Melanosporum Research Articles

Peat-based amendment of soils reduces the complexity of the volatile profile in cultivated black truffles.

Truffle cultivation is evolving rapidly and new agronomic practices such as 'truffle nests' (localized peat amendments of the orchard soil) are being developed. Truffle nests improve the shape of truffles and their depth in the soil and reduce the occurrence of insect damage but have also raised concerns about their impact on the ripeness and maturity of the harvested truffles. In this study, the effect of the nests on the volatile organic compounds profile and the aromatic profile of black truffles was evaluated, as well as the existence of perceptible sensorial differences in truffles. For this, truffles growing in nests were compared with truffles growing in the bulk soil of the same host tree. Gas chromatography showed that nest truffles had a less complex volatile organic compound profile than bulk-soil truffles. Olfactometry indicated that nest truffles were associated with higher modified frequency values of odorants corresponding to sulfur-containing compounds. Despite this, sensory evaluation with consumers could not clearly show that nest truffles can be distinguished sensorially from bulk-soil truffles. The results prove that soil conditions can influence the aromatic profile of truffles and thus suggest the possibility of managing truffle aroma using agronomic practices. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Application of Quantitative Magnetic Resonance Imaging (QMRI) to Evaluate the Effectiveness of Ultrasonic Atomization of Water in Truffle Preservation.

Truffles of the Tuber genus (Pezizales, Ascomycetes) are among the most valuable and expensive foods, but their shelf life is limited to 7-10 days when stored at 4 °C. Alternative preservation methods have been proposed to extend their shelf life, though they may alter certain quality parameters. Recently, a hypogeal display case equipped with an ultrasonic humidity system (HDC) was developed, extending the shelf life to 2-3 weeks, depending on the truffle species. This study assesses the efficacy of HDC in preserving Tuber melanosporum and Tuber borchii ascomata over 16 days, using quantitative magnetic resonance imaging (QMRI) to monitor water content and other parameters. Sixteen T. melanosporum and six T. borchii ascomata were stored at 4 °C in an HDC or a static fridge (SF) as controls. QMRI confirmed that T. borchii has a shorter shelf life than T. melanosporum under all conditions. HDC reduced the rate of shrinkage, water, and mass loss in both species. Additionally, the Apparent Diffusion Coefficient (ADC), longitudinal relaxation time (T1), and transverse relaxation time (T2), which reflect molecular changes, decreased more slowly in HDC than SF. QMRI proves useful for studying water-rich samples and assessing truffle preservation technologies. Further optimization of this method for industrial use is needed.

Black and white truffles as a source of neuroactive biogenic amines

This study reports the first data about content of serotonin, histamine, and dopamine in black and white truffles fruiting body based on UHPLC-ESI-QqQ-MS/MS. Here we have demonstrated that black (Tuber macrosporum) and white (T. magnatum) truffles are a source of biogenic amines. Also, we demonstrated the absence of correlation between the quality of truffles and the content of biogenic amines.

Effect of Different Coating Materials on Black Périgord Truffle (Tuber melanosporum) Aroma Profile and Its Shelf Life

AbstractBlack Périgord truffles (Tuber melanosporum) are the highest-priced edible fungus in the world due to their unique flavour, rarity, short growing season, difficulty in mass cultivating, and short shelf-life. The current industrial practices have not been effective in extending truffle shelf-life while preserving its aroma profiles. This study aimed to determine the effectiveness of several preservation methods on Australian-grown black Périgord truffles, which include assessing the changes in the volatile organic compounds (VOCs) of truffles treated with edible coatings, antimicrobial agents, or antioxidants such as chitosan, gum Arabic, kafirin, natamycin, tocopherol, vitamin C, and citric acid at the interval of 0, 7, and 14 days of storage. The study also aimed to assess the capability of gamma-cyclodextrin (γ-CD) in encapsulating truffle VOCs at the intervals of 0, 14, and 28 days of storage. Among all the edible coatings, chitosan-treated truffles had the least change in VOCs, especially the black truffle aroma volatile markers, 2-methyl-1-butanal, 2,4-dithiapentane, and dimethyl sulphide. Chitosan also resulted in no significant changes (P < 0.05) in the carbon dioxide emissions of truffle. The PCA plots showed that chitosan-coated samples displayed the least changes. The sole application of antimicrobial agents or antioxidants was ineffective in delaying the deterioration process. On the other hand, the results show that γ-CD was able to encapsulate 30 truffle’s VOCs, which included 3-methyl-1-butanal, 2-methyl-1-butanol, dimethyl sulphide, and 2,4-dithiapentane with no significant changes over the storage period.

Comparative Study of Mechanical and Biological Pretreatment for Releasing Spores of Black Truffle Tuber aestivum

It is well known that the number of true truffles in the wild is decreasing. The aim of the study was to develop an effective, simple and affordable method of asci disruption to release black truffle spores. It was shown that the spore release can be achieved by different ways, such as mechanical or biological destruction. Mechanical homogenization of fruiting bodies using an immersion blender in tandem with a ball mill was shown to be effective and led to destruction of at least 85% of asci and release of spores. Also, the first approach we applied was the biological method of spore activation performed by African and grape snails. As a result of digestion of truffle fruiting bodies, the spores not only lost their protective shells, but also changed their morphology, which promoted their germination in vitro. The spores obtained using these two methods are capable of forming mycelial hyphae on nutrient media. The results of our study can be used to prepare inoculum of Tuber spp. and to obtain their pure cultures in agriculture.

Volatilome changes during black truffle (Tuber melanosporum) ontogeny

The aroma is critical in the reproductive biology of truffles and in their commercial quality. However, previous research has almost exclusively focused on characterizing ripe ascocarps. We characterized the volatilome of the highly-prized black truffle (Tuber melanosporum) ascocarps from July, in an early development stage, to March, in the late harvesting season, and investigated the relationships among aroma, ascocarp growth and morphogenetic development. The aroma profile was analyzed using a head space gas chromatography technique coupled with mass spectrometer. Seventy-one volatile compounds were identified and three development stages were clearly distinguished according to the volatile profile. In unripe ascocarps of July-September, the profile was dominated by methanethiol (19 %), 4-penten-2-ol (11 %) and acetone (11 %), the monthly mean weight of ascocarps ranged 2–20 g, and morphogenetic stages 4-6a were prevalent. In unripe ascocarps of October-December, the most abundant volatiles were 4-penten-2-ol (21 %), methanethiol (20 %) and ethanol (13 %), the monthly mean ascocarp weight ranged 28–43 g, and morphogenetic stages 6a, 6b-c were prevalent. In ripe ascocarps (December-March), the most abundant volatiles were 4-penten-2-ol (17 %), dimethyl sulfide (16 %) and ethanol (10 %), ascocarp weight did not increase significantly, and 6b-c was practically the sole morphogenetic stage. Thirty volatiles were associated to one of these three development stages. Amongst those with higher occurrence, 4-penten-2-ol, dimethyl sulfide, ethyl acetate, 2-pentanol and 2-butanone were associated to ripe truffles, whereas methanethiol, isobutyl isobutyrate, butanedione and 3-methylanisole were associated to unripe truffles.

Elaboration and Characterization of Novel Kombucha Drinks Based on Truffles (Tuber melanosporum and Tuber aestivum) with Interesting Aromatic and Compositional Profiles.

The organoleptic and bioactive properties of truffles place these fungi as interesting materials for use in the of design functional foods based on fruiting bodies outside commercial standards. Moreover, kombucha beverages have become more popular in the Western world, leading to novel drinks using alternative substrates instead of tea leaves. In this work, two truffle species (Tuber melanosporum, TMEL; Tuber aestivum, TAES) and three different symbiotic consortia of bacteria and yeasts (SCOBYs: SC1, SC2, and SC3) were tested. Fermentation (21 days) was monitored in terms of physicochemical (pH, viscosity), biochemical (total carbohydrates, alcohol, soluble proteins, phenolic compounds), and sensory attributes (volatile organic compounds, VOCs). The obtained pH ranges were adequate, alcohol levels were undetectable or very low, and sugar content was lower than in traditional kombuchas or other beverages. In most cases, the usual bottling time could be applied (7-10 days), although longer fermentations are recommended (14 days) to reach higher protein and phenolic compounds contents. Truffle kombuchas produced up to 51 volatile organic compounds (alcohols, acids, esters, ketones, and aldehydes, among others), with TMEL showing a more complex profile than TAES. During the first week, acidic compound production was observed, especially acetic acid. Similar behavior in the VOC profile was reported with different SCOBYs.

Transcriptome analysis reveals the effect of cold storage time on the expression of genes related to oxidative metabolism in Chinese black truffle.

Chinese black truffle (Tuber indicum) is a hypogenous fungus of great value due to its distinctive aroma. In this study, both transcriptome and physicochemical analyses were performed to investigate the changes of nutrients and gene expression in truffle fruiting bodies during cold storage. The results of physicochemical analysis revealed the active metabolism of fruiting bodies in cold storage, showing the decreased contents of protein and soluble sugar, the variations in both polyphenol oxidase activity and total phenol content, and the detrimental effect of reactive oxygen species production caused by heavy metals (cadmium and lead) in truffles. Transcriptome analysis identified a total of 139,489 unigenes. Down-regulated expression of genes encoding the catalase-like domain-containing protein (katE), glutaredoxin protein (GRX), a copper/zinc superoxide dismutase (Sod_Cu), and aspartate aminotransferase (AAT) affected the degradation metabolism of intracellular oxides. Ribulose-5-phosphate-3-epimerase (RPE) was a key enzyme in response to oxidative stress in truffle cells through the pentose phosphate pathway (PPP). A total of 51,612 simple sequence repeats were identified, providing valuable resources for further genetic diversity analysis, molecular breeding, and genetic map-ping in T. indicum. Transcription factors GAL4 and SUF4-like protein were involved in glucose metabolism and histone methylation processes, respectively. Our study provided a fundamental characterization of the physicochemical and molecular variations in T. indicum during the cold storage at 4°C, providing strong experimental evidence to support the improvement of storage quality of T. indicum.

Odour Fingerprints of Black (Tuber mesentericum) and Bianchetto (Tuber borchii) Truffles from Different Areas of the Campania Region

Odour Fingerprints of Black (Tuber mesentericum) and Bianchetto (Tuber borchii) Truffles from Different Areas of the Campania Region

Changes in Black Truffle (Tuber melanosporum) Aroma during Storage under Different Conditions.

The enticing aroma of truffles is a key factor for their culinary value. Although all truffle species tend to be pricy, the most intensely aromatic species are the most sought after. Research into the aroma of truffles encompasses various disciplines including chemistry, biology, and sensory science. This study focusses on the chemical composition of the aroma of black truffles (Tuber melanosporum) and the changes occurring under different storage conditions. For this, truffle samples were stored under different treatments, at different temperatures, and measured over a total storage time of 12 days. Measurements of the truffle aroma profiles were taken with SPME/GC-MS at regular intervals. To handle the ample data collected, a systematic approach utilizing multivariate data analysis techniques was taken. This approach led to a vast amount of data which we made publicly available for future exploration. Results reveal the complexity of aroma changes, with 695 compounds identified, highlighting the need for a comprehensive understanding. Principal component analyses offer initial insights into truffle composition, while individual compounds may serve as markers for age (formic acid, 1-methylpropyl ester), freshness (2-Methyl-1-propanal; 1-(methylthio)-propane), freezing (tetrahydrofuran), salt treatment (1-chloropentane), or heat exposure (4-hydroxy-3-methyl-2-butanone). This research suggests that heat treatment or salt contact significantly affects truffle aroma, while freezing and cutting have less pronounced effects in comparison. The enrichment of compounds showing significant changes during storage was investigated with a metabolomic pathway analysis. The involvement of some of the enriched compounds on the pyruvate/glycolysis and sulfur pathways was shown.

Black Truffle Aroma Evaluation: SPME-GC-MS vs. Sensory Experts.

Nowadays, the truffle aroma attribute is not included as a quality parameter in the current recommendation that explains the truffle quality (UNECE standard 53 FFV3) and establishes the truffle commercial categories. However, the aroma is the main reason why truffles are worldwide appreciated. Indeed, more than 30 aromatic molecules compose it, and this is the reason why the human evaluation and identification of these odorants, without previous training, is quite subjective. Analytical techniques such as gas chromatography techniques, however, can establish an aromatic profile and detect potential aromatic markers. In this study, 16 tasting experts were trained to make more objective the truffle aroma evaluation and odorants identification. For this, a comparison between solid-phase microextraction gas chromatography coupled with mass spectrometry (SPME-GC-MS) and sensory expert evaluation was carried out in six sessions during different harvesting times in the black truffle season (December, January, and February). Both techniques were able to separate truffles depending on the harvesting time. Also, a list of volatile organic compounds related to the aromatic attributes was reported. This information will help to provide a more objective T. melanosporum truffle sensory evaluation.

Habitat is more important than climate for structuring soil fungal communities associated in truffle sites

The ectomycorrhizal fungi Tuber melanosporum Vittad. and Tuber aestivum Vittad. produce highly valuable truffles, but little is known about the soil fungal communities associated with these truffle species in places where they co-occur. Here, we compared soil fungal communities present in wild and planted truffle sites, in which T. melanosporum and T. aestivum coexist, in Mediterranean and temperate regions over three sampling seasons spanning from 2018 to 2019. We showed that soil fungal community composition and ectomycorrhizal species composition are driven by habitat type rather than climate regions. Also, we observed the influence of soil pH, organic matter content and C:N ratio structuring total and ectomycorrhizal fungal assemblages. Soil fungal communities in wild sites revealed more compositional variability than those of plantations. Greater soil fungal diversity was found in temperate compared to Mediterranean sites when considering all fungal guilds. Ectomycorrhizal diversity was significantly higher in wild sites compared to plantations. Greater mould abundance at wild sites than those on plantation was observed while tree species and seasonal effects were not significant predictors in fungal community structure. Our results suggested a strong influence of both ecosystem age and management on the fungal taxa composition in truffle habitats.

Attraction of entomopathogenic nematodes to black truffle and its volatile organic compounds: A new approach for truffle beetle biocontrol

The European truffle beetle, Leiodes cinnamomeus, is the most important pest in black truffle (Tuber melanosporum) plantations. Entomopathogenic nematodes (EPNs) are a promising biological control agents against L. cinnamomeus. EPNs may employ multiple sensory cues while seeking for hosts, such as volatile organic compounds (VOCs) and CO2 gradients. We report for the first time the attraction of EPNs to truffle fruitbodies, and identified some VOCs potentially playing a key role in this interaction. We conducted olfactometer assays to investigate the attraction behavior of Steinernema feltiae and Steinernema carpocapsae towards both T. melanosporum fruitbodies and larvae of L. cinnamomeus. Subsequently, a chemotaxis assay using agar plates was performed to determine which of the 14 of the main VOCs emitted by the fruitbodies attracted S. feltiae at low (0.1 %) and high (mg/100 g truffle) concentrations. Both EPN species were attracted to mature fruitbodies of T. melanosporum, which may enhance the likelihood of encountering L. cinnamomeus during field applications. L. cinnamomeus larvae in the presence of truffles did not significantly affect the behavior of EPNs 24 h after application, underscoring the importance of the chemical compounds emitted by truffles themselves. Chemotaxis assays showed that four long-chain alcohol compounds emitted by T. melanosporum fruitbodies attracted S. feltiae, especially at low concentration, providing a first hint in the chemical ecology of a little-studied ecological system of great economical value. Further studies should be conducted to gain a finer understanding of the tritrophic interactions between T. melanosporum, EPNs, and L. cinnamomeus, as this knowledge may have practical implications for the efficacy of EPNs in the biological control of this pest.

Potential of a smart gelatine hydrogel-based package to extend fresh black truffle (Tuber melanosporum) shelf-life preserving its aroma profile

Many post-harvest technologies have been applied to prolong the shelf life of black truffles; however, most of them negatively modify the genuine fresh truffle aroma. A novel edible gelatine hydrogel-based package (GHP) was tested to extend the shelf life of truffles and trap the aromatic compounds released by fresh truffles. First, the key physicochemical properties and microbiological and sensory attributes of the materials were characterised. The aromatic profile (by gas chromatography and a trained panel) and microbiological populations were monitored for 35 days in truffles stored in GHP and gelatine. Truffle preservation in GHP was compared to that in microperforated modified atmosphere packages (MAP) and that in macroperforated packages (C). The gelatine hydrogel exhibited extraordinarily high oxygen permeability and maintained a low microorganism load during storage (<4 log CFU·g−1 after 28 days). Some key volatile organic compounds were detected in the gelatine after seven days. Gelatine from GHP reduced microbial growth in truffles compared to the C and MAP conditions at day 21. Firmness loss at day 28 was an indicator of spoilage behaviour; therefore, 21 days was selected as the shelf-life extension of truffles under GHP, obtaining fresh truffles with high quality and an edible gelatine hydrogel with truffle aroma.

Effect of Bacteria Inoculation on Colonization of Roots by Tuber melanosporum and Growth of Quercus ilex Seedlings.

Tuber melanosporum is an ascomycete that forms ectomycorrhizal (ECM) symbioses with a wide range of host plants, producing edible fruiting bodies with high economic value. The quality of seedlings in the early symbiotic stage is important for successful truffle cultivation. Numerous bacterial species have been reported to take part in the truffle biological cycle and influence the establishment of roots symbiosis in plant hosts and the development of the carpophore. In this work, three different bacteria formulations were co-inoculated in Quercus ilex L. seedlings two months after T. melanosporum inoculation. At four months of bacterial application, the T. melanosporum ECM root tip rate of colonization and bacterial presence were assessed using both morphological and molecular techniques. A 2.5-fold increase in ECM colonization rate was found in the presence of Pseudomonas sp. compared to the seedlings inoculated only with T. melanosporum. The same treatment caused reduced plant growth either for the aerial and root part. Meanwhile, the ECM colonization combined with Bradyrhizobium sp. and Pseudomonas sp. + Bradyrhizobium sp. reduced the relative density of fibrous roots (nutrient absorption). Our work suggests that the role of bacteria in the early symbiotic stages of ECM colonization involves both the mycorrhizal symbiosis rate and plant root development processes, both essential for improve the quality of truffle-inoculated seedlings produced in commercial nurseries.

Retracted: Black Truffle Extract Exerts Antidiabetic Effects through Inhibition of Inflammation and Lipid Metabolism Regulation.

[This retracts the article DOI: 10.1155/2022/6099872.].

A way to mechanically activate a suspension of black truffle spores

True truffles, or fungi of the genus Tuber, are among the most valuable, rare, expensive, and mysterious fungi on the planet. The reproduction of truffle fungi in the wild is a complex process that depends on many factors, for example, the presence of suitable host trees, as well as the optimal parameters of temperature, moisture, soil pH, soil symbionts, and other organisms. Successful production and reproduction of truffles require a large number of conditions to be met, primarily the release of their spores from ascii, germination under favorable conditions, and distribution in the environment. The present study aims to develop a method for mechanically breaking ascii in order to release the spores of black truffle. It is shown that homogenization by means of an immersion blender together with a vibrating chopper is a highly effective way to break ascii. The microscopic examination of the specimens at all stages of the experiment revealed that this method leads to the breakdown of at least 85% of ascii and the release of truffle spores. The use of centrifugation approaches enables the separation of black truffle spores from hyphal debris, fungal symbionts, and various associated organisms. Such spores can form mycelium on growth media. The study results can be used to prepare Tuber sp. inoculum and obtain their axenic cultures. Thus, the introduction of methods for releasing spores from ascii improves the efficiency of black truffle cultivation.

Niche construction by two ectomycorrhizal truffle species (Tuber aestivum and T. melanosporum)

Niche construction by biota has been frequently reported for animals and plants, but fungi have received less attention. For mycorrhizal fungi, mutualistic niche construction has been proposed where partners construct niches for each other. However, few eco-evolutionary mechanisms leading to mutualistic symbiotic feedback have been described. Here we report niche construction by two ectomycorrhiza-forming species of truffles, Tuber aestivum and T. melanosporum. The soils of 263 spots of these Tuber, which create bare patches (brûlés), have been monitored (9–10 years) in natural forests to analyze whether they meet the criteria to satisfy the mutualistic niche construction process. Soil habitat modification by these Tuber can be seen in their brûlés, where the vegetation of arbuscular mycorrhizal plants and soil organisms are largely suppressed. Tuber melanosporum brûlés were smaller more productive, and with lower vegetation cover than T. aestivum. Interactions among soil carbonates, pH, and brûlé size are associated with increased carpophore productivity, indicating positive feedback between habitat modification and fungal fitness. Both species modified the habitat differentially. Tuber aestivum brûlés had higher pH and lower total carbonate than soils outside. Tuber melanosporum brûlés had lower total carbonate and higher exchangeable Ca2+ and active carbonate than soils outside. In conclusion, brûlés show a mutualistic niche construction process because modifies their soil environment and increases the fitness of both Tuber spp. Enhanced carbonate weathering by truffles resulted in lower soil quality with a negative impact on plants, indicating that mutualistic niche construction does not equally benefit both partners.

From Nontargeted to Targeted Analysis: Feature Selection in the Differentiation of Truffle Species (Tuber spp.) Using 1H NMR Spectroscopy and Support Vector Machine.

The price of different truffle types varies according to their culinary value, sometimes by more than a factor of 10. Nonprofessionals can hardly distinguish visually the species within the white or black truffles, making the possibility of food fraud very easy. Therefore, the identification of different truffle species (Tuber spp.) is an analytical task that could be solved in this study. The polar extract from a total of 80 truffle samples was analyzed by 1H NMR spectroscopy in combination with chemometric methods covering five commercially relevant species. All classification models were validated applying a repeated nested cross-validation. In direct comparison, the two very similar looking and closely related black representatives Tuber melanosporum and Tuber indicum could be classified 100% correctly. The most expensive truffle Tuber magnatum could be distinguished 100% from the other relevant white truffle Tuber borchii. In addition, signals for a potential Tuber borchii and a potential Tuber melanosporum marker for targeted approaches could be detected, and the corresponding molecules were identified as betaine and ribonate. A model covering all five truffle species Tuber aestivum, Tuber borchii, Tuber indicum, Tuber magnatum, and Tuber melanosporum was able to correctly discriminate between each of the species.

European Tuber melanosporum plantations: adaptation status in Hungary, mycorrhizal level, and first ascocarp detection in two truffle orchards

Tuber melanosporum is one of the most economically important truffle species. Besides harvesting from its natural habitats, this truffle can also be extensively grown through artificial cultivation. However, the natural habitat of T. melanosporum has drastically declined, and the demand for the truffle in society is rapidly increasing. Therefore, enhancing production in truffle orchards by seeking new places for the establishment and regularly monitoring its adaptability might be an effective method for ensuring the sustainable productivity of the species. As a truffle science, recent information is important to further success in the growth of this truffle species. This study reports mycorrhization level and ascocarp production in two truffle plantations in Hungary. The estimated mycorrhization levels of the host plants were 43.36% in Biatorbágy and 42.93% in Jászszentandrás plantations. In March 2020, the 6-year-old and 18-year-old T. melanosporum plantations yielded around 100 g and 980 g of ascocarps, respectively. In general, adaptation of mycorrhizal seedlings in Hungary may become more effective as present management practices improve.