Detection of summer truffle (Tuber aestivum Vittad.) in ectomycorrhizae and in soil using specific primers

Abstract

Tuber aestivum is becoming an important commodity of great economical value in some European countries. At the same time, it is a highly protected organism in other countries, where it needs careful treatment. A reliable method of detection in roots and soil is thus needed for assessment of geographic distribution, ecological studies and inoculation efficiency testing in man-made experiments. A PCR-based method of detection of T. aestivum using specific primers was therefore developed. A pair of PCR primers Tu1sekvF/Tu2sekvR selective for T. aestivum and some genotypes of Tuber mesentericum was designed on the basis of the known internal transcribed spacer T. aestivum sequences. TaiI restriction cleavage was then used to distinguish the two species. The selectivity of the designed primer pair was evaluated using DNA extracted from specimens of a further 13 Tuber spp. Subsequently, the selectivity and robustness to false-positive results with nontarget DNA of the designed primers was compared with two other primer pairs (UncI/UncII and BTAE-F/BTAEMB-R). The occurrence of T. aestivum in soil and ectomycorrhizae collected in its native habitat has been successfully detected using the designed primers and nested PCR. The method is reliable and thus suitable for detection of T. aestivum in the field.

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

“…mycelium in soil can be considered crucial for monitoring the status of a cultivated truffle orchard before ascoma production [21]. It is also a powerful tool for assessing truffle presence in natural forests in those countries where ascoma harvesting is forbidden [22] or where all truffle collectors have open access to forests and woodlands [1]. This is particularly important for T. magnatum as the truffle production sites, in natural truffières, are dispersed and not visible to the naked eye, unlike black truffles ( T. melanosporum and T. aestivum ) which produce burnt areas (called “brûlée” in France, “bruciate” or “pianello” in Italy) around the productive trees where grass development is inhibited [1].…”

Plant, Soil and Microbes

“…Distributed in almost all Europe (between 37° and 57° N), North Africa, Korea and China (Weden et al 2004 ;Granetti et al 2005 ;Hall et al 2007 ;Jeandroz et al 2008 ;Milenkovic et al 2009 ;Yun and Liu 2009 ;Chevalier 2010 ;Streiblova et al 2010 ;Gryndler et al 2011 ;Gogan et al 2012 ;Salerni et al 2013 ;Azul et al 2014 ;Gezer et al 2014 Globose ascoma, often lobbed or irregular, often with a depressed base, but without cavities, 2-3(-7) cm in diameter (Fig. 2a ).…”

Isolation and Identification of Allelochemicals from Ascocarp of Tuber Species

“…Distributed in almost all Europe (between 37° and 57° N), North Africa, Korea and China (Weden et al 2004 ;Granetti et al 2005 ;Hall et al 2007 ;Jeandroz et al 2008 ;Milenkovic et al 2009 ;Yun and Liu 2009 ;Chevalier 2010 ;Streiblova et al 2010 ;Gryndler et al 2011 ;Gogan et al 2012 ;Salerni et al 2013 ;Azul et al 2014 ;Gezer et al 2014 ).…”