Sergio Sánchez

Truffles in the genus Tuber (Pezizales) are among the few ectomycorrhizal taxa successfully cultivated worldwide. Australia has recently become one of the top producers of Tuber melanosporum , a high-valued black truffle native to Europe. Truffle co-introductions in the Southern Hemisphere with their Northern Hemisphere plant symbionts are hypothesized to benefit from reduced ectomycorrhizal competition and natural enemies. In this study, we tested whether T. melanosporum in Australia experiences competition release by sampling soils and truffles across France, Spain, Italy, and Australia. Fungal and prokaryotic communities in truffle plantations were assessed in native (European) and non-native (Australian) habitats through ITS and 16S rDNA amplicon sequencing from soil and truffles. Community composition was primarily structured by site of origin and secondarily by presence of brûlé, vegetation-free area induced by truffle production of plant-growth inhibiting compounds, with significant interactions indicating site-dependent brûlé effects. European soils showed higher fungal richness outside the brûlé, with higher evenness overall and for ectomycorrhizal fungi only outside brûlé. T. melanosporum showed higher abundance in Australia, with significant differences restricted to outside the brûlé. Overall, ectomycorrhizal fungi in European soils had more than four times the taxa and higher diversity compared to Australian soils. Among the main competitors, Tomentella , Inocybe , and Trichophaea co-dominated in Europe, versus Scleroderma , Hebeloma , and Tarzetta in Australia. Despite differences in soil microbiomes, bacterial communities within T. melanosporum truffle ascocarps were strikingly similar across sites and continents and were dominated by Bradyrhizobium . Despite high site-level variation, our results support the competition release hypothesis, with reduced enemies benefiting T. melanosporum colonization outside the brûlé in Australia. IMPORTANCE: This study provides the first cross-hemisphere analysis of the truffle microbiome, comparing native and non-native soils and truffles from Europe and Australia. We demonstrate that the remarkable success of Tuber melanosporum cultivation in Australia is compatible with ecological release from competitors, which favors its development outside the brûlé. At the same time, we reveal striking cross-hemispheric similarities in truffle-associated bacterial communities, consistently dominated by Bradyrhizobium . These findings highlight both the novelty and transcontinental relevance of our work, offering new perspectives on fungal ecology and truffle cultivation.

As truffle cultivation expands, growers empirically develop new agronomic management practices aimed at promoting truffle growth such as “truffle nes

The success of truffle cultivation is especially dependent on the quality of truffle-mycorrhized seedlings, which are typically produced in nurseries under aseptic conditions to avoid root colonization by undesired ectomycorrhizal fungi. However, such practices may also eliminate beneficial microorganisms that could support truffle symbiosis and improve seedling quality. In this study, twelve endophytic bacterial and fungal strains, isolated from the Tuber melanosporum environment (gleba tissue, mycorrhizae and truffle brûlé), were tested for their effect on T. melanosporum mycorrhization levels in inoculated Quercus ilex seedlings under nursery conditions. Co-inoculation with a strain of Agrobacterium tumefaciens significantly enhanced root colonization by T. melanosporum, supporting its potential role as mycorrhizal helper bacterium. In contrast, a strain of Trichoderma harzianum negatively affected mycorrhization. The remaining strains did not show significant effects on seedling mycorrhization or seedling growth. Our findings support the hypothesis that specific bacterial strains associated with truffles can act as mycorrhizal helper bacteria, highlighting the potential for co-inoculation strategies to enhance quality of truffle-inoculated seedlings in nurseries. However, further research is needed to gain a deeper understanding of the interactions within the mycorrhizosphere that could contribute to improving nursery seedling quality.

Black truffle, Tuber melanosporum Vittad., production is increasing due to an improvement in cultivation management and to the demand for this highly appreciated fungus. However, this intensification of truffle cultivation has led to the appearance of problems related to pest incidence. Specifically, the truffle beetle, Leiodes cinnamomeus (Panzer, 1793) (Coleoptera: Leiodidae), causes significant losses in black truffle marketability. However, its biology is still poorly known, and no effective agro‐ecological methods exist to mitigate its damage to the truffles. This study aimed at assessing the population dynamics of L. cinnamomeus over four seasons (2019–2023) in an orchard located in NE Spain and relating these dynamics to weather variables and damage to truffle fruit bodies. Moreover, we described the diversity of arthropods captured in the traps in search of potential natural enemies of this beetle. The maximum population peak was observed in November, except for a single season in which it occurred in December. Moreover, the sex ratio was balanced (0.54 on average), but it varied over the growing season and among years. Significant and positive relationships of the population density of truffle beetles with air temperature and relative humidity were observed. The number of beetles per trap and day was strongly linked to heat accumulation. Finally, the Carabid Percus (Pseudopercus) patruelis (L. Daufour, 1820) was identified as a natural enemy of L. cinnamomeus. These results could be used in the future for monitoring and predicting truffle beetle populations.

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.

The fungal biodiversity associated with a stable plant community appears to vary from year to year. To analyse the annual behaviour in the formation of fruiting bodies, a statistical study of the data obtained for 4 years (2009–2012) in an apparently undisturbed Mediterranean ecosystem dominated by the holm oak (Quercus ilex L. subsp. ballota (Desf.) Samp.), located in the Midwest of the Iberian Peninsula, was carried out. These data were related to the main meteorological variables. The 150 species collected showed a significant annual, monthly, and weekly difference in their fruiting during the collection period. All this implies a variation in the annual fungal fruiting which can modify the moment of when maximum peaks of fruiting appear, their duration, and the number of species that compose them. In addition, the results make it possible to establish an annual behaviour pattern, with sporocarp formation throughout the year and four fruiting groups (two of them in the dry season). They also allow for inferring a possible response to climate change, with a delay in the fruiting of the autumn-winter group and earlier fruit bearing in the winter-spring group.

Sustainable agriculture has drawn attention to the maintenance and enhancement of soil health. However, research on soil quality has been carried out mainly in field crops and, to a lesser extent, in mature orchards, neglecting the relevance of assessing the soil quality status in the first years of tree plantations. Therefore, the aim of this study was to assess the soil quality index of young almond orchards located in marginal lands and managed under different practices. The survey was carried out in the Teruel Province (Northeast Spain), in three almond orchards: Alacón (2 years old, 0.75 ha, rainfed, conventionally managed), San Martín (1 year old, 0.4 ha, irrigated, organically managed), and Valdealgorfa (6 years old, 0.2 ha, rainfed, organically managed). The composite soil samples were taken from three spots within each orchard. To determine the soil quality index, four main soil functions were considered: filtering and buffering, nutrient supply, water relations, and crop limitation. The soil quality indices were 0.55, 0.75, and 0.54 for Alacón, San Martín, and Valdealgorfa orchards, respectively. These values suggested that the evaluated soils are adequate for almond production, although they require management actions to improve their quality (for instance, the application of organic amendments) and increase the sustainability of these agroecosystems. Furthermore, this work provides a framework for the assessment of the soil quality in tree orchards at a young stage.