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Amanita beckeri (Amanitaceae, Agaricales), a rare and little-known species of section Vaginatae — the first record in Bulgaria
B. Assyov1, F. Bozok2, M. Yarar3, H. Ta§km4
'Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Gagarin Str., 1113 Sofia, Bulgaria 2Department of Biology, Faculty of Arts and Science, Osmaniye Korkut Ata University,
80000 Osmaniye, Turkiye 3Department of Biotechnology, Institute of Natural and Applied Sciences, Cukurova University,
01330 Adana, Turkiye
4Department of Horticulture, Faculty of Agriculture, Cukurova University, 01330 Adana, Turkiye Corresponding author: B. Assyov, contact@boletales.com
Abstract. Amanita beckeri was described six decades ago, but still remains among the less known and least featured in the mycological literature species of the genus in Europe. The authors present concise description and illustrations of a collection of the species from the Balkan Peninsula, which identity is supported by analysis of rITS sequences. Comparison with similar brownish-colored ring-less amanitas is presented and an account of the current knowledge about its distribution is included.
Keywords: Amanita brunneoconulus, Amanitopsis, biogeography, Southeastern Europe, taxonomy.
Первая находка в Болгарии Amanita beckeri (Amanitaceae, Agaricales) — редкого и малоизвестного вида секции Vaginatae
Б. Асьов1, Ф. Бозок2, М. Ярар3, Х. Ташкын4
1Институт биоразнообразия и экосистемных исследований, Болгарская академия наук, ул. Майора Юрия Гагарина, д. 2, 1113 София, Болгария
2Кафедра биологии, Факультет искусства и науки, Университет Османии Коркут Ата,
80000 Османия, Турция 3Кафедра биотехнологии, Институт естественных и прикладных наук, Университет Чукурова,
01330 Адана, Турция 4Кафедра садоводства, Факультет сельского хозяйства, Университет Чукурова,
01330 Адана, Турция Автор для переписки: Б. Асьов, contact@boletales.com
Резюме. Amanita beckeri был описан шесть десятилетий назад, но до сих пор остается одним из малоизвестных и редко представленных в микологической литературе видов рода в Европе. Приведены подробное описание и иллюстрации образца этого вида с Балканского полуострова, идентичность которого подтверждена молекулярно-филогенетическим анализом
https://doi.org/1031111/nsnr/2022.56.2287
287
ДНК-последовательностей региона rITS. Представлено сравнение с другими безкольцевыми мухоморами коричневатого цвета, а также дано обобщение сведений о распространении вида.
Ключевые слова: Amanita brunneoconulus, Amanitopsis, биогеография, таксономия, Юго-восточная Европа.
Amanita Pers. is a genus with worldwide distribution, comprising a large number of species, among which both deadly poisonous and highly prized edible representatives are known (Tulloss, Rodriguez-Caycedo, 2011; Cui et al., 2018; Loizides et al., 2018). Stemming from the understanding of the importance of the genus, no less than twenty robust works were produced since the beginning of the 20th century in Europe alone, while the number of papers dealing with individual problems or species is practically impossible to recount (e. g., the bibliographic accounts in Fraiture, 1993; Neville, Pou-marat, 2004, 2009; Hanss, Moreau, 2020; Assyov et al., 2021).
The first molecular study including European members of Amanita was published in the end of the 20th century (Weiß et al, 1998), but this work focused on resolving the infrageneric classification of the genus and obtained limited number of sequences from European materials. Molecular assessments in different problematic groups followed suit (Gonzalez et al, 2002; Moreno et al, 2008; Justo et al, 2010; Vizzini et al, 2012, 2016, 2020; Malysheva, Kovalenko, 2015; Loizides et al, 2018; Riccioni et al, 2019; Hanss, Moreau, 2020; Crous et al, 2021; Sevcikova et al, 2021). It was not until recently however, when systematic DNA studies in section Vaginatae Quel. began, this grouping being recognized as most troublesome by several European authors (Fraiture, 1993; Malysheva, Kovalenko, 2015; Hanss, Moreau, 2020). Since the first decade of this century the interest in section Vaginatae increased considerably and a number of publications employing molecular techniques was published, solving numerous problems in this undisputedly perplexed group (Malysheva, Kovalenko, 2015; Vizzini et al, 2016; Loizides et al, 2018; Hanss, Moreau, 2020; Crous et al, 2021; Sevcikova et al, 2021). Among those latter studies the one of Hanss and Moreau (2020) ought to be highlighted in particular, for it released sequences of type materials for a number of species, facilitating further research into the European Vaginatae.
A number of new species were introduced throughout the late 20th and the beginning of the 21st century within section Vaginatae in Europe (e. g., Huijsman, 1962a, b; Piane, 1972; Lanne, 1979; Bas, 1982; Knudsen, Borgen, 1987; Contu, 1988, 1998, 1999a, b; Contu, Pacioni, 1998; Consiglio, Contu, 1999; Neville et al, 2000; Tulloss, Traverso, 2001; Neville, Poumarat, 2007, 2009). While some of them were shown to be conspecific by recent DNA studies, for others the self-standing status is now confirmed (Vizzini et al, 2016; Hanss, Moreau, 2020). Among these, A. beckeri Huijsman was described over half a century ago (Huijsman 1962a, b), but despite its outstanding morphological features that make it easily recognizable, it remains one of the least known species of the group of the ringless amanitas in Europe, with a few descriptions in the pertinent literature (Huijsman 1962a, b; Sebek, 1985; Tulloss, 1994; Macau, Perez-De-Gregorio, 2011). During their taxonomic studies in Amanita on the Balkans and Turkey, the authors were
provided with one excellently documented specimen, which molecular and morphological assessment confirmed it as belonging to this elusive species. In an attempt to add to the scanty knowledge of A. beckeri, this collection is presented herein.
Materials and Methods
The specimen was documented in fresh state by the collector and desiccated for permanent preservation in the Mycological Collection of the Institute of Biodiversity and Ecosystem Research of the Bulgarian Academy of Sciences (SOMF). The colors in the macroscopic description match as closely as possible the color standards in the "Flora of British Fungi Colour Identification Chart" (Colour..., 1969) and are followed by color code, beginning with the abbreviation "BFF". Color names without such code are vernacular designations.
Microscopic study was held with an AmScope T360B compound light microscope with AmScope MU900 digital camera. Examination was performed on dried samples, using 5% KOH with addition of Congo red in ammonia as to highlight the observed micromorphological features. Microscopic measurements were read by Piximetre v. 5.10 (Piximetre., 2022) from calibrated digital images. In the description below, the dimensions of basidiospores are obtained from mounts in 5% KOH and are presented by the minimum and maximum values for length, width and spore quotient (length/width ratio — Q), followed by the average values of length (L), width (W ) and quotient (Qav); "n =" denotes the number of spores measured, "m =" stands for the number of collections studied and "p" indicates the number of basidiomata from which the measurements were obtained. The remaining microscopic structures are presented with their minimum and maximum values, based on 15 measurements. Melzer's reagent after the recipe in Kirk et al. (2008) was used to assess the amyloidity of basi-diospores and to test the K-K-reaction (Kotilova-Kubickova, 1982; Neville, Pouma-rat, 2004). Drawings of microscopic features were prepared by tracing objects from microphotographs onto semi-transparent paper.
The methods for total genomic DNA extraction, amplification and sequencing of the rITS region followed the protocol described in Bozok et al. (2020). The obtained forward and reverse reads were assembled into consensus sequence and edited upon necessity with Sequencher version 5.4.5 (Gene Codes, Ann Arbor, MI, USA).
The dataset for phylogenetic analysis included the sequence of A. beckeri generated in this study, as well as 29, derived from previously published works (Malysheva, Kovalenko, 2015; Vizzini et al, 2016; Hanss, Moreau, 2020) or unpublished accessions available on GenBank and UNITE databases. It was composed, aligned and visually checked in BioEdit Sequence Alignment Editor software (Hall, 1999). Multiple sequence alignment was carried out by MUSCLE 3.8.31 (Edgar, 2004). A sequence of Amanita submembranacea (Bon) Groger was selected as an outgroup, taking into consideration the phylogenetic inference in Hanss, Moreau (2020). The phylogenetic analysis was performed on the internet portal Phylogeny.fr (Dereeper et al, 2008; Phylo-genyFr, 2022). Maximum likelihood phylogenetic analysis used PhyML 3.0 (Guindon,
Gascuel, 2003; Guindon et al, 2010) with HKY85 substitution model. The Shimodai-ra-Hasegawa version of the approximate likelihood-ratio test (SH-aLRT) was used for calculation of the branch support (Anisimova et al, 2011). The phylogenetic tree was drawn in TreeDyn 198.3 (Chevenet et al, 2006) and its final technical formatting was performed in InkScape ver. 1.0.2-2 (InkScape..., 2022).
Results
Amanita beckeri Huijsman ex Huijsman, 1962, Bull. Soc. Mycol. France 78: 217. = Amanita beckeri Huijsman, 1962 [1961], Bull. Soc. Mycol. France 77: 349. nom. illeg. (lacking Latin diagnosis and type not indicated, ICN art. 39.1 and art. 40.1). = Ama-nitopsis beckeri (Huijsman) Bon, 1975, Bull. Mens. Soc. Lin. Lion 44(6): 180. = Ama-nitopsis beckeri (Huijsman) Wasser, 1988, Ukr. Bot. Zhurn. 45(6): 77. comb. superfl. (an isonym without nomenclatural status, ICN art. 6.3, note 2). ? = Amanita brunneoconulus Bas et Gröger, in Bas, 1982, Persoonia 11(4): 432.
(Figs 1, 2)
Fig. 1. Macromorphological features of Amanita beckerii (SOMF 30422) — basidiomata in situ. A — habitus; B — detail of pileal surface; C — details of the upper stipe and hymenophore; D — details of the lower stipe and volva. Figures not to scale. Photographs by P. Nedelev.
Pileus up to 8 cm across, applanate, with or without a low umbo, milky coffee (BFF28), pale clay pink (BFF30), vinaceous buff (BFF31), clay buff (BFF32), hazel (BFF27) or snuff brown (BFF17), sometimes with darker center and slight zona-tion; surface smooth, somewhat viscid, covered to a different extent by buff (BFF52), vinaceous buff (BFF31) or cinnamon (BFF10) verrucose remnants of the general veil, which may be washed away in wet weather; margin flat, striate 1/3 to 1/2 of the pileal radius. Lamellae white, moderately crowded, free; edge dentate, brownish; lamellulae 0-3 between lamellae, truncate. Stipe up to 10 cm long and up to 3 times longer than the pileal diameter, cylindrical or slightly enlarged towards the base, hollow, in places the cavity filled with cottony substance (pith); surface whitish, covered with brownish (cinnamon — BFF10, brick — BFF15, to vinaceous buff — BFF31) zig-zag-like markings, particularly in the upper 2/3 or 3/4, towards the base usually more or less smooth; volva friable (type I), in the shape of rounded cup-like enlargement and a belt of tissue at the subbulbous stipe base, off-white or brownish, sometimes with adhering brownish veil remnants similar to the ones on the pileus. Context white. Odour inconspicuous.
Fig. 2. Microscopic features of Amanita beckeri (SOMF 30422). A — basidiospores; B — basidia and basidioles with elements of the underlying subhymenium; C — marginal cells from lamellar edge; D — thromboplerous hyphae from stipe context; E — elements of volva (inner stratum); F — acrophysalides from cottony context of the stipe cavity;
G — acrophysalides from stipe context. Scale bars: A, B, C — 10 |m; D, E, F, G — 30 |m.
Basidiospores 9.4-11.6 x 9.3-11.8 |m, Q = 1-1.18, Lav = 10.9-11 |m, Wav = 10.410.6 |m, Qav = 1.04-1.05 (n = 60, m = 1, p = 2), globose to subglobose, rarely broadly ellipsoid, thin-walled, inamyloid, usually with a single large guttule, rarely pluriguttu-late or with granular content. Basidia 37.9-53.4 x 8-14.3 |m, generally 4-spored, but 2- or 1-spored also present, clavate to elongate clavate, thin-walled; sterigmata generally up to 4 |m, by exception up to 6 |m long. Lamellar edge sterile; marginal cells pyriform, 14.2-23.1 x 7.8-13.5 |m, or spheropendunculate, 9.9-15 |m across, thin-walled, hyaline. Hymenial trama not revived successfully; hymenopodium apparently not present; subhymenium cellular. Pileipellis an ixotrichodermium of filamentous, 2.3-5.1 |m wide, yellowish in KOH, interwoven hyphae. Universal veil on lower stipe (volva): on the outer side composed of 2-4.4 |m wide, undifferentiated, frequently branching, filamentous hyphae and abundant thin-walled, spherical, subspherical or ovoid inflated elements with sizes 36.5-65.1 x 31-55.8 |m; on the inner side as the exterior, but some broadly clavate inflated elements also present; thromboplerous hy-phae not seen in the tissues of either surface; filamentous hyphae in both strata fragile, easily crushed in microscopic mounts. Universal veil remnants on pileus not present in the studied basidiomata. Stipe context rather compact and difficult to disentangle, longitudinally acrophysalidic, filamentous undifferentiated hyphae abundant, 4.3-8.2 |m wide; acrophysalides up to 214 x 35.2 |m, thin-walled, narrowly clavate to clavate; thromboplerous hyphae uncommon, flexuous, with straw-colored content in KOH, thin-walled; cottony substance (pith) from the stipe cavity composed of interwoven, flexuous, filamentous undifferentiated hyphae 4.6-6.6 |m wide; acrophysalides scarce, narrowly to broadly clavate or sometimes ovoid, thin-walled, up to 93.9 x 52.5 |m; thromboplerous hyphae scarce, flexuous, with straw-colored content in KOH, thin-walled. Clamp connections not observed in any tissue. K-K reaction negative.
Specimen examined: Bulgaria, Vraca Province, Stara Planina Mts, Vrachanski Balkan Nature Park, Trudovashkata Cheshma water well above Zgorigrad village, 43°11'01.5"N, 23°29'59.6"E, elev. 775 m, forest of Fagus sylvatica L. and Carpinus betulus L., on limestone, 4 VII 2018, P. Nedelev, SOMF 30422 (GenBank 0N287053).
Discussion
The initial BLAST search of the sequence from the Bulgarian specimen returned as closest matches two sequences of Amanita beckeri (MN490686, MN490691), generated by Hanss, Moreau (2020). The pairwise comparison of the ITS1-5.8S-ITS2 region resulted in 100% identity of the three accessions. Further phylogenetic analysis of sequences belonging to Clade 5 as defined in Hanss and Moreau (2020) inferred the tree presented in Fig. 3. The analysis resolved six major, well-supported lineages apart of A. friabilis (P. Karst.) Bas, which position remains unresolved. The six clades correspond to the results of the same work and were treated informally as stirpes by Hanss and Moreau (2020). These are: 1) stirps Populiphila, including A. beckeri, A. popu-liphila Tulloss et E. Moses, A. protecta Tulloss et G. Wright and A. simulans Contu; 2) stirps Betulae, with A. betulae Neville et Poumarat and A. 'kryorhodon' Tulloss et
Kudzma (nom. illeg., nom. prov.); 3) stirps Coryli, comprising A. alba sensu UNITE, A. 'albiceps' Lamoureux (nom. illeg., nom. prov.), A. 'chiricahuana' Tulloss (nom. illeg., nom. prov.), A. cistetorum Contu et Pacioni, A. coryli Neville et Poumarat and A. 'lu-zernensis' Tulloss et coll. (nom. illeg., nom. prov.); 4) stirps Lividopallescens, including A. lividopallescens (Gillet) Bigeard et H. Guill.; 5) stirps Mairei, with A. brunneofuli-ginea f. ochraceopallida (Contu) Hanss and A. mairei Foley; 6) stirps Albogrisescens, encompassing A. albogrisescens Contu and A. alseides Hanss. The obtained here sequence of A. beckeri nested together with the two French sequences of this species in a highly supported clade (branch support 98%), thus confirming the identity of the Bulgarian specimen.
The macroscopic features of the Bulgarian collection compare well to the original description and the validating diagnosis (Huijsman 1962a, b), and its microscopic details are congruent with the type study, published by Tulloss (1994). The basidiospores dimensions from our SDecimen comDare well to the data, obtained from the holot.vDe
96
96
93
84
-KU248104 Amanita friabilis
100 I-KX834255 Amanita simulans (neotype) 'MN490696 Amanita simulans NR154689 Amanita populiphila (holotype)
NR154690 Amanita protecta (holotype) MN490686 Amanita beckeri (France) MN490691 Amanita beckeri (France) 'ON287053 Amanita beckeri (Bulgaria)
i-KX270308 Amanita 'kryorhodon'
'—MN490668 Amanita betulae (holotype) -MF422621 Amanita 'luzernensis'
98
Amanita beckeri
89r MN490667 Amanita coryli (holotype) r~lMN490699 Amanita coiyli
KX834235 Amanita cistetorum
89
93
С
■ KX834236 Amanita cistetorum
97
MN493559 Amanita alba sensu UNITE 'UDB002307 Amanita alba sensu UNITE -KX219634 Amanita 'chiricahuana'
100
75
KP224316 Amanita 'albiceps'
_iKX834245 Amanita lividopallescens (epitype)
'MN490679 Amanita lividopallescens 77|MN490653 Amanita brunneofuliginea (forma) S3
76
92
MN490688 Amanita brunneofuliginea (forma) r- KX834248 Amanita mairei
1MN490695 Amanita mairei 98 iMN490649 Amanita alseides (holotype) 'MN490678 Amanita alseides
90
iMN490654 Amanita albogrisescens 1MN490665 Amanita albogrisescens
■ KM658295 Amanita submembranacea
0.0 5
Fig. 3. Phylogenetic tree of ITS sequences of Clade 5 (Hanss, Moreau, 2020) of Amanita section Vaginatae, obtained by analysis in PhyML. Branch support values obtained by SH-aLRT-test are shown next to branches as percent; only values above 50% are annotated. The sequence generated in this work is shown in boldface. The clade corresponding to stirps Populiphila is highlighted in grey.
(Tulloss, 1994). Notable difference from the findings of the latter study is the observed by us cellular subhymenium, while in the holotype it was found to have different structure, apparently corresponding to the ramose type (Tulloss, 1994). This difference may be attributed to the early stages of maturity of the studied here basidiomata (cf. Hanss, Moreau, 2020), which is also corroborated by the high abundance in the mounts of immature basidia, notable quantity of such with fewer than four sterigmata, as well as the well-preserved structure of the sterile lamellar edge. Further observations of this character on fresh and differently aged material thus may be worthy. Kotilová-Kubicková (1982) reported positive K-K reaction in A. beckeri. We could not confirm this peculiarity in any part of basidiomata in our specimen and urge other researchers studying Amanita to test this reaction on molecularly supported specimens and report it, as to accumulate data for future judgement of its application in taxonomy of the genus. As of present, merely a few reliable color illustrations of A. beckeri exist in the mycological literature and for such, apart of the present work, the reader may consult Galli (2001: 104, bottom photo) and Macau and Pérez-De-Gregorio (2011: 93). Some microscopic details are illustrated in Tulloss (1994).
Amanita brunneoconulus Bas et Groger was described on materials from Germany (Bas, 1982). It was accepted as self-standing species by Fraiture (1993), who on the other hand placed A. beckeri in the A. lividopallescens complex. The high degree of similarity of A. beckeri and the former species was already discussed and it was suggested that they may be conspecific (Macau, Pérez-De-Gregorio, 2011). Indeed, the morphological features of the two entities overlap considerably, prompting such consideration. To the arguments in Macau and Pérez-De-Gregorio (2011) we shall add the resemblance observed in the subhymenium of our specimens and the data presented in the original description of A. brunneoconulus. On the other hand in the vol-val tissues of the Bulgarian specimen no thromboplerous hyphae were observed, while these are allegedly present in A. brunneoconulus (Bas, 1982), although it is unknown if this difference could be deemed conclusive. Should the two entities are proven con-taxic, A. brunneoconulus will appear to be posterior synonym of A. beckeri. As far as sequences from type materials of the two names are not available so far, the decision is better postponed until the alleged synonymy could be backed up by molecular data, preferably obtained from the respective holotypes.
Of the phylogenetically closely related European species, A. beckeri has to be compared to A. simulans, a species known for its macroscopic variability, sometimes featuring greyish brown pileal colors and velar remnants on pileus that may turn in times yellowish buff (Vizzini et al., 2016). Amanita simulans however ought to be distinguished confidently by the obviously saccate (albeit fragile), often orange rusty spotted and greying volva, which although with common inflated elements, is predominantly composed by filamentous undifferentiated hyphae (Vizzini et al, 2016).
Among the species with semifriable volva, the most similar to A. beckeri is probably the distantly related A. alesides, for this recently described taxon is now well-known as having high chromatic variability and specimens with brownish-colored pilei are
likely not uncommon (Hanss, Moreau, 2020; Assyov et al., 2021; Kibby, Rogerson, 2021). Amanita alseides should be nevertheless easily distinguished from the species presented here as its velar remnants tend to become greyish with age, its stipe surface was never observed so far to develop dark zig-zag-like markings and its basidio-spores are with different quotient ratio, which average values range between 1.12 and 1.17 (Hanss, Moreau, 2020; Assyov et al, 2021; Kibby, Rogerson, 2021; Nicoletti et al, 2021).
Another species showing some similarity to A. beckeri is A. ceciliae (Berk. et Broome) Bas, which resembles the former macroscopically by the friable, darkening veil, leaving numerous remnants on the pileal surface, as well as by the dark zig-zaglike markings developing often on stipe surface. In A. ceciliae however the veil remnants, volva, stipe ornaments and often also lamellae turn greyish with age (Fraiture, 1993; Breitenbach, Kranzlin, 1995; Galli, 2001; Krieglsteiner, 2003; Vesterholt, 2008; Kibby, 2012), features that are sharply distinct from the ones observed in A. beckeri. Besides, A. ceciliae along with the next species features elements with brownish pigment in the veil (Hanss, Moreau 2020).
Among the species with greying volva and veil with predominantly spherical elements one must also take into account A. submembranacea, which often has pilei colored in different shades of brown, although this species has distinctly saccate, semifria-ble, greying volva and greying remnants of veil on pileus, when these latter are present (Fraiture, 1993; Breitenbach, Kranzlin, 1995; Galli, 2001; Krieglsteiner, 2003; Vester-holt, 2008), obviously different from the one seen in A. beckeri. This makes the confusion between the two highly unlikely, moreover A. submembranacea to our experience shows marked preference for occurrence in acidophilic coniferous forests, while the known collections of A. beckeri are from calcareous soil and more or less thermophilous habitats with broadleaf trees (Sebek, 1985; Tulloss, 1994; Galli, 2001; Krieglsteiner, 2003; Macau, Pérez-De-Gregorio, 2011).
Amanita spadicea Pers. is among the morphologically highly similar species to A. beckeri. This is primarily due to the brownish colored pilei in the former, combined with stipe with brownish zig-zag-like markings. It is however distinguished easily due to the well-developed, membranaceous, white, vaginate volva (Boccardo et al, 2008; Consiglio, Pierotti, 2012), decisively different from the rudimentary volva seen in the latter.
Amanita fulva Fr. and A. fulvoides Neville et Poumarat may possibly resemble superficially A. beckeri on account of the variable brownish coloration of pilei, as well as lamellae, which feature darker edge. They should be easily set apart due to their volva, which is vaginate, not friable, and develops rusty spots with age (Fraiture, 1993; Breitenbach, Kranzlin, 1995; Galli, 2001; Krieglsteiner, 2003; Vesterholt, 2008; Neville, Poumarat, 2009). Microscopically the volval tissues in those two species are composed predominantly of filamentous hyphae, mixed with some inflated elements (Fraiture, 1993; Breitenbach, Kranzlin, 1995; Neville, Poumarat, 2009), while in A. beckeri they are composed mainly of sphaerocytes.
Amanita betulae and A. coryli are further species that may also resemble to certain extent A. beckeri, due to the pileal coloration, volva and velar remnants on pileus surface which tend to develop yellowish buff color, and the stipe surface at least sometimes featuring ochraceous tinted ornaments (Neville, Poumarat, 2009). Those two equally scarcely presented in the mycological literature species however have volva of completely different architecture, composed primarily of undifferentiated filamentous hyphae and few inflated elements (Neville, Poumarat, 2009).
Amanita dryophila Consiglio et Contu, yet another little-known species, has brownish colored pilei and friable universal veil with numerous inflated elements (Consiglio, Contu, 1999). It could be separated from A. beckeri by the shape of the basi-diospores, which in the former are mainly broadly ellipsoid, with average quotient of 1.18 (Consiglio, Contu, 1999; Boccardo et al., 2008).
Amanita beckeri was described from France (Huijsman, 1962b) and so far available sequences in public databases originate exclusively from this country. Records exist also from neighboring Switzerland (Senn-Irlet et al, 2007). The species was documented also from a couple of localities in Spain (for discussion on the Spanish findings consult Macau, Perez-De-Gregorio, 2011). In the more northern western European countries it was found only in Denmark (Vesterholt, 2008) and Germany (Krieglsteiner, 2003). It was listed from Italy (Merlo, Traverso, 1983; Galli, 2001), but not all illustrated collections in those works appear to be typical and some of them might belong to other taxa (part of those inconsistencies were noted previously; cf. Macau, Perez-De-Grego-rio, 2011; Kibby, 2012). In the Mediterranean Europe the species was recorded also from Cyprus (Loizides, 2011), but it was later noted that this record may in fact belong to different species (Loizides et al, 2018). In Central Europe A. beckeri was reported from several localities in Austria (Krieglsteiner, 2003), Czech Republic (Sebek, 1985), Hungary (Siller, Vasas, 1995), Slovakia (Sebek, 1985) and Slovenia (Jurc et al, 2005). Further east, a record exists also from Romania (Pal-Fam, 2006). In the Balkan Peninsula, the fungus was previously mentioned from Croatia (Mesic, Tkalcec, 2002) and Serbia (Lukic, 2008, 2013), but judging from the published illustrations, the Serbian collections seems to represent other, likely unrelated taxa. In this paper we provide the first molecularly supported Balkan record of the species. The presented here Bulgarian collection appears to be the easternmost so far known finding of A. beckeri in Europe.
Acknowledgments
The financial support from the Bulgarian Academy of Sciences and the Scientific and Technological Research Council of Turkiye (Bilateral grant agreement between BAS and TUBlTAK, project number 118Z640) is gratefully acknowledged. The authors present their thanks to Mr. Pavel Nedelev (Sofia, Bulgaria) for providing the specimen and relevant documentation on the finding discussed in this study, as well as for granting permission to use his photographs, shown on the plate with the macro-morphological features of A. beckeri. They are also indebted to Mr. Gabriele Cacialli (Livorno, Italy) for the kindly offered help with missing literature.
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