Development and validation of a real-time PCR assay for detection and quantification of Tuber magnatum in soil

Abstract

BackgroundTuber magnatum, the Italian white truffle, is the most sought-after edible ectomycorrhizal mushroom. Previous studies report the difficulties of detecting its mycorrhizas and the widespread presence of its mycelium in natural production areas, suggesting that the soil mycelium could be a good indicator to evaluate its presence in the soil. In this study a specific real-time PCR assay using TaqMan chemistry was developed to detect and quantify T. magnatum in soil. This technique was then applied to four natural T. magnatum truffières located in different regions of Italy to validate the method under different environmental conditions.ResultsThe primer/probe sets for the detection and quantification of T. magnatum were selected from the ITS rDNA regions. Their specificity was tested in silico and using qualitative PCR on DNA extracted from 25 different fungal species. The T. magnatum DNA concentration was different in the four experimental truffières and higher in the productive plots. T. magnatum mycelium was however also detected in most of the non-productive plots. Ascoma production during the three years of the study was correlated with the concentration of T. magnatum DNA.ConclusionsTaken together, these results suggest that the specific real-time PCR assay perfected in this study could be an useful tool to evaluate the presence and dynamics of this precious truffle in natural and cultivated truffières.

Ecosystems supporting Tuber magnatum Pico production in Serbia experience specific soil environment seasonality that may facilitate truffle lifecycle completion

“…Long lasting problems of detecting white truffle mycelia or ECM in the soil were solved very recently with development of efficient protocols based on molecular methods (Murat et al, 2005;Zampieri et al, 2010;Iotti et al, 2012), but with puzzling results. Often, mycelia or ECM could not be detected even in highly productive sites, while they were simultaneously detected in unproductive areas (Murat et al, 2005;Zampieri et al, 2010;Iotti et al, 2012;Leonardi et al, 2013). Apparently, mycelia or ECM presence is no clear sign of site productivity and ascocarp appearance remains the most trustful and undeniable evidence of the truffle life cycle completion in situ.…”

How Ectomycorrhizae Structures Boost the Root System?

“…(Gryndler et al 2013), Tuber melanosporum Vittad. Zampieri et al 2012), and Tuber magnatum Picco (Iotti et al 2012).…”

Plant, Soil and Microbes

“…It is not possible to cultivate successfully all the species in truffl e beds (Iotti et al 2012 ;Reyna and Garcia-Barreda 2014 ). Aside from the importance of site suitability and seedling quality, one critical factor in the precocity and regularity of truffl e production has proven to be correct management of the truffl e beds.…”