhead, Vilgalys, and Moncalvo	X	▐	–	ND	JX118664	JX118817
Coprinopsis cinerea (Schaeff.) Redhead, Vilgalys, and Moncalvo	X	♠	–	M	MF161131	KM272007
Coprinopsis ephemeroides (DC.) G. Moreno	X	♠	–	ND	–	–
Coprinopsis friesii (Quél.) P. Karst.	X	♣	–	ND	–	FN396191
Coprinopsis gonophylla (Quél.) Redhead, Vilgalys & Moncalvo	X	♠	–	ND	MH856188	MH867714
Coprinopsis lagopides (P. Karst.) Redhead, Vilgalys & Moncalvo	X	♠	–	ND	MN892574	AF041488
Coprinopsis lagopus (Fr.) Redhead, Vilgalys & Moncalvo	X	♠ ▐ ♣	–	ND	MH856194	MH867720
Coprinopsis macrocephala (Berk.) Redhead, Vilgalys & Moncalvo	X	♠ ♣	–	ND	FN396126	FN396175
Coprinopsis marcescibilis (Britzelm.) Örstadius & E. Larss.	X	♠ ▐	–	ND	ON952484	FM876278
Coprinopsis martinii (P.D. Orton) Redhead, Vilgalys & Moncalvo	X	♣	–	ND	GU234126	–
Coprinopsis nivea (Pers.) Redhead, Vilgalys & Moncalvo	X	♠	–	ND	HQ847032	HQ847117
Coprinopsis patouillardii (Quél.) Gminder	X	♠	–	ND	FN396150	FN396197
Coprinopsis picacea (Bull.) Redhead, Vilgalys & Moncalvo	X	♠	–	S	JN943110	JQ045885
Coprinopsis sclerotiger (Watling) Redhead, Vilgalys & Moncalvo	X	♠	–	ND	MF161091	MF161132
Coprinopsis scobicola (P.D. Orton) Redhead, Vilgalys & Moncalvo	X	♠ ▐	–	ND	HQ847021	HQ847106
Coprinopsis urticicola (Berk. & Broome) Redhead, Vilgalys & Moncalvo	X	▐ ♣	–	ND	MH300615	HQ847101
Coprinus comatus (O.F. Müll.) Pers.	(considered edible in Europe and also cultivated)	♠	–	SM	MH817141	MH997559
Coprinus sterquilinus (Fr.) Fr.	X	♠	–	ND	MH854689	AF041530
Cortinarius bivelus (Fr.) Fr.	X	☼	–	ND	FR852016	–
Cortinarius caesiocortinatus Jul. Schäff.	X	☼	–	ND	FR852020	–
Cortinarius casimirii (Velen.) Huijsman	X	☼	–	ND	FR851999	–
Cortinarius causticus Fr.	X	☼	–	ND	FJ157016	FJ157016
Cortinarius cinnabarinus Fr.	X	☼	–	ND	KC842405	KC842476
Cortinarius cinnamomeus (L.) Gray		☼	–	ND	NR_131816	KC842483
Cortinarius cotoneus Fr.	X	☼	–	ND	KC842423	KC842493
Cortinarius decipiens Fr.	X	☼	–	ND	HE687043	–
Cortinarius diasemospermus Lamoure	X	☼	–	ND	HE687042	–
Cortinarius erumpens Rob. Henry	○	☼	–	ND	–	–
Cortinarius ferrugineovelatus Kytöv., Liimat. & Niskanen	○	☼	–	ND	NR_131875	MK277631
Cortinarius fluryi (M.M. Moser) M.M. Moser	○	☼	–	ND	–	–
Cortinarius hildegardiae Schmidt-Stohn, Brandrud & Dima	○	☼	–	ND	MT535704	MT554297
Cortinarius hinnuleus Fr.	X	☼	–	ND	AY083183	AF388779
Cortinarius infractus (Pers.) Fr.	X	☼	–	ND	NR_130225	KC842497
Cortinarius olivaceofuscus Kühner		☼	–	ND	AY669585	MK277762
Cortinarius paracephalixus Bohus	X	☼	–	ND	KR080708	–
Cortinarius parvannulatus Kühner	X	☼	–	ND	HE687041	–
Cortinarius persoonianus Bidaud	○	☼	–	S	MT535741	MT554330
Cortinarius pluviorum Jul. Schäff. ex M.M. Moser	X	☼	–	ND	FJ157038	FJ157038
Cortinarius uraceonemoralis Niskanen, Liimat., Dima, Kytöv., Bojantchev & H. Lindstr.	○	☼	–	ND	NR_131836	MK277663
Cortinarius valgus Fr.	X	☼	–	ND	MT935583	–
Cortinarius vernus H. Lindstr. & Melot	X	☼	–	ND	MW263848	MW263545
Cortinarius vespertinus (Fr.) Fr.	X	☼	–	ND	KC842457	KC842527
Cortinarius vibratilis (Fr.) Fr.	X	☼	–	ND	KC842440	KC842510
Cortinarius violaceus (L.) Gray		☼	–	W	NR_173726	MK277758
Craterellus cinereus (Pers.: Fr.) Maire		☼	–	ND	–	–
Craterellus cornucopioides (L.) Pers.		☼	–	SM	JF907967	MN227282
Craterellus tubaeformis (Fr.) Quél.		☼	–	S	HM468493	MF797698
Crepidotus applanatus (Pers.) P. Kumm.	X	▐	–	ND	MH855941	MH867439
Crepidotus caspari Velen.	X	▐	–	ND	MW722982	AF205678
Crepidotus cesatii (Rabenh.) Sacc.	X	▐	–	ND	JF907962	MK277881
Crepidotus crocophyllus (Berk.) Sacc.	X	▐	–	ND	FJ596825	AF367939
Crepidotus mollis (Schaeff.) Staude	X	▐	–	ND	AM882996	AM882996
Crepidotus subverrucisporus Pilát	X	▐	–	ND	MT535745	AF367948
Crinipellis scabella (Alb. & Schwein.) Murrill	X	♠	–	ND	MH857177	MH868716
Cuphophyllus virgineus (Wulfen) Kovalenko		♠	–	ND	MT535688	MT554284
Cyclocybe cylindracea (DC.) Vizzini & Angelini		▐	–	MW	ON952480, ON952485	ON930146
Cystoderma aureum (Matt.) Kühner & Romagn.	X (generally edible, but some health problems described after eating)	♠	–	ND	MH864957	MH876401
Deconica coprophila (Bull.) P. Karst.	X	♠	–	ND	MH855878	MH867388
Deconica crobula (Fr.) Romagn.	X	♠	–	ND	MT535747	MH867478
Delicatula integrella (Pers.) Fayod	X	▐	–	ND	MZ159362	MK277924
Dermoloma cuneifolium (Fr.) Singer ex Bon	X	♠	–	ND	MW193843	–
Echinoderma asperum (Pers.) Bon		♠	–	W	MH856136	MH867652
Entoloma clypeatum (L.) P. Kumm.	(frequently eaten in the Czech Republic, considered poisonous in Chinab)	☼	–	ND	KC710059	KC710136
Entoloma griseoluridum (Kühner) M.M. Moser	X	♠	–	ND	–	–
Entoloma griseorubellum (Lasch) Kalamees & Urbonas	○	♠	–	ND	–	–
Entoloma hirtipes (Schumach.) M.M. Moser	X	♠	–	ND	MN088710	MN088715
Entoloma incanum (Fr.) Hesler	X	♠	–	ND	OK161249	OK161276
Entoloma majaloides P.D. Orton	X	♠	–	ND	MW633049	MW633049
Entoloma mammosum (L.) Hesler	○	♠	–	ND	–	–
Entoloma niphoides Noordel.	X	☼	–	ND	JF907999	FJ794075
Entoloma rhodopolium (Fr.) P. Kumm.		♠	–	ND	LN850497	LN850705
Entoloma sericellum (Fr.) P. Kumm.	X	♠	–	ND	KC898453	GQ289190
Entoloma sinuatum (Bull. ex Pers.) P. Kumm.		☼	–	ND	KC710116	KC710154
Entoloma subcollariatum (Kühner) Bon	○	♠	–	ND	MH453494	–
Entoloma vernum S. Lundell		♠	–	ND	MF476911	MF487802
Flammula alnicola (Fr.) P. Kumm.	(considered edible in the Czech Republic)	♠	–	ND	MH862103	MH873792
Flammulaster erinaceellus (Peck) Watling	X	▐	–	ND	MF755278	EF537889
Flammulaster ferrugineus (Maire) Watling	X	▐	–	ND	MF039253	–
Flammulaster gracilis (Quél.) Watling	X	▐	–	ND	–	–
Flammulaster granulosus (J.E. Lange) Watling	X	♠	–	ND	–	–
Flammulina velutipes (Curtis) Singer		▐	Desjardin et al. (2008)	SM	MT535715	MT554307
Galerina hypnorum (Schrank) Kühner		♠	–	ND	OL771728	MK299406
Galerina marginata (Batsch) Kühner		▐	–	ND	MK346203	MK346279
Galerina mniophila (Lasch) Kühner	X	♠	–	ND	AJ585456	AJ871514
Galerina pumila (Pers.) M. Lange	X	♠	–	ND	AJ585477	AJ871546
Galerina sphagnorum (Pers.) Kühner	X	♠	–	ND	AJ585455	AJ871510
Gymnopilus penetrans (Fr.) Murrill	X (considered inedible in the Czech Republic)	▐	–	W	KR011987	KR011988
Gymnopilus spectabilis (Weinm.) A.H. Sm.	X (considered inedible in the Czech Republic)	▐	–	S	MT535703	MT554296
Gymnopus androsaceus (L.) J.L. Mata & R.H. Petersen	X	▐ ♣	–	ND	MH857176	MH868715
Gymnopus aquosus (Bull.) Antonín & Noordel.		♠	–	ND	MT535700	MT554293
Gymnopus brassicolens (Romagn.) Antonín & Noordel.	X	▐ ♠	–	ND	MZ088117	MK278106
Gymnopus dryophilus (Bull.) Murrill	(considered poisonous in China)	♠	–	W	MH589967	MH589985
Gymnopus erythropus (Pers.) Antonín, Halling & Noordel.		▐	–	ND	JX536136	AY207167
Gymnopus foetidus (Sowerby) J.L. Mata & R.H. Petersen	X	▐	–	ND	KY026682	KY026682
Gymnopus fusipes (Bull.) Gray	X (only young basidiomata edible)	▐	–	W	KY026727	KY026727
Gymnopus hybridus (Kühner & Romagn.) Antonín & Noordel.	X	♠	–	ND	MT535705	MT554299
Gymnopus inodorus (Pat.) Antonín & Noordel.	X	▐	–	ND	–	–
Gymnopus terginus (Fr.) Antonín & Noordel.	X	♠	–	ND	–	MK278118
Hebeloma birrus (Fr.) Gillet	X	☼	–	ND	JF908029	–
Hebeloma crustuliniforme (Bull.) Quél.		☼	–	ND	MH856151	MH867674
Hebeloma hiemale Bres.	X	☼	–	ND	KT591536	KT591556
Hebeloma incarnatulum A.H. Sm.	X	☼	–	ND	KX687211	–
Hebeloma mesophaeum (Pers.) Quél.	X	☼	–	ND	NR_173705	MK880553
Hebeloma sinapizans (Paulet) Gillet		☼	–	M	KT591542	KT591562
Hemimycena cucullata (Pers.) Singer	X	▐ ♠ (most frequently wood-inhabiting)	–	ND	–	–
Hodophilus hymenocephalus (A.H. Sm. & Hesler) Birkebak & Adamčík	X	♠	–	ND	DQ484066	DQ457679
Hohenbuehelia atrocoerulea (Fr.) Singer	X	▐	–	ND	KU355304	KU355389
Hohenbuehelia auriscalpium (Maire) Singer	X	▐	–	ND	MT525860	MT534052
Hohenbuehelia petaloides (Bull.) Schulzer		▐	–	ND	NR_173155	KU355402
Homophron spadiceum (P. Kumm.) Örstadius & E. Larss.	X	▐	–	ND	MK968340	MN028523
Hydropus marginellus (Pers.) Singer	X	▐	–	ND	DQ490627	DQ457674
Hygrocybe acutoconica (Clem.) Singer		♠	–	ND	OK157438	MK278174
Hygrocybe chlorophana (Fr.) Wünsche		♠	–	ND	JF908052	MK278164
Hygrophorus eburneus (Bull.) Fr.		☼	–	S	MK088116	AF430279
Hygrophorus mesotephrus Berk. & Broome		☼	–	ND	MT981695	–
Hygrophorus persoonii Arnolds		☼	–	ND	MN243172	KF291213
Hymenopellis radicata (Relhan) R.H. Petersen		▐	–	M	MZ159452	MT554280
Hypholoma capnoides (Fr.) P. Kumm.		▐	–	W	FJ596780	AY207211
Hypholoma fasciculare (Huds.) P. Kumm.		▐	–	SM	MT535706	MT554300
Hypholoma lateritium (Schaeff.) P. Kumm.	X	▐	–	SM	MH856121	MH866989
Hypholoma radicosum J.E. Lange	X (considered edible in China)	▐	–	ND	–	DQ071685
Hypholoma subericaeum (Fr.) Kühner	X	♠	–	ND	–	MK278215
Hypsizygus ulmarius (Bull.) Redhead		▐	–	S	AY265850	AF042584
Infundibulicybe geotropa (Bull. ex DC.) Harmaja	(considered poisonous in China)	♠	–	MW	KT122792	KT122793
Infundibulicybe gibba (Pers.) Harmaja		♠	–	ND	MH856103	MZ719010
Infundibulicybe trulliformis (Fr.) Gminder	X	♠	–	ND	JF907809	–
Inocybe amethystina Kuyper		☼	–	ND	HE687066	–
Inocybe asterospora Quél.		☼	–	ND	HM060326	HM060325
Inocybe castaneicolor A. La Rosa, Bizio, Saitta & Tedersoo		☼	–	ND	KY213954	KY213954
Inocybe cincinnata (Fr.) Quél.		☼	–	ND	MG489949	KC305372
Inocybe corydalina Quél.		☼	–	ND	MH216083	MH220259
Inocybe decemgibbosa (Kühner) Vauras		☼	–	ND	HE687073	–
Inocybe flocculosa Sacc.		☼	–	ND	LT716045	KY418861
Inocybe geophylla (Sowerby) P. Kumm.		☼	–	ND	KY990536	JN974951
Inocybe godeyi Gillet		☼	–	ND	FN550897	FN550897
Inocybe hirtella Bres.		☼	–	ND	EU523581	EU307822
Inocybe huijsmanii Kuyper		☼	–	ND	FR852248	–
Inocybe ionolepis Cullington & E. Larss.c	○	☼	–	ND	FR852270	–
Inocybe langei R. Heim		☼	–	ND	HE687072	JN974962
Inocybe leptocystis G.F. Atk.		☼	–	ND	AM882801	AM882801
Inocybe lilacina (Peck) Kauffman		☼	–	ND	KY990528	KY990484
Inocybe mixtilis (Britzelm.) Sacc.		☼	–	ND	HQ586870	HQ641113
Inocybe mystica Stangl & Glowinski		☼	–	ND	NR_158509	–
Inocybe napipes J.E. Lange		☼	–	ND	KP308784	KP170955
Inocybe paludinella (Peck) Sacc.		☼	–	ND	JF908135	–
Inocybe praetervisa Quél.		☼	–	ND	KY033785	KY033785
Inocybe pusio P. Karst.		☼	–	ND	FR852266	AY388643
Inocybe subnudipes Kühner		☼	–	ND	FN550925	FN550925
Inocybe tabacina Furrer-Ziogas		☼	–	ND	HQ586865	HQ641106
Inocybe terrifera Kühner		☼	–	ND	–	–
Inosperma adaequatum (Britzelm.) Matheny & Esteve-Rav.		☼	–	ND	NR_153149	JQ815407
Inosperma bongardii (Weinm.) Matheny & Esteve-Rav.		☼	–	ND	FN550943	FN550943
Inosperma cookei (Bres.) Matheny & Esteve-Rav.		☼	–	ND	AM882956	AM882956
Inosperma erubescens (A. Blytt) Matheny & Esteve-Rav.		☼	–	ND	AM882951	AM882951
Inosperma maculatum (Boud.) Matheny & Esteve-Rav.		☼	–	ND	MH578017	MT228862
Laccaria amethystina Cooke		☼	–	W	KU685654	KU685797
Laccaria bicolor (Maire) P.D. Orton		☼	–	ND	KM067831	KU685788
Laccaria laccata (Scop.) Cooke		☼	–	W	KM067835	KU685859
Laccaria tortilis (Bolton) Cooke		☼	–	ND	MG519533	MG519576
Lacrymaria lacrymabunda (Bull.) Pat.	X	♠	–	ND	MK968341	MN031155
Lactarius acris (Bolton) Gray	X	☼	–	ND	JQ446084	JQ446156
Lactarius circellatus Fr.	X	☼	–	ND	FR852038	JN388995
Lactarius deliciosus (L.) Gray		☼	–	MW	KJ769672	KF133305
Lactarius fulvissimus Romagn.	X	☼	–	ND	FR852027	–
Lactarius rubrocinctus Fr.	X	☼	–	ND	UDB005472 (UNITE)	–
Lactarius scrobiculatus (Scop.) Fr.	X	☼	–	ND	KX441098	KX441345
Lactarius serifluus (DC.) Fr.		☼	–	ND	KT165294	–
Lactarius subdulcis (Pers.) Gray	X	☼	–	ND	KX395722	MH872686
Lactarius tabidus Fr.		☼	–	ND	KT165309	JN389012
Lactarius zonarius (Bull.) Fr.	(edible in Iran, inedible in Europe)	☼	–	ND	FR852035	MT747331
Lactifluus glaucescens (Crossl.) Verbeken	X	☼	–	ND	MT535681	MT554278
Lactifluus piperatus (L.) Roussel	(edible after special preparation)	☼	–	SM	KF220122	KF220215
Lactifluus vellereus (Fr.) Kuntze	X (reported as edible in Turkey by Dogan and Aydin (2013); as inedible in Europe by Heleno et al. (2012))	☼	–	SM	KF220123	KF220216
Lactifluus volemus (Fr.) Kuntze		☼	–	W	JQ753936	JQ348387
Lentinellus cochleatus (Pers.) P. Karst.		▐	–	ND	AF506417	AF506417
Lentinellus ursinus (Fr.) Kühner	(inedible in Europe)	▐	–	ND	MH857168	MH868705
Lentinellus vulpinus (Sowerby) Kühner & Maire	X	▐	–	ND	AY513230	–
Lentinus cyathiformis (Schaeff.) Bres.	(inedible in Europe)	▐	–	ND	KM411461	KM411477
Lentinus lepideus (Fr.) Fr.	(inedible in Europe)	▐	–	M	KM411454	KM411478
Lentinus sajor-caju (Fr.) Fr.		▐	–	MW	OL771751	OL771792
Lentinus strigosus Fr.	X	▐	–	ND	KM411451	KM411468
Lentinus tigrinus (Bull.) Fr.		▐	–	SM	ON952481	MT554282
Lepiota anthomyces (Berk. & Broome) Sacc.	○	♠	–	ND	–	–
Lepiota brunneoincarnata Chodat & C. Martin		♠	–	ND	MK651615	MK685374
Lepiota castanea Quél.		♠	–	ND	MK685380	MK651688
Lepiota cristata (Bolton) P. Kumm.	X (considered poisonous in China)	♠	–	ND	LT716026	KY418841
Lepiota echinella Quél. & G.E. Bernard	X	♠	–	ND	AY176366	AY176367
Lepiota felina (Pers.) P. Karst.		♠	–	ND	MK685381	MK278264
Lepiota helveola Bres.		♠	–	ND	MH979466	–
Lepiota leprica (Berk. & Broome) Sacc.	○	♠	–	ND	–	–
Lepiota lilacea Bres.		♠	–	ND	AY176379	AY176380
Lepiota metulispora (Berk. & Broome) Sacc.	○	♠	–	ND	EU681778	MK651673
Lepiota micropholis (Berk. & Broome) Sacc.	X	♠	–	ND	–	–
Lepiota subalba Kühner ex P.D. Orton	X	♠	–	ND	AY176489	–
Lepiota subincarnata J.E. Lange		♠	–	ND	U85329	U85294
Lepista irina (Fr.) H.E. Bigelow	(edible/inedible in the Europe)	♠	–	ND	MH862098	MH873787
Lepista nuda (Bull.) Cooke		♠	–	SM	KU215619	DQ071713
Lepista saeva (Fr.) P.D. Orton		♠	–	ND	MK785234	MH878430
Leratiomyces squamosus (Pers.) Bridge & Spooner	X	♠	–	ND	MH043620	MH036179
Leucoagaricus americanus (Peck) Vellinga		♠	–	ND	MT573394	AF482891
Leucoagaricus badhamii (Berk. & Broome) Singer		♠	–	ND	GQ329056	–
Leucoagaricus carneifolius (Gillet) Wasser	X	♠	–	ND	–	–
Leucoagaricus holospilotus (Berk. & Broome) Bon	○	♠	–	ND	–	–
Leucoagaricus leucothites (Vittad.) Wasser	(reported edible in Turkey by Aslim and Ozturk (2011); edible but sometimes caused health problems)	♠	–	SM	MT535726	MT554316
Leucoagaricus nympharum (Kalchbr.) Bon		♠	–	ND	EU416310	EU416311
Leucoagaricus roseoalbus (Henn.) Heinem.	○	♠	–	ND	–	–
Leucoagaricus serenus (Fr.) Bon & Boiffard		♠	–	ND	AY176420	AF482893
Leucocoprinus birnbaumii (Corda) Singer		♠	–	ND	MH861267	MH873036
Leucocoprinus brebissonii (Godey) Locq.		♠	–	ND	AF482859	AY176446
Leucocoprinus cepistipes (Sowerby) Pat.	X	♠	–	ND	LT716023	KY418838
Leucocoprinus magnusianus (Henn.) Singer	○	♠	–	ND	–	–
Leucocybe candicans (Pers.) Vizzini, P. Alvarado, G. Moreno & Consiglio		♠	–	ND	KJ681027	KJ681051
Leucocybe houghtonii (W. Phillips) Halama & Pencakowski	X	♠	–	ND	KY474108	–
Leucopaxillus compactus (P. Karst.) Neuhoff	X	♠	–	ND	–	–
Leucopaxillus giganteus (Sowerby) Singer	(edible in Europe, considered poisonous in China)	♠	–	M	JQ639151	JQ639152
Leucopaxillus pinicola J. Favre	○	♠	–	ND	–	–
Lyophyllum atratum (Fr.) Singer	X	♠	–	ND	KJ461896	KJ461895
Lyophyllum baeospermum Romagn.	X	♠	–	ND	–	–
Macrocybe gigantea (Massee) Pegler & Lodge		♠	–	S	MG867660	AF042591
Macrolepiota excoriata (Schaeff.) Wasser	(edible in Europe)	♠	–	MW	U85313	U85278
Macrolepiota mastoidea (Fr.) Singer		♠	–	MW	HM125532	MH867678
Macrolepiota permixta (Barla) Pacioni		♠	–	ND	HQ412661	–
Macrolepiota procera (Scop.) Singer	(poisonous in China)	♠	–	SM	ON952483	AM946456
Mallocybe dulcamara (Pers.) Vizzini		☼	–	ND	HQ604787	EU569836
Mallocybe terrigena (Fr.) Matheny, Vizzini & Esteve-Rav.	X (edible in China)	☼	–	ND	AM882864	AM882864
Marasmiellus candidus (Fr.) Singer	X	▐	–	ND	MT573397	MH867503
Marasmiellus confluens (Pers.) J.S. Oliveira	X	♠	–	ND	–	–
Marasmiellus peronatus (Bolton) J.S. Oliveira	X (considered edible/poisonous in China)	♠	–	S	AY256706	–
Marasmiellus ramealis (Bull.) Singer	X	▐	–	ND	KY404985	KY404980
Marasmius atrorubens (Berk.) Mont.	X	♠	–	ND	KP635207	KP635160
Marasmius corrugatiformis Singer	X	♠	–	ND	KX148981	–
Marasmius epiphyllus (Pers.) Fr.	X	♣	–	ND	JN943599	JN941147
Marasmius favoloides Henn.	X	♠	–	ND	–	–
Marasmius ferrugineus Berk. & M.A. Curtis	X	♠	–	ND	–	–
Marasmius haematocephalus (Mont.) Fr.	X	♠	–	ND	KX148986	EF160083
Marasmius oreades (Bolton) Fr.		♠	–	SM	LT716048	KY418864
Marasmius rotula (Scop.) Fr.	X	▐	–	ND	JN943598	JN941146
Marasmius rubroflavus (Theiss.) Singer	X	♠	–	ND	–	–
Marasmius wynneae Berk. & Broome		♠	–	ND	FJ904979	MH868580
Megacollybia platyphylla (Pers.) Kotl. & Pouzar	X	▐	–	ND	MT535698	MT554291
Melanoleuca cognata (Fr.) Konrad & Maubl.		♠	–	ND	JX429190	JX429180
Melanoleuca exscissa (Fr.) Singer		♠	–	S	MT535742	MT554331
Melanoleuca graminicola (Velen.) Kühner & Maire		♠	–	ND	JN616438	–
Melanoleuca grammopodia (Bull.) Fayod		♠	–	ND	JF908351	MH868277
Melanoleuca strictipes (P. Karst.) Jul. Schäff.		♠	–	ND	JX429116	JX429162
Melanoleuca subpulverulenta (Pers.) Métrod Note: synonym to M. friesii (Bres.) Bon (Antonín et al., 2022) but identification probably tentative.		♠	–	ND	JN616473	–
Montagnea arenaria (DC.) Zeller		♠	–	ND	NR_173482	MK278380
Montagnea haussknechtii Rabenh.	X	♠	–	ND	–	–
Mycena acicula (Schaeff.) P. Kumm.	X	♣ ▐	–	ND	MW540677	MK278389
Mycena clavicularis (Fr.) Gillet	X	♣	–	ND	MW540674	AF042637
Mycena crocata (Schrad.) P. Kumm.	X	♣	–	ND	JF908492	MH868172
Mycena filopes (Bull.) P. Kumm.	X	♣	–	ND	OM473731	–
Mycena galericulata (Scop.) Gray		▐	–	ND	DQ404392	MH866154
Mycena galopus (Pers.) P. Kumm.	X (edible in China)	♣	Treu and Agerer (1990)	ND	FR846482	AY207250
Mycena haematopus (Pers.) P. Kumm.	X	▐	Bermudes et al. (1992)	ND	LT716053	KY418869
Mycena inclinata (Fr.) Quél.	X	▐	Desjardin et al. (2008)	ND	MK532829	MK278392
Mycena metata (Fr.) P. Kumm.	X	♣	–	ND	MZ315004	–
Mycena pearsoniana Dennis ex Singer		♠	–	ND	FN394614	FN394633
Mycena pelianthina (Fr.) Quél.		♠	–	ND	FN394549	FN394626
Mycena polygramma (Bull.) Gray	X	▐	Treu and Agerer (1990)	ND	MH856239	MH867768
Mycena pura (Pers.) P. Kumm.		♠	Treu and Agerer (1990)	ND	KF913023	FN394630
Mycena rapiolens J. Favre	X	♣	–	ND	–	–
Mycena sanguinolenta (Alb. & Schwein.) P. Kumm.	X	♣	Desjardin et al. (2008)	ND	MH856662	AY207257
Mycena xantholeuca Kühner	X	♣	–	ND	MT535719	MT554310
Mycenastrum corium (Guers.) Desv.	X (inedible in Europe)	♠	–	S	MH855530	–
Mycenella salicina (Velen.) Singer	X	♠	–	ND	JF908497	DQ071720
Mycetinis alliaceus (Jacq.) Earle		▐	–	ND	MH856155	MH867679
Mycetinis scorodonius (Fr.) A.W. Wilson & Desjardin	X	♣ ▐	–	ND	MH856330	MH867884
Myxomphalia maura (Fr.) Hora	X	♠	–	ND	MH856673	MH868189
Neofavolus suavissimus (Fr.) J.S. Seelan, Justo & Hibbett	X	▐	–	ND	KM411460	KM411476
Neolentinus adhaerens (Alb. & Schwein.) Redhead & Ginns	X	▐	–	ND	HM536096	KJ141188
Omphaliaster asterosporus (J.E. Lange) Lamoure	X	♠	–	ND	MZ159333	–
Omphalina mutila (Fr.) P.D. Orton	X	♠	–	ND	FJ770399	–
Omphalina pyxidata (Bull.) Quél.	X	♠	–	ND	MF319071	MF318927
Omphalotus olearius (DC.) Singer		▐	Kotlobay et al. (2018)	SM	AF525061	AF042010
Ossicaulis lignatilis (Pers.) Redhead & Ginns		▐	–	ND	DQ825426	AF261397
Ossicaulis salomii Siquier & Bellanger	○	▐	–	ND	MT535738	MT554327
Panaeolus acuminatus (P. Kumm.) Quél.		♠	–	ND	MH856251	MH867783
Panaeolus campanulatus (L.) Quél.		♠	–	ND	JF908522	–
Panaeolus fimicola (Fr.) Quél.		♠	–	ND	JF908519	MK278431
Panaeolus olivaceus F.H. Møller		♠	–	ND	MH285992	MK278433
Panaeolus papilionaceus (Bull.) Quél.		♠	–	ND	MH100681	MK278435
Panaeolus plantaginiformis (Lebedeva) E.F. Malysheva	○	♠	–	ND	MK397579	MK397601
Panaeolus rickenii Hora		♠	–	ND	JF908523	–
Panaeolus semiovatus (Sowerby) S. Lundell & Nannf.	X	♠	–	ND	MH856675	MH868191
Panaeolus speciosus P.D. Orton		♠	–	ND	–	–
Panaeolus teutonicus Bride & Métrod		♠	–	ND	–	–
Panellus stipticus (Bull.) P. Karst.	X	▐	Kotlobay et al. (2018) (chemiluminescence: Shimomura, 1991)	ND	MH855557	MH867062
Panus conchatus (Bull.) Fr.		▐	–	M	OL477381	OL477382
Paragymnopus perforans (Hoffm.) J.S. Oliveira	X	♣	–	ND	MH856221	AJ406586
Paralepista flaccida (Sowerby) Vizzini		♠	–	M	MZ159662	MZ675572
Parasola auricoma (Pat.) Redhead, Vilgalys & Hopple	X	♠ ▐	–	ND	MH855972	MH867468
Parasola hemerobia (Fr.) Redhead, Vilgalys & Hopple	X	♠ ▐	–	ND	FM163189	FM160720
Parasola leiocephala (P.D. Orton) Redhead, Vilgalys & Hopple	X	♠ ▐	–	ND	JN943113	JQ045887
Parasola miser (P. Karst.) Redhead, Vilgalys & Hopple	X	♠	–	ND	KY928619	KY928638
Parasola plicatilis (Curtis) Redhead, Vilgalys & Hopple	X	♠	–	ND	KY928625	KY928643
Paraxerula caussei (Maire) Petersen		▐	–	ND	OL770198	AM946473
Phaeomarasmius erinaceus (Fr.) Scherff. ex Romagn.	X	▐	–	M	MH856667	MH868183
Phaeonematoloma myosotis (Fr.) Bon	X	♠	–	ND	AF195599	AF195599
Phellorinia herculeana (Pers.) Kreisel	X	♠	–	ND	JX984569	–
Phloeomana speirea (Fr.) Redhead	X	▐	–	ND	MH856159	MK278448
Pholiota adiposa (Batsch) P. Kumm.		▐	–	SM	MT535689	MT554285
Pholiota astragalina (Fr.) Singer	X	▐	–	ND	MT187979	MT228845
Pholiota gummosa (Lasch) Singer	X	▐	–	ND	MH861987	MH873679
Pholiota highlandensis (Peck) A.H. Sm. & Hesler	X	♠	–	ND	MH348872	MH867483
Pholiota jahnii Tjall.-Beuk. & Bas	X	▐	–	ND	MT535737	MT554326
Pholiota populnea (Pers.) Kuyper & Tjall.-Beuk.	X	▐	–	ND	MG735315	–
Pholiota scamba (Fr.) M.M. Moser	X	▐	–	ND	JF908585	–
Pholiota spumosa (Fr.) Singer	X	▐	–	ND	MN209776	MN251159
Pholiota squarrosa (Oeder) P. Kumm.	(edible but very tough; considered poisonous in China)	▐	–	ND	MN209778	MN251161
Pholiota squarrosoides (Peck) Sacc.	X (considered edible/poisonous in China)	▐	–	ND	JF908591	AF261641
Pholiotina aporos (Kits van Wav.) Clémençon	X	♠	–	ND	JX968260	JX968376
Pholiotina arrhenii (Fr.) Singer	X	♠	–	ND	JX968261	JX968377
Pholiotina striipes (Cooke) M.M. Moser	X	♠	–	ND	JX968150	JX968267
Pholiotina vexans (P.D. Orton) Bon	X	♠	–	ND	JX968265	JX968380
Phyllotopsis nidulans (Pers.) Singer		▐	–	W	MF686492	AF042578
Pleurotellus chioneus (Pers.) Kühner	X	▐	–	ND	–	–
Pleurotus calyptratus (Lindblad ex Fr.) Sacc.		▐	–	W	EU424283	EU365640
Pleurotus cornucopiae (Paulet) Rolland		▐	–	SM	MT535734	MT554324
Pleurotus djamor (Rumph. ex Fr.) Boedijn		▐	–	SMW	EU424306	EU365661
Pleurotus dryinus (Pers.) P. Kumm.		▐	–	M	EU424292	MH872241
Pleurotus elongatipes Peck	○	▐	–	ND	–	–
Pleurotus eryngii (DC.) Quél.		♠	–	SMW	MT535679	MT554276
Pleurotus fossulatus (Cooke) Sacc.		▐	–	W	HM998828	U04136
Pleurotus nebrodensis (Inzenga) Quél.		♠	–	SM	HM998835	EU365659
Pleurotus ostreatus (Jacq.) P. Kumm.		▐	–	SMW	ON952482	MT554286
Pleurotus pulmonarius (Fr.) Quél.		▐	–	SMW	LT716061	KY418877
Pluteus aurantiorugosus (Trog) Sacc.	X	▐	–	ND	JF908613	AF261579
Pluteus cervinus (Schaeff.) P. Kumm.		▐	–	SM	MT535687	MT554283
Pluteus chrysophaeus (Schaeff.) Quél.	X	▐	–	ND	MH010881	MH010881
Pluteus cinereofuscus J.E. Lange	X	▐	–	ND	MH595963	MK278491
Pluteus depauperatus Romagn.	X	▐	–	ND	–	–
Pluteus exiguus (Pat.) Sacc.	X	▐	–	ND	FJ774083	–
Pluteus leoninus (Schaeff.) P. Kumm.		▐	–	ND	HM562045	HM562234
Pluteus nanus (Pers.) P. Kumm.	X	▐	–	ND	MH595974	MK278504
Pluteus pellitus (Pers.) P. Kumm.		▐	–	ND	HM562037	HM562225
Pluteus petasatus (Fr.) Gillet		▐	–	ND	HM562038	HM562224
Pluteus punctipes P.D. Orton	X	▐	–	ND	–	–
Pluteus romellii (Britzelm.) Sacc.		▐	–	ND	HM562062	HM562238
Pluteus salicinus (Pers.) P. Kumm.	(considered edible in China)	▐	–	ND	HM562051	HM562233
Pluteus semibulbosus (Lasch) Gillet	X	▐	–	ND	KR022021	MH867792
Pluteus thomsonii (Berk. & Broome) Dennis	X	▐	–	ND	HM562053	HM562230
Pluteus umbrosus (Pers.) P. Kumm.		▐	–	ND	HM562140	HM562232
Pogonoloma macrocephalum (Schulz.) Sánchez-García	○	♠	–	ND	MH595847	KJ417209
Psathyrella bivelata Contu	X	♠	–	S	MT535693	MT554288
Psathyrella clivensis (Berk. & Broome) P.D. Orton	X	♠ ▐	–	ND	DQ389683	DQ389683
Psathyrella fatua (Fr.) Konrad & Maubl.	X	▐	–	ND	MT535695	MT554290
Psathyrella hellebosensis D. Deschuyteneer & A. Melzer	X	♠	–	ND	MT535716	MT554308
Psathyrella laevissima (Romagn.) Singer	X	▐	–	ND	–	–
Psathyrella microrhiza (Lasch) Konrad & Maubl.	X	♠ ▐	–	ND	MH856265	MH867801
Psathyrella multipedata (Peck) A.H. Sm.	X	♠ ▐	–	ND	GQ249282	GQ249291
Psathyrella obtusata (Fr.) A.H. Sm.	X	♠ ▐	–	ND	MH860428	–
Psathyrella pennata (Fr.) A. Pearson & Dennis	X	♠	–	ND	AM712259	AM712259
Psathyrella piluliformis (Bull.) P.D. Orton		▐	–	ND	FN396136	FN396185
Psathyrella prona (Fr.) Gillet	X	♠ ▐	–	ND	MH856268	MH867805
Psathyrella pseudogracilis (Romagn.) M.M. Moser	X	▐	–	ND	MH856200	MH867728
Psathyrella spadiceogrisea (Schaeff.) Maire	X	♠ ▐	–	ND	MK045737	MK045738
Psathyrella squamosa (P. Karst.) A.H. Sm.	X	♠ ▐	–	ND	AM712250	AM712250
Psathyrella tephrophylla (Romagn.) M.M. Moser	X	♠ ▐	–	ND	AM712270	AM712270
Pseudoclitocybe cyathiformis (Bull.) Singer		♠	–	ND	MT535721	MT554311
Pseudosperma perlatum (Cooke) Matheny & Esteve-Rav.		☼	–	ND	JQ408767	JQ319698
Pseudosperma rimosum (Bull.) Matheny & Esteve-Rav.		☼	–	ND	MF278770	EU600853
Psilocybe atrobrunnea (Lasch) Gillet	X	♠	–	ND	HG423575	HG423577
Psilocybe cyanescens Wakef.		♠	–	ND	NR_111478	NG_069074
Psilocybe serbica M.M. Moser & E. Horak		♠	–	ND	MF958473	MF958467
Resupinatus applicatus (Batsch) Gray	X	▐	–	ND	NR_171800	NG_075208
Rhodocollybia maculata (Alb. & Schwein.) Singer	X (edible in China)	♠	–	ND	MH857674	MH869212
Rhodocollybia prolixa (Fr.) Antonín & Noordel.	X	▐	–	ND	–	MK278563
Rhodotus palmatus (Bull.) Maire	X (maybe poisonous; edibility considered unknown by Heleno et al. (2012))	▐	–	MW	MK287617	MK287618
Russula acetolens Rauschert		☼	–	ND	–	–
Russula alutacea (Pers.) Fr.	(considered poisonous in China)	☼	–	SM	JF908676	–
Russula anthracina Romagn.		☼	(chemiluminescence: Bondar et al., 2012; Gitelson et al., 2012)	S	MW172321	MW182481
Russula atropurpurea Peck [non R. atropurpurea (Krombh.) Britzelm. (= R. undulata Velen.)]		☼	–	ND	JF908691	KU237550
Russula atrorubens Quél.	X	☼	–	ND	KX579812	KX812877
Russula brunneoviolacea Crawshay		☼	–	ND	AM113956	–
Russula carminipes J. Blum		☼	–	ND	–	KU237523
Russula claroflava Grove		☼	–	ND	KT933997	KT933858
Russula cyanoxantha (Schaeff.) Fr.		☼	(chemiluminescence: Bondar et al., 2012; Gitelson et al., 2012)	S	MW646981	MW646993
Russula delica Fr.		☼	(chemiluminescence: Bondar et al., 2012; Gitelson et al., 2012)	W	KX812842	KX812864
Russula emetica (Schaeff.) Pers.	(European R. emetica extremely pungent and inedible/toxic; reported by Kaewnarin et al. (2016) as popular edible mushroom in Thailand)	☼	–	M	KX813352	KX812896
Russula emeticolor J. Schaeffer		☼	–	S	MT535680	MT554277
Russula farinipes Romell	X (edible/poisonous in China)	☼	–	ND	KY800361	KU237561
Russula foetens Pers.	X (may cause health problems)	☼	(chemiluminescence: Gitelson et al., 2012)	ND	KT934016	KT933877
Russula graveolens Romell		☼	–	ND	KU205298	–
Russula grisea Fr.		☼	–	ND	MT738286	MT738262
Russula heterophylla (Fr.) Fr.		☼	–	ND	AF418609	AF325309
Russula integra (L.) Fr.		☼	–	M	KY582682	KX812899
Russula ionochlora Romagn.		☼	–	ND	MW683795	KU237508
Russula lilacea Quél.		☼	–	ND	JN944005	JN940592
Russula luteotacta Rea	X (edible/poisonous in China)	☼	–	ND	JF908652	KU237512
Russula nigricans Fr.		☼	–	S	–	–
Russula ochroleuca Pers.		☼	(chemiluminescence: Bondar et al., 2012; Gitelson et al., 2012)	ND	HM189900	KU237519
Russula ochroleucoides Kauffman	X	☼	–	ND	–	–
Russula olivacea Pers.		☼	–	M	AF418635	KU237492
Russula pectinata Fr.	X	☼	–	ND	MW355005	–
Russula pectinatoides Peck	X (edible only when very young)	☼	–	ND	EU598185	KU237462
Russula persicina Krombh.	X	☼	–	ND	HE687094	KU237494
Russula perlactea Murrill	○	☼	–	ND	–	–
Russula puellaris Fr.		☼	–	ND	AF418628	KU237515
Russula queletii Fr.		☼	–	ND	KT934007	KT933868
Russula risigallina (Batsch) Sacc.		☼	–	ND	JF908685	–
Russula romellii Maire		☼	–	ND	KT933987	KT933848
Russula rosea Pers.		☼	–	S	JN944003	JN940602
Russula silvestris (Singer) Reumaux	X	☼	–	ND	KX579800	–
Russula solaris Ferd. & Winge	X	☼	–	ND	AF418627	JN940606
Russula sororia (Fr.) Romell	X	☼	–	ND	KF318053	–
Russula torulosa Bres.	X	☼	–	ND	MZ005531	–
Russula versicolor Jul. Schäff.		☼	–	ND	JN944009	JN940594
Russula veternosa Fr.	X	☼	–	ND	FR852104	AF325321
Russula vinosopurpurea Jul. Schäff.	X	☼	–	ND	FR852115	–
Russula virescens (Schaeff.) Fr.		☼	–	SMW	AY061727	AF041548
Russula xerampelina (Schaeff.) Fr.		☼	–	W	AY061734	AF218542
Saproamanita codinae (Maire) Redhead, Vizzini, Drehmel & Contu		♠	–	ND	–	MK277524
Sarcomyxa serotina (Pers.) V. Papp	○	▐	–	ND	MH856703	MH868220
Simocybe centunculus (Fr.) P. Karst.	X	▐	–	ND	MT535746	KT715786
Sphagnurus paluster (Peck) Redhead & V. Hofst.	X	♠	–	ND	KP192547	MH873802
Strobilurus esculentus (Wulfen) Singer		♠	–	W	MH014049	AY207299
Strobilurus tenacellus (Pers.) Singer	X	♠	–	ND	MF063166	MF063102
Stropharia aeruginosa (Curtis) Quél.	X (edible in the Czech Republic)	♠	–	ND	MW492534	MW492637
Stropharia coronilla (Bull.) Quél.		♠	–	ND	MH856747	MH868269
Stropharia melanosperma (Bull.) Quél.		♠	–	ND	–	–
Tricholoma acerbum (Bull.) Quél.	X (inedible in the Czech Republic)	☼	–	M	MH628231	MK278598
Tricholoma argyraceum (Bull.) Gillet		☼	–	ND	GU060278	MK278614
Tricholoma caligatum (Viv.) Ricken		☼	–	ND	KU058510	KU058548
Tricholoma cingulatum (Almfelt ex Fr.) Jacobasch		☼	–	ND	MH620781	AY207308
Tricholoma equestre (L.) P. Kumm.	(considered poisonous last years, but may include more species)	☼	–	SM	EU186278	AM946471
Tricholoma fulvum (Fr.) Bigeard & H. Guill.	X (inedible in the Czech Republic)	☼	–	ND	KU058514	KU058552
Tricholoma lascivum (Fr.) Gillet	(considered edible in China)	☼	–	ND	LT000131	–
Tricholoma orirubens Quél.		☼	–	ND	DQ389734	DQ389734
Tricholoma psammopus (Kalchbr.) Quél.	X	☼	–	ND	AF377241	–
Tricholoma robustum (Alb. & Schwein.) Ricken	X (inedible in southern Europe)	☼	–	ND	AB699669	–
Tricholoma scalpturatum (Fr.) Quél.	(considered poisonous in China)	☼	–	ND	JN389305	JN389350
Tricholoma sulphureum (Bull.) P. Kumm.		☼	–	M	AY462032	AY462040
Tricholoma terreum (Schaeff.) P. Kumm.		☼	–	MW	EU653301	EU653305
Tricholoma ustale (Fr.) P. Kumm.	X	☼	–	M	LC574882	AY207306
Tricholoma ustaloides Romagn.	X	☼	–	ND	LT000094	–
Tricholoma vaccinum (Schaeff.) P. Kumm.	X	☼	–	ND	AF062628	GQ289219
Tricholomopsis formosa (Murrill) Singer	X	▐	–	ND	–	–
Tubaria confragosa (Fr.) Harmaja	X	▐	–	ND	MF039262	EF051053
Tubaria conspersa (Pers.) Fayod	X	♠ ▐	–	ND	MF039274	AF205692
Tubaria furfuracea (Pers.) Gillet		♠ ▐	–	W	MZ159504	MH867638
Tubaria pallidospora J.E. Lange	X	♠ ▐	–	ND	–	–
Volvariella iranica (Fallahyan) Szczepka	○ (probably inedible)	▐	–	ND	–	–
Volvariella bombycina (Schaeff.) Singer	X	▐	–	W	MT913623	MK278653
Volvariella hypopithys (Fr.) Shaffer	X	♠	–	ND	MN738658	MN738582
Volvariella murinella (Quél.) M.M. Moser ex Dennis, P.D. Orton & Hora	X	♠	–	ND	MK412400	MK278657
Volvariella pusilla (Pers.) Singer	X	♠	–	ND	HM246494	MK278658
Volvariella volvacea (Bull.) Singer		♠	–	M	OM417506	OM373623
Volvopluteus gloiocephalus (DC.) Vizzini, Contu & Justo		♠	–	M	MN738645	MN738593
Xerula pudens (Pers.) Singer		▐	–	S	MT535743	MT554333
Zhuliangomyces ochraceoluteus (P.D. Orton) Redhead		♠	–	ND	MT863767	MT862275

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GenBank accessions in bold were generated in this study. (For species synonyms and distribution data of the species consult Ghobad-Nejhad et al., 2020).

^aNew to Iran. Voucher: Iran, Tehran, Latmalkan, on soil, IV.2021, Ghobad-Nejhad 4404.

^bInformation on edibility of the species in China from Wu et al. (2019).

^cOccurrence in Iran shown by Crous et al. (2020).

Two concatenated datasets of nLSU+ITS were constructed, one representing the taxa belonging to the order Agaricales (dataset 1), and the other dataset for taxa of Cantharellales, Polyporales, and Russulales (dataset 2). Contumyces rosellus, the single Iranian agaric Hymenochaetales, was used as an outgroup for both datasets.

Sequences were aligned using MUSCLE (Madeira et al., 2019). To optimize the alignment, problematic columns were reduced with Noisy 1.5.12 (Dress et al., 2008) and were further identified and removed after careful visual inspection. Special attention was paid to excluding the poorly aligned columns of the ITS region and keeping the finely aligned parts. (Sequences of Amanita eliae, Mycena xantholeuca, Pluteus semibulbosus, and Tricholoma ustale were deleted from the final dataset due to poor alignment.)

Antioxidant properties

Antioxidant properties of the species were obtained via published references as well as own experiments performed in the present study (Tables 1, ​,2;2; Supplementary Table 1). A thorough literature survey was performed to extract and summarize the available data on the antioxidant properties of the agaric species. Published references were searched via Google Scholar, PubMed, and other standard repositories. Each literature was scrutinized carefully, avoiding poor quality and ambiguous data. Disqualified literature, unpublished data, and papers published in non-standard journals were removed from our analyses. In total, ca. 300 literature were surveyed and ca. 170 references were cited in this work and in Supplementary material. The majority of studies reported the antioxidant potential as EC₅₀ values, i.e., half maximal effective concentration, based on DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2, 2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) assays. To have an approximate comparison of the antioxidant potential of the species, we tentatively categorized the EC₅₀ values as strong, moderate, and weak. For this, the EC₅₀ values less than 1mg/ml were considered as “strong” (S), EC₅₀ values ranging from 1 to 10 mg/ml as “moderate” (M), and EC₅₀ values more than 10mg/ml were tentatively considered as “weak” (W) antioxidants (Table 1). For several species, we found different EC₅₀ values reported in different studies. We preferred to keep the data as is for any future reference so that we assigned more than one code to classify the antioxidant potential of these species (e.g., SM standing for strong to moderate). (In a number of studies the antioxidant potential had been expressed only as radical scavenging activity% (RSA %). For these, the RSAs >80% were hesitantly considered as strong, RSA 50%–80% as moderate, and RSA<50% were tentatively considered as weak, paying careful attention also to the values from the antioxidant standards; see Supplementary Table 1). In the cases where the antioxidant potential values were contrasting in different studies, we preferred to keep the data as is for any future reference, and therefore the antioxidant potential of the corresponding species are shown here with the combined codes SM, MW, and SMW, where applicable (Table 1).

Table 2

The EC₅₀ values and the percentage of radical scavenging activity (RSA) obtained by ABTS assays in this study.

Species	Voucher	RSA % at different concentrations					EC50 mg/ml
		0.01mg/ml	0.025mg/ml	0.05mg/ml	0.075mg/ml	0.1mg/ml
Agaricus arvensis	Ghobad-Nejhad 4295	14.22	22.07	32.35	43.63	54.92	0.10 ± 0.003
Agaricus bitorquis	Ghobad-Nejhad 4284	16.60	22.75	35.67	47.58	59.10	0.08 ± 0.003
Agaricus iodosmus	Ghobad-Nejhad 4277	26.85	29.78	37.33	43.88	50.43	0.09 ± 0.021
Agaricus pseudopratensis	Ghobad-Nejhad 4278	38.94	39.54	43.20	45.86	48.13	0.11 ± 0.004
Agrocybe dura	Ghobad-Nejhad 4299	26.32	27.48	30.41	31.34	33.27	0.31 ± 0.004
Armillaria mella	Ghobad-Nejhad 4394	14.25	15.25	16.79	18.53	20.11	0.55 ± 0.016
Cantharellus alborufescens	Ghobad-Nejhad 4408	11.45	12.07	12.83	13.79	14.66	1.09 ± 0.06
Coprinopsis atramentaria	Ghobad-Nejhad 4406	7.54	9.42	12.15	15.68	18.81	0.34 ± 0.021
Coprinus comatus	Ghobad-Nejhad 4407	8.17	10.15	13.45	16.74	19.03	0.34 ± 0.028
Cortinarius persoonianus	Ghobad-Nejhad 4206	17.44	19.24	23.25	25.25	28.26	0.27 ± 0.019
Gymnopilus spectabilis	Ghobad-Nejhad 4207	18.76	21.13	25.62	29.91	33.20	0.20 ± 0.001
Hymenopellis radicata	Ghobad-Nejhad 4204	20.19	20.70	21.50	22.09	22.78	1.05 ± 0.06
Hypholoma fasciculare	Ghobad-Nejhad 4201a	29.47	35.57	48.41	60.25	72.09	0.05 ± 0.007
Lentinus tigrinus	Ghobad-Nejhad 4397	7.14	8.09	9.68	11.26	12.16	0.75 ± 0.01
Leucoagaricus leucothites	Ghobad-Nejhad 4279 4276	19.46	20.80	22.90	25.10	27.50	0.35 ± 0.001
Melanoleuca exscissa	Ghobad-Nejhad 4375	28.27	29.61	31.50	34.07	36.90	0.24 ± 0.009
Pholiota aurivella	Ghobad-Nejhad 600	38.83	39.48	40.61	41.54	42.84	0.26 ± 0.016
Pleurotus cornucopiae	Ghobad-Nejhad 4308	24.75	25.60	27.23	28.77	30.32	0.41 ± 0.001
Pleurotus eryngii	Ghobad-Nejhad 1068	32.30	33.14	35.26	37.11	38.40	0.26 ± 0.011
Pleurotus ostreatus	Ghobad-Nejhad 4403	6.66	8.20	10.58	12.85	15.33	0.46 ± 0.003
Pluteus cervinus	Ghobad-Nejhad 4271	13.14	18.16	27.11	34.96	44.83	0.11 ± 0.002
Psathyrella bivelata	Ghobad-Nejhad 4303 4310	36.34	37.11	38.40	39.69	40.18	0.31 ± 0.062
Russula emeticcolor	Ghobad-Nejhad 4149	44.45	45.19	46.42	47.66	48.01	0.14 ± 0.028
Xerula pudens	Sohrabi 30619	31.78	33.55	35.80	37.55	39.55	0.22 ± 0.002
Trolox							0.023 ± 0.011

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Dried basidiomata from 24 species were sampled and examined for their antioxidant potential via ABTS assay following Re et al. (1999). Voucher samples were deposited at the Iranian Cryptogamic Herbarium (ICH) herbarium (acronym by Index Herbariorum) or at MG personal collection. The ABTS solution (7mM) was prepared in 2.45mM potassium sulfate and was kept at room temperature in the dark for 16h. The mixture was then mixed with phosphate-buffered saline (PBS) as a control, and the absorbance reached 0.7±0.02 at 734nm. The extract samples with final concentrations of 0.01, 0.025, 0.05, 0.075, and 0.1mg/ml were mixed with 980μl of ABTS solution. The absorbance at 734nm was measured after 6min. The percentage of radical scavenging activity was calculated by the following equation, where A stands for absorbance (Öztürk et al., 2011):

$$\begin{array}{l}\mathrm{Scavenging\; activity}\left(\%\right)\\ =\left(\mathrm{A}\mathrm{control}-\mathrm{A}\mathrm{sample}\right)/\mathrm{A}\mathrm{control}\times 100\end{array}$$

The EC₅₀ values were obtained through interpolation from linear regression analysis (Supplementary Figure 1). Trolox was used as a positive control at different concentrations (0.005, 0.01, 0.015, 0.02, 0.025, and 0.03mg/ml).

Results

The results of our survey on the resources of agaric species in Iran are summarized in Table 1. Altogether, 558 agaric species from five orders were surveyed for their resources of edible and poisonous species, their ecological guilds, bioluminescence, and antioxidant potential. The two species Conocybe olivaceopileata and Inocybe ionolepis were added here to the Iranian mycota (see Table 1).

Phylogeny

The Agaricales dataset consisted of 428 taxa and 1,341 characters of which, 243 characters were constant, 144 variable, and 954 characters were informative. The best-fit evolutionary model suggested by MrModeltest was GTR+I+G for each of the LSU and ITS partitions. The Agaricales phylogram is shown in Figure 1. Nineteen families were phylogenetically retrieved with moderate to good posterior probabilities (PPs) and were shown in colored boxes, while the rest of the taxa were incertae sedis or received low to moderate branch support.

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Figure 1

Phylogram from the combined nLSU+ITS sequence dataset representing the phylogenetic relationships of Iranian Agaricales. Posterior probabilities (PPs)≥0.8 are shown as light lilac dots on the nodes. Terminals in red are species with antioxidant activity and the letters inside brackets are the tentative antioxidant codes: S, strong; M, moderate; W, weak (see the text for full details).

The species with antioxidant data were distributed in all the families shown in colored boxes except for the two families Entolomataceae and Inocybaceae (Figure 1). For some families such as Bolbitiaceae, Marasmiaceae, and Tubariaceae, there was only a single species with antioxidant activity, while other families such as Agaricaceae, Psathyrellaceae, and Pleurotaceae contained several antioxidant species.

Dataset 2 (Cantharellales, Polyporales, Russulales) consisted of 71 taxa and 1,528 characters of which, 279 characters were constant, 347 variable, and 902 characters were informative. The best-fit evolutionary model as suggested by MrModeltest was GTR+G for each of the LSU and ITS partitions. The phylogram obtained from the analyses of dataset 2 is presented in Figure 2. The orders Polyporales, Russulales, and Cantharellales were retrieved as moderate to well-supported monophyletic clades (PPs 0.75, 0.94, and 1.00, respectively). (For Polyporales, the two families Panaceae and Polyporaceae were not retrieved. Moreover, Panellus stipticus found a position close to the outgroup Contumyces rosellus, and we could not solve this.) The species with antioxidant data were distributed within the three orders in the phylogram (Figure 2). Out of six Cantharellales agaric members in Iran (Ghobad-Nejhad et al., 2020), four species have antioxidant data (Figure 2; Craterellus cinereus had no good LSU/ITS, so is missing in the phylogeny here). Polyporales has nine agaric species in Iran, four of which possess antioxidant activity (Figure 2). Russulales has 60 agaric species in Iran (Ghobad-Nejhad et al., 2020) from which, 16 species have antioxidant data (Figure 2; Table 1). Contumyces rosellus is the only Hymenochaetales agaric in Iran and has no antioxidant data.

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Figure 2

Phylogram from the combined nLSU+ITS sequence dataset representing phylogenetic relationships of the Iranian Cantharellaeles, Russulales, and Polyporales. Posterior probabilities are shown below branches. Terminals in red are species with antioxidant activity and the letters inside brackets are the tentative antioxidant codes: S, strong; M, moderate; W, weak (see the text for full details).

Altogether, there were 50 agaric species lacking both ITS and LSU sequences and so did not appear in the phylogenetic analyses (Table 1); these species also lacked antioxidant data, except for Russula nigricans which was scored as a “strong” antioxidant species (Table 1).

Edibility, ecological guild, and luminescence

Results of the survey on edibility, ecological guilds, and luminescence of agaric species occurring in Iran are shown in Table 1 and Figures 3, ​,4.4. It is revealed that about 189 species of agarics in Iran can be classified as edible, 128 species as poisonous, and 271 species as inedible (Table 1; Figure 3). Moreover, 10 species can be assigned as edible only if well-cooked, whereas the edibility of 30 species is uncertain or unknown.

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Figure 3

The number of Iranian agaric species in each edibility category.

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Figure 4

The number of Iranian agaric species in each ecological guild.

Concerning ecological guilds, our results show that about 254 species of agarics in Iran are soil saprotrophic, 172 species ectomycorrhizal, 146 species wood-inhabiting, 18 species leaf/litter-inhabiting, and nine species are parasitic (Table 1; Figure 4). Parasitic species include Armillaria borealis, A. cepistipes, A. gallica, A. mellea, Collybia tuberosa, Pleurotus eryngii, P. nebrodensis which are sapro-parasitic, Gymnopus fusipes which is wood-inhabiting parasitic, and Asterophora lycoperdoides which grows on basidiomata of Lactarius and Russula species (Table 1).

Among 558 agaric species in Iran, 19 species are categorized as luminescent (Table 1). These include Armillaria (four spp.), Collybia tuberosa, Flammulina velutipes, Mycena (six spp.), Omphalotus olearius, Panellus stipticus, and Russula (five spp.). The six species with chemiluminescence include Russula anthracina, R. cyanoxantha, R. delica, R. foetens, R. ochroleuca, as well as Panellus stipticus.

Antioxidant potential

Results of our survey on the antioxidant potential of agaric species occurring in Iran are shown in Table 1 and Figure 5 (see also Supplementary Table 1 for details on the antioxidant potential of the species and the corresponding references). According to the results, antioxidant activity data is available for 113 species phylogenetically distributed in four orders (Agaricales, Cantharellales, Russulales, Polyporales) and 21 agaric families including 17 families in the Agaricales (Strophariaceae, Hymenogastraceae, Tubariaceae, Bolbitiaceae, Cortinariaceae, Lyophyllaceae, Tricholmataceae, Psathyrellaceae, Agaricaceae, Omphalotaceae, Marasmiaceae, Physalacriaceae, Amanitaceae, Pluteaceae, Mycenaceae, Pleurotaceae, Hydnangiaceae; Figure 1), as well as Hydnaceae (=Cantharellaceae), Russulaceae, Polyporaceae, and Panaceae (Figure 2, families not shown on the tree). However, 445 species still lack information on their antioxidant potential (for a handful of species, the available antioxidant values in the literature had been expressed only by other methods such as TEAC and FRAP; as far as these cases were very few, they were not taken into account here, to keep the rest of the data comparable). The antioxidant potential of 24 species was assayed in this study and their EC₅₀ values are reported in Table 2. Species assayed for the first time in this study included: Agaricus iodosmus, A. pseudopratensis, Agrocybe dura, Cantharellus alborufescens, Cortinarius persoonianus, Hymenopellis radicata, Melanoleuca exscissa, Psathyrella bivelata, Russula emeticolor, and Xerula pudens (Table 2; Supplementary Table 1). In general, the EC₅₀ values of the agaric species ranged between 0.0015mg/ml (for Psathyrella candolleana) up to 31.42mg/ml (for Tricholoma terreum).

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Figure 5

Categorization of antioxidant potential of Iranian agaric species. S, strong; M, moderate; W, weak; ND, not determined. See the text for full details.

Among the 113 species having antioxidant data, 27 species could roughly be classified as “strong,” 25 species as “strong to moderate,” 27 species as “moderate,” nine species as “moderate to weak,” and 20 species could be tentatively regarded as “weak” antioxidants (Figure 5). Some of the species in the S category are Agaricus arvensis, Agrocybe dura, Amanita rubescens, Candolleomyces candolleanus, Clitocybe nebularis, Coprinellus micaceus, Coprinopsis picacea, Craterellus tubaeformis, Gymnopilus spectabilis, Hygrophorus eburneus, Hypsizygus ulmarius, Macrocybe gigantean, Marasmiellus peronatus, Mycenastrum corium, Russula anthracina, Russula cyanoxantha, Russula emeticolor, Russula nigricans, Russula rosea, and Xerula pudens (Table 1).

The overall phylogenetic distribution of the agaric species with antioxidant data is shown in Figures 1, ​,2.2. Russula nigricans was the only species in our dataset with antioxidant data but lacked LSU/ITS DNA sequences in GenBank, so could not be used in our phylogenetic analyses.

Discussion

In this study, we comprehensively investigated the resources of agarics in Iran. Indeed, no published data have yet been available on number of recorded edible, poisonous, and other agarics in Iran, so the present work fills in these gaps. It is shown that there are currently about 189 edible, 128 poisonous, 254 soil saprotrophic, 172 ectomycorrhizal, 146 wood-inhabiting, 18 leaf/litter-inhabiting, 9 parasitic, and 19 luminescent agaric species in the country. The two species Conocybe olivaceopileata and Inocybe ionolepis were newly added to the Iranian mycota, new DNA sequences were obtained from Iranian samples, and the first phylogenetic reconstruction was provided for agarics of Iran. Evidently, this work is not final and therefore further studies of Iranian fungal diversity would add new species to the list presented here. About 500 agaric species belonging to the five orders Agaricales, Cantharellales, Polyporales, Russulales, and Hymenochaetales were phylogenetically analyzed based on nLSU+ITS sequence datasets. Thorough analyses with additional gene regions and vouchered samples must be utilized in the future to resolve the phylogenetic relationships of Iranian agarics. Yet, the preliminary phylogenetic analysis of agaric species presented here would help to inspire the investigation of many taxa in need of taxonomic revision. Phylogeny backbones can be used for visualization of the phylogenetic distribution of species possessing particular characteristics, herein, antioxidant potential, but also other features in the future. For instance, phylogenetic assessments have been used to screen the pleuromutilin-producing basidiomycete species (Hartley et al., 2009), fungal strains capable of degrading industrial compounds (Navarro et al., 2021), or other natural products (Adamek et al., 2019).

For a few species, the edibility assignment was based on own observation in Iran, but as stated earlier, most of the species were categorized based on available knowledge on central and southern European species. (It might be relevant to note that a number of previous studies have shown a high similarity of the Iranian mycota to that of Europe, e.g., Ghobad-Nejhad et al., 2012; Ghobad-Nejhad and Bernicchia, 2019.) Basically, edibility assignments should always be regarded with caution and it is generally recommended to avoid consuming raw or insufficiently identified mushrooms. There are still noticeable gaps in the knowledge of edible/poisonous mushrooms identification in Iran and the level of education, public awareness, and citizen science is far from medium standards. Concerning usage of edible fungi in Iran, published references are lacking, and our available data is fragmentary. In the reports and statistics on mushroom poisoning in Iran, there is no proper documentation of the species involved or at best, the species are only ambiguously characterized (Kiarsi et al., 2019).

The present work calculated as many as about 172 ectomycorrhizal agaric species for Iran. Ectomycorrhizal fungi are essential components of forest ecosystems to supply the symbiont trees with water and nutrients such as phosphorus and nitrogen, and therefore are highly important in forest sustainability (Varma and Hock, 2013). A large number of ectomycorrhizal agarics are also edible and may be harvested in the wild for culinary use, so they are in need of immediate conservation actions (Vaario and Matsushita, 2021); this is the case, especially with the Cantharellus species in northern Iran (Parad et al., 2018, 2020).

In this study, we listed 146 wood-inhabiting agaric species for Iran. There have been several studies on the diversity and taxonomy of wood-inhabiting aphyllophoroid fungi in Iran (e.g., Hallenberg, 1981; Ghobad-Nejhad and Hallenberg 2012; Amoopour et al., 2016; Ghobad-Nejhad and Langer, 2017; Nazari Mahroo et al., 2018; Ghobad-Nejhad and Bernicchia, 2019) but agarics growing on wood in Iran have not been studied systematically. Wood rotting fungi play a key role in terrestrial carbon cycling and have high potential in biotechnology, enzyme industry, biorefinery, and bioremediation of waste material and recalcitrant compounds (Gadd, 2001; Nguyen et al., 2018; Mäkelä et al., 2021). While Polyporales members are best known for their wood decomposition ability, genomic studies have revealed that several Agaricales taxa have evolved the enzymatic machinery comparable to the white-rot Polyporales (Floudas et al., 2020; Ruiz-Dueñas et al., 2020).

Another aspect surveyed in this study for the Iranian agaric species was bioluminescence. Bioluminescence, i.e., the ability of organisms to emit visible light, has been developed independently in the evolution of different organisms. Concerning fungi, 109 fungal taxa are known to exhibit bioluminescence all of which (except one Xylariales) are white-spored saprotrophic Basidiomycota distinguished in four phylogenetic lineages (Chew et al., 2015; Ke and Tsai, 2022) all sharing the same type of luciferin and luciferase (Oliveira et al., 2012). Interestingly, it has been shown that luminescence could be linked to the antioxidant/radical scavenging defense mechanism against some environmental stress factors (Vydryakova and Bissett, 2016; Oba et al., 2017). Moreover, the fungal bioluminescence capacity can be used in environmental biomonitoring of metals or organic compounds and to develop toxicity tests (Ke and Tsai, 2022).

In this work, a thorough survey was done to reveal the antioxidant potential of 558 agaric species and a new approach was used to combine antioxidant data with phylogeny of the species. Ten species were subjected to antioxidant analyses for the first time, belonging to the genera Agaricus, Agrocybe, Cantharellus, Cortinarius, Hymenopellis, Melanoleuca, Psathyrella, Russula, and Xerula. ABTS assay is one of the most frequently used method for quantification of antioxidant activity of mushrooms. Numerous antioxidant assays have been introduced which are usually classified into two groups based on the mechanism of action: single electron transfer and hydrogen atom transfer (Tan and Lim, 2015; Xiao et al., 2020). Compared to other methods, ABTS has the advantage of involving more or less both mechanisms (Prior et al., 2005). Yet, more examinations are required to fully investigate the antioxidant capacity of the species studied here, and to quantify and characterize the underlying bioactive compounds. Here, we could resume antioxidant data for 20% of agaric species (113 spp.), but noted that 80% of the species (445 spp.) have no antioxidant data. This is noteworthy compared to the fact that antioxidant tests are among the most popular bioactivity assays and it may show that macrofungi have remained little studied in this regard. The highest antioxidant capacities (the lowest EC₅₀ values) were shown by the species categorized as S (27 spp.) and then as SM (25 spp.; Table 1; Figure 5). As noted earliers, in several cases, various EC₅₀ values had been reported in different studies for some species, so that we assigned more than one code for them. We emphasize that such classification is approximate and for detailed comparisons, more precise methods are recommended to be applied. For five species, the EC₅₀ measures ranged significantly, in a way that the code assignment could only be expressed as SMW: Pleurotus djamor, P. eryngii, P. ostreatus, P. pulmonarius, and Russula virescens. Of course, differences in the solvents, standards, modifications in the assays procedures, and even identification issues can account for the different measures under the same species name. Ideally, the identity of the voucher specimens should be fully characterized and the species should be assayed with exactly the same procedure so as to be able to have the best quality comparisons. In general, for the studies where both ABTS and DPPH assays had been conducted, ABTS values seemed to slightly outperform the DPPH values, showing lower EC₅₀ measures. Many of the species in the S or “strong” antioxidant category are edible: Agaricus arvensis, Agrocybe dura, Amanita rubescens, Candolleomyces candolleanus, Clitocybe nebularis, Craterellus tubaeformis, Hygrophorus eburneus, Hypsizygus ulmarius, Macrocybe gigantea, Russula anthracina, R. cyanoxantha, R. emeticolor, R. nigricans, R. rosea, and Xerula pudens (Table 1). Oxidative stress is the root of a cascade of numerous acute and chronic human diseases (Kosanic et al., 2013). Diets rich in natural antioxidants enforce the native defense system and protect against oxidative damage (Ferreira et al., 2009). Mushroom species that are edible and possess high level of biological activities with perspectives on promoting human health are considered noteworthy candidates for developing functional foods and nutra-pharmaceutical products (Kozarski et al., 2015; Lu et al., 2020; Niego et al., 2021; Shaffique et al., 2021; El Sheikha, 2022). It is evident that thorough analyses are needed to fully characterize the mycochemical constitutes of such species and their various bioactivities.

Our results pave the avenue for advanced studies on edible, poisonous, saprotrophic, ectomycorrhizal, wood-inhabiting, parasitic, luminescent, and antioxidant species of agarics of Iran. Twenty percent of the Iranian agaric species possess antioxidant activity, phylogenetically distributed in four orders and 21 agaric families. About 5% of the antioxidant species can be considered strong antioxidants, many of which are also edible and could be utilized for the development of functional foods. Various edible agaric species are grown commercially in the world, while only 1–2 are commonly grown in Iran (personal comm.). Ectomycorrhizal and wood-inhabiting species are important components of forest sustainability. Forests in Iran are very scanty, comprising less than 10% of the total country area, and are on the verge of severe depletion due to numerous anthropological and environmental threats. Yet, Iranian old-growth forests, categorized as part of the northern hemisphere glacial refugia (Ghobad-Nejhad et al., 2012, 2020), harbor a rich reservoir of agaric fungi with diverse characteristics and beneficial aspects. Resources of Iranian agarics provide valuable opportunities for biotechnology and mycochemistry, and should be regarded for preservation and habitat conservation. Our preliminary phylogenetic trees would guide the selection of agaric taxa to be examined in the future for taxonomic revisions, biotechnological applications, and applied phylogeny studies. The thorough survey of antioxidant data of 558 agaric species would provide the state of the knowledge on agarics examined so far and the remaining gaps to be filled in the future.

Data availability statement

The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Supplementary material.

Author contributions

MG-N conceptualized and designed the study, performed the molecular study and provided the first draft. VA contributed to the trait assignments. MG-N, VA, and EL wrote the manuscript. MG-N and MM performed the experiments. All authors contributed to the article and approved the submitted version.

Funding

This work was supported by the Center for International Scientific Studies & Collaboration (CISSC), Ministry of Science, Research, and Technology of Iran. The studies of VA were enabled by the support provided to the Moravian Museum by the Ministry of Culture of the Czech Republic as part of its long-term conceptual development program for research institutions (DKRVO, ref. MK000094862).

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

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Footnotes

References