Overview of molecular methods for quick identification of Tuber species

Truffles represent a resource of great value all over the world. The various species are widespread and occupy various habitats. Methods to identify their value in terms of marketing, cultivation, and the ecological–forestal field are needed. The traditional classification of Tuber species is based on their morphology; nevertheless, the macroscopic and microscopic characteristics of the fruiting body, spores, and ectomycorrhizae may be absent or insufficiently discriminating. This problem is amplified when dealing with valuable truffle species, which are often replaced by truffles with lower organoleptic qualities and commercial value. Today, molecular biology can help to correctly identify them through several widely used methods that are fast, specific, and sensitive. Molecular methods are mostly PCR-based, with primers designed on various targets, genes, or regions widely variable between species but sufficiently conserved within the same species. The DNA regions analyzed for identifying truffles include ribosomal internal transcribed spacer (ITS), ribosomal large subunit (LSU), β-tubulin, and EF-α elongation factor. This review summarizes the main and most reliable DNA-based molecular methods for identifying Tuber species. The methods are listed schematically and clearly for quick consultation and use in all areas of study in which monitoring, and verification of the species is essential

Molecular Biology and Innovative Bacterial Application to Understanding Truffle Ecosystem Dynamics

Truffles are edible ectomycorrhizal fungi of the Tuber genus with considerable economic value. Studying valuable Tuber species is essential not only for their commercial potential but also to understand the little-known aspects of their biology. Today, climate changes and anthropogenic activities strongly affect forest natural production. Consequently, most truffles harvested worldwide are now produced in orchards, where seedlings are inoculated under controlled nursery conditions. Truffles play a central role in the agro-economy of the Marche region. This region boasts diverse natural habitats that provide ideal growing conditions for various truffle species, including the prized white truffle (Tuber magnatum) and the black truffle (Tuber melanosporum). In addition, truffle cultivation represents a significant economic activity in the Marche region, particularly in marginal areas with limited agricultural opportunities. The “Angellozzi Tartuficoltura” is a family company that has refined the knowledge and improved techniques for truffle cultivation, significantly contributing to the richness and uniqueness of the truffle industry in the region. “Angellozzi Tartuficoltura” partnered in this research thesis, which was funded by the Marche Region under the “Progetto di Dottorato Innovativo a caratterizzazione industriale” (POR Marche- FSE 2014/2020). The thesis aims to understand the truffle ecosystem, involving microbiology, botany, ecological-forestry, and molecular biology. The research leads to finding new strategies to improve and increase the production of natural truffle ground or truffle orchards. The strategy proposed consists in the use of bacteria, that take part in the symbiotic relationship between truffles and plants. Numerous studies have revealed their potential roles in affecting the aroma and fitness of the plant, promoting plant and mycelium growth, protecting against pathogens, fixing nitrogen, and modulating spore germination and hyphal growth. The research was conducted in the nursery and the field. We analyzed the effects of Pseudomonas spp. and Bradyrhizobium spp. on stimulating the mycorrhization process and the quality of Quercus ilex seedlings mycorrhized with T. melanosporum. Additionally, we evaluated the influence of Bradyrhizobium japonicum on the quantity of T. magnatum mycelia in the soil of a natural truffle ground in Sant’Angelo in Vado (PU). These findings could provide useful insights for developing innovative cultivation strategies using bacterial inoculation in greenhouses and fields, thereby supporting the regional economy by improving truffle production.Truffles are edible ectomycorrhizal fungi of the Tuber genus with considerable economic value. Studying valuable Tuber species is essential not only for their commercial potential but also to understand the little-known aspects of their biology. Today, climate changes and anthropogenic activities strongly affect forest natural production. Consequently, most truffles harvested worldwide are now produced in orchards, where seedlings are inoculated under controlled nursery conditions. Truffles play a central role in the agro-economy of the Marche region. This region boasts diverse natural habitats that provide ideal growing conditions for various truffle species, including the prized white truffle (Tuber magnatum) and the black truffle (Tuber melanosporum). In addition, truffle cultivation represents a significant economic activity in the Marche region, particularly in marginal areas with limited agricultural opportunities. The “Angellozzi Tartuficoltura” is a family company that has refined the knowledge and improved techniques for truffle cultivation, significantly contributing to the richness and uniqueness of the truffle industry in the region. “Angellozzi Tartuficoltura” partnered in this research thesis, which was funded by the Marche Region under the “Progetto di Dottorato Innovativo a caratterizzazione industriale” (POR Marche- FSE 2014/2020). The thesis aims to understand the truffle ecosystem, involving microbiology, botany, ecological-forestry, and molecular biology. The research leads to finding new strategies to improve and increase the production of natural truffle ground or truffle orchards. The strategy proposed consists in the use of bacteria, that take part in the symbiotic relationship between truffles and plants. Numerous studies have revealed their potential roles in affecting the aroma and fitness of the plant, promoting plant and mycelium growth, protecting against pathogens, fixing nitrogen, and modulating spore germination and hyphal growth. The research was conducted in the nursery and the field. We analyzed the effects of Pseudomonas spp. and Bradyrhizobium spp. on stimulating the mycorrhization process and the quality of Quercus ilex seedlings mycorrhized with T. melanosporum. Additionally, we evaluated the influence of Bradyrhizobium japonicum on the quantity of T. magnatum mycelia in the soil of a natural truffle ground in Sant’Angelo in Vado (PU). These findings could provide useful insights for developing innovative cultivation strategies using bacterial inoculation in greenhouses and fields, thereby supporting the regional economy by improving truffle production

GTPases in Hyphal Growth

Hyphal growth is a fundamental event in filamentous fungi, in that they grow by means of apical expansion of tube-shaped cells (hyphae). This event requires vesicular trafficking, reorganization of the cytoskeleton, construction of the cell wall. The underlying molecular mechanism it is complex and diverse. GTPases play important roles as molecular switches, passing from the active-GTP bound state to the inactive-GDP bound state. These proteins turn various pathways on or off and are crucial regulators of hyphal growth, septation, cell wall synthesis and the exocytic and endocytic pathway. In particular, the roles of Ras, Arf, Rab, Rho have been deepened