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A new Chinese truffle species, Tuber sinomacrosporum belongs to the Macrosporum group was illustrated and described, it is characterized by brownish-red ascoma, prosenchymatous peridium, dark brown gleba, 1(2) spored asci and large ellipsoid ascospores with 2–6 μm high reticulate-alveolate. Molecular analysis of the internal transcribed spacers (ITS) of the nuclear ribosomal DNA supports this species is distinct from the known species within the Macrosporum group. Phylogenetic relationships and morphological differences between this new species and its related species are discussed.

Tuber melanorufum L. Fan sp. nov. (FIG. 2) MycoBank:—MB 846497 Diagnosis:—differed from other species by the large red-brown to blackish ascomata covered with distinctively fine warts and the ascospores ornamented with crowded spine-reticulations. Etymology:— melano, latin, means black; rufum, latin, means red. Referring to the color of ascomata that are red-brown when young and change to blackish to black with age. Typification:— CHINA. Yunnan Province: Huize County, in soil under a Pinus forest, 31 Oct 2020, collected by Jinzhong Cao (BJTC FAN1273). Description:— Ascomata 1.6–4.5 cm diam, irregularly globose to subglobose, reddish (#f18f5f) to red brown (#800000) when young, then changing blackish to black (#460707) with age, surface covered with distinctly fine warts, often with some superficial and pale colored furrows. Odor light, favour not recorded. Gleba pale when young, brown (#4e0000) to coffee brown (#670000) at maturity, but never black, marbled with narrow and numerous whitish veins. Peridium 350–400 μm thick, two-layered: outer layer 200–250 μm thick including warts of 30–50 μm high, pseudoparenchymatous, composed of subglobose to irregularly shaped cells of 5–15 × 5–20 μm wide, with slightly thickened walls, brown walls at outermost cells; inner layer 150–200 μm thick, composed of interwoven hyphae 2.5– 7.5 μm wide, hyaline thin-walled. Asci subglobose or ovoid, 65–75 × 55–62.5 μm, hyaline, thick-walled of 2.5–3μm, 1–4(–5)-spored, sessile or with a short stalk at maturity. Ascospores ellipsoid to oblong-ellipsoid, usually narrowed in one end, yellow brownish (#a36e12) at maturity, ornamented with densely spine-reticulations of 2.3–3(–5) μm high, negative in Melzer’s reagent, excluding ornamentation, in 1-spored asci 25–57.8 × 15–28.5 μm, Q = 1.24–2.57 (Qm =1.91 ± 0.66) (n = 30), in 2-spored asci 26–51 × 13.6–25.6μm, Q = 1.41–2.62 (Qm =2.02 ± 0.6) (n = 30), in 3-spored asci 22.1–43.9 × 13.3–23.2 μm, Q = 1.18–2.31 (Qm = 1.75 ± 0.56) (n = 30), in 4(–5)-spored asci 17.9–40 × 11.7–21.2 μm, Q = 1–2.45 (Qm = 1.73 ± 0.73) (n = 30). Habit, habitat and distribution: —hypogeous, gregarious, in soil under Pinus forest in Yunnan province, China. Currently known only from China. Specimens examined. CHINA. Yunnan Province: Huize County, in soil under forest of Pinus sp., 21 Oct 2020, collected by Jinzhong Cao (BITC FAN1274, BJTC 1276, BJTC FAN1287)., Published as part of Fu, Hao-Yu & Fan, Li, 2023, Tuber melanorufum and T. tomentellum, two new truffle species from China, pp. 178-190 in Phytotaxa 594 (3) on pages 182-186, DOI: 10.11646/phytotaxa.594.3.2, http://zenodo.org/record/7901081

Tuber melanorufum and T. tomentellum spp. nov. are described and illustrated in this paper based on the morphological and phylogenetic data. Tuber melanorufum is diagnosed by the red-brown to blackish ascomata covered with distinctly fine warts and the ellipsoid to long ellipsoid ascospores ornamented with very crowded spine-reticulations. Tuber tomentellum is characterized by the large ascospores and two types of hairs arising from the outer cells of the peridium. Phylogenetic analyses place T. melanorufum and T. tomentellum in the Rufum and Puberulum clades, respectively.

Tuber tomentellum L. Fan & H.Y. Fu sp. nov. (FIG. 3) MycoBank:—MB 846498 Diagnosis:—differed from other species by the large ascospores reaching 78.55 μm in length and the numerous hyphoid or seta-like hairs on the peridial surface of ascomata. Etymology:— tomentellum, tomemtose, referring to the ascomatal surface covered with short woolly hairs. Typification: — CHINA. Yunnan Province: Kunming City, in soil under a Pinus forest, Oct 2020, collected by Jinzhong Cao (BJTC FAN1340) Description:— Ascomata 1.3–2.2 cm, yellow brown (#f4d28d) to grey brown (#654321), surface even and finely tomemtose, with an indistinctly basal depression. Odor mild, flavour not recorded. Gleba brown (#7c2f20) at maturity, marbled with large and rare white veins. Peridium 200–300 μm thick, two-layered: outer layer 150–200 μm thick, pseudoparenchymatous, composed of subglobose to irregularly shaped cells of 7.5–32.5 × 5–25 μm, thin-walled, light brown (#f5dbc2) toward outside surface; inner layer 75–150 μm thick, composed of interwoven hyphae 4–7.5 μm wide, hyaline, thin-walled, septate. Hairs arising from outermost cells of the peridium, two types, hyphoid, flexible, and having obtuse ends, or seta-like, erect, and having sharp ends, 30–90 × 2.5–4 μm, hyaline, slightly thick-walled, 1–3-septa. subglobose or ovoid, 80– 110 × 50–75 μm, hyaline, thin-walled, 1–3-spored, sessile. Ascospores ellipsoid to broadly ellipsoid, brown (#a52a2a) at maturity, reticulate, negative in Melzer’s reagent, excluding ornamentation, in 1-spored asci 30–78.5 × 25.7–46 μm, Q = 1.07–1.96 (Qm =1.52 ± 0.45) (n = 30), in 2-spored asci 31.1–64.5 × 26.2–41.4 μm, Q = 1.1–1.75 (Qm =1.43 ± 0.33) (n = 30), in 3-spored asci 29.6–62 × 24.2–38.9 μm, Q = 1.07–1.76 (Qm = 1.42 ± 0.35) (n = 30), reticulum 4–6 μm high, 6–7 meshes across spore width. Habit, habitat and distribution: —hypogeous, gregarious, in soil under a Pinus forest in Yunnan Province, China. Currently, only known from China. Additional specimens examined: — CHINA. Yunnan Province: Dali City, Xiangyun County, in soil under forest of Pinus sp., 21 Oct 2020, Collected by Baoxiang Shi (BITC FAN1330); Kunming City, in soil under a Pinus forest, Oct 2020, collected by Baoxiang Shi (BJTC FAN1346).

Tuber sinomacrosporum S. P. Wan, & F. Q. Yu, sp. nov. (FIGURE 2) MycoBank: 846502 Typification: CHINA. Yunnan Province, Dali City (25° 38′ N , 100° 68′ E), in humic soil under a pure Pinus forest, at about 2090 m, 26 December 2020, wsp1043-8, YNAU016 (GenBank ITS number = OK625303). Diagnosis: Tuber sinomacrosporum is characterized by the brownish-red ascoma, mostly 1(2)-spored asci, large and ellipsoid ascospores. Etymology: referring to a Chinese species have large spores. Description: Ascoma 2.2 × 1.35 cm in diam, oval-shaped or irregular, light brown, partial brown to brownishred when fresh, becoming brownish-red when dried, surface partially covered with yellow tomentose. Odor pleasant. Peridium 108.0–148.0 μm thick, one layer, prosenchymatous, composed of yellowish to transparent cells and interwoven hyphae, cells (1.0–)2.2–5.7(–7.8) × (1.0–)1.7–4.5(–5.4) μm, hyphae 1.3–2.8 μm in diam. Gleba solid, blackish at maturity, marbled with different whitish to greyish veins continuous with the peridium; composed of hyaline, interwoven, thin-walled hyphae, 0.7–2.4 μm broad at the septa, the cells subglobose to globose, 1.1–7.3(– 10.5) × 0.9–5.7(–8.6) μm. Asci 114.0–143.0(–158.0) × 86.0–117.0(–124.0) μm, ellipsoid, rarely pyriform or irregular, hyaline, sessile or with a short or tall stalk 7.5–49.0 μm, 1-spored, rarely 2-spored. Ascospores ellipsoid, sometimes oblong elliptical and broadly ellipsoid, hyaline when young, becoming brownish-red to brown at maturity; excluding alveolar ornamentation, in 1-spored asci (80.0–)89.0–105.0(–109.0) × (55.0–)61.0–76.0(–78.0) μm (Q = 1.32–1.53, Q m = 1.43 ± 0.08) (n = 120), in 2-spored (51.0–)55.0–78.0(–84.0) × (32.0–)37.0–55.0(–57.0) μm (Q = 1.25–1.65, Q m = 1.4 ± 0.16) (n = 30). Ornamented with alveolate reticulum, 2–6 μm deep, constituted of irregularly polygonal meshes, reticulum with 3–7 meshes along the spore length and 3–7 across. Distribution and habitat: Hypogeous, in soil under pure stand of Pinus forest in Yunnan Province, China. Known only from China., Published as part of Wan, Shan-Ping, Liu, Wei, Cui, Meng-Jin, Wang, Rui, Wang, Yu-Yun, Liu, Mei-Ju, Wang, Rui- Xue, Yu, Cheng-Jin & Yu, Fu-Qiang, 2023, Morphological and molecular evidence support a new truffle, Tuber sinomacrosporum, in Macrosporum group, pp. 283-291 in Phytotaxa 591 (4) on pages 286-288, DOI: 10.11646/phytotaxa.591.4.5, http://zenodo.org/record/7835575

An optimized PLE method was applied to several truffle species using three different solvent mixtures to obtain bioactive enriched fractions. The pressurized water extracts contained mainly (1 ? 3),(1 ? 6)-?-D-glucans, chitins, and heteropolymers with galactose and mannose in their structures. The ethanol extracts included fatty acids and fungal sterols and others such as brassicasterol and stigmasterol, depending on the species. They also showed a different fatty acid lipid profile depending on the solvent utilized and species considered. Ethanol:water extracts showed interesting lipids and many phenolic compounds; however, no synergic extraction of compounds was noticed. Some of the truffle extracts were able to inhibit enzymes related to type 2 diabetes; pressurized water extracts mainly inhibited the ?-amylase enzyme, while ethanolic extracts were more able to inhibit ?-glucosidase. Tuber brumale var. moschatum and T. aestivum var. uncinatum extracts showed an IC50 of 29.22 mg/mL towards ?-amylase and 7.93 mg/mL towards ?-glucosidase. Thus, use of the PLE method allows o bioactive enriched fractions to be obtained from truffles with antidiabetic properties. This research was funded by the fellowship Ibercaja-CAI Estancias de Investigación number CA 1/20. The research leading to these results has received funding from the European Union under “Horizon 2020—the Framework Programme for Research and Innovation (2014–2020)”. Grant Agreement of the Project: “Innovation in truffle cultivation, preservation, processing and wild truffle resources management”, INTACT, Project number 101007623. truffles β-D-glucans ergosterol fatty acids antidiabetic amylase glucosidase Published

We describe two new Japanese truffle species, Tuber iryudaense and Tuber tomentosum, based on molecular and morphological analyses. Both species are clearly distinguishable from other Tuber species by the ocher tomentose mycelium covering the ascoma surfaces. Tuber iryudaense has one-spored asci that each contain a large (68-97 × 51-80 µm), reddish-brown ascospore; these microscopic characters are similar to those of closely related Chinese species, T. calosporum, T. gigantosporum, T. glabrum, T. monosporum, and T. sinomonosporum. Tuber tomentosum forms one to four ascospores per ascus with a reddish-brown color similar to that of ascospores of T. macrosporum and T. canaliculatum, although their spores are much larger than those of T. tomentosum (27‒64 × 26‒55 µm). Molecular phylogenetic analyses based on ribosomal internal transcribed spacer and partial 28S nuc rDNA sequences support that both species are distinct within the Macrosporum group.

A new hypogeous species, Choiromyces cerebriformis sp. nov. is described and illustrated from Yunnan province, China. Both morphological evidence and sequence analyses of the internal transcribed spacer region (ITS1-5.8S-ITS2) support the species as new to science. C. cerebriformis differs from other Choiromyces species in having ascomata with larger lobes and light orange-brown gleba, and globose ascospores with short stick-like sparse spines ornamentation. Morphological differences and genetic distances with the similar species C. helanshanensis and C. alveolatus are discussed. A phenotypic key including related species is provided.

A new black truffle species with a basal cavity, Tuber melanoexcavatum, is identified and described. The new species is morphologically similar to T. pseudobrumale, T. pseudoexcavatum, and T. mesentericum in having a basal cavity. However, T. melanoexcavatum differs from T. pseudobrumale and T. pseudoexcavatum in having black pyramidal warts and 5 to 8 spored asci, and differs from T. mesentericum by having spiny ascospores and 1 to 5 spored (rarely 6 spored) asci. Phylogenetic analysis indicates that T. melanoexcavatum is most closely related to T. pseudobrumale, but the two still clearly differentiate. Tuber melanoexcavatum grows as a mycorrhizal association with Cyclobalanopsis glaucoides. Tuber melanoexcavatum is considered an edible black truffle species and is indigenous to China.

The Chinese black truffle Tuber indicum is the main commercial truffle species in China and has strong potential for cultivation. Most studies have focused so far on the production and planting of mycorrhizal seedlings of tree species native to China. Here, we selected four exotic tree species, Quercus pubescens, Q. ilex, Q. palustris and Pinus pinea and inoculated axenically germinated seedlings, five replicates per tree species, with T. indicum spore suspension. As shown by morphological, anatomical and molecular analyses, mycorrhizae were successfully synthesized under greenhouse conditions from 6 months after inoculation and the mycorrhization was stable for at least 24 months in the glasshouse environment. Despite slight morphological variations, T. indicum mycorrhizae were similar on all tree species, i.e. swollen, red-brownish with long hyaline emanating hyphae showing right-angle ramifications. Our observations confirmed the similarity of T. indicum mycorrhizae with those of T. melanosporum regardless of the geographic origin of host trees. Four out of five T. indicum-inoculated pine seedlings were cross-contaminated by T. borchii that was inoculated to another group of P. pinea seedlings raised in the same glasshouse. This is the first study to document the mycorrhization of exotic tree species by T. indicum in China and the first report for Q. palustris. With two successful out of five inoculated seedlings, Q. palustris was less receptive to T. indicum mycorrhization than the other two oak species. Further work is needed to assess whether the T. indicum symbioses obtained here are maintained after planting in the field and if ascomata can be produced by exotic tree species under Chinese environmental conditions.

The Australian genus Reddellomyces comprises several closely related symbiotic ascomycetes with chambered gleba (solid ptychothecium), ornamented ascospores and cylindrical paraphyses that are sometimes difficult to identify based on morphological aspects. Several Reddellomyces species are spread with their Australian host plants (Eucalyptus and Acacia) worldwide including Mediterranean areas. Specimens of an unusual truffle were collected from Acacia saligna stands in Tunisian semi-arid lands and eucalypt forest of Antalya (Turkey) dominated by E. camaldulensis Dehnh. and E. grandis W. Hill. Based on morphological features, these fungi are affiliated to Reddellomyces genus, differing from other species of the genus mainly based on ascospore size. Further analysis based on nuclear rDNA LSU and ITS and digital RFLP of the ITS sequenced amplicons with selected restriction endonucleases showed the unique identity of Tunisian specimens within the genus Reddellomyces. Our results showed also that at least six Reddellomyces species are present including two unnamed species restricted to Australia.

Tuber songlu S.P. Wan, sp. nov. (Fig. 4) MycoBank: MB 839734 Typification: CHINA. Yunnan Province, Huize County (29.103��E, 25.56��N), in humic soil under a pure Pinus armandii forest, at about 2415 m, 14 August 2016, wsp695, HKAS 95771 (GenBank Acc. No.: ITS = KX904883, LSU = KY 013656). Diagnosis: Tuber songlu differs from other species by its whitish ascomata, dense spine-like dermatocystidia, ellipsoid ascospores and 1���4 spored asci. Etymology: Refers to the local name of the fungus. Description: Ascomata 1���2 cm in diam, globose to subglobose or irregular, white, light reddish brown when fresh, becoming brown when dried. Peridium 130���500 ��m thick, densely pubescent, one layer, prosenchymatous, composed of subglobose to subangular cells, (1���) 1.5���18(���23) �� (1���) 1.5���7(���8.5) ��m, hyaline. Gleba solid, brown when mature, marbled with brownish yellow veins, composed of hyaline, interwoven, thin-walled cylindrical hyphae, 1.5���8.5 ��m broad at the septa, and interwoven, inflated hyphae 2���8.5 ��m broad. Dermatocystidia, straight or bent, obtuse or apiculate at the tip, up to 230 ��m long, 7 ��m in diam, multiseptate, hyaline to whitish. Asci (63���) 72���96 �� 54���75 (���81) ��m, globose to subglobose, pyriform, ellipsoid or irregular, hyaline, sessile or with a short stalk, thin-walled 1���2 ��m thick, 1���4 spored. Ascospores, ellipsoid, sometimes broadly ellipsoid, subglobose, hyaline when young, becoming brown at maturity; excluding their alveolate-reticulate ornamentation, in 1-spored asci (41���) 42.5���51.5 (���56) �� (26.5���) 32.5���40 (���41) ��m (Q = 1.10���1.70), in 2-spored (22���) 24���42 (���46) �� (18���) 19���34 ��m (Q = 1.10���1.51), in 3-spored (20���) 26���33 (���35) �� 19 (���21)���29 (���30) ��m (Q = 1.13���1.24), and in 4-spored (19���) 20���27.5 (���29) �� 15���17 (���27) ��m (Q = 1.1���1.2); reticulum with 3���8 meshes along the spore length and 3���8 across. The alveolar walls up to 1���7 ��m tall. Distribution and habitat: Hypogeous, in soil under a pure forest of Pinus armandii in Yunnan Province, China. Known only from China. Other material examined: CHINA. Yunnan Province, Province, Huize County, in soil under a pure forest of P. armandii, December 2016, wsp701, HKAS 95777 (GenBank Acc. No.: ITS = KX904884, LSU = KY 013657); wsp749, HKAS 95851 (GenBank Acc. No.: ITS = KX904886, LSU = KY 013658)., Published as part of Wan, Shanping, Liu, Jianwei, Huang, Lanlan, Qin, Xiaomin, Liu, Wei & Yu, Fuqiang, 2021, Tuber qujingense and T. songlu, two new species from Yunnan, China, pp. 248-256 in Phytotaxa 527 (4) on pages 253-254, DOI: 10.11646/phytotaxa.527.4.2, http://zenodo.org/record/5766179

Tuber qujingense and T. songlu are described as new species based on both phylogenetic and morphological analyses. Phylogenetically, these two species are clearly distinct from described species of Tuber. Morphologically, these two species have whitish smooth ascomata, pubescent and prosenchymatous peridium and 1–4 spored asci; however, T. qujingense is characterized by its fusiform ascospores and taller alveolar walls (4.5–11 µm) while T. songlu has larger ellipsoid ascospores and lower alveolar walls (1–7 µm).

Tuber qujingense S. P. Wan, sp. nov. (Fig. 3) MycoBank: MB 839733 Typification: CHINA. Yunnan Province, Huize County (18.103��E, 23.26��N), in humic soil under a pure Pinus armandii forest, at about 2400 m, 12 August 2016, wsp721, HKAS 95823 (GenBank Acc. No.: ITS = KX904885, LSU = KY 013659). Diagnosis: Tuber qujingense differs from related species by its greyish white ascomata, brown snowflake-shaped gleba, prosenchymatous peridium, fusiform ascospores and 1���4 spored asci. Etymology: Refers to the location of the type collection. Description: Ascoma 2.5 cm in diam, subglobose or irregular, greyish white when fresh, becoming brown when dried. Peridium 200���500 ��m thick, smooth to pubescent, one layer, prosenchymatous, composed of big, subglobose to subangular cells, (1���) 2���33 (���34.5) �� (1���) 1.5���22 (���33) ��m, light earthy yellow. Gleba solid, brownish purple when mature, marbled with white veins, composed of hyaline, interwoven, thin-walled hyphae, 1.5���6 ��m, and cylindrical, inflated hyphae 2.8���50 �� 2.8���43 ��m. Dermatocystidia or setae, straight or bent, obtuse or apiculate at the tip, up to 110 ��m long, 6.5 ��m in diam, septate, hyaline to whitish. Asci (40���) 51���80 �� (30���) 31���60 ��m, globose to subglobose, pyriform, ellipsoid or irregular, hyaline, sessile or with a short stalk, thin-walled 1���2 ��m thick, 1���4 spored. Ascospores, fusiform, ellipsoid, sometimes broadly ellipsoid, subglobose, hyaline when young, becoming brown at maturity; excluding the alveolate-reticulate ornamentation, in 1-spored asci (35���)37���48(���50) �� (23���) 25���30(���32) ��m (Q = 1.45���1.65), in 2-spored (22���) 28���40(���41) �� (14���) 18-27 ��m (Q = 1.41���1.7), in 3-spored (20���)3 8���18(���20) �� 15���19(��� 24) ��m (Q = 1.23���1.75), and in 4-spored (17���) 20���34(���36) �� 14���21(���22) ��m (Q = 1.14���1.72); reticulum with 3���10 meshes along the spore length and 3���8 across. The alveolar walls up to 4.5���11 ��m tall. Distribution and habitat: Hypogeous, in soil under pure stand of P. armandii in Yunnan Province, China. Known only from China., Published as part of Wan, Shanping, Liu, Jianwei, Huang, Lanlan, Qin, Xiaomin, Liu, Wei & Yu, Fuqiang, 2021, Tuber qujingense and T. songlu, two new species from Yunnan, China, pp. 248-256 in Phytotaxa 527 (4) on pages 251-252, DOI: 10.11646/phytotaxa.527.4.2, http://zenodo.org/record/5766179

Th e true truffl es, Tuber aestivum Vittad., T. borchii Vittad., T. magnatum Picco and T. melanosporum Vittad., are among the most studied fungal species; they also have a high economic value due to their special aromatic and nutritional properties that make them a much sought delicacy. Despite this, their identifi cation has been based on morphological and then molecular characters in the absence of reference type specimens. Although long of scientifi c, commercial and regulatory use, these four scientifi c names are at risk due to a lack of nomenclatural priority. To provide the scientifi c community with reference voucher samples and to initiate nomenclatural proposals for the recognition of their status as conserved names, three collections from sites mentioned by their authors (Picco and Vittadini) are proposed as epitypes for Tuber aestivum, T. borchii and T. melanosporum, and one as a neotype for T. magnatum. Th e type of each name is described morphologically and molecularly characterized with the sequences of three markers: ITS, ?-tubulin, elongation factor 1?. Th e taxonomy and nomenclature of each species are discussed. Th e conservation of the names Tuber aestivum against the previous homonymous Tuber aestivum (Wulfen) Spreng. and the competing name Tuber blotii Eudes-Desl., T. magnatum against Tuber griseum Borch ex Pers., and T. melanosporum against Tuber nigrum Bull. will be proposed. Th e name Tuber borchii has no previous synonyms and therefore it is legitimate and does not require conservation.

Tuber borchii Vittad. (Figs 2; 4B; 5C, D) Monographia Tuberacearum: 44 (1831). MYCOBANK NUMBER. — MB 118774. GENBANK. — MZ423174 (nrITS), MZ458418 (nr β- tubulin), MZ458422 (nrEF 1-α). LECTOTYPE OF TUBER BORCHII VITTAD. — Vittadini 1831 tab. I (presented with the indication XX), fig. III E-F here reprinted in Fig. 2. (here designated; MycoBank Typification number: MBT 10001893). EPITYPE OF TUBER BORCHII VITTAD. — Italy. Lombardy, Varzi, Castello Oramala, sub Quercus pubescens Willd., 6.III.2019,44°50’28”N, 9°11’32”E, twenty ascomata, leg. Stefano Seghezzi, det. Giovanni Pacioni (epi-, AQUI[AQUI10151] here designated MycoBank Typification number: MBT 10001895). DESCRIPTION Ascomata Hypogeous globose, lobate or irregular, up to 7 cm in diameter, rarely larger, sometimes with a flat base without excavation, surface of peridium pubescent, whitish (FCF3CF light grayish yellow, F9E79F very soft yellow, F0B27A soft orange), yellowish-gray, ocher, brownish, sometimes with reddish spots, smooth when fully mature, still pubescent in the depressions. Gleba Firm from whitish (D5DBdB light grayish green, EC7063 soft red, 797D7F dark grayish blue) to ocher with reddish hues, becoming greyish or reddish-brown by maturity, with ramified-anastomoses and thin sterile veins whitish in youth. Later ochraceous or reddish, marbled with wide, anastomosing veins which arise from various places of the peridium. Odor Pleasant, then strongly garlic sulfurous; taste strong. Peridium 150-600 Μm thick, exoperidium pseudoparenchymatous, 100-300 Μm thick with textura globosa, formed of swollen cells ranging from suglobose to irregularly ellipsoid 10-50 × 8-24 Μm, yellowish but strongly coloured on the surface, with walls of collenchymatic type thickened up to 4 Μm, surface with tapered hyaline thick-walled, one to three celled cystidia up to 100 Μm long, up to 8 Μm wide at the inflated base; endoperidium plectenchymatous 60-450 Μm wide with intertwined, mainly periclineal, hyaline hyphae 20-24 × 5-6 Μm, with thin walls, mixed with a few rounded cells with slightly thickened walls; sterile veins emerge from the endoperidium to penetrate the gleba from various parts of the peridium. Asci Globose to subglobose, 60-100 × 50-80 Μm, with walls 1-1.5 Μm to 5 Μm thick, sessile or short stalked, croziered at the foot, containing 1-4 spores. Ascospore Subglobose to broadly ellipsoid, Q 1.06-1.46, 20-55 × 18-42Μm excluding ornamentation, inversely proportional in size to the number of spores in the ascus, at first translucent, then becoming light yellow to yellowish brown, sometimes with a reddish hue, regularly alveolate with mostly hexagonal but sometimes rectangular or elongated alveolae numbering 4-11 along the spore length and 3-9 across the spore width, 4-10 Μm wide but usually near 7 Μm, and 3-5 Μm high. Endosporium Often two-layered in KOH. Glebal Hyphae hyaline, 4-10 × 20-32 Μm HABITAT. — Species naturally present in all Europe to the southern Scandinavian peninsula, from Portugal to the Caucasus and Anatolia, under broadleaved and coniferous trees, mainly sandy soil, commonly from late autumn to late spring (Ceruti et al., 2003; unpublished data) NOMENCLATURE AND TAXONOMY There may be a precedent synonym for this species: Tuber albidum Picco, Melethemata inauguralia: 79 (1788) [MB 227650] which however is unavailable, unpriorable, because of the sanctioned homonym Tuber albidum Fr. (1823) [MB 227428], which refers to a different species. This Fries’ taxon is difficult to identify because of its diagnosis “ verrucis exasperatum, albidum ”. Vittadini (1831: 40, Obs.II) wrote about it: ‘the whitish external color in warty Tuber is unknown to me, I suspect. So the (species) named albidum is doubtful. (Obs. II. Tuber albidum Fr. … Color externus albidus in Tuberibus muricatis mihi prorsus extraneus, suspectus. Hinc albidi nomen ambiguum.)’. Perhaps due to a ‘ lapsus calami’, Tuber albidum Fr. is a questionable species that does not even today find a match among the known species of Tuber where all the warty peridium species are black or deep brown (Bonito et al. 2013). Fries (1823: 291) claims to have examined a dried sample (vs, vidi siccam), but no specimen of Tuber albidum was found in the Herbarium of Fries (UPS-FRIES) or a drawing of this taxon among Fries documents. From the Fries’ description it is understood that it could refer to Tuber aestivum and two references (Cesalpino 1583: 613, and Micheli 1729: 221) could confirm it, while a third (Gleditsch 1753: 257) ‘ Lycoperdon globosum, subterraneum...’ could describe a Rhizopogon. In this situation, the most appropriate solution is to propose the rejection of the name Tuber albidum Fr. in application of Art. 56.1 of the Shenzhen Code (Turland et al., 2018) and a proposal will be made to that effect. The older name Tuber albidum Picco (1788) in addition to being unavailable (Mycobank http://www.mycobank.org/ Biolomics.aspx?Table=Mycobank&Rec=28125&Fields=All) is a poorly defined and variously interpreted Tuber species that can include several whitish species such as T. maculatum Vittad., T. rapiodorum Tul. & C.Tul., T. dryophilum Tul. & C.Tul. and T. puberulum Berk. & Broome, etc. (Halász et al. 2005). The previous neotypification of T. borchii proposed by Mello et al. (2000) cannot be accepted because of formal and substantive reasons: 1) presence of an existent possible lectotype [tab. I, fig.III E-F inVittadini (1831) here designated]; 2) the probable loss of the three collections indicated as ‘neotype’, lacking however of any collecting data. They were sent by Vittadini to the Tulasne brothers (Tulasne & Tulasne 1851: 146) and preserved originally in PC (Cryptogamic Herbarium of Muséum National d’Histoire Naturelle, Paris), and later retained in TO (Mattirolo’s Herbarium); 3) the fact that the proposed neotype was not sequenced. Hence there is a need to propose an epitype using a specimen with well-defined collection data and molecular characterization. Bonuso et al. (2010) hypothesized the presence of two cryptic species (referred to as haplotype 1 and haplotype 2) within Italian populations of T. borchii. The T. borchii epitype designated here belongs to haplotype 1., Published as part of Leonardi, Marco, Iotti, Mirco, Mello, Antonietta, Vizzini, Alfredo, Paz-Conde, Aurelia, Trappe, James & Pacioni, Giovanni, 2021, Typification of the four most investigated and valuable truffles: Tuber aestivum Vittad., T. borchii Vittad., T. magnatum Picco and T. melanosporum Vittad., pp. 149-170 in Cryptogamie, Mycologie 20 (9) on pages 154-160, DOI: 10.5252/cryptogamie-mycologie2021v42a9, http://zenodo.org/record/7826657, {"references":["VITTADINI C. 1831. - Monographia tuberacearum. F. Rusconi, Milan, 88 p.","CERUTI A., FONTANA A. & NOSENZO C. 2003. - Le Specie Europee del Genere Tuber. Museo Regionale di Scienze Naturali, Torino.","BONITO G., SMITH M. E., NOWAK M., HEALY R. A., GUEVARA G., CAZARES E., KINOSHITA A., NOUHRA E. R, DOMINGUEZ L. S., TEDERSOO L., MURAT C., WANG Y., MORENO B. A., PFISTER D. H., NARA K., ZAMBONELLI A., TRAPPE J. M. & VIL- GALYS R. 2013. - Historical biogeography and diversification of truffles in the Tuberaceae and their newly identified southern hemisphere sister lineage. PLoS ONE 8: e 52765. https: // doi. org / 10.1371 / journal. pone.","FRIES E. 1823. - Systema Mycologicum sistens fungorum ordines, genera et species. Officina Berlingiana, Lundae. https: // doi. org / 10.5962 / bhl. title. 5378","CESALPINO A. 1583. - De Plantis Libri XVI. Giorgio Marescotti, Firenze.","MICHELI P. A. 1729. - Nova plantarum genera iuxta Tournefortii methodum disposita quibus plantae 1900 recensentur. Bernardi Paperinii, Firenze.","GLEDITSCH J. G. 1753. - Methodus fungorum exhibens genera, species et varietates cum charactere. Sumtibus Scholae Realis, Berlin.","TURLAND N. J., WIERSEMA J. H., BARRIE F. R., GREUTER W., HAWK- SWORTH D. L., HERENDEEN P. S., KNAPP S., KUSBER W. H., LI D. Z., MARHOLD K., MAY T. W., MCNEILL J., MONRO A. M., PRADO J., PRICE M. J. & SMITH G. F. (eds) 2018. - International Code of Nomenclature for algae, fungi, and plants (Shenzhen Code) adopted by the 19 th International Botanical Congress Shenzhen, China, July 2017. Regnum Vegetabile 159. Glashutten, Koeltz Botanical Books. https: // doi. org / 10.12705 / Code. 2018","PICCO V. 1788. - Melethemata Inauguralia. Ioan. Mich. Briolus: Turin, 283 p.","HALASZ K., BRATEK Z., SZEGO D., RUDNOY S., RACZ I., LASZ- TITY D. & TRAPPE J. M. 2005. - Tests of species concepts of the small, white, European group of Tuber spp. based on morphology and rDNA ITS sequences with special reference to Tuber rapaeodorum. Mycological Progress 4: 281 - 290. https: // doi. org / 10.1007 / s 11557 - 006 - 0132 - 6","MELLO A., VIZZINI A., LONGATO S., ROLLO F., BONFANTE P. & TRAPPE J. M. 2000. - Tuber borchii versus Tuber maculatum: neotype studies and DNA analyses. Mycologia 92: 326 - 331. http: // doi. org / 10.1080 / 00275514.2000.12061163","TULASNE L. - R. & TULASNE C. 1851. - Fungi hypogaei: Histoire et monographie des champignons hypoges. F. Klincksieck, Paris.","BONUSO E., ZAMBONELLI A., BERGEMANN S. E., IOTTI M. & GAR- BELOTTO M. 2010. - Multilocus phylogenetic and coalescent analyses identify two cryptic species in the Italian bianchetto truffle, Tuber borchii Vittad. Conservation Genetics 11: 1453 - 1466. https: // doi. org / 10.1007 / s 10592 - 009 - 9972 - 3"]}

Tuber melanosporum Vittad. (Figs 2D; 3G, H; Appendix 2) Monographia Tuberacearum: 36 (1831). Tuber nigrum Bull., Herbier de la France 8: t.356 (1788). (MB 204568). MYCOBANK NUMBER. — MB 192144. GENBANK. — MZ423176 (nrITS), MZ458420 (nr β- tubulin), MZ458424 (nrEF 1-α). LECTOTYPE OF TUBER MELANOSPORUM VITTAD. — Vittadini 1831: tab. II fig. III; here reprinted in Fig. 3. (here designated; MycoBank Typification number: MBT 10001899): EPITYPE OF TUBER MELANOSPORUM VITTAD. — Italy. Lombardy, Monza, Parco Villa Reale, 1.II.2019, sub Carpinus betulus and Tilia cordata, 45°35’39”N, 9°16’25”E, six ascomata, legit Stefano Seghezzi, det. Giovanni Pacioni (epi-, AQUI[AQUI 10152] here designated; MycoBank Typification number: MBT 10001900). Authentic Vittadini specimens examined: K(M) 254905, 254904 e 254904, TO e UPS (F‒628213). DESCRIPTION Ascomata Hypogeous, globose or irregular, sometimes lobed, rarely more than 10 cm broad, surface of peridium reddish brown then reddish black or brownish black (#330000 very dark red), covered with pyramidal warts 2-5 mm at the base and 0.5-2.5 mm tall, depressed at the apex and with 4-6 sides joined with radial edges. Gleba Firm and compact, from reddish-gray to black-purple or black with violet hues (3B2512-3B3938 dark shade orange to 26211D-2B2017 very dark orange shade); sterile veins thin, much branched, whitish or reddish when exposed to the air. Odor Intense, complex, pleasant, the taste with a bitter aftertaste. Peridium 300-450 Μm thick; exoperidium pseudoparenchymatous, 70-15 Μm thick, the outer layer 40-80 Μm thick, of globose to sub-globose sclerenchymatous pigmented cells, with very thickened walls and the cellular lumen reduced or absent, 12-20 × 7-12 Μm, limited to the superficial part of the warts underlaid by a palisade layer 33-65 Μm thick of elongated cells (16-26 × 5-6.5 Μm) with the major axis perpendicular to the surface just below the exoperidium; endoperidium 240-350 Μm thick, pale with a pseudoparenchymatous mixture of cells 4-11 × 2-7 Μm and ranging from subglobose, cuboid, polygonal, cylindric or irregularly swollen, elongated at one end, pale with collenchymatic walls 2-3 Μm thick. The palisade structure is not always visible, depending on the cut, generally within the outermost layer of sclerenchymatous cells lies a homogeneous pseudoparenchymatous layer of endoperidium. Asci Globose to subglobose, 90-145 × 70-125 Μm, with walls up to 4 Μm, sometimes thicker in the final stages VI of ripening (Zarivi et al. 2014), short stalked or thick, sessile with a crozier at the base; containing 1-6 spores. Ascospores Spiny, ellipsoid, Q 1.28-1.56, 20-56 × 14-36 Μm excluding ornamentation, inversely proportional in size to the number of spores in the ascus, opaque when ripe, intense brown, blackish, with black-dark brownish spines, short, robust and rigid, sometimes curved, 1.5-3.0 (-4) Μm long to 2 Μm wide at the base, usually separated but sometimes connected at the base to form short crests, occurring at a density of 11-13 spines per 100 Μm 2. Gleba Hyphae hyaline, 14-36 × 6-8 Μm, at full maturity brownish. HABITAT. — mainly under broadleaf trees and shrubs, especially thermophilic oaks, but also European hop-hornbeam (Ostrya carpinifolia) and hazelnuts (Corylus avellana), rare under pines or other conifers. Its natural environments are extended in calcareous soils in southern Europe from the Iberian Peninsula to France and Italy, rarer in the Balkans and western Anatolia, late autumn and winter (Le Tacon 2017)., Published as part of Leonardi, Marco, Iotti, Mirco, Mello, Antonietta, Vizzini, Alfredo, Paz-Conde, Aurelia, Trappe, James & Pacioni, Giovanni, 2021, Typification of the four most investigated and valuable truffles: Tuber aestivum Vittad., T. borchii Vittad., T. magnatum Picco and T. melanosporum Vittad., pp. 149-170 in Cryptogamie, Mycologie 20 (9) on pages 161-162, DOI: 10.5252/cryptogamie-mycologie2021v42a9, http://zenodo.org/record/7826657, {"references":["VITTADINI C. 1831. - Monographia tuberacearum. F. Rusconi, Milan, 88 p.","ZARIVI O., CESARE P., RAGNELLI A. M., AIMOLA P., LEONARDI M., BONFIGLI A., COLAFARINA S., POMA A. M., MIRANDA M. & PACIONI G. 2014. - Validation of reference genes for quantitative real-time PCR during Tuber melanosporum developmental stages. Phytochemistry 116: 78 - 86. https: // doi. org / 10.1016 / j. phytochem. 2015.02.024","LE TACON F. 2017. - Les Truffes. AgroParisTech: Nancy."]}

Tuber aestivum Vittad. (Figs 3; 4A; 5A, B; Appendix 1) Monographia Tuberacearum: 38 (1831). MYCOBANK. — MB 218597. GENBANK. — MZ423173 (nrITS), MZ458417 (nr β- tubulin), MZ458421 (nrEF 1-α). LECTOTYPE OF TUBER AESTIVUM VITTAD. — Vittadini 1831: tab. II fig. IV C, D (here designated; MycoBank Typification number: MBT 10001890; Fig. 3). EPITYPE TUBER AESTIVUM VITTAD. — Italy. Lombardy, Monza, Parco Villa Reale, sub Tilia cordata Mill., 8.IX.2019, 45°37’09”N, 9°16’48”E, five ascomata, leg. Stefano Seghezzi, det. Giovanni Pacioni (epi-, AQUI[AQUI 10150], here designated; MycoBank Typification number: MBT 10001891). Authentic Vittadini specimens examined: K(M) 254890, TO and UPS (F-628206). DESCRIPTION Ascomata Hypogeous, globose or irregular, up to 10 cm in diameter or more, rarely with a flat or slightly basal excavation, surface of peridium black to very dark gray (#000000 to 333333) in both mature and immature specimen, covered with pyramidal warts polygonal at the base, with 4-7 sides, 3-13 mm wide × 0.5- 5.0 mm high, generally acute but often depressed or concave, with sharp edges or with longitudinal radial fissures, surface of the warts typically with evident parallel transverse streaks. Gleba Firm and compact, whitish (FAE5D3 light greyish orange) to more or less brown (E59866 soft orange, 6E2C00, very dark orange, brown tone) in relation to season, habitat or state of ripeness, with abundant ramified and thin, sterile, white veins not discolouring when exposed to the air. Odor Weak, similar to stigmas of maize in the whitish gleba of not quite mature specimens, then more and more intense and pleasant like the flavour, which resembles that of nuts, mainly hazelnuts. Peridium 200-480 Μm thick, exoperidium pseudoparenchymatous (100- 180 Μm), almost opaque in section, composed of globosepolygonal cells, 7-20 Μm wide, with reduced cell lumen and deep brown walls up to 4 Μm thick and fused with those of adjacent cells; endoperidium 100-300 Μm thick, pseudoparenchymatous in the interior part of the endoperidium, the cells hyaline and smaller with a confusing pattern in contact with a layer of intertwined thin (2-3 Μm) hyphae that extend into sterile veins of the gleba. Asci Globose to subglobose, 60-120 × 55-85Μm, with walls usually up to 3 Μm thick, rarely to 10 Μm, shortly stalked or sessile, 60-100 × 50-80Μm, with basal crozier, containing 1-6 spores. Ascospores Subglobose to broadly ellipsoid, Q 1.21-1.37, 20-45 × 18-35 Μm excluding ornamentation, inversely proportional in size to the number of spores in the ascus, light brown reticulate, meshes 4-10 Μm wide × 3-7 Μm high, 3-5 across the spore width and often with an incomplete secondary crest inside, and with the top of the ornamentations sometimes hooked at the apex of the reticular knots. Endosporium Often two-layered in KOH. Glebal Hyphae hyaline, 2-8 Μm wide. HABITAT. — Europe from southern Europe to Ireland, Great Britain and Sweden (Gotland Isle) and East to the Caucasus Mountains, North Maghreb (rare), near and north middle Asia to Turkmenistan / Iran borders. Under broadleaved and coniferous trees and shrubs; its ascomata develop a bit throughout the year, commonly from summer to late autumn (Molinier et al. 2016; unpublished data). NOMENCLATURE AND TAXONOMY Although the nomenclature of Tuber aestivum is seriously compromised due to its frequent use in the wrong sense, the name Tuber aestivum proposed by Vittadini (1831) clearly connected to Micheli’s taxon (1729). It is so well characterized morphologically that past mycologists and traders have always preferred it to valid antecedent synonyms such as Tuber blotii Eudes-Desl. (1824), Mémoires de la Société Linnéenne de Calvados 1824: 47 (1824) [MB 102874] (Maire 1930). However, the early legitimate homonym Tuber aestivum (Wulfen) Spreng. 1827 [MB 218548] exists; consequently Tuber aestivum Vittad. is illegitimate (Art. 53.1). The basionym of this earlier homonyn is Lycoperdon aestivum Wulfen, in Jacquin, Collnea bot. 1 (2): 349 (1787) [1786] [MB 162907]. The Wulfen’s description (Appendix 3) clearly identifies the fungus as a Rhizopogon for which there is the sanctioned name Rhizopogon aestivus (Wulfen) Fr. (1823). Tulasne & Tulasne (1851) included Rhizopogon aestivus as a synonym of their Rhizopogon rubescens, whose current name is R. roseolus (Corda) Th.Fr., and as such sometimes subsequently considered to include as synonymous the names Lycoperdon aestivum or Tuber aestivum (Wulfen) Spreng. (Saccardo 1888). Martin (1996) examined all the specimens labeled as R. aestivus present in the main European herbaria, determining them as R. roseolus. Wulfen himself concludes that it is not the species described by Micheli both for size and for the very black external surface with large pyramidal warts, smell and taste. In fact, Lycoperdon aestivum had a smooth peridium, furfuraceous at best, thin and often cracked, whitish then reddish, like a potato, slightly brownish and finally blackish [‘ extus sordide albidum, & ex albido obsolete subrubescens, instar pomorum terrae (Solanum tuberosum L.), dein dilute fuscens, nigrans denique; glabrum… cuticula tenerrima, hinc inde disrupta, & exstans, squamules exhiberet furfuraceus …], spongy gleba cellulose-porous at first white then almost ashy, brownish, blackish (Caro intus spongiosa, subcoriacea, prima alba, tum obsolete, & triste ex albido subcinerascens, fuscescens, subnigricans… celluloso-porosa), without marked smell and without taste (‘ Odor recentium nullus, aut certe debilissimus & subnauseosus. Sapor nullus, nisi imaginarius’). No specimens of this species have been found in the herbaria of Wulfen (W), Fries (UPS) and KPJ Sprengel (VH/de), for this reason we selected as neotype of Lycoperdon aestivum a specimen of Rhizopogon roseolus collected in Carinthia. (Austria), place of origin of Lycoperdon aestivum. A proposal will be written to conserve the name Tuber aestivum Vittad. and reject the legitimate early homonym Tuber aestivum (Wulfen) Spreng. and the early synonym Tuber blotii Eudes-Desl. The proposal to retain the name Rhizopogon roseolus (Corda) Th. Fr. against Rhizopogon aestivus (Wulfen) Fr. should also be considered. NEOTYPE OF LYCOPERDON AESTIVUM WULFEN. — Austria. Carinthia, St. Mergereten, Gotschich, leg. Irmegard Krisai Greilhuber, 9.IX.1998, sub nomine Rhizopogon roseolus (neo-, WU[WU 25744], here designated; MycoBank Typification number: MBT 10001892)., Published as part of Leonardi, Marco, Iotti, Mirco, Mello, Antonietta, Vizzini, Alfredo, Paz-Conde, Aurelia, Trappe, James & Pacioni, Giovanni, 2021, Typification of the four most investigated and valuable truffles: Tuber aestivum Vittad., T. borchii Vittad., T. magnatum Picco and T. melanosporum Vittad., pp. 149-170 in Cryptogamie, Mycologie 20 (9) on pages 151-154, DOI: 10.5252/cryptogamie-mycologie2021v42a9, http://zenodo.org/record/7826657, {"references":["VITTADINI C. 1831. - Monographia tuberacearum. F. Rusconi, Milan, 88 p.","PAOLOCCI F., RUBINI A., RICCIONI C., TOPINI F. & ARCIONI S. 2004. - Tuber aestivum and Tuber uncinatum: two morphotypes or two species? FEMS Microbiology Letters 235: 109 - 115. https: // doi. org / 10.1111 / j. 1574 - 6968.2004. tb 09574. x","ROUX C., SEJALON DELMAS N., MARTINS M., PARGUEY LEDUC A., DARGENT R. & BECARD G. 1999. - Phylogenetic relationships between European and Chinese truffles based on parsimony and distance analysis of ITS sequences. FEMS Microbiology Letters 180: 147 - 155. https: // doi. org / 10.1111 / j. 1574 - 6968.1999. tb 08789. x","BONUSO E., ZAMBONELLI A., BERGEMANN S. E., IOTTI M. & GAR- BELOTTO M. 2010. - Multilocus phylogenetic and coalescent analyses identify two cryptic species in the Italian bianchetto truffle, Tuber borchii Vittad. Conservation Genetics 11: 1453 - 1466. https: // doi. org / 10.1007 / s 10592 - 009 - 9972 - 3","MELLO A., GARNERO L., MEOTTO F. & BONFANTE P. 1998. - Specific primers for rapid typing of Tuber borchii mycorrhizal roots. Acta Horticultura e 457: 229 - 234. https: // doi. org / 10.17660 / ActaHortic. 1998.457.28","MELLO A., CANTISANI A., VIZZINI A. & BONFANTE P. 2002. - Genetic variability of Tuber uncinatum and its relatedness to other black truffles. Environmental Microbiology 4: 584 - 594. https: // doi. org / 10.1046 / j. 1462 - 2920.2002.00343. x","MELLO A., MURAT C., VIZZINI, A., GAVAZZA, V., & BONFANTE, P. 2005. - Tuber magnatum Pico, a species of limited geographical distribution: its genetic diversity inside and outside a truffle ground. Environmental Microbiology 7: 55 - 65. https: // doi. org / 10.1111 / j. 1462 - 2920.2004.00678. x","RICCIONI C., BELFIORI B., RUBINI A., PASSERI V., ARCIONI S. & PAOLOCCI F. 2008. - Tuber melanosporum outcrosses: analysis of the genetic diversity within and among its natural populations under this new scenario. New Phytologist 180: 466 - 478. https: // doi. org / 10.1111 / j. 1469 - 8137.2008.02560. x","RUBINI A., PAOLOCCI F., GRANETTI B. & ARCIONI S. 1998. - Single step molecular characterization of morphologically similar black truffle species. FEMS Microbiology Letters 164: 7 - 12. https: // doi. org / 10.1111 / j. 1574 - 6968.1998. tb 13060. x","STOBBE U., BUNTGEN U., SPROLL L., TEGEL W., EGLI S. & FINKA S. 2012. - Spatial distribution and ecological variation of rediscovered German truffle habitats. Fungal Ecology 5: 591 - 599. https: // doi. org / 10.1016 / j. funeco. 2012.02.001","BENUCCI G. M. N., CSORBAI A. G., BACIARELLI FALINI L., MAROZZI G., SURIANO E., SITTA N. & DONNINI D. 2016. - Taxonomy, Biology and Ecology of Tuber macrosporum Vittad. and Tuber mesentericum Vittad., in ZAMBONELLI A., IOTTI M., MURAT C. (eds) True Truffle (Tuber spp.) in the World. Soil Biology, vol 47. Springer, Cham. https: // doi. org / 10.1007 / 978 - 3 - 319 - 31436 - 5 _ 5","BONITO G. M., GRYGANSKYI A. I. P., TRAPPE J. M. & VILGALYS R. 2010. - A global meta-analysis of Tuber ITS rDNA sequences: species diversity, host associations and long-distance dispersal. Molecular Ecology 19: 4994 - 5008 https: // doi. org / 10.1111 / j. 1365 - 294 X. 2010.04855. x","MURAT C., VIZZINI A., BONFANTE P. & MELLO A. 2005. - Morphological and molecular typing of the below-ground fungal community in a natural Tuber magnatum truffle-ground. FEMS Microbiology Letters 245: 307 - 313. https: // doi. org / 10.1016 / j. femsle. 2005.03.019","TEDERSOO L., HANSEN K., PERRY B. A. & KJOLLER R. 2006. - Molecular and morphological diversity of pezizalean ectomycorrhiza. New Phytologist 170: 581 - 596. https: // doi. org / 10.1111 / j. 1469 - 8137.2006.01678. x","BONITO G., SMITH M. E., NOWAK M., HEALY R. A., GUEVARA G., CAZARES E., KINOSHITA A., NOUHRA E. R, DOMINGUEZ L. S., TEDERSOO L., MURAT C., WANG Y., MORENO B. A., PFISTER D. H., NARA K., ZAMBONELLI A., TRAPPE J. M. & VIL- GALYS R. 2013. - Historical biogeography and diversification of truffles in the Tuberaceae and their newly identified southern hemisphere sister lineage. PLoS ONE 8: e 52765. https: // doi. org / 10.1371 / journal. pone.","HALASZ K., BRATEK Z., SZEGO D., RUDNOY S., RACZ I., LASZ- TITY D. & TRAPPE J. M. 2005. - Tests of species concepts of the small, white, European group of Tuber spp. based on morphology and rDNA ITS sequences with special reference to Tuber rapaeodorum. Mycological Progress 4: 281 - 290. https: // doi. org / 10.1007 / s 11557 - 006 - 0132 - 6","BOUTAHIR S., IOTTI M., PIATTONI F. & ZAMBONELLI A. 2013. - Morphological and molecular characterization of Tuber oligospermum mycorrhizas. African Journal of Agricultural Research 8: 4081 - 4087. https: // doi. org / 10.5897 / AJAR 2013.7354","BROCK P. M., DORING H. & BIDARTONDO M. I. 2009. - How to know unknown fungi: the role of a herbarium. New Phytologis t 181: 719 - 724. https: // doi. org / 10.1111 / j. 1469 - 8137.2008.02703. x","IOTTI M., AMICUCCI A., STOCCHI V. & ZAMBONELLI A. 2002. - Morphological and molecular characterization of mycelia of some Tuber species in pure culture. New Phytologist 155: 499 - 505. https: // doi. org / 10.1046 / j. 1469 - 8137.2002.00486. x","IOTTI M., AMICUCCI A., BONITO G., BONUSO E., STOCCHI V. & ZAMBONELLI A. 2007. - Selection of a set of specific primers for the identification of Tuber rufum: a truffle species with high genetic variability. FEMS Microbiology Letters 277: 223 - 231. https: // doi. org / 10.1111 / j. 1574 - 6968.2007.00963. x","WANG Y., TAN Z. M., ZHANG D. C., MURAT C., JEANDROZ S. & LE TACON F. 2006 a. - Phylogenetic relationships between Tuber pseudoexcavatum, a Chinese truffle, and other Tuber species based on parsimony and distance analysis of four different gene sequences. FEMS Microbiology Letters 259: 269 - 281. https: // doi. org / 10.1111 / j. 1574 - 6968.2006.00283. x","WANG Y., TAN Z. M., ZHANG D. C., MURAT C., JEANDROZ S. & LE TACON F. 2006 b. - Phylogenetic and populational study of the Tuber indicum complex. Mycological Research 110: 1034 - 1045. https: // doi. org / 10.1016 / j. mycres. 2006.06.013","MOLINIER V., PETER M., STOBBE U. & EGLI S. 2016. - The Burgundy truffle (Tuber aestivum syn. uncinatum): a truffle species with a wide range over Europe. in ZAMBONELLI A., IOTTI M. & MURAT C. (eds) True Truffles (Tuber spp.) in the World. Soil Biology 47. Springer, Cham: Switzerland: 33 - 47. https: // doi. org / 10.1007 / 978 - 3 - 319 - 31436 - 5 _ 3","MAIRE R. 1930. - Sur le Tuber blotii E. Deesl. Bulletin Societe Mycologique de France 46: 149 - 150.","TULASNE L. - R. & TULASNE C. 1851. - Fungi hypogaei: Histoire et monographie des champignons hypoges. F. Klincksieck, Paris.","SACCARDO P. A. 1888. - Sylloge Fungorum omnium hucusque cognitorum. Vol. VII (I). Typ. Seminarii, Padua. https: // doi. org / 10.5962 / bhl. title. 5371","MARTIN M. P. 1996. - The genus Rhizopogon in Europe. Societat Catalana de Micologia: Barcelona, 173 p."]}

Tuber magnatum Picco (Figs 4C; 5E, F) Melethemata inauguralia: 79 (1788). Tuber griseum Borch ex Pers., Synopsis Methodica Fungorum: 127 (1801); sanct. Fr., Systema Mycologicum 2 (2): 292 (1823). (MB 179335). MYCOBANK NUMBER. — MB 184470. GENBANK. — MZ423175 (nrITS), MZ458419 (nr β- tubulin), MZ458423 (nrEF 1-α). NEOTYPE OF TUBER MAGNATUM PICCO. — Italy. Piedmont, Montechiaro D’Asti, Loc. Seria, 45°00’30”N, 8°06’45”E, lowland grove with Quercus robur L. and Populus tremula L., under Q. robur, 29.IX.2019, four ascomata, leg. Pino Panzini, det. Alfredo Vizzini (neo-, TO[HG3458] here designated; MycoBank Typification number: MBT 10001896). DESCRIPTION Ascomata Hypogeous, globose to lobate or flat, up to 10-15 cm broad and more rarely with a detachable basal mycelial cluster.Peridium whitish (FFFFF0 ivory), yellowish, pale ocher (FFD700 gold, DAA520 goldenrod) more or less greyish (DCDCDC gainsboro, D3D3D3 light gray) sometimes with greenish or reddish hues; smooth surface finely grainy under a lens. Gleba Whitish (FFFFF0 ivory), yellowish or pale ocher, brownish (8B4513 saddle brown, A0522D sienna, F2F2F2) with greyish tones, often with reddish purple spots; veins thin, whitish, branched, anastomosing, sometimes forming white gangliform thickenings. Odor Pleasant, penetrating, of garlic or strong cheese; taste pleasant. Peridium 200-500 Μm thick, entirely pseudoparenchymatous basically of globular cells; exoperidium 80-110 Μm in the outer layer with globose cells having walls up to 4 Μm thick and slightly yellow; peridial surface with infrequent stocky, cylindrical-clavate dermatocystidia 14-20 × 8-10Μm; endoperidium 120-380 Μm thick, of thin-walled globose and polygonal cells 6.0-36 × 6-23 Μm with rare cylindrical cells 6-9 × 14-20 Μm. Asci Globose, short-stalked or sessile, 60-90 × 40-70Μm (Fig. 2F), with walls up to 4 Μm thick and a basal crozier, containing 1-4 spores. Ascospores Alveolate, globose to broadly ellipsoid, Q 1.05-1.33, 20-50 × 15-42 Μm excluding ornamentation, inversely proportional in size to the number of spores in the ascus, yellowish, pale ocher or sometimes light yellow-brown; reticulum with 1-3 regular or coarsely irregular, 3-6 sided alveoli 10-22 Μm wide and 4-5 (-8) Μm high, sometimes with ridges within the alveolae. Endosporium Often two-layered in KOH. Glebal Hyphae hyaline, 4-8 Μm broad, in the external veins with cylindrical or clavate-elongated cells mixed with subglobose cells 5-42 × 4-16 Μm. HABITAT. — moist (mesophilic) woods mainly with broad-leaved trees and conifers such as Abies alba Mill., on sandy-silty soils in southern Europe (Italy and a few neighbouring French and Swiss areas, the Balkan-Pannonia region, Northern Anatolia), from late summer to early winter (Belfiori et al. 2020; unpublished data). It has also been reported in Thailand, but vouchers have not been made available (Suwannarach et al., 2017). NOMENCLATURE AND TAXONOMY The name most frequently used for this species is Tuber magnatum Picco (1788) for which a neotype is herein designated. However, a competing name exists, namely Tuber griseum Borch ex Pers., validated by Persoon (1801) and sanctioned by Fries (1823), for which a lectotype and epitype are also herein designated. The epitype is represented by a T. magnatum with a greyish peridium. Borch’s or Picco’s authentic specimens were not available for either T. griseum or T. magnatum. LECTOTYPE OF TUBER GRISEUM. — Truffe grise De Borch, Lettres sur les Truffes du Piémont, 1 st figure (1780); here reprinted in Appendix 4 (here designated; MycoBank Typification number: MBT 10001897). EPITYPE OF TUBER GRISEUM. — Italy. Piedmont, Monte Magno, 44°59’02”N, 8°19’35”E, under Quercus robur, 07.XII.1998, one fragment, leg. Virgilio Gavazza, det. Alfredo Vizzini (epi-, TO [HG 3557] here designated; MycoBank Typification number: MBT 10001898). A proposal will be written to conserve the name T. magnatum and reject the sanctioned name T. griseum., Published as part of Leonardi, Marco, Iotti, Mirco, Mello, Antonietta, Vizzini, Alfredo, Paz-Conde, Aurelia, Trappe, James & Pacioni, Giovanni, 2021, Typification of the four most investigated and valuable truffles: Tuber aestivum Vittad., T. borchii Vittad., T. magnatum Picco and T. melanosporum Vittad., pp. 149-170 in Cryptogamie, Mycologie 20 (9) on pages 160-161, DOI: 10.5252/cryptogamie-mycologie2021v42a9, http://zenodo.org/record/7826657, {"references":["BELFIORI B., D'ANGELO V., RICCIONI C., LEONARDI M., PAOLOCCI F., PACIONI G. & RUBINI A. 2020. - Genetic structure and phylogeography of Tuber magnatum populations. Diversity 12: 44. https: // doi. org / 10.3390 / d 12020044","PICCO V. 1788. - Melethemata Inauguralia. Ioan. Mich. Briolus: Turin, 283 p.","PERSOON C. H. 1801. - Synopsis methodica Fungorum. H. Dieterich: Gottingen.","FRIES E. 1823. - Systema Mycologicum sistens fungorum ordines, genera et species. Officina Berlingiana, Lundae. https: // doi. org / 10.5962 / bhl. title. 5378"]}

Tuber mixtecorum is described as a new species from Mexico based on morphological and molecular evidence. This species belongs to the Maculatum clade and is characterized by its subtomentose to glabrous, whitish to pale brown peridium, dark brown gleba, ellipsoid reticulate-alveolate ascospores (15–33.9 × 11.3–23.3 µm), and the putative mycorrhizal association with Quercus castanea in Quercus-Pinus dry mixed forests.

Yuan, Tian-Jun, Raspé, Olivier, Li, Yong-Jun, Wang, Li, Su, Kai-Mei, Wang, Yun, Su, Hong-Yan, Xiong, Hai-Kuang, Li, Shu-Hong (2021): Choiromyces cerebriformis (Tuberaceae), a new hypogeous species from Yunnan, China. Phytotaxa 482 (3): 251-260, DOI: 10.11646/phytotaxa.482.3.3, URL: http://dx.doi.org/10.11646/phytotaxa.482.3.3

Choiromyces cerebriformis T. J. Yuan, S. H. Li & Y. Wang, sp. nov. Fig. 2a���f MycoBank: MB837904. Diagnosis: Choiromyces cerebriformis differs from similar species in having ascomata up to 10 cm in diameter, with larger and deeper lobes and light orange-brown gleba. Holotype: CHINA, Yunnan Province, Diqing Autonomous Prefecture, Shangri-La County, Three dam township, Haba village, 27��21���6.3647���N, 100��06��� 34.6895���E, elevation 4005 m, under Abies delavayi Franch. and Rhododendron simsii Planch. in mixed evergreen hill forest, with soil types mainly red and brown soils, 24 Sep. 2020, collected by X. H. Wang (KUN-HKAS 107566, holotype; YAAS 8890, isotype). Etymology: cerebriformis refers to the aspect of ascomata which looks like a brain. Ascomata hypogeous, up to 10 cm in diam, knobby, with deep lobes; surface glabrous, whitish when young, then pale yellow to reddish-brown with dark spots at maturity (Fig. 2b, c). Peridium 210���280 ��m thick, plectenchymatous, with two layers, the outer layer 60���80 ��m thick, reddish-brown, the inner layer 150���200 ��m thick, hyaline with larger cells (25���40 ��m diam, wall 0.5���2.0 ��m thick; Fig. 2d). Gleba (Fig. 2d, e) solid, labyrinthoid, light orange-brown, marbled with greyish white veins. Odor strong, pleasant. Asci hyaline, clavate to sac-like, 8-spored, (75���)75���85.5��� 96(���96) �� (42���)43���54���65(���66) ��m (N = 20). Ascospores (N = 60) (17.5���)17.8���21.3���24.8(���25.0) �� (17.5���)17.6��� 21.0���24.4(���24.5) ��m, Q = (1.00���)0.97���1.02���1.06(���1.12), globose to subglobose, pale yellow-brown, ornamented with straight or slightly curved stick-like sparse spines, 2���3 ��m tall, with truncate or rounded tips (Fig. 2e���g). Habitat, phenology, and distribution: Growing under Abies delavayi Franch., Rhododendron simsii Planch. and Pinus yunnanensis in mixed evergreen hill forest, in late autumn. So far found in Diqing and Yongren counties, Yunnan, China. Other material examined: CHINA, Yunnan Province, Chuxiong Autonomous Prefecture, Yongren County, 26.0810��N 101.6632��E, elevation 1,834 m, under Pinus yunnanensis in mixed evergreen hill forest, with soil types mainly red and brown soils, 01 Nov. 2019, collected by H. K. Xiong. (YAAS TJ16-1 and YAAS TJ16-2). Remarks: Choiromyces cerebriformis is characterized by hypogeous, glabrous, pale yellow or white, irregularly knobby, much lobed ascomata, and globose spores ornamented with conspicuous, straight or slightly curved spines with truncate or rounded tips when observed with SEM. This species resembles white Tuber truffles with white ascomata and sinuous veins in gleba. However, it is different from those Tuber species and from other Choiromyces species by its globose spores ornamented with conspicuous spines and small pits. C. cerebriformis can be further distinguished from C. meandriformis, which has paler gleba, and spores with much taller spines (Csorbain�� et al. 2009; Moreno et al. 2012). C. echinulatus Trappe & Marasas (Ferdman et al. 2005), differs from C. cerebriformis by spores with straight, obtuse spines �� 2 ��m high (spines are 2���3 ��m high in C. cerebriformis). Moreover, C. echinulatus has tissues with greatly inflated cells, which is characteristic of all desert truffles. C. cerebriformis differs from C. alveolatus (Liu et al. 1990) in having larger asci (75���96 �� 42���66) ��m, mostly with 8 spores, compared to 40���85 �� 30���55 ��m asci, with (2���) 4���6 spores in the latter species. C. magnusii (Moreno et al. 2012) spores ornamentation is verrucose-reticulated with a minutely-meshed reticulum, which is clearly different from C. cerebriformis. C. helanshanensis (Chen et al. 2016) differs from C. cerebriformis by its coarsely knobby ascomata 2���8 cm diam (ascomata up to 10 cm and with larger, deeper lobes in C. cerebriformis) and ascosporal ornamentation as conspicuous dense spines (as short stick-like sparse spines in C. cerebriformis)., Published as part of Yuan, Tian-Jun, Rasp��, Olivier, Li, Yong-Jun, Wang, Li, Su, Kai-Mei, Wang, Yun, Su, Hong-Yan, Xiong, Hai-Kuang & Li, Shu-Hong, 2021, Choiromyces cerebriformis (Tuberaceae), a new hypogeous species from Yunnan, China, pp. 251-260 in Phytotaxa 482 (3) on page 256, DOI: 10.11646/phytotaxa.482.3.3, http://zenodo.org/record/5419405, {"references":["Csorbaine, A. G., Illyes, Z., Dimeny, J., Merenyi, Z. & Bratek, Z. (2009) Choiromyces meandriformis and Mattirolomyces terfezioides: peculiar truffles with new perspectives. Micologia Italiana 38 (1): 21 - 28.","Moreno, G., Alvarado, P. & Manjon, J. L. (2012) Phylogenetic affiliation of (Choiromyces magnusii and C. venosus Tuberaceae ascomycota) from Spain. Mycological Progress 11 (2): 463 - 471. https: // doi. org / 10.1007 / s 11557 - 011 - 0762 - 1","Ferdman, Y., Aviram, S., Rothbejerano, N., Trappe, J. M. & Kaganzur, V. (2005) Phylogenetic studies of Terfezia pfeilii and Choiromyces echinulatus (Pezizales) support new genera for southern African truffles: Kalaharituber and Eremiomyces. Mycological Research 109 (2): 237 - 245. https: // doi. org / 10.1017 / S 0953756204001789","Liu, B., Tao, K. & Chang, M. C. (1990) New species and new records of hypogeous fungi from china III. Mycosystema 9 (1): 25 - 30.","Chen, J., Sun, L. H., Su, Y., Zhao, W. Q. & Liu, P. G. (2016) Choiromyces helanshanensis sp. nov., a new species of subterranean truffle from China. Mycoscience 57: 279 - 286. https: // doi. org / 10.1016 / j. myc. 2016.04.001"]}

Pecan trees are known to form ectomycorrhizae with a wide range of fungi, including commercial truffles that can be cultivated in agroforestry systems. Here, we investigated whether pecans can be successfully mycorrhized by Tuber spp. and whether the mycorrhization affects growth parameters and nutrient content of two productive pecan cultivars under southern subtropical conditions. We also sought to define procedures of mycorrhization adapted to the subtropics. The seedlings of two pecan cultivars (Barton and Importada) were inoculated with both Tuber aestivum and T. brumale and grown in a nursery for 12 months. The levels of ectomycorrhizal colonization and the morphological and chemical parameters of the seedlings were analyzed. The proposed mycorrhization protocol was successful, as all truffle-inoculated seedlings exhibited a high percentage of mycorrhized fine roots, on average 70% with T. aestivum and 88% with T. brumale. In both pecan cultivars, mycorrhization by the Tuber species reached levels at which all seedlings would qualify as suitable for commercialization. Notably, the cultivar Barton inoculated with T. brumale exhibited higher total biomass accumulation and growth of roots and shoots. Shoot and root parameters did not show nutrient deprivation. The nutrient status after one year of cultivation in a nursery also supported that the applied protocol was adequate to maintain healthy mycorrhized seedlings with suitable nutritional status for out-planting of seedlings in a commercial plantation. To our knowledge, this is the first study on the mycorrhization of pecan seedlings with European truffles under southern subtropical conditions. Our results suggested that the tested cultivars have a high potential for truffle production considering the necessary substrate, site, and management adaptations for selected commercial truffle species and the requirements for pecan orchards.