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The kingdom Fungi comprises one of the most diverse groups of living organisms. They are numerous, ubiquitous and undertake many roles both independently and in association with other organisms. Fungi display a wide diversity of forms... more
The kingdom Fungi comprises one of the most diverse groups of living organisms. They are numerous, ubiquitous and undertake many roles both independently and in association with other organisms. Fungi display a wide diversity of forms also mirrored by functional diversity and play such a dominant role in human society that they are arguably biotechnologically the most important group of organisms. Ex situ conservation of fungi, not only guarantees correct management and conservation of biodiversity, but also its exploitation in different fields. This article describes the major biological resource centres dealing with fungi in Italy and gives several examples of fungal exploitation in different fields of applications.
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Artemisia petrosa subsp. eriantha is a protected species whose essential oil is of great interest mainly for liqueur industry; it was micropropagated by in vitro culture technique followed by transfer to field. The content and the... more
Artemisia petrosa subsp. eriantha is a protected species whose essential oil is of great interest mainly for liqueur industry; it was micropropagated by in vitro culture technique followed by transfer to field. The content and the characteristics of the essential oils from regenerated plants collected after 1
and 2 years of field growing and from regenerated plants grown ‘in situ’ were assessed by gas chromatography/mass spectrometry (GC/MS) and compared with those ofwild plants. Results indicate that in vitro propagated plants produce oils rich in sesquiterpenes. Forty seven compounds were identified, the main constituents being α- and β-thujone, whose content increased with the plant age. Quantitative, but not qualitative variations were observed both in wild and in micropropagated plants in relation with plant age and environmental factors. Thus, micropropagation provides plants suitable for the industrial exploitation of this species.
and 2 years of field growing and from regenerated plants grown ‘in situ’ were assessed by gas chromatography/mass spectrometry (GC/MS) and compared with those ofwild plants. Results indicate that in vitro propagated plants produce oils rich in sesquiterpenes. Forty seven compounds were identified, the main constituents being α- and β-thujone, whose content increased with the plant age. Quantitative, but not qualitative variations were observed both in wild and in micropropagated plants in relation with plant age and environmental factors. Thus, micropropagation provides plants suitable for the industrial exploitation of this species.
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The kingdom Fungi comprises one of the most diverse groups of living organisms. They are numerous, ubiquitous and undertake many roles both independently and in association with other organisms. Fungi display a wide diversity of forms... more
The kingdom Fungi comprises one of the most diverse groups of living organisms. They are numerous, ubiquitous and undertake many roles both independently and in association with other organisms. Fungi display a wide diversity of forms also mirrored by functional diversity and play such a dominant role in human society that they are arguably biotechnologically the most important group of organisms. Ex situ conservation of fungi, not only guarantees correct management and conservation of biodiversity, but also its exploitation in different fields. This article describes the major biological resource centres dealing with fungi in Italy and gives several examples of fungal exploitation in different fields of applications.
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The genus Amanita Pers. ex Fr. in Rome neighbourood. — The presence of the genus Amanita in Rome neighbourood, small district, with vegetation and soil diversified, has been studied. The list includes 30 entities (A. caesarea, A.... more
The genus Amanita Pers. ex Fr. in Rome neighbourood. — The presence of the genus Amanita in Rome neighbourood, small district, with vegetation and soil diversified, has been studied. The list includes 30 entities (A. caesarea, A. argentea°, A. nivalis, A. vaginata, A. vaginata var. cinerea°, A. fulva, A. crocea°, A. umbrinolutea°, A. lividapallescens°, A. strangulata, A. junquillea, A. eliae°, A. muscaria, A. pantherina, A. alba, A. phalloides, A. phalloides var. alba, A. verna, A. virosa, A. citrina, A. citrina var. alba°, A. porphyria°, A. rubescens, A. spissa. A. excelsa, A. aspera, A. vittadini, A. codinae°, A. strobiliformis, A. boudieri°), 11 new also for latium (°), among these A. codinae and A. vaginata var. cinerea hitherto not found in Italy. At last, the genus in whole region is examined.
Translated by Simon Pleasance. Giovanni Pacioni, The Macdonald Encyclopedia of Mushrooms and Toadstools ISBN 0 356 07913 9. Price £7.95 (but already available at a reduced price). (English edition), Macdonald and Co. (Publishers) Ltd., London and Sydney (1985), p. 512 ISBN 0 356 07913 9. Price £7...more
The present work studies the expression of tyrosinase (monophenol:diphenol oxygen oxidoreductase, EC 1.14.18.1) during the development of the black truffle Tuber melanosporum Vittad., an ectomycorrhizal fungus of great biological and... more
The present work studies the expression of tyrosinase (monophenol:diphenol oxygen oxidoreductase, EC 1.14.18.1) during the development of the black truffle Tuber melanosporum Vittad., an ectomycorrhizal fungus of great biological and economic interest. As widely reported in the literature, melanins and the enzymes that synthesize them, are of paramount importance in fungal development and sexual differentiation. Tyrosinase and laccase are the enzymes that produce melanins from monophenols and diphenols. We have detected tyrosinase expression from the stage of free living mycelium, through the mychorrizal stage and the six fruit body developmental stages by measuring the levels of tyrosinase mRNA by quantitative PCR (q-PCR), spectrophotometry, histochemistry, immunohistochemistry and electrophoresis. Tyrosinase is always expressed, from the free living mycelium to the ripe fruit body developmental stages, when it is very low. The switching off of the tyrosinase gene during T. melanosporum development when the fruit body is ripe and no more cell walls are to be built is discussed in relation of thioflavour production. Specific primers, prepared from the cloned T. melanosporum tyrosinase cDNA were used for the q-PCR and the deduced aminoacid sequences of the CuA and CuB binding sites were compared to those of various ascomycetes and basidiomycetes.Tuber melanosporum fruit body develops from the ectomycorrhized Corylus avellana plants. The occurrence of melanin in the truffle hyphae, spores and fruit body is evident.► In truffle development Tmeltyr gene is switched on but off when the spores are ripe. ► Tmeltyr tyrosinase is close to that of Coccidioides immitis. ► T. melanosporum tyrosinase activity and protein occur in fruiting body fertile veins.
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Artemisia petrosa subsp. eriantha is a protected species whose essential oil is of great interest mainly for liqueur industry; it was micropropagated by in vitro culture technique followed by transfer to field. The content and the... more
Artemisia petrosa subsp. eriantha is a protected species whose essential oil is of great interest mainly for liqueur industry; it was micropropagated by in vitro culture technique followed by transfer to field. The content and the characteristics of the essential oils from regenerated plants collected after 1 and 2 years of field growing and from regenerated plants grown ‘in situ’ were assessed by gas chromatography/mass spectrometry (GC/MS) and compared with those of wild plants. Results indicate that in vitro propagated plants produce oils rich in sesquiterpenes. Forty seven compounds were identified, the main constituents being α- and β-thujone, whose content increased with the plant age. Quantitative, but not qualitative variations were observed both in wild and in micropropagated plants in relation with plant age and environmental factors. Thus, micropropagation provides plants suitable for the industrial exploitation of this species.
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The relationship between bacteria and the sporocarpic structures of truffles of the genus Tuber has been studied by means of scanning electron microscopy. Pieces of the truffles, critical-point dried then broken by traction, showed... more
The relationship between bacteria and the sporocarpic structures of truffles of the genus Tuber has been studied by means of scanning electron microscopy. Pieces of the truffles, critical-point dried then broken by traction, showed bacteria on hyphae of the ‘venae externae’ as well as inside the mature asci. Possibly the bacteria initially aid development of sporocarpic hyphae through the interchange of metabolites and may subsequently be associated with activation of ascospores.
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Quinonoid and polyphenolic biopolymers are described as the most important constituents of truffle pigments. Both alkali-soluble and insoluble fractions of peridium and gleba tissues of truffles of the genus Tuber have been investigated.... more
Quinonoid and polyphenolic biopolymers are described as the most important constituents of truffle pigments. Both alkali-soluble and insoluble fractions of peridium and gleba tissues of truffles of the genus Tuber have been investigated. All the data collected suggest that these black pigments are allomelanins of polyketide origin.
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In the present article, magnetic resonance imaging spectroscopy (MRI) was used to study fresh black truffle, with the aim of elucidating the internal structure, the effects of fungal invasion, and physical changes occurring in... more
In the present article, magnetic resonance imaging spectroscopy (MRI) was used to study fresh black truffle, with the aim of elucidating the internal structure, the effects of fungal invasion, and physical changes occurring in post-harvest. MRI is a non-destructive and non-invasive analytical technique offering the almost unique opportunity of studying foodstuff while leaving it intact, without any sample preparation. The internal morphology and modifications induced by external agents (e.g. fungal invasion, diseases, dehydration) are some of the aspects addressed by means of this innovative analytical tool.
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Interesting Italian entomogenous Fungi: I. Paecilomyces fumosoroseus, Cordyceps memorabilis.—Two entomogenous fungi, collected in Italy, are described. Paecilomyces fumoso-roseus (Wize) Brown et Smith is a species new for the Italian... more
Interesting Italian entomogenous Fungi: I. Paecilomyces fumosoroseus, Cordyceps memorabilis.—Two entomogenous fungi, collected in Italy, are described. Paecilomyces fumoso-roseus (Wize) Brown et Smith is a species new for the Italian flora, while Cordyceps memorabilis is reported for the first time after its description by Cesati.
The malate dehydrogenase (MDH; EC 1.1.1.37; L-malate-NAD+-oxidoreductase) activities of truffles of the genus Tuber (Tuber melanosporum Vittad., Tuber brumale Vittad., Tuber aestivum Vittad., Tuber magnatum Pico, Tuber rufum Pico) have... more
The malate dehydrogenase (MDH; EC 1.1.1.37; L-malate-NAD+-oxidoreductase) activities of truffles of the genus Tuber (Tuber melanosporum Vittad., Tuber brumale Vittad., Tuber aestivum Vittad., Tuber magnatum Pico, Tuber rufum Pico) have been characterized with regard to the Km and Vmax values in the direct and reverse reactions. The isoelectrofocusing has revealed bands showing pI values ranging from pH 5.85 to 7.8. The MDH of T. melanosporum has been partially purified by hydroxyapatite treatment, DEAE-cellulose and Sephadex G-75 columns. With the partially purified T. melanosporum MDH activity polyclonal anti-T. melanosporum MDH antibodies have been prepared and used to localize MDH in the mycorrhizae and ascocarps of T. melanosporum. These antibodies inhibit T. melanosporum MDH activity as well as that of T. magnatum but not that of rabbit liver; this supports the specificity of the MDH antibodies used to localize MDH in truffle tissues.
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The filametous ascomycete Tuber borchii is a plant-symbiotic ectomycorrhizal microrganism with an high value due to the production of hypogeous fruitbodies (truffles). The present work was undertaken to develop a procedure for the release... more
The filametous ascomycete Tuber borchii is a plant-symbiotic ectomycorrhizal microrganism with an high value due to the production of hypogeous fruitbodies (truffles). The present work was undertaken to develop a procedure for the release of T. borchii viable protoplasts from Tuber mycelium, isolate ATTC 96540; several factors which affect the isolation, morphology and viability were examined and developed in order to improve applications of T. borchii protoplasts in morphological, biochemical and genetic investigations (protoplast fusion or transformation). Functional delivery of liposome content into T. borchii protoplasts has also been examined with a cytotoxic ribosome inactivator as saporin. T. borchii protoplasts incubation/fusion with saporin containing liposomes were made to demonstrate the absence of cell wall of 16 days cultured protoplasts.
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A 1-year time series of fungal spore concentrations has been used to calibrate an artificial neural network for the estimation of Alternaria and Pleospora concentrations associated to observed meteorological variables. Analysis of the... more
A 1-year time series of fungal spore concentrations has been used to calibrate an artificial neural network for the estimation of Alternaria and Pleospora concentrations associated to observed meteorological variables. Analysis of the results revealed that the daily average values of these meteorological variables are suitable to predict with high confidence the number of fungal spores that are actually observed. The calibrated neural network has also been used randomizing each single input parameter in order to evaluate which meteorological variable contributes more to the formation and the depletion of the selected fungal spores. Emphasis is given to the possibility of using the proposed model for operational activities, predicting the future spore concentrations on the basis of meteorological forecasts.
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Tuber spp. are ectomycorrhizal ascomycetes that produce subterranean ascomata known as truffles. Truffles can be regarded as complex microhabitats hosting bacteria and yeasts. In this paper we show that guest filamentous fungi are also... more
Tuber spp. are ectomycorrhizal ascomycetes that produce subterranean ascomata known as truffles. Truffles can be regarded as complex microhabitats hosting bacteria and yeasts. In this paper we show that guest filamentous fungi are also associated to truffle ascomata, regardless of the Tuber spp., and report the morpho-molecular characterization of seven truffle-hosted mycelia isolated from healthy and intact Tuber ascomata. Some of these isolates were shown to be related to the fungal endophytes of plants. Interestingly, the truffle-hosted mycelia grew stuck to the hyphal wall of their partner when co-cultivated with the Tuber borchii mycelium, but not when co-cultivated with the test species Agaricus macrosporus. The present data suggest that guest filamentous fungi can be added to the list of truffle-interacting microorganisms.
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Truffles live in an environment poor in oxygen, and are a good model for investigation of enzymatic adaptation to microaerobic conditions. We studied the antioxidant and glutathione dependent enzymatic endowment of Tuber truffles.... more
Truffles live in an environment poor in oxygen, and are a good model for investigation of enzymatic adaptation to microaerobic conditions. We studied the antioxidant and glutathione dependent enzymatic endowment of Tuber truffles. Superoxide dismutase, catalase, glutathione peroxidase Se dependent, glutathione reductase, glyoxalase 1 and glyoxalase 2 are expressed and correlate with the microaerobic metabolism, growth rate and mycorrhizal symbiosis of truffles. A very low or undetectable glutathione S-transferase activity was found. For comparison the same enzyme activities were investigated in Agaricus bisporus which is epigeous and non-mycorrhizal.
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Research Interests: Kinetics, Melanin, Enzyme Inhibitors, Biological Sciences, Copper, and 17 moreSulfides, Methane, Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis, Levodopa, Enzyme, Substrate Specificity, Catechols, Agaricus, Molecular weight, Species Specificity, Cell Wall, Tyrosinase, Melanine, Ascomycota, Oxidation-Reduction, Tyrosine, and Mimosine
Specimens from several Italian populations ascribable to nine taxa of the section Dapetes of the genus Lactarius (Russulales, Basidiomycota) were subjected to electrophoretical analysis of the gene-enzyme systems from eleven loci. Results... more
Specimens from several Italian populations ascribable to nine taxa of the section Dapetes of the genus Lactarius (Russulales, Basidiomycota) were subjected to electrophoretical analysis of the gene-enzyme systems from eleven loci. Results show that the four taxa characterised by carrot-red latex (Lactarius deliciosus complex) are genetically well separated, whereas the five taxa with vinaceous-red latex (L. sanguifluus complex) fall into two groups. Accordingly, only one of the two conflicting conceptions of Lactarius vinosus present in the literature may be regarded as a good species while the other, which is genetically identical, has to be considered as a morphological form of L. sanguifluus.
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A new entomogenous Ascomycete found in cave: Cordyceps riverae.—A new species of entomogenous Cordyceps, belonging to Subgenus Racemella (Ces.) Sacc., found on an adult Lepidotteron in a cave near L'Aquila (Abruzzo, Italy) is described.... more
A new entomogenous Ascomycete found in cave: Cordyceps riverae.—A new species of entomogenous Cordyceps, belonging to Subgenus Racemella (Ces.) Sacc., found on an adult Lepidotteron in a cave near L'Aquila (Abruzzo, Italy) is described. The characters of this species, named Cordyceps riverae, are compared with those of other similar species of Cordyceps.
Mycorrhizal fungi belonging to the genus Tuber produce, after the establishment of a productive interaction with a plant host, hypogeous fruitbodies of great economic value known as ‘‘truffles’’. This review summarizes the state of art on... more
Mycorrhizal fungi belonging to the genus Tuber produce, after the establishment of a productive interaction with a plant host, hypogeous fruitbodies of great economic value known as ‘‘truffles’’. This review summarizes the state of art on life cycle, genetic, and biotechnological investigations of Tuber spp. The ascocarp formation in truffles is a consequence of the activation of the sexual phase of the biological cycle. The formation of a dikaryotic secondary mycelium and the karyogamy in the ascal cell (followed by meiosis with ascospores formation) have been hypothesized by several authors but some doubts yet arise from the Tuber cycle by considering that a series of abnormalities have been pointed out in respect to other Ascomycetes. It is unclear if binucleated hyphal cells are derived from the fusion of mononucleated cells belonging to mycelia from different mating types or from one only. According to the karyotypes of Tuber melanosporum, Tuber magnatum, and Tuber borchii, the numbers of hyphal chromosomes suggest a chromosome number of eight (2n); these values are in the range of those of several Ascomycetes and observed for Tuber aestivum (2n=10). The importance and growth in interest during the last years in the fungi protoplasts isolation and transformation techniques can be related to current developments in Tuber genetics and biotechnology. T. borchii could be transformed through liposome-mediated delivery of genetic material as mycelial protoplasts isolation and fusion with liposomes has already been established. On the other hand, Agrobacterium-mediated transformation has been successfully established for T. borchii.
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Chromosome number is generally considered to be the most accurate and direct measure of ploidy. In fungi, however, counting of chromosomes is difficult and inaccurate because of their small size. To date truffles have been characterized... more
Chromosome number is generally considered to be the most accurate and direct measure of ploidy. In fungi, however, counting of chromosomes is difficult and inaccurate because of their small size. To date truffles have been characterized using molecular approaches (analysis of DNA and PCR-based techniques) in order to find out differences among species, but not by a cytogenetical approach. Although the small size of truffle chromosomes is of some hindrance for a cytogenetic study, in the present work Tuber aestivum chromosome counts were determined in metaphase configurations from haploid nuclei of ascospores by aceto-orcein staining. Nuclear chromosome number and topography were also evaluated by propidium iodide DNA staining using confocal microscopy. Tuber aestivum was found to possess a basic number of 5 or 6 chromosomes, medium length ≤0.95 μm. The karyology of ascospores during their developmental stages was also investigated.
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The taxonomy of some species hitherto referred to Isaria is revised. Hirsutella eleutheratorum (Nees ex S. F. Gray) Petch is redescribed and H. entomophila Pat., originally named the type species of Hirsutella Pat., is regarded as a... more
The taxonomy of some species hitherto referred to Isaria is revised. Hirsutella eleutheratorum (Nees ex S. F. Gray) Petch is redescribed and H. entomophila Pat., originally named the type species of Hirsutella Pat., is regarded as a synonym. Isaria eleutheratorum var. racemosa Auersw. is identified with Tilachlidiopsis racemosa Keissler. The new genus Troglobiomyces is described with the type species Troglobiomyces guignardii (Maheu) Pacioni (syn. Isaria guignardii Maheu).
This paper reports the effect of the tyrosinase (monophenol o-diphenol:oxygen oxidoreductase; EC 1.14.18.1) inhibitors diethyldithiocarbamate (DETC), L-tropolone, kojic acid, phenylthiourea (PTU) and L-mimosine on the in vitro growth of... more
This paper reports the effect of the tyrosinase (monophenol o-diphenol:oxygen oxidoreductase; EC 1.14.18.1) inhibitors diethyldithiocarbamate (DETC), L-tropolone, kojic acid, phenylthiourea (PTU) and L-mimosine on the in vitro growth of Tuber borchii (a white truffle) mycelium. A significant inhibitory effect on mycelium growth was observed for DETC, PTU and L-tropolone (0% growth compared to control at 100 μg ml−1 DETC, PTU and L-tropolone and at 10 μg ml−1 DETC and L-tropolone). As a comparison the action of the same inhibitors was also tested on the growth and pigmentation of the mould Cladosporium sphaerospermum. In the presence of CuSO4 10−6 M T. borchii mycelium acquired pigmentation (as rounded aggregates compared to control revealed by SEM microscopy). Tyrosinase activity in the extract from T. borchii mycelium (18-day culture) was detected spectrophotometrically.
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White truffles, Tuber magnatum Pico and Tuber excavatum Vitt. are both albino tyrosinase positive fungi. Interestingly, while both l-tyrosine-3-monooxygenase and l-3,4-dihydroxyphenylalanine oxidase activities of T. excavatum Vitt.... more
White truffles, Tuber magnatum Pico and Tuber excavatum Vitt. are both albino tyrosinase positive fungi. Interestingly, while both l-tyrosine-3-monooxygenase and l-3,4-dihydroxyphenylalanine oxidase activities of T. excavatum Vitt. decrease versus time after decompartmentalization, those of T. magnatum Pico increase. The Km of both species for l-tyrosine are lower than those for l-3,4-dihydroxyphenylalanine as substrates, while the Vmax with l-tyrosine are very much lower than those with l-3,4-dihydroxyphenylalanine. No change of Km versus time was found, only Vmax change, thus tentatively the inactivation or activation found in T. excavatum Vitt. and T. magnatum Pico tyrosinase activity versus time may be ascribed either to the binding or release of an inhibitor or to the release or binding of an activator, respectively. l-Tyrosine 3-monooxygenase and l-3,4 dihydroxyphenylalanine oxidase co-localize as shown histochemically.
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Several species of Tuber (Ascomycotina, Tuberales) produce a ‘burned’ area around their symbiotic plants as a result of a phytotoxic action. The volatile substances from Tuber sporophores, truffles, have been previously identified by gas... more
Several species of Tuber (Ascomycotina, Tuberales) produce a ‘burned’ area around their symbiotic plants as a result of a phytotoxic action. The volatile substances from Tuber sporophores, truffles, have been previously identified by gas chromatography coupled to gas mass spectrometry and then tested for their toxic effects on higher plants as well as soil micro-organisms. All tested organisms appeared to suffer a great inhibition when exposed to some of these substances evaporated into desiccators. Three aldehydes (2-methyl propanal, 2-methyl butanal and 3-methyl butanal) produced these effects on the two soil fungi tested (Penicillium vinaceum and Aspergillus alliaceus). In addition to some of these, two alcohols (2-methyl butanol and 3-methyl butanol) are also active on plants (Triticum vulgare and Lens culinaris). 2-Methyl butanol generally affects plants to a high degree, but the strongest effects to microfungi are caused by 3-methyl butanal. However, a strain of Pseudomonas isolated from truffle surface tolerated much higher concentrations of the three aldehydes. The substances affecting the soil micro-aerobic environment, where seeds germinate and roots live, are normal compounds of fungal metabolism produced via pyruvate pathway. Their toxic action could be a consequence of alterations caused both to the cellular membrane and to nucleic acids. The new term ‘hydnosphere’ has been proposed to define this peculiar case of soil relationships between truffle and neighbouring coenosis.
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Odour composition of some taxa related to Tuber melanosporum has been studied by gas chromatography coupled to a mass spectrometer (GC-MS). Although qualitative differences have been observed, the relative amounts of substances present... more
Odour composition of some taxa related to Tuber melanosporum has been studied by gas chromatography coupled to a mass spectrometer (GC-MS). Although qualitative differences have been observed, the relative amounts of substances present may be more significant for their chemical taxonomy. A concept of the Tuber odour according to its ecological function is discussed
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Page 1. SIMON & SCHUSTER'S GUIDE TO MUSHROOMS Page 2. Page 3. Page 4. Page 5. Simon & Schuster's Guide to MUSHROOMS By Giovanni Pacioni US Editor: Gary Lincoff A FIRESIDE BOOK PUBLISHED BY SIMON & SCHUSTER INC. Page... more
Page 1. SIMON & SCHUSTER'S GUIDE TO MUSHROOMS Page 2. Page 3. Page 4. Page 5. Simon & Schuster's Guide to MUSHROOMS By Giovanni Pacioni US Editor: Gary Lincoff A FIRESIDE BOOK PUBLISHED BY SIMON & SCHUSTER INC. Page 6. ...
Volatile substances, identified as components of the truffle odour, were tested as to their ability to attract truffle-eating insects. Arthropods were trapped by the thousand in all traps, but the truly mycetophilous insects, as well as... more
Volatile substances, identified as components of the truffle odour, were tested as to their ability to attract truffle-eating insects. Arthropods were trapped by the thousand in all traps, but the truly mycetophilous insects, as well as those species strictly bound to truffles in their biology, appear clearly attracted only by dimethyl sulphide.
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Eleven enzymes from 139 specimens of thirteen Italian populations of Tuber magnatum were analysed with multilocus horizonfal starch gel electrophoresis. The research was carried out to obtain insight into the genetic variability of this... more
Eleven enzymes from 139 specimens of thirteen Italian populations of Tuber magnatum were analysed with multilocus horizonfal starch gel electrophoresis. The research was carried out to obtain insight into the genetic variability of this species across its geographic range. All the gene-enzyme systems scored appear to be fixed in homozygosity in accordance with what is hitherto known in the genus Tuber; nine seemed monomorphic, whereas two, that is MDH-1 and ME-2, showed three alleles each. These results indicated a self-reproductive system and the low genetic variability is in agreement with the restricted endemism of the white truffle. The distribution of the electrophoretic types is discussed as a basis for further molecular applications.