RESUMO
Bacterial growing resistance to antibiotics poses a critical threat to global health. This study investigates, for the first time, the antibiofilm properties of Vicia ervilia agglutinin (VEA) from six different V. ervilia accessions against pathogenic bacteria, and the yeast Candida albicans. In the absence of antimicrobial properties, purified VEA significantly inhibited biofilm formation, both in Gram-positive and Gram-negative bacteria, but not in C. albicans. With an inhibitory concentration ranging from 100 to 500 µg/ml, the VEA antibiofilm activity was more relevant against the Gram-positive bacteria Streptococcus aureus and Staphylococcus epidermidis, whose biofilm was reduced up to 50% by VEA purified from accessions #5 and #36. VEA antibiofilm variability between accessions was observed, likely due to co-purified small molecules rather than differences in VEA protein sequences. In conclusion, VEA seed extracts from the accessions with the highest antibiofilm activity could represent a valid approach for the development of an effective antibiofilm agent.
Assuntos
Antibacterianos , Biofilmes , Candida albicans , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Testes de Sensibilidade Microbiana , Biofilmes/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Antibacterianos/farmacologia , Lectinas de Plantas/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Sementes/químicaRESUMO
In order to investigate the intraspecific diversity of wild Humulus lupulus (hop) in Central Italy, 12 populations were evaluated for their genetic polymorphism by means of 13 SSR loci together with six commercial cultivars as a reference. High levels of polymorphism were found across the populations, being 140 the number of multilocus genotypes over 159 samples analyzed. Moreover, the observed heterozygosity was higher than expected in most of the populations. High levels of gene flow were thus envisaged to occur within and among wild populations, and our sampling strategy allowed us to gain insights on the propagation modes of this species, i.e. clonal versus sexual propagation. Nevertheless, a genetic structure of populations with at least five genetically different clusters was disclosed. Private alleles were observed in both wild and cultivated hops. Chemical analysis of bittering and aromatic quality of female flowers from a subset of 8 wild populations revealed a high variability among plants, especially for essential oil components. Overall, the high variability of wild accessions here examined represent a valid source to be exploited in future breeding programs for new or improved hop cultivars development.
Assuntos
Humulus/genética , Fluxo Gênico/genética , Variação Genética/genética , Humulus/fisiologia , Itália , Repetições de Microssatélites/genética , Polimorfismo Genético/genética , Reprodução/genética , Reprodução Assexuada/genéticaRESUMO
Ectomycorrhizal ascomycetes belonging to the genus Tuber produce edible fruiting bodies known as truffles. Tuber aestivum, in particular, is a fungus appreciated worldwide and has a natural distribution throughout Europe. Most of the molecular studies conducted on this species have been focused on the question as to whether or not T. aestivum and the morphologically similar T. uncinatum are conspecific. Conversely, only a handful of studies have assessed the level and distribution of genetic diversity and occurrence of phylogeographic patterns in this species. Here, we analyzed the genetic diversity of T. aestivum over a wide geographic range, performing an extensive sampling of specimens from Turkey, which is novel, to the best of our knowledge. We compared the internal transcribed spacer (ITS) profiles of 45 samples collected in different Turkish areas with those of 144 samples from all over Europe. We identified 63 haplotypes, 32 of which were exclusively present in Turkey. The majority of these haplotyes were also population specific. Haplotype network analysis and statistical tests highlighted the presence of a genetic structure and phylogeographic pattern, with three spatially distinct genetic clusters (northeastern Europe, southern Europe, and Turkey), with Turkey representing a diversity hotspot. Based on these results, we hypothesize the presence of glacial refugia for T. aestivum in Turkey, whereas European populations likely experienced a population bottleneck. The possible occurrence of cryptic species among Turkish T. aestivum samples also emerged. Our results are of practical relevance for the marketing of T. aestivum truffles and mycorrhizal seedlings and the preservation of the biodiversity of this species.
Assuntos
DNA Fúngico/genética , DNA Ribossômico/genética , Variação Genética , Micorrizas/genética , Polimorfismo Genético , DNA Intergênico/genética , Haplótipos , Micorrizas/classificação , Filogenia , Filogeografia , TurquiaRESUMO
Tuberaceae is one of the most diverse lineages of symbiotic truffle-forming fungi. To understand the molecular underpinning of the ectomycorrhizal truffle lifestyle, we compared the genomes of Piedmont white truffle (Tuber magnatum), Périgord black truffle (Tuber melanosporum), Burgundy truffle (Tuber aestivum), pig truffle (Choiromyces venosus) and desert truffle (Terfezia boudieri) to saprotrophic Pezizomycetes. Reconstructed gene duplication/loss histories along a time-calibrated phylogeny of Ascomycetes revealed that Tuberaceae-specific traits may be related to a higher gene diversification rate. Genomic features in Tuber species appear to be very similar, with high transposon content, few genes coding lignocellulose-degrading enzymes, a substantial set of lineage-specific fruiting-body-upregulated genes and high expression of genes involved in volatile organic compound metabolism. Developmental and metabolic pathways expressed in ectomycorrhizae and fruiting bodies of T. magnatum and T. melanosporum are unexpectedly very similar, owing to the fact that they diverged ~100 Ma. Volatile organic compounds from pungent truffle odours are not the products of Tuber-specific gene innovations, but rely on the differential expression of an existing gene repertoire. These genomic resources will help to address fundamental questions in the evolution of the truffle lifestyle and the ecology of fungi that have been praised as food delicacies for centuries.
Assuntos
Ascomicetos/genética , Genoma Fúngico , Características de História de Vida , Micorrizas/genética , Simbiose , Ascomicetos/fisiologia , DNA Fúngico/análise , Micorrizas/fisiologia , Filogenia , Análise de Sequência de DNARESUMO
Truffles are the fruiting structures of ascomycetes in the genus Tuber. Because of their economic importance, truffles have been cultivated for many years using artificially inoculated host plants. Nevertheless, the life cycle and reproductive mode of Tuber spp. are still poorly understood. In filamentous ascomycetes, sexual reproduction is genetically controlled by the mating-type (MAT) locus. Among Tuber spp., the MAT locus has been recently characterized in the black truffles Tuber melanosporum and Tuber indicum. Here, by using sequence information derived from these species and from a Tuber borchii expressed sequence tag (EST) showing similarity to the mat1 gene of Alternaria brassicicola, we embarked on a chromosome-walking procedure to sequence the complete MAT region of T. borchii. This fungus produces highly commercialized whitish truffles and represents a model species for addressing basic questions concerning the life cycle of Tuber spp. We show that T. borchii is heterothallic, as its MAT locus is organized into two idiomorphs, each harbored by different mycelial strains. The alignment of the MAT locus from black truffles and T. borchii reveals that extensive sequence rearrangements and inversions occurred between these species. Moreover, by coupling mating-type analyses to karyological observation, we show that mycelia isolated from ascocarps and mycorrhizae are formed by homokaryotic hyphae.
Assuntos
Ascomicetos/fisiologia , Proteínas Fúngicas/metabolismo , Sequência de Aminoácidos , Ascomicetos/genética , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Reprodução , Especificidade da EspécieRESUMO
Major breakthroughs in our understanding of the life cycles of the symbiotic ascomycetes belonging to the genus Tuber have occurred over the last several years. A number of Tuber species produce edible fruiting bodies, known as truffles, that are marketed worldwide. A better understanding of the basic biological characteristics of Tuber spp. is likely to have tremendous practical relevance for their cultivation. Tuber melanosporum produces the most valuable black truffles and its genome has been recently sequenced. This species is now serving as a model for studying the biology of truffles. Here, we review recent progress in the understanding of sexual reproduction modalities in T. melanosporum. The practical relevance of these findings is outlined. In particular, the discoveries that T. melanosporum is heterothallic and that strains of different mating types compete to persist on the roots of host plants suggest that the spatial and temporal distributional patterns of strains of different mating types are key determinants of truffle fructification. The spatial segregation of the two mating types in areas where T. melanosporum occurs likely limits truffle production. Thus, host plant inoculation techniques and agronomic practices that might be pursued to manage T. melanosporum orchards with a balanced presence of the two mating partners are described.
Assuntos
Ascomicetos/fisiologia , Micorrizas/fisiologia , Agricultura , Ascomicetos/genética , Ascomicetos/crescimento & desenvolvimento , Carpóforos/crescimento & desenvolvimento , Carpóforos/fisiologia , Micorrizas/genética , Micorrizas/crescimento & desenvolvimento , Reprodução/genética , Microbiologia do SoloRESUMO
Tuber spp. are filamentous ascomycetes which establish symbiosis with the roots of trees and shrub species. By virtue of this symbiosis they produce hypogeous ascocarps, known as truffles. Filamentous ascomycetes can reproduce by homothallism or heterothallism depending on the structure and organization of their mating type locus. The first mating type locus in a truffle species has been recently characterized in Tuber melanosporum and it has been shown that this fungus, endemic in Europe, is heterothallic. The availability of sequence information for T. melanosporum mating type genes is seminal to cloning their orthologs from other Tuber species and assessing their reproductive mode. Here we report on the organization of the mating type region in T. indicum, the black truffle species present in Asia, which is the closest relative to T. melanosporum and is characterized by an high level of morphological and genetic variability. The present study shows that T. indicum is also heterothallic. Examination of Asiatic black truffles belonging to different genetic classes, sorted according to the sequence polymorphism of the internal transcribed spacer rDNA region, has revealed sequence variations and rearrangements in both coding and non-coding regions of the mating type locus, to suggest the existence of cryptic species within the T. indicum complex. The presence of transposable elements within or linked to the mating type region suggests a role of these elements in generating the genotypic diversity present among T. indicum strains. Overall, comparative analyses of the mating type locus have thus allowed us to tackle taxonomical and phylogenetic issues within black truffles and make inferences about the evolution of T. melanosporum-T. indicum lineage. Our results are not only of fundamental but also of applied relevance as T. indicum produces edible fruit bodies that are imported also into Europe and thus may represent a biological threat for T. melanosporum.
Assuntos
Ascomicetos/genética , DNA Fúngico/genética , Variação Genética , Filogenia , Reprodução/genética , Especificidade da EspécieRESUMO
The genetic structure of ectomycorrhizal (ECM) fungal populations results from both vegetative and sexual propagation. In this study, we have analysed the spatial genetic structure of Tuber melanosporum populations, a heterothallic ascomycete that produces edible fruit bodies. Ectomycorrhizas from oaks and hazels from two orchards were mapped and genotyped using simple sequence repeat markers and the mating type locus. The distribution of the two T. melanosporum mating types was also monitored in the soil. In one orchard, the genetic profiles of the ascocarps were compared with those of the underlying mycorrhizas. A pronounced spatial genetic structure was found. The maximum genet sizes were 2.35 and 4.70 m in the two orchards, with most manifesting a size < 1 m. Few genets persisted throughout two seasons. A nonrandom distribution pattern of the T. melanosporum was observed, resulting in field patches colonized by genets that shared the same mating types. Our findings suggest that competition occurs between genets and provide basic information on T. melanosporum propagation patterns that are relevant for the management of productive truffle orchards.
Assuntos
Ascomicetos/genética , Repetições de Microssatélites , Variação Genética , Genética Populacional , Itália , Micorrizas/genética , Plantas Comestíveis/genética , Quercus/microbiologia , Reprodução Assexuada/genética , Microbiologia do SoloRESUMO
Truffles are hypogeous ectomycorrhizal (EM) fungi belonging to the genus Tuber. Although outplanting of truffle-inoculated host plants has enabled the realization of productive orchards, truffle cultivation is not yet standardized. Therefore, monitoring the distribution of fungal species in different truffle fields may help us to elucidate the factors that shape microbial communities and influence the propagation and fruiting of Tuber spp. In this study, we compared the fungal biodiversity in cultivated and natural Tuber melanosporum truffle fields located in Central Italy. To this end, ectomycorrhizas (ECM) and soil samples were molecularly analyzed, and an inventory of the fungi associated with Quercus pubescens plants colonized by T. melanosporum, Tuber aestivum or Tuber brumale was compiled. T. melanosporum and T. aestivum were dominant on the cultivated plants, and the number of EM species was markedly lower in the cultivated sites than in the natural sites. However, in the same site, EM biodiversity was higher in T. brumale-colonized plants than in T. melanosporum-colonized plants. These results suggest that different Tuber spp. may have different competitive effects on the other mycobionts. Additionally, in keeping with our previous findings, we found that the number of T. melanosporum genotypes recovered from the soil samples was higher than that of the underlying ECM.
Assuntos
Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Micorrizas/classificação , Micorrizas/isolamento & purificação , Quercus/microbiologia , Microbiologia do Solo , Ascomicetos/genética , Ascomicetos/crescimento & desenvolvimento , Biodiversidade , Técnicas de Cultura , Itália , Dados de Sequência Molecular , Micorrizas/genética , Micorrizas/crescimento & desenvolvimentoRESUMO
Belowground ectomycorrhizal communities are often species rich. Characterization of the ectomycorrhizas (ECMs) underneath native truffle areas and/or cultivation sites is particularly relevant to identifying fungal species that might interfere with or promote truffle propagation and fruiting. Fungal identification at the genus/species level can now be achieved by combining detailed morphological and anatomical descriptions with molecular approaches. In a survey of the mycorrhizal biodiversity of Tuber melanosporum orchards and inoculated host plants in nurseries, we repeatedly sampled ECMs with morphological features resembling those of the ECMs widely known as the AD type. Despite the fact that the AD type is regarded as one of the most competitive fungal species towards Tuber spp., its taxonomical rank has yet to be resolved. By analyzing the 28S and internal transcribed spacer (ITS) rDNA regions, here, we show that AD-type ECMs result from host plant colonization by the pyronemataceous species Trichophaea woolhopeia. Further to this, the 28S and ITS phylogenetic trees built from the AD-type ECMs analyzed sustain the hypothesis that T. woolhopeia is a species complex.
Assuntos
Ascomicetos/classificação , Ascomicetos/crescimento & desenvolvimento , Biodiversidade , Micorrizas/classificação , Micorrizas/crescimento & desenvolvimento , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Genes de RNAr , Dados de Sequência Molecular , Micorrizas/genética , Filogenia , RNA Fúngico/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNARESUMO
⢠The genome of Tuber melanosporum has recently been sequenced. Here, we used this information to identify genes involved in the reproductive processes of this edible fungus. The sequenced strain (Mel28) possesses only one of the two master genes required for mating, that is, the gene that codes for the high mobility group (HMG) transcription factor (MAT1-2-1), whereas it lacks the gene that codes for the protein containing the α-box- domain (MAT1-1-1), suggesting that this fungus is heterothallic. ⢠A PCR-based approach was initially employed to screen truffles for the presence of the MAT1-2-1 gene and amplify the conserved regions flanking the mating type (MAT) locus. The MAT1-1-1 gene was finally identified using primers designed from the conserved regions of strains that lack the MAT1-2-1 gene. ⢠Mating type-specific primer pairs were developed to screen asci and gleba from truffles of different origins and to genotype single ascospores within the asci. These analyses provided definitive evidence that T. melanosporum is a heterothallic species with a MAT locus that is organized similarly to those of ancient fungal lineages. ⢠A greater understanding of the reproductive mechanisms that exist in Tuber spp. allows for optimization of truffle plantation management strategies.
Assuntos
Ascomicetos/genética , Genes Fúngicos , Genes Fúngicos Tipo Acasalamento , Primers do DNA , Reprodução/genética , Simbiose/genéticaRESUMO
⢠In light of the recent finding that Tuber melanosporum, the ectomycorrhizal ascomycete that produces the most highly prized black truffles, is a heterothallic species, we monitored the spatial distribution of strains with opposite mating types (MAT) in a natural truffle ground and followed strain dynamics in artificially inoculated host plants grown under controlled conditions. ⢠In a natural truffle ground, ectomycorrhizas (ECMs), soil samples and fruit bodies were sampled and genotyped to determine mating types. Simple sequence repeat (SSR) markers were also used to fingerprint ECMs and fruit bodies. The ECMs from nursery-inoculated host plants were analysed for mating type at 6 months and 19 months post-inoculation. ⢠In open-field conditions, all ECMs from the same sampling site showed an identical mating type and an identical haploid genotype, based on SSR analysis. Interestingly, the gleba of fruit bodies always demonstrated the same genotype as the surrounding ECMs. Although root tips from nursery-grown plants initially developed ECMs of both mating types, a dominance of ECMs of the same MAT were found after several months. ⢠The present study deepens our understanding of the vegetative and sexual propagation modes of T. melanosporum. These results are highly relevant for truffle cultivation.
Assuntos
Ascomicetos/genética , Genes Fúngicos Tipo Acasalamento , Variação Genética , Genoma Fúngico , Genótipo , Micorrizas/genética , Betulaceae/microbiologia , Biodiversidade , Carpóforos , Haploidia , Quercus/microbiologia , Sequências Repetitivas de Ácido Nucleico , Microbiologia do SoloRESUMO
The Périgord black truffle (Tuber melanosporum Vittad.) and the Piedmont white truffle dominate today's truffle market. The hypogeous fruiting body of T. melanosporum is a gastronomic delicacy produced by an ectomycorrhizal symbiont endemic to calcareous soils in southern Europe. The worldwide demand for this truffle has fuelled intense efforts at cultivation. Identification of processes that condition and trigger fruit body and symbiosis formation, ultimately leading to efficient crop production, will be facilitated by a thorough analysis of truffle genomic traits. In the ectomycorrhizal Laccaria bicolor, the expansion of gene families may have acted as a 'symbiosis toolbox'. This feature may however reflect evolution of this particular taxon and not a general trait shared by all ectomycorrhizal species. To get a better understanding of the biology and evolution of the ectomycorrhizal symbiosis, we report here the sequence of the haploid genome of T. melanosporum, which at approximately 125 megabases is the largest and most complex fungal genome sequenced so far. This expansion results from a proliferation of transposable elements accounting for approximately 58% of the genome. In contrast, this genome only contains approximately 7,500 protein-coding genes with very rare multigene families. It lacks large sets of carbohydrate cleaving enzymes, but a few of them involved in degradation of plant cell walls are induced in symbiotic tissues. The latter feature and the upregulation of genes encoding for lipases and multicopper oxidases suggest that T. melanosporum degrades its host cell walls during colonization. Symbiosis induces an increased expression of carbohydrate and amino acid transporters in both L. bicolor and T. melanosporum, but the comparison of genomic traits in the two ectomycorrhizal fungi showed that genetic predispositions for symbiosis-'the symbiosis toolbox'-evolved along different ways in ascomycetes and basidiomycetes.
Assuntos
Ascomicetos/genética , Evolução Molecular , Genoma Fúngico/genética , Simbiose/genética , Carboidratos , Elementos de DNA Transponíveis/genética , Carpóforos/metabolismo , Genes Fúngicos/genética , Genômica , Haploidia , Dados de Sequência Molecular , Análise de Sequência de DNA , Enxofre/metabolismoRESUMO
Tuber melanosporum is an ectomycorrhizal ascomycete producing edible ascocarps. The prevalent view is that this species strictly selfs, since genetic analyses have never detected heterozygotic profiles in its putatively diploid/dikaryotic gleba. The selfing model has also forged the experimental approaches to assess the population genetic variability. Here, the hypothesis that T. melanosporum outcrosses was tested. To this end, SSR (simple sequence repeats) and ITS (internal transcribed spacer) markers were employed to fingerprint asci and the surrounding gleba within single ascocarps. The distribution of genetic variability was also investigated at different geographical levels using single (SSR and ITS) and multilocus (AFLP, amplified fragment length polymorphism) markers. It is shown that T. melanosporum outcrosses since asci display additional alleles besides those present in the surrounding, uniparental, gleba. Furthermore, SSR and AFLP data reveal a high rate of intrapopulation diversity within samples from the same ground and root apparatus and the highest rate of genetic variability within the southernmost populations of the distributional range. These data call for a profound re-examination of T. melanosporum mating system, life cycle and strategies for managing man-made plantations. They also strongly support the idea that the last glaciation restricted the species distribution to the Italian and Spanish peninsulas.
Assuntos
Ascomicetos/genética , Cruzamentos Genéticos , DNA Fúngico , Variação Genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados/métodos , DNA Intergênico , DNA Ribossômico , Genética Populacional , Haplótipos , Endogamia , Repetições Minissatélites , Micorrizas/genética , Polimorfismo de Nucleotídeo Único , Reprodução/genéticaRESUMO
Retrotransposons are suitable targets for developing molecular markers for population genetics studies. Transposable elements have not yet been isolated from ectomycorrhizal fungi of the genus Tuber. In this paper, we report on the isolation and characterization of Tmt1, an LTR-retrotransposon from Tuber melanosporum. The Tmt1 sequence shows relatedness to Ty3/gypsy retrotransposons from which it differentiates for the presence of a dUTPase extra-domain between protease and reverse transcriptase. Phylogenetic analyses suggest a horizontal transfer of the dUTPase gene (dut) from a fungal host genome. The presence of non-identical LTRs and degenerate ORFs substantiate an ancient integration of Tmt1 in T. melanosporum genome. Furthermore, transcripts analyses proved an inactive status of Tmt1, whereas Southern analysis showed that Tmt1 is a repetitive T. melanosporum species-specific element. Tmt1-based markers will help us to gain more insights into population biology in this fungal species.
Assuntos
Ascomicetos/genética , DNA Fúngico/isolamento & purificação , Retroelementos/genética , Elementos de DNA Transponíveis/genética , DNA Fúngico/classificação , Marcadores Genéticos , Genoma Fúngico/genética , Dados de Sequência Molecular , Filogenia , Pirofosfatases/genética , DNA Polimerase Dirigida por RNA/genética , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Sequências Repetidas TerminaisRESUMO
Below-ground ectomycorrhizal communities are often species-rich, and monitoring their dynamics is important to understand the conditions that promote truffle fructification. Characterization of the different ectomycorrhizas (ECM) at the species level can now be achieved by combining detailed morphological and anatomical descriptions with molecular approaches. Following this strategy, we have characterized ectomycorrhizal biodiversity in an artificial Tuber melanosporum plantation. Although the plantation was unproductive, T. melanosporum mycorrhizas were the most present and two Tuber-like mycorrhizal morphotypes, named ECMm1 and ECMm3, showing distinctive features were found. Internal transcribed spacer (ITS) phylogenetic analysis demonstrated that ECMm3 is related to the Tuber rufum/Tuber ferrugineum species complex, whereas ECMm1 shows the highest ITS similarity with Tuber scruposum and fungi-colonizing Epipactis roots. The results presented here provide more insights into genetic variability, mycorrhizal morphology, and below-ground distribution of fungi associated with artificial truffle plantations.
Assuntos
Agricultura , Basidiomycota , Biodiversidade , Micorrizas/genética , Micorrizas/isolamento & purificação , Micorrizas/citologia , Filogenia , Microbiologia do SoloRESUMO
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.
Assuntos
Ascomicetos/crescimento & desenvolvimento , Repetições de Microssatélites/genética , Micorrizas/fisiologia , Quercus/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Ascomicetos/genética , Ascomicetos/fisiologia , Cruzamentos Genéticos , DNA Fúngico/análise , DNA Fúngico/isolamento & purificação , Haploidia , Reação em Cadeia da Polimerase , Esporos Fúngicos/genética , Esporos Fúngicos/fisiologiaRESUMO
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.
Assuntos
Ascomicetos/classificação , Ascomicetos/crescimento & desenvolvimento , DNA Fúngico/análise , Variação Genética , Técnicas de Tipagem Micológica , Filogenia , Simbiose , Ascomicetos/genética , Croácia , Itália , Desequilíbrio de Ligação , Repetições de Microssatélites/genética , EslovêniaRESUMO
Tuber spp. are ectomycorrhizal fungi that establish symbioses with shrubs and trees. Because of their different smell and taste, Tuber uncinatum and Tuber aestivum are two truffle morphotypes with a different market value, but whether or not T. uncinatum and T. aestivum are different taxa is still an open debate among mycologists. In order to identify molecular keys characterizing both T. aestivum and T. uncinatum morphotypes, ITS/RFLPs analyses were carried out on a large collection of samples from all over Italy and from other European countries, followed by a study of the phylogenesis of ITS, beta-tubulin and EF 1-alpha genes, on representative samples. The present study provides compelling evidence that: (i) T. uncinatum and T. aestivum belong to the same species, (ii) neither morphotype presents a specific molecular fingerprint, but they may even share identical alleles at any of the loci analysed; (iii) T. aestivum is most likely under a selfing reproductive mode. Our findings suggest that ecological, rather than genetic causes may account for differences in sporal morphology, taste and smell between T. aestivum and T. uncinatum truffles.