Some species in the genus Peziza develop in extreme cold environments, at the margins of melting snow. In this work, we characterized the genetic structure of nivicolous Peziza populations in Campo Pericoli, a glacial basin on the Gran Sasso massif, the highest peak of the Apennines (central Italy). A hundred ascomata collected in 2020-2021 were analysed using multigene sequencing (ITS, LSU, beta-tubulin, RPB2). Four topographical variables (elevation, slope, aspect, topographic wetness index) were recorded for each fruiting point and correlated with genetic diversity. Two main clades, attributable to P. heimii and P. nivalis, were inferred through clustering and phylogenetic reconstruction. No significant differences in topographical variables were found between clades, but kernel density revealed distinct spatial distributions. However, considering the global distribution of these species, P. nivalis appears to exhibit broader ecological adaptability than P. heimii. Climate warming could affect competition between these species, potentially favoring P. nivalis over P. heimii.

Truffles in the genus Tuber are hypogeous fungi living in symbiosis with the roots of many trees and shrubs. Within the genus Tuber, the Melanosporum clade includes some commercial species of great economic interest that are cultivated worldwide. At least four species of the clade have been described in Europe: Tuber melanosporum, T. brumale, T. crytpobrumale and T. petrophilum. Recent phylogenetic studies have also shown the existence of cryptic species in this clade. While analysing ascomata of T. brumale from Bulgaria, we found distinctive macro and micro-morphological features in some specimens. Phylogenetic analysis conducted on the internal transcribed spacer (ITS), the large subunit (LSU) rDNA and the protein kinase C like (PKC) sequences showed that these specimens form a monophyletic, well-supported taxon within the Melanosporum clade, closely related to T. brumale complex and T. petrophilum. Morphological and molecular analyses supported the recent proposal of the new species T. thracicum.

Typification plays a crucial role in mycological taxonomy as it ensures nomenclatural stability and anchors species names to clearly defined reference material. This is particularly critical when original material is incomplete, poorly preserved, or lacks molecular data, since reliable reference points are essential for accurate species delimitation and consistent phylogenetic interpretation. Here, typifications are proposed for two nivicolous Peziza species. For Galactinia nivalis R. Heim & L. Rémy, a lectotype is designated from the original illustrations, and the holotype of Peziza fortoulii Donadini, a later synonym, serves as a supporting epitype. For Peziza heimii Pfister, the holotype in the MNHN herbarium (PC) is unsuitable for genetic characterization, and thus a supporting epitype is designated. In both cases, sequences from the epitypes are already available. These typifications strengthen nomenclatural stability and provide a robust foundation for future taxonomic, morphological, and molecular studies on nivicolous taxa within Peziza.

The aim of this study was to investigate the species composition of insects inhabiting the fruit- ing bodies of Burgundy truffle Tuber aestivum in Polish forests. Field work was carried out in 2016-2017 at four natural truffle sites in southern Poland. The fruiting bodies were searched with the help of a trained truffle dog or collected randomly. Adult insects were captured direct- ly from the fruiting bodies using an exhaustor. Fruit bodies that were possibly inhabited by insect larvae were collected, brought to the laboratory and placed in special containers to devel- op them. Insects were reared in a biological chamber with constant parameters. Adult insects collected from the field and reared in the laboratory were identified using morphological meth- ods. During the study, 584 Burgundy truffle sporocarps were found and examined (364 in 2016 and 220 in 2017). In 2016, 90 truffles were infested by insects (about 24.7%), while in 2017, 93 fruiting bodies were damaged, representing 42.2% of the harvest. A total of 330 insect speci- mens belonging to 21 species were identified. Some of them can cause significant losses in truf- fle plantations. Others can be considered as indicators of the presence of truffles in the forest environment. During the study, 236 specimens of Diptera were obtained from breeding, and a total of 86 imagines of Coleoptera and 8 imagines of Hymenoptera were collected directly from the fruiting bodies. Ant larvae were also observed in the truffles but were not collected for breeding. The assemblage of insects inhabiting truffles was identified, with two visibly dominant species: Cheilosia soror fly from the Syrphidae family and truffle beetle Colenis immunda from Leiodidae family. Among other common insect species, the flies Suillia affinis and Suillia pallida and the beetles Leiodes cinnamomea and Leiodes oblonga were observed. For the first time, a case of colonization of truffle fruiting bodies by ants Myrmica rubra has been described. The pio- neering research carried out in Polish forests has helped to significantly increase knowledge about truffle-inhabiting insects, their occurrence and biology, which is necessary to reduce the damage they cause to truffle production.

Tuber wenchuanense ascomata (Ascomycota, Pezizales), a species originally described from Sichuan (China), were found in the Tatra Mountains in southern Poland. The purpose of this work was to (i) report and assess the first case of the holarctic natural distribution of a Tuber species, (ii) amend the original description of the species, (iii) summarize data on its host plants and (iv) describe its ectomycorrhiza. Specimens of Tuber wenchuanense from the Tatra Mountains were studied morphologically and molecularly. The ectomycorrhiza of this truffle with Picea abies was described for the first time. The distribution of T. wenchuanense, which is reconstructed based on sequences deposited in the publicly available nucleotide sequence databases, makes it the first holarctic Tuber species and the one with the northernmost habitat. In fact, its habitat is confined mainly to mountain coniferous forests and alpine and arctic tundra; although, according to known observations, the fruiting bodies of T. wenchuanense can be produced only under conifers. Based on the sequences of the internal transcribed spacer, this species appears to have low genetic variability over the entire distribution range. The phylogenetic tree showed that some of the unidentified phylotypes from the Rufum clade found by other researchers belong to T. wenchuanense. The ecological implications of these findings are discussed.

Truffles in the genus Tuber host and feed a wide community of organisms including insects, bacteria, and fungi. Among microorganisms, truffle-inhabiting fungi (TIF) have been poorly investigated and only a limited number of yeast and filamentous species have been identified. In this work we isolated and barcoded culturable TIF from 16 ascomata of T. melanosporum, T. aestivum and T. borchii of different provenances. Eighteen strains were identified a species level, four at genus level and one at order level. Penicillium was the most represented genus with five species and the only TIF genus isolated from T. melanosporum ascomata. Six TIF had colonies with a yeast morphology, including two species of Candida. Five species (Leohumicola sp., Knufia tsunedae, Hypocreales sp., Sagenomella cf. verticillata, Kondoa sp.) had rare ITS sequences compared to those deposited in GenBank. Except Peniophora cinerea, all TIF identified in this work have never been reported in Tuber ascomata until now. In-depth investigations on TIF and their host specificity are needed to evaluate the effects of these fungi on the truffle life cycle and the quality of ascomata.

Tuber borchii is an edible ectomycorrhizal mushroom of considerable economic value. Its cultivation has become popular in recent years, but there are few studies on the factors affecting its productivity. In this work, the ascoma production and the ectomycorrhizal (ECM) community of a T. borchii plantation, established in an intensive farming area where this truffle is not naturally present, were studied. Tuber borchii production drastically declined from 2016 to 2021, and ascomata of other Tuber species (T. maculatum and T. rufum) were found from 2017. Molecular characterization of ectomycorrhizae carried out in 2016 identified 21 ECM fungal species, of which T. maculatum (22%) and Tomentella coerulea (19%) were the most abundant. Tuber borchii ectomycorrizae (16%) were almost entirely confined to the fruiting points. The diversity and structure of the ECM community on Pinus pinea were significantly different from those observed on hardwood trees. The obtained results suggest that T. maculatum (a native of the study site) tends to replace T. borchii through a mechanism of competitive exclusion. Although T. borchii cultivation is possible in suboptimal environments, particular care should be taken to limit competition with ECM fungi more suitable for local conditions.

Many countries regulate the names of commercially important truffles, such as France (Accord Interprofessionnel Truffes Fraîches. 1996, https://www.legifrance.gouv.fr/jorf/id/JORFTEXT000000563056, https://www.interfel.com/wp-content/uploads/2017/10/accord-interprofessionnel-truffe.pdf), Italy (Legge 16 December 1985 n.752, https://www.normattiva.it/uri-res/N2Ls?urn:nir:stato:legge:1985;752), Spain (Decreto 1688/1972, https://boe.es/diario_boe/txt.php?id=BOE-A-1972-986; Real Decreto 30/2009, https://www.boe.es/diario_boe/txt.php?id=BOE-A-2009-1110), and New Zealand (Fresh Truffles for Human Consumption, HIS.FP.TRUFFLE 2 October 2018, https://www.mpi.govt.nz/dmsdocument/16813-Fresh-Truffles-for-Consumption-Import-Health-Standard). These regulations all concern the names of the most important species of truffles that have long been referred to by the following scientific and common names: Tuber aestivum Vittad. as summer truffle or truffe noire de Bourgogne, Tuber magnatum Picco as Italian white truffle or alba white truffle, and Tuber melanosporum Vittad. as black truffle or Truffe du Périgord. In reviewing the scientific names of these fungi, we have discovered several competing names. To maintain stability in communication about these commercially important fungi, we propose to conserve the commonly used scientific names against competing earlier or sanctioned names. The starting point for recognizing priority of publication of names has changed over the decades and is now determined by the latest International Code of Nomenclature for algae, fungi, and plants (ICN; Turland & al. in Regnum Veg. 159. 2018). Although the starting date for names of all fungi is now 1 May 1753 with the publication of Linnaeus's Species Plantarum, from the Brussels Rules (Briquet, Règles Int. Nomencl. Bot., ed. 2. 1912) until the Sydney Code (Voss & al. in Regnum Veg. 111. 1983), exceptions to this starting point were made for names in certain fungal groups adopted by Christiaan Persoon in 1801 and Elias Fries between 1821 and 1832. Although the starting date has been 1753 since 1983, names published in these works by Persoon and Fries (see Art. F.3.1 of the ICN) have been sanctioned, giving them precedence over earlier names that were not sanctioned (Art. F.3.2). Although some names were published in the genus Tuber prior to Fries, many were sanctioned by Fries (Syst. Mycol. 2: 289–293. 1823) and thus these names have priority over the earlier ones. Several names currently applied to Tuber species in Europe were published by Vittadini (Monogr. Tuberac. 1831), who did not always accept the names used by Fries (l.c.). Rather Vittadini (l.c.) applied names of his own or others to the same Friesian species. Yet the names in Tuber used by Vittadini (l.c.) are the ones commonly used over the past 190 years, whereas those published earlier and/or sanctioned by Fries (l.c.) have generally been ignored. Thus, we formally propose herein the conservation of the following three names: T. aestivum Vittad., T. magnatum Picco, and T. melanosporum Vittad. and the rejection of their competing names. Their typification as well as their taxonomic and nomenclatural situation were recently clarified by Leonardi & al. (in Cryptog. Mycol. 42: 149–170. 2021) with T. blotii typified herein. (2867) Tuber aestivum Vittad., Monogr. Tuberac.: 38. 1831, nom. cons. prop. Typus: [icon in] Vittadini, Monogr. Tuberac.: t. 2, fig. 4. 1831. MBT 10001890. Epitypus (vide Leonardi & al. in Cryptog. Mycol. 42: 151. 2021): Italy, Lombardy, Monza, Parco Villa Reale, sub Tilia cordata Mill., 8 Sep 2019, Seghezzi (AQUI No. 10150). MBT 10001891. (H) Tuber aestivum (Wulfen) Spreng., Syst. Veg. 4(1): 416. 1827 (ante 7 Jan) (Lycoperdon aestivum Wulfen in Jacquin, Collectanea 1: 349. Jan–Sep 1787), nom. rej. prop. Neotypus (vide Leonardi & al. in Cryptog. Mycol. 42: 154. 2021): Austria. Carinthia, St. Margareten, Gotschuchen, 8 Sep 1998, Krisai-Greilhuber, sub nomine Rhizopogon roseolus (WU-MYC No. 0025744). MBT 10001892. (=) Tuber blotii Eudes-Desl. in Mém. Soc. Linn. Calvados 1824: 47. 1824, nom. rej. prop. Lectotypus (hic designatus): [icon] “Tuber Blotii” in Mém. Soc. Linn. Calvados 1824: fig. 1–3. 1824. MBT 10005854. Tuber aestivum Vittad. (Monogr. Tuberac.: 38. 1831) has been used legally and commercially for this species of prized, edible truffle, commonly known as the summer truffle, found throughout Europe. One earlier name represents the same species, and an earlier homonym competes for use. However, the Vittadini name has been widely applied to this species and thus should be conserved. The name Tuber aestivum Vittad. was accepted by Tulasne & Tulasne (Fung. Hypog.: 137–138. 1851), who identified it as the truffle that in France was commonly called “truffle de la Saint-Jean”, providing an unmistakable illustration (l.c.: t. 7, fig. 3). Since then, in Italy and France, the most important countries in the production and trade of truffles, this species was indicated with the Vittadini name (Ferry de la Bellone, La Truffe: 37. 1888; Chátin, Truffe, ed. 2: 62. 1892; Mattirolo in Mem. Reale Accad. Sci. Torino, ser. 2, 53: 339. 1903). In other European countries, mycologists who dealt with truffles or edible mushrooms used the Vittadini name, as in Denmark (Lange, Danish Hypog. Macromycetes. 1956), Germany (Hesse, Hypog. Deutschl. 2: 14. 1894; Fischer in Rabenh. Krypt.-Fl., ed. 2, 1(5): 37. 1897), Great Britain (Berkeley & Broome in Ann. Mag. Nat. Hist., ser. 2, 7: 183. 1851; Hawker in Philos. Trans., Ser. B 237: 495. 1954), and Hungary (Hollós, Magyarorsz. Földalatti Gombái: 36. 1911). In addition, Frank (in Ber. Deutsch. Bot. Ges. 3: 145, t. 10, fig. 7. 1885) described for the first time the phenomenon of mycorrhizal symbiosis using the name Tuber aestivum Vittad. for a mycorrhiza of this truffle formed with Fagus sylvatica. Leonardi & al. (in Cryptog. Mycol. 42: 151. 2021) lectotypified Tuber aestivum Vittad. by the illustration cited above. Tuber blotii is a name that was described and illustrated by Eudes-Deslongchamps in 1824. Its original material was studied by Maire (in Bull. Soc. Linn. Normandie, sér. 6, 4: viii–ix. 1910), who confirmed its identity as T. aestivum Vittad. The samples were probably lost during World War II. The building of the Institute Botanique de Caen was destroyed (https://www.repository.naturalis.nl/document/571005), and the remains of the collections were sent to the National Museum of Natural History (MNHN) in Paris, where this material could not be found. Consequently, the illustration from the original publication is designated herein as lectotype. Maire (in Bull. Soc. Mycol. France 46: 150. 1930) recognized the priority of T. blotii but preferred to use the name T. aestivum Vittad. because the binomial T. blotii had not been used except as a synonym of T. aestivum. The name T. blotii has never been used either commercially or in scientific research. Leonardi & al. (l.c.) examined the taxonomic situation of Lycoperdon aestivum Wulfen (in Jacquin, Collectanea 1: 349. 1787) and concluded that the fungus described was a species of Rhizopogon. Fries (Syst. Mycol. 2: 294. 1823) also considered L. aestivum to be a Rhizopogon, which he called Rhizopogon aestivus, thus sanctioning it. However, Sprengel (Syst. Veg. 4(1): 416. 1827) proposed the combination Tuber aestivum with the synonymy: “In Carinthia (Lycoperdon aestivum Wulff., Rhizopogon Fr.).” Thus, Tuber aestivum Vittad. is a later homonym (Art. 53.1). Tulasne & Tulasne (l.c.) included Rhizopogon aestivus as a synonym of their R. rubescens, the current name of which is R. roseolus (Corda) Th. Fr. Although several scientific publications of recent years have referred to the summer truffle as ‘T. aestivum (Wulf.) Spreng.’ instead of T. aestivum Vittad., this is erroneous because the basionym L. aestivum Wulfen refers to a Rhizopogon. Of these two scientific names that compete for use, Tuber aestivum is the more widely used as measured by a Restricted Google Scholar Search: T. blotii = 15, T. aestivum = 2830; and in GenBank: T. blotii = 0, T. aestivum = 4192, including data on the whole genome (Murat & al. in Nature, Ecol. Evol. 2: 1956–1965. 2018). (2868) Tuber magnatum Picco, Meleth. Inaugural.: 79. 1788, nom. cons. prop. Typus (vide Leonardi & al. in Cryptog. Mycol. 42: 160. 2021): Italy, Piedmont, Montechiaro D'Asti, Loc. Seria, with Quercus robur L. and Populus tremula L., 29 Sep 2019, Panzini (TO No. HG 3458). MBT 10001896. (=) Tuber griseum Pers., Syn. Meth. Fung.: 127. 1801, nom. sanct. (Fries, Syst. Mycol. 2: 292. 1823), nom. rej. prop. Lectotypus (vide Leonardi & al. in Cryptog. Mycol. 42: 161. 2021): [icon in] Borch, Lett. Truffes Piémont: fig. [1] A–G. 1780. MBT 10001897. Epitypus (vide Leonardi & al. in Cryptog. Mycol. 42: 161. 2021): Italy, Piedmont, Monte Magno, 7 Dec 1998, under Quercus robur, Gavazza (TO No. HG 3557). MBT 10001898. The Italian white truffle, also called the Piedmont white truffle, has long been known as Tuber magnatum. Vittadini (Monogr. Tuberac.: 42. 1831) applied the name T. magnatum Picco (Meleth. Inaugural.: 79. 1788) for this species and listed T. griseum as a synonym. Leonardi & al. (in Cryptog. Mycol. 42: 160. 2021) neotypified T. magnatum by the recently collected specimen cited above. Tuber griseum was sanctioned by Fries (Syst. Mycol. 2: 292. 1823); thus, this name T. griseum has precedence. Tuber magnatum has been widely used, while T. griseum remains obscure as evidenced by a Restricted Google Scholar Search: T. magnatum = 2320, T. griseum = 18; and GenBank: T. magnatum = 1233, including data on the whole genome (Murat & al. in Nature, Ecol. Evol. 2: 1956–1965. 2018), T. griseum = 0. It is proposed that the name Tuber magnatum be conserved against Tuber griseum (2869) Tuber melanosporum Vittad., Monogr. Tuberac.: 36. 1831, nom. cons. prop. Typus: [icon in] Vittadini, Monogr. Tuberac.: t. 2, fig. 3. 1831. MBT 10001899. Epitypus (vide Leonardi & al. in Cryptog. Mycol. 42: 161. 2021): Italy, Lombardy, Monza, Parco Villa Reale, sub Carpinus betulus and Tilia cordata, 1 Feb 2019, Seghezzi (AQUI No. 10152). MBT 10001900. (=) Tuber nigrum Bull., Herb. France 8(85–86): t. 356. 1788, nom. rej. prop. Lectotypus (vide Leonardi & al. in Cryptog. Mycol. 42: 162. 2021): [icon] “La Truffe noire” in Bulliard, Herb. France: t. 356. 1788. MBT 10001901. Epitypus (vide Leonardi & al. in Cryptog. Mycol. 42: 162. 2021): France. Lot 46090 Bellefont–La Rauze, 14 Dec 2019, Q. pubescens, Sourzat (AQUI No. 10208). MBT 10001902. Tuber melanosporum Vittad. (Monogr. Tuberac.: 36. 1831) is the scientific name that has been applied legally and commercially to this species, commonly known as the black truffle. The name was lectotypified by Leonardi & al. (in Cryptog. Mycol. 42: 161. 2021) by the illustration cited above. It competes with one other scientific name published earlier but rarely used. This first name applied to the black truffle was Tuber nigrum Bull. (Herb. France 8(85–86): t. 356. 1788). Vittadini (l.c.: 36) recognized the characteristic blackish spiny spores of T. melanosporum, thereby differentiating it from T. brumale Vittad. and the other warty black peridium species. Among these two names, T. melanosporum Vittad. is the more commonly used (Restricted Google Scholar Search: T. melanosporum = 5860, T. nigrum = 114; and GenBank: T. melanosporum = 103,946, including data on the whole genome [Martin & al. in Nature 464: 1033–1038. 2010], T. nigrum = 6). Thus, it is proposed that the name Tuber melanosporum be conserved against the earlier name T. nigrum. (2870) Tuber albidum Fr., Syst. Mycol. 2: 291. 1823, nom. rej. prop. Typus: non designatus. Fries (Syst. Mycol. 2: 291. 1823) published and thus sanctioned the name Tuber albidum with the diagnosis “verrucis exasperatum, albidum”, listing as synonyms “Tuber albidum Cæsalp. 616. Tuber aestivum pulpa subobscura &c. Mich. gen. p. 221 n. 2. Lycop. globosum, subsubterraneum &c. Gled. meth. p. 157.” Based on Fries's diagnosis, this name could refer to T. aestivum Vittad. (Monogr. Tuberac.: 38. 1831) and two references (Cesalpino, Pl. Libri XVI: 613–614. 1583 [not “616”]; Micheli, Nov. Pl. Gen.: 221. 1729) confirmed this. On the other hand, a third reference (Gleditsch, Meth. Fung.: 157. 1753) included in its diagnosis: “Lycoperdon globosum, subterraneum, solidum et scabrum” and in the subsequent discussion “[…] cortex albidus, eminentiis non nihil exasperatus […] substantia interior […] per maturitatem plus minus spongiosa, laxa, mollis, sicca et pulposa evadit”, suggesting that this name describes a Rhizopogon. Vittadini (l.c.: 38–39) proposed the name T. aestivum listing “Tub. albidum? Fries” as a synonym but he wrote in a commentary on the name (Vittadini, l.c.: 40): “Color externus albidus in Tuberibus muricatis mihi prorsus extraneus, suspectus. Hinc albidi nomen ambiguum.” We concur with Vittadini that T. albidum Fr. is a questionable species that does not agree with any known species of Tuber in which the warty peridium is black or deep brown (Bonito & al. in PLoS ONE 8(1): e52765. 2013, https://doi.org/10.1371/journal.pone.0052765). In attempting to typify T. albidum, Leonardi & al. (in Cryptog. Mycol. 42: 160. 2021) searched for the dried sample mentioned by Fries (l.c.: 291), but no specimen of T. albidum was found in the herbarium of Fries (UPS-FRIES) nor was a drawing of this taxon. Thus, we propose the rejection of the name Tuber albidum Fr. in application of Art. 56.1 of the Shenzhen Code. (2871) Tuber cibarium Bull., Hist. Champ. France: 74. 1791, nom. sanct. (Fries, Syst. Mycol. 2: 290. 1823), nom. rej. prop. Typus: non designatus. Bulliard (Hist. Champ. France.: 74. 1791) proposed the name Tuber cibarium for truffles having a warty black peridium. Within T. cibarium he recognized four main varieties, the first of which was listed as “1. Tub. cibarium nigrum, vid. Tuber nigrum, tab. 356.” The name T. cibarium Bull. was applied to the black truffle in the early literature (Withering, Bot. Arr. Brit. Pl., ed. 2, 3(2): 458. 1792; Sibthorp, Fl. Oxon.: 398. 1794; Sowerby, Col. Fig. Engl. Fung.: t. 309. 1800; Turpin, Dict. Sci. Nat. Pl. Bot. [1] Vég. Acot.: t. 46. 1821). The name T. cibarium, which would otherwise be a superfluous and illegitimate name for T. nigrum Bull., was sanctioned by Fries (Syst. Mycol. 2: 290. 1823) in close accordance with the treatment by Bulliard (l.c.). Bulliard gave the specific character of the species as “La Truffe comestible est la seule espèce de ce genre dont la surface soit comme verruqueuse, ou relevée de petites éminances à peu près prismatiques.” He included under this name not only the numerous species of black truffles now known in Europe (T. aestivum, T. brumale, T. macrosporum Vittad., T. melanosporum, T. mesentericum Vittad., T. bituminatum Berk. & Broome, T. malençonii Donadini & al., and T. suave Pacioni & M. Leonardi) but also unidentifiable white or greyish truffles. Given the confusion concerning the name, we propose the rejection of the name Tuber cibarium Bull. in application of Art. 56.1 of the Shenzhen Code. GP, https://orcid.org/0000-0002-0201-5789 JMT, https://orcid.org/0000-0003-2032-1904 ML, https://orcid.org/0000-0003-3502-4232 AYR, https://orcid.org/0000-0002-8191-2663

Truffles in the genus Tuber produce subterranean fruiting bodies that are not able to actively discharge their spores in the environment. For this reason, truffles depend on mycophagous animals for reproduction. Fungus consumption (mycophagy) is a behaviour typical of both vertebrates and invertebrates. Mammals, especially rodents, are the most studied group of mycophagists and have been found to consume a great variety of fungi. Among invertebrates, mycophagy is documented in arthropods, but rarely in molluscs. In our study we assessed the effect on the morphology and mycorrhizal colonization of Tuber aestivum spores after passage through the gut of slugs (Deroceras invadens) and, for comparison, of a house mouse (Mus musculus). Light, scanning electron and atomic force microscopy revealed that the digestion, especially by slugs, freed spores from the asci and modified their morphology. These are believed to be the reasons why we observed an improvement in oak mycorrhization with the slug and rodent ingested spores in comparison to a fresh spore inoculation. We also demonstrated by molecular barcoding that slugs' guts sampled on a Tuber melanosporum truffle ground contain spores from this species and Tuber brumale, further suggesting that some invertebrates are efficient Tuber spore dispersers.

The aim of this study was to enable searches for truffles (Tuber spp.), particularly the Burgundy truffle (T. aestivum Vittad.), to be carried out in forests based on a method that has been constantly developed since 2007 by the Forest Research Institute. The method is termed "Virtual Truffle Hunting" and it takes 12 parameters into account: bedrock, soil pH, Ca+ and CaCO3 content in soil, C/N ratio, soil structure, altitude of terrain, type of forest site, forest structure, the Burgundy truffle host trees, and the presence of particular species including orchids and insects. A simple "Virtual Truffle Hunting" software has also been developed, which makes the use of the method easy, fast, and effective. This method is to ascertain the truffle potential for all areas in which digital maps are not available. In 2015, the method was tested in 20 sites, representing forests in 5 Polish macroregions. Hunting for hypogeous fungi was conducted from June to October with the help of trained dogs. Thanks to this method, 14 new truffle sites were found. The knowledge of environmental conditions conducive to the Burgundy truffle growth enabled us to form an effective tool in order to identify new sites of truffle presence.

Tuber mesentericum is an edible European black truffle, apparently easy to recognize, but showing a high degree of genetic variability. In this study, we performed an integrative taxonomic assessment of the T. mesentericum complex, combining a multilocus phylogeographic approach with morphological analyses, and including authentic specimens of Vittadini, and Berkeley and Broome. We performed maximum likelihood phylogenetic analyses, based on single and concatenated gene datasets (ITS rDNA, β-tubulin, elongation factor 1-α), and including all available sequences from previous studies. Phylogenetic analyses consistently recovered three reciprocally monophyletic and well-supported clades: clade I, with a wide range across Europe; clade II, specimens collected mainly in the Iberian, Italian, and Balkan peninsulas; and clade III, specimens collected almost exclusively in central Italy. Genetic distance between clades ranged from 10.4% to 13.1% at the ITS region. We also designed new primer pairs specific for each phylogenetic lineage. Morphology of spores, asci, and peridium were investigated on specimens representing the three lineages. Macro- and micromorphological analyses of ascomata revealed only a few, but not diagnostic, differences between the three phylogenetic lineages, thus, confirming that they are morphologically cryptic. By studying authentic specimens of Vittadini, and Berkeley and Broome, it was possible to identify the three clades as T. mesentericum, Tuber bituminatum, and Tuber suave sp. nov., and to designate an epitype for T. mesentericum s.s. and a lectotype for T. bituminatum. Future investigations on volatile organic compound (VOC) composition are needed to define the aroma repertoires in this species complex.

The true truffles, 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 identification has been based on morphological and then molecular characters in the absence of reference type specimens. Although long of scientific, commercial and regulatory use, these four scientific names are at risk due to a lack of nomenclatural priority. To provide the scientific 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. The type of each name is described morphologically and molecularly characterized with the sequences of three markers: ITS, beta-tubulin, elongation factor 1 alpha. The taxonomy and nomenclature of each species are discussed. The 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. The name Tuber borchii has no previous synonyms and therefore it is legitimate and does not require conservation.

Perhaps with the exception of a small number of cultivated saprobic species, the true truffles (Tuber spp.) command the attention of more scientists than other species of mushroom. Many thousands of scientific papers have been devoted to topics ranging among taxonomy; aroma; molecular makeup; the food, pharmaceutical, and cosmetics industries; and perhaps, at the bottom of the list, their cultivation. The genus Tuber is widespread in the northern hemisphere and with the recent exploration of new areas and the application of molecular identification methods, the number of proposed species is growing almost exponentially. The past 25 years in particular have seen a huge expansion in the cultivation of Tuber melanosporum, T. borchii, and T. aestivum in the Southern Hemisphere and a flurry of papers dealing with various molecular aspects of the truffles. This has resulted in a greater understanding of how to grow truffles in areas which would have been considered totally unsuitable in the Northern Hemisphere, and a depth of knowledge that early 1970s/1980s' researchers would never have conceived such as the cultivation of T. melanosporum on heavily limed, naturally very acidic soils. Our current review looks at the state of knowledge in the fields of biodiversity and ecology, biotechnological applications, and the cultivation of true truffles.

Arbutus unedo (the strawberry tree) is a Mediterranean shrub which forms arbutoid mycorrhizae with a variety of Asco- and Basidiomycetes. After the discovery of the mycorrhizal symbiosis between A. unedo and Tuber borchii, in this study, arbutoid mycorrhizae were synthetized in greenhouse with Tuber aestivum and Tuber melanosporum. Six months after inoculation, both species colonized the roots of all inoculated A. unedo seedlings, but mature mycorrhizae were only observed after 12 months. Ultrastructure analysis of Tuber arbutoid mycorrhizae was described for the first time, showing, as observed in typical endosymbiosis, a rearrangement of host cells and the creation of an interface compartment with both truffle species. Immunolabelling experiments suggested that pectins are not present in the interface matrix surrounding the intracellular hyphae. Thus, the ability to establish symbiosis with A. unedo seems to be a common feature in the genus Tuber, opening up the possibility to use this plant for mycorrhization with valuable truffles. This could represent an important economic opportunity in Mediterranean areas by combining the production of truffles, edible fruits and valued honey.

Aflatoxins are mycotoxins produced by some Aspergillus species. They are remarkably toxic and included among the most known carcinogenic substances. Article 139 of the Italian presidential decree of 30 June 1965, no. 1204 and subsequent amendments, includes hepatocellular carcinoma due to exposure to aflatoxin B1 the list of diseases for which reporting is mandatory. Aflatoxins could contaminate food of plant and animal origin, therefore the areas of work most at risk are the agro-food and, in general, all the activities carried out in humid and poorly ventilated areas, subject to fungal contamination. Although the procedures for controlling aflatoxins in food are now fairly well established, there is still no full knowledge of the risk of workplace exposure. A review of the risks relating to exposure to aflatoxin of workers in the agri-food sector is presented here based on bibliographic data and surveys conducted in some Italian regions by local health authorities and by the National Institute for Insurance against Accidents at Work. To protect the health of workers in the sectors considered, we therefore consider it necessary to evaluate the presence of aflatoxins and inform workers about the application of good prevention practices and the use of adequate personal protective equipment.

Coprophilous fungi are specialized microorganisms, playing key roles in ecosystems and in several other contexts, whose protection requires more substantial efforts. This paper aims to highlight and discuss valuable and critical aspects faced during the process of the threat status assessment ofPoronia punctata, providing inspirations for future conservation actions.

The ectomycorrhizal fungus Tuber magnatum produces the white truffle appreciated worldwide for its unique aroma. With respect to other Tuber spp. of economic interest, T. magnatum presents a narrower geographical range. This species has, in fact, long been considered endemic to Italy. However, over the last few decades several reports have documented the presence of white truffles in different Mediterranean countries and in particular in various areas of south-east Europe. In this study, samples from several Pannonian and Balkan countries such as Hungary, Serbia, Romania, Bulgaria and Greece have been collected and genotyped with microsatellite markers and the data merged with those available for Italian populations. Our objectives were to test whether Italian and south-east European populations are differentiated and to evaluate the genetic diversity of T. magnatum all over its distributional range. We show the genetic structure of T. magnatum populations with the differentiation of four main groups: northern Italy, central-northern Italy, southern Italy and the Balkan/Pannonian region. The present study allowed us to refine the evolutionary history of T. magnatum and track the possible post-glacial expansion route of this species. The assessment of T. magnatum’s genetic structure is not only of scientific relevance, but it is also important for the conservation and market traceability of this prestigious fungus.

Recent collections of Tuber malacodermum from Spain, Corsica, and Mexico were compared in an integrative morphological and molecular phylogenetic framework, including study of type material. Phylogenetic analyses of nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) and nuc 28S rDNA (28S) regions showed that specimens from Spain and Corsica form a monophyletic group closely related to T. melosporum and T. rufum, whereas Mexican specimens form a clade within the T. lyonii species complex. Peridium and ascospore morphology contribute clear morphological distinctions among specimens from Spain and Corsica, Mexico, and the type specimen of T. malacodermum. Based on results of the morphological and molecular phylogenetic analyses, we assigned the specimens from Spain and Corsica to Tuber pustulatum, sp. nov., and the Mexican specimens to Tuber theleascum, sp. nov. We restrict T. malacodermum to the sole type material. Formal descriptions and illustrations of these taxa are provided.

The paper presents a review on health hazards involving operators that cultivate and handle mushrooms. Intensive production of edible and medicinal fungi is a common agricultural management all over the world and involves a number of operators whose health must be safeguarded due the number of risks. Growth techniques and environmental conditions for producing and handling mushrooms expose workers to several types of allergens that possibly cause occupational diseases. Namely, mushroom workers suffer from allergic pulmonary diseases and, more rarely, from forms of contact dermatitis. The cause of these clinical manifestations may be found in the exposure to several factors, such as the peculiar production conditions leading to the presence of many allergens (bacteria, moulds, mycotoxins, endotoxins) and the direct contact of workers with some fungal species, which are themselves allergens.