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Amanita pseudovalens (Neville & Poumarat) R.Arraiano-Castilho, A.C.Silva, C.Vila-Viçosa, M.R.Castro, L.Morgado & P.Oliveira var. pseudovalens comb. nov., stat. nov. Basionym: Amanita curtipes f. pseudovalens Neville & Poumarat, Fungi Europaei 9: 656 (Neville & Poumarat 2004). SPECIMENS EXAMINED. — France. Landes, 15.V.1993, Mesplède, G74; 19.V.1965, J. Beller échantillon 2937, MPU1. Morocco. Larache, 04. V.1964, G. Malençon échantillon 5225, MPU 2 (DNA only). MYCOBANK. — MB 845581. NOTES This taxon corresponds to the French specimens (see Discussion for other possible locations). The basionym diagnosis (Neville & Poumarat 2004) remains unaltered for this variety (Table 6)., Published as part of Arraiano-Castilho, Ricardo, Silva, Ana Cristina, Vila-Viçosa, Carlos, Castro, Mário Rui, Morgado, Luís Neves & Oliveira, Paulo, 2022, The Amidella clade in Europe (Basidiomycota: Amanitaceae): clarification of the contentious Amanita valens (E. - J. Gilbert) Bertault and the importance of taxon-specific PCR primers for identification, pp. 139-157 in Cryptogamie, Mycologie 20 (6) on page 144, DOI: 10.5252/cryptogamie-mycologie2022v43a6, http://zenodo.org/record/7829510, {"references":["NEVILLE P. & POUMARAT S. 2004. - Amaniteae: Amanita, Limacella & Torrendia. Candusso, Alassio, 1120 p. (Fungi Europaei 9)."]}

Amanita pseudovalens var. tartessiana R.Arraiano-Castilho, A.C.Silva, C.Vila-Viçosa, M.R.Castro, L.Morgado & P.Oliveira, var. nov. HOLOTYPE. — Portugal. Alentejo, Beja district, Odemira, Luzianes A, 12.IV.2015, A. C. Silva, Ode12 (holo-, PO [PO-F2143]). ISOTYPE. — Portugal. Alentejo, Beja district, Odemira, Luzianes A, 21. III.2015, A. C. Silva, Ode02 (iso-, PO [PO-F2133]). ADDITIONAL SPECIMENS EXAMINED. — Portugal. Alentejo, Beja district, Odemira, Luzianes A, 21.III.2015, A. C. Silva, Ode01; Ode03; 28.III.2015, A. C. Silva, Ode04; 04.IV.2015, A. C. Silva, Ode08; Luzianes B, 04.IV.2015, A. C. Silva, Ode05; Ode06; Ode07; 12.IV.2015, A. C. Silva, Ode10; Ode11; Portalegre district, Portalegre, 27.IV.2015, N. Alegria, Ode16; Évora district, Portel, 07.IV.2010, R. Arraiano-Castilho, P01 (for basidiospores only). ETYMOLOGY. — The epithet refers to the ancient Tartessian civilization (ТαΡΤησσός) located in the south-west of the Iberian Peninsula (Celestino Pérez & López-Ruiz 2016). PHENOLOGY. — Late winter and spring. HABITAT. — Mediterranean, in association with Cistus spp., typically on compact, acidic and eroded soils, corresponding to regressive shrubland stages of evergreen oak forests (Fig. 5). DISTRIBUTION. — Portugal. Reported from the NUTS III regions of Alentejo Litoral, Baixo Alentejo, Alentejo Central and Alto Alentejo. MYCOBANK. — MB 845582. INDEX FUNGORUM. — IF 559871. NOTES This taxon corresponds to the Portuguese specimens described in this study. It differs from the autonym by its habitat (on acidic schist soils, with Cistus spp.), the ellipsoid to oblong, infrequently subcylindric basidiospores, and longer basidia (Table 6). Such differences are not considered to be at the rank of form. A species rank is currently not supported, due to the lack of genetic resolution of the nrDNA sequences. Both taxa share, aside from the characters in common with other species of series Amidella, the vernal fruiting season, the medium to small size, and the indistinct odour (Table 6), and can be confirmed with the Aps diagnostic PCR primers described in Table 2. DESCRIPTION To describe the new species, we used fresh materials obtained from sites A and B near Luzianes in spring 2015 (Ode01- 12 except Ode09) and the basidiospores from a collection in the Portel county (P01, near Monte Novo, spring 2010) and another from São Mamede Park near Monte Carvalho, Portalegre county (Ode16, spring 2015). All specimens used in this description were deposited in the PO herbarium (see Material and Methods). Pileus Flat to slightly depressed, convex at the margin, expanding to a diameter of 7.5 cm. Most collections whitish in situ, turning rose/ochre with either aging, handling, or scratching. A pale grey plaque from the universal veil is frequently present. Some collections present brownish scales close to the margin (Fig. 3B, C). Margin thinly appendiculate. Hymenophore Adnexed ascending, white, turning rose/ochre with either aging, bruising or scratching, with lamellulae. Stipe Almost cylindrical,slightly tapering toward the apex,non-bulbous, base obconical. Concolorous with the pileus, with a very fugacious annulus (Fig. 3H). A scale covering can be seen below the annulus region (Fig. 3A; C; E; F). Height not longer than the diameter of the expanded pileus, thickness 2.2 cm at the most. Veil Universal veil leaving a sac-like thick volva with a lobed margin, pale grey, with an internal ridge raised in contact with the stipe (Fig. 3H); often it remains also as a single pale grey plaque on the pileus. Partial veil leaving a fugacious non-membranous annulus at roughly two-thirds of the stipe height, and narrow remnants on the pileus margin. Context Concolorous with the surface, homogeneous, relatively compact, non-putrescent. Odour indistinct. Reaction with 10% FeSO 4 on rehydrated samples from the stipe develops an immediate change to greenish grey that lasts a few minutes. Basidiospores White, amyloid, ellipsoid to oblong, average length 11.78 µm, average width 6.97 µm, average length/width ratio (Q) 1.696 (Table 5), overlapping the A. ponderosa sporograph but not the one for A. curtipes f. pseudovalens (Fig. 4). Due to the lack of spore print, collections Ode01 and Ode16 were not included in the summary calculations. Statistical testing rejected the hypothesis of homogeneity among the collections included in the summary statistics, for all three variables (Appendices 4; 5). Indeed, the heterogeneity among collections was the rule (Table 5; Appendix 8): Ode10 had longer spores and higher Q, bordering on standard A. pseudovalens comb. nov., stat. nov. limits; Ode08 had wider spores and lower Q, even more than A. ponderosa, while Ode11 had spores of average Q values but small size. Nested ANOVA (within and between sites) also suggested heterogeneity within sites for the three variables, but only for length and width between sites.Site A collections have on average significantly higher values (Appendix 6). Basidia Clavate, with 4 sterigmata, base unclamped, average length 56.9 µm (equal to the median), range 41.0-73.4 µm, n = 123. The measurements were obtained from collections Ode02b, Ode05, Ode06a, Ode07, Ode08, Ode10, Ode11 and Ode12, revealing a normal distribution of the global data (Shapiro-Wilk’s W = 0.990, P = 0.555). On average, similar basidia sizes were observed across all collections, although Ode11 had a higher average length of 63.9 ± 5.0 µm (Appendix 7). Universal veil Sagital sections (Ode02, Ode07, Ode11, Ode12) revealed a thin outer layer, 80-100 µm deep, composed of slightly interwoven, longitudinally oriented hyphae, very compact and 10 µm wide (as measured in transversal sections), from which thinner hyphae, very loose, projected outwards (not found in Ode11). Many of the latter hyphae had a slightly widened, bulbous termination. No clamp connections were observed. The remainder of the structure was composed of more loosely packed longitudinal, slightly wavy hyphae, with very conspicuous lacunae interpreted as remnants of larger inflated, ellipsoid to oblong, hyphal elements. Measurement of these lacunae in Congo Red SDS stained sections (Ode02, Ode07, n = 41), under low magnification, gave an estimate of 40-71 µm length (average 54 µm, C. V. 17%) by 20-46 µm width (average 33 µm, C. V. 16%)., Published as part of Arraiano-Castilho, Ricardo, Silva, Ana Cristina, Vila-Viçosa, Carlos, Castro, Mário Rui, Morgado, Luís Neves & Oliveira, Paulo, 2022, The Amidella clade in Europe (Basidiomycota: Amanitaceae): clarification of the contentious Amanita valens (E. - J. Gilbert) Bertault and the importance of taxon-specific PCR primers for identification, pp. 139-157 in Cryptogamie, Mycologie 20 (6) on pages 144-146, DOI: 10.5252/cryptogamie-mycologie2022v43a6, http://zenodo.org/record/7829510, {"references":["TAMURA K. & NEI M. 1993. - Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution 10: 512 - 526. https: // doi. org / 10.1093 / oxfordjournals. molbev. a 040023"]}

IDENTIFICATION KEY TO AMANITA PERS. SERIES AMIDELLA (E.- J.GILBERT) NEVILLE & POUMARAT 1. Pileus remaining convex for most of the development of the basidiome, sometimes with an umbo, diameter notably smaller than the length of the developed stipe, covered at least on the margin with heterogeneous scales from the inner layer of the universal veil, which become brown like the exposed context, stipe base bulbous, margin of the volva leaning towards the stipe, context discoloration moderate to intense. Rare occurrences, widely spaced in time (A. lepiotoides)............................................................................................................ 2 — Pileus becoming flat then depressed at the centre, diameter about the same as the stipe length, generally without scales, stipe slightly or not bulbous, margin of the volva free, pale context discoloration, rose then ochre. Occurrence generally annual........................................................................................................................ 3 2. Context discoloration intense, reddish then dark brown, stipe base distinctly bulbous, basidiospores mostly ellipsoid (Qm A. lepiotoides f. lepiotoides Barla — Context discoloration moderate, rose then ochre, stipe base slightly bulbous, basidiospores mostly oblong and cylindrical (Qm> 1.65).................................................. A. lepiotoides f. subcylindrospora Neville & Poumarat 3. Occurrence in Autumn, on siliceous sandy soil............................................................ A. curtipes E.- J.Gilbert — Occurrence from late winter (rarely in January) to spring, also early summer............................................... 4 4. Heavy habit, semi-hypogeous, typically with an earthy odour, associated with Cistus, basidiospores mostly ellipsoid to oblong (1.60 A. ponderosa f. ponderosa Malençon & R.Heim — Medium-sized (pileus diameter generally less than 10 cm)............................................................................ 5 5. On siliceous sandy soil, associated or not with Cistus, pines, etc., basidia with average length Cistus, basidia with average length> 55 µm............................................................................................................................................ 7 6. Relatively small (pileus diameter usually around 5 to 6 cm and below 8 cm), taste of the context indistinct, basidiospores mostly oblong (1.8 A. curtipes E.- J.Gilbert — Medium-sized (pileus diameter usually between 7 and 10 cm), taste can be slightly of hazelnut, then bitter, basidiospores mostly cylindrical (2.0 A. pseudovalens var. pseudovalens comb. nov., stat. nov. 7. Odour and/or taste typically distinct, semi-hypogeous habit (A. ponderosa).................................................. 8 — Odour indistinct, aftertaste slightly bitter, epigeous habit, pileus margin occasionally with brown scales.................................................................................................................... A. pseudovalens var. tartessiana var. nov. 8. Basidiospores broadly oblong (Qm = 1.60 – 1.80), strictly thermophile......................................................................................................................................................... A. ponderosa f. ponderosa Malençon & R.Heim — Basidiospores long-ellipsoid (Qm = 1.45 – 1.55), also in cooler areas...................................................................................................................................................... A. ponderosa f. valens (Gilbert) Neville & Poumarat Only the European taxa within Amanita series Amidella are considered. Parts of the key are from the work of Neville & Poumarat (2004)., Published as part of Arraiano-Castilho, Ricardo, Silva, Ana Cristina, Vila-Viçosa, Carlos, Castro, Mário Rui, Morgado, Luís Neves & Oliveira, Paulo, 2022, The Amidella clade in Europe (Basidiomycota: Amanitaceae): clarification of the contentious Amanita valens (E. - J. Gilbert) Bertault and the importance of taxon-specific PCR primers for identification, pp. 139-157 in Cryptogamie, Mycologie 20 (6) on page 150, DOI: 10.5252/cryptogamie-mycologie2022v43a6, http://zenodo.org/record/7829510, {"references":["NEVILLE P. & POUMARAT S. 2004. - Amaniteae: Amanita, Limacella & Torrendia. Candusso, Alassio, 1120 p. (Fungi Europaei 9)."]}

COMPARISON WITH AMANITA CURTIPES F. PSEUDOVALENS AND AMANITA PONDEROSA When comparing the descriptions of all European taxa of series Amidella, based on Neville & Poumarat (2004), as shown in Table 6, the Portuguese specimens of Amanita pseudovalens comb. nov., stat. nov. did not conform with the description for the genetically indistinct A. curtipes f. pseudovalens (Neville & Poumarat 2004), diverging in their soil and vegetation affinities and in the microscopy. Therefore, we understand them as representing a separate taxon, however at infraspecific level. Moreover, Table 6 shows that the tartessiana variety resembles A. ponderosa in almost every aspect – the obvious difference being the so far unknown occurrence of large and heavy specimens in the former – being, by comparison, much less similar to the conspecific A. pseudovalens comb. nov., stat. nov. collected in France., Published as part of Arraiano-Castilho, Ricardo, Silva, Ana Cristina, Vila-Viçosa, Carlos, Castro, Mário Rui, Morgado, Luís Neves & Oliveira, Paulo, 2022, The Amidella clade in Europe (Basidiomycota: Amanitaceae): clarification of the contentious Amanita valens (E. - J. Gilbert) Bertault and the importance of taxon-specific PCR primers for identification, pp. 139-157 in Cryptogamie, Mycologie 20 (6) on page 146, DOI: 10.5252/cryptogamie-mycologie2022v43a6, http://zenodo.org/record/7829510, {"references":["NEVILLE P. & POUMARAT S. 2004. - Amaniteae: Amanita, Limacella & Torrendia. Candusso, Alassio, 1120 p. (Fungi Europaei 9)."]}