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

BMC Microbiology

Methodology Article

Alessandra Zambonelli¹  Mirco Iotti¹  Elena Baraldi¹  Elena Salerni²  Marco Leonardi³  Marilena Oddis³ 

[1] Dipartimento di Protezione e Valorizzazione Agroalimentare, Alma Mater Studiorum Università di Bologna, via Fanin 46, 40127, Bologna, Italy;Dipartimento di Scienze Ambientali “G. Sarfatti”, Università degli Studi di Siena, via Mattioli 4, 53100, Siena, Italy;Dipartimento di Scienze Ambientali, Università dell’Aquila, via Vetoio, Coppito 1, 67100, L’Aquila, Italy;

关键词: Real-time PCR;    Taq-man probe;    Tuber magnatum;    Soil DNA extraction;    ITS primers;    Truffle production;   

DOI  :  10.1186/1471-2180-12-93

 received in 2011-12-22, accepted in 2012-05-14,  发布年份 2012

来源: Springer

【 摘 要 】

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.

【 授权许可】

Unknown   
© Iotti et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.