Identification of ectomycorrhizal fungi of the genus <i>Tuber</i> by species‐specific ITS primers

Amicucci, Antonella; Zambonelli, Alessandra; Giomaro, Giovanna; Potenza, Lucia; Stocchi, Vilberto

doi:10.1046/j.1365-294x.1998.00357.x

Cited by 113 publications

(70 citation statements)

References 9 publications

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“…The specific PCR (polymerase chain reaction) was carried out using Tuber species-specific ITS (internal transcribed spacer) primers for the identification of T. magnatum, T. dryophilum, T. borchii and T. brumale. 23,24 Tuber mesentericum and T. aestivum, for which the specific primers are not known, were identified by the sequence analyses of the ribosomal DNA (rDNA) of the ITS regions. The ITS regions were amplified with the primers ITS1/ITS4.…”

Section: Experimental Truffle Species Identificationmentioning

confidence: 99%

Solid‐phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles

Gioacchini

Menotta

Bertini

et al. 2005

Rapid Comm Mass Spectrometry

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This study describes a rapid method to identify different truffle species by analysis of their volatile compound fraction using static headspace solid-phase microextraction gas chromatography/mass spectrometry. The volatile organic compounds (VOCs) were extracted using a new 2-cm 50/30 mm DVB/CAR/PDMS fiber placed for 10 min in the headspace of the truffle sample with the vial maintained at 208C (in a thermostatically controlled analysis room). The mass spectra of the VOC chromatograms were represented as 'fingerprints' of the analysed samples. Next, stepwise factorial discriminant analysis afforded a limited number of characteristic fragment ions that allowed a classification of the truffle species studied. This new method provides an effective approach to rapid quality control and identification of truffle species by analysis of their volatile fraction. Moreover, this method offers the advantage of minimizing thermal, mechanical, and chemical modifications of the truffles, thereby reducing the risk of analytical artifacts.

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Section: Experimental Truffle Species Identificationmentioning

confidence: 99%

Solid‐phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles

Gioacchini

Menotta

Bertini

et al. 2005

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

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“…The reproductive structures of these species in pure cultures have not been reported, and axenic spore germination remains an unresolved problem (26). Furthermore, the mating-type genes have never been characterized in truffles.Molecular markers are being developed to type each truffle species to overcome the difficulty of identifying these species solely on their morphological traits (1,8,11,17,18,19,22,23). By combining molecular markers with an appropriate sampling strategy, we may be able to critically evaluate the truffle reproductive system and life cycle even without reproducing the entire life cycle in the lab.…”

mentioning

confidence: 99%

Reevaluation of the Life Cycle of Tuber magnatum

Paolocci

Rubini

Riccioni

et al. 2006

Appl Environ Microbiol

115

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Tuber spp. are ectomycorrhizal ascomycetes that produce ascocarps known as truffles. Basic aspects of Tuber biology have yet to be fully elucidated. In particular, there are conflicting hypotheses concerning the mating system and the ploidy level of the mycorrhizal and truffle hyphae. We used polymorphic microsatellites to compare the allelic configurations of asci with those from the network of the surrounding hyphae in single Tuber magnatum truffles. We then used these truffles to inoculate host plants and evaluated the microsatellite configurations of the resulting mycorrhizal root tips. These analyses provide direct evidence that T. magnatum outcrosses and that its life cycle is predominantly haploid. In addition to its scientific significance, this basic understanding of the T. magnatum life cycle may have practical importance in developing strategies to obtain and select nursery-produced mycorrhizal plants as well as in the management of artificial plantations of this and other Tuber spp.Tuber spp. are Ascomycetes fungi that establish an ectomycorrhizal symbiosis with trees and shrubs. As a result of this mutualistic symbiosis, ascocarps known as truffles are produced. Some Tuber spp. produce edible truffles that, given their distinctive taste and aroma, are highly valued by gourmets. Research on these fungi has focused on promoting the cultivation of these fungi to meet increasing worldwide demand and to provide replacements for the catastrophic decline in their natural production (10). Truffle cultivation is no longer an agronomic practice confined to Europe, where the most profitable species are endemic. Truffle plantations have been established in various countries worldwide, including New Zealand and Israel. Nevertheless, the understanding of many basic aspects of truffle biology is still in its infancy, and the ecological requirements for some of these species are still not known. One of the most elusive goals has been discerning the reproductive system of Tuber spp. The reproductive structures of these species in pure cultures have not been reported, and axenic spore germination remains an unresolved problem (26). Furthermore, the mating-type genes have never been characterized in truffles.Molecular markers are being developed to type each truffle species to overcome the difficulty of identifying these species solely on their morphological traits (1,8,11,17,18,19,22,23). By combining molecular markers with an appropriate sampling strategy, we may be able to critically evaluate the truffle reproductive system and life cycle even without reproducing the entire life cycle in the lab. To date, Tuber melanosporum Vittad. and Tuber magnatum Pico, the finest black and white truffle species, respectively, have been regarded as selfing species. When codominant markers were evaluated, heterozygous ascocarps were not detected (2,3,7,15,16). These studies proceed from the assumption that the ascocarps are diploid (dikaryotic) structures.We recently used simple sequence repeat (SSR) markers and a large survey of...

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“…Molecular markers have been developed to type most of the economically important truffle species (2,10,15,19,23,24,25,33,34), but studies of the environmental and molecular determinants for different portions of the life cycle generally are lacking. The spatial distribution and ecological requirements of these symbiotic fungi vary by species.…”

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confidence: 99%

Genetic and Phylogeographic Structures of the Symbiotic Fungus Tuber magnatum

Rubini

Paolocci

Riccioni

et al. 2005

Appl Environ Microbiol

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The quality and market price of truffles vary with the species and, traditionally, the place of origin. The premium species Tuber magnatum produces white truffles and has a patchy distribution restricted to Italy and some Balkan areas. We used polymorphic microsatellites to evaluate 316 specimens grouped into 26 populations sampled across the species' geographic range to determine if natural populations of T. magnatum are genetically differentiated. We found that the southernmost and the northwesternmost populations were significantly differentiated from the rest of the populations. The simple sequence repeat data also could be used to make inferences about the postglacial T. magnatum expansion pattern. This study is the first to identify a genetic and phylogeographic structure in T. magnatum. The presence of a genetic structure can be of practical interest in tracing truffle populations according to their geographic origin for marketing strategies. Evidence for extensive outcrossing in field populations of T. magnatum also is provided for the first time.Tuber spp. are hypogeous ascomycetes that grow in ectomycorrhizal symbioses with some shrub and tree species and produce ascomata, known as truffles. Some truffles are edible and appreciated worldwide. Their quality and market price depend on the species and, traditionally, the place of origin. Natural truffle production has declined dramatically over the past century (12). This decline and the flourishing truffle market have encouraged large-scale programs for growing these fungi through the planting of nursery-produced mycorrhizal trees. These programs have been widely developed for some species, e.g., Tuber melanosporum Vittad. and Tuber uncinatum Chatin, and most of the commercial demand for these truffles is satisfied by these artificial plantations rather than natural field collections (12). How the deliberate introduction of foreign strains into native populations has affected truffle production and biodiversity remains unknown.The complexity of the truffle life cycle, the difficulties of growing these fungi under controlled conditions, and the lack of reliable phenotypic markers to differentiate morphologically similar species have been major obstacles to understanding the distribution, propagation, and fructification of these hypogeous fungi. Molecular markers have been developed to type most of the economically important truffle species (2,10,15,19,23,24,25,33,34), but studies of the environmental and molecular determinants for different portions of the life cycle generally are lacking. The spatial distribution and ecological requirements of these symbiotic fungi vary by species. Some species are widely distributed and have pronounced morphological and molecular variability (9,11,22,25,27,32), while others have a more restricted distribution and little intraspecific polymorphism in either morphological or genetic traits (3,4,8,9,26). Tuber magnatum Pico, which produces white ascomata, is harvested only in Italy and some countries on the Balkan Peninsul...

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Identification of ectomycorrhizal fungi of the genus Tuber by species‐specific ITS primers

Cited by 113 publications

References 9 publications

Solid‐phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles

Solid‐phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles

Reevaluation of the Life Cycle of Tuber magnatum

Genetic and Phylogeographic Structures of the Symbiotic Fungus Tuber magnatum

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