Truffle (
Tuber melanosporum
Vitt.) culture is an agroforestry sector in Chile of increasing interest due to the high prices that
truffles fetch in the national market and the recent evidence that its commercial production is possible in Chilean climatic and
soil conditions. In this study, the efficiency of three methods of DNA extraction from a mix of 5 g of soil and roots from both
nursery and field plants of
Quercus ilex
L. mycorrhized with
T. melanosporum were evaluated, and a simple and reproducible
protocol was established. Detection of
T. melanosporum was performed by the technique of cleaved amplified polymorphic
sequence (CAPS) from amplicons generated with the primers ADL1 (5'-GTAACGATAAAGGCCATCTATAGG-3') and
ADL3 (5'-CGTTTTTCCTGAACTCTTCATCAC-3`), where a restriction fragment of 160 bp specific for
T. melanosporum
was generated, which allows the discrimination of this species from the rest of the species belonging to the
Tuber sp.
genus. Direct detection of
T. melanosporum in one step was also obtained by polymerase chain reaction (PCR) from
total DNA isolated from mycorrhized roots and with the primers ITSML (5'-TGGCCATGTGTCAGATTTAGTA-3') and
ITSLNG (5'-TGATATGCTTAAGTTCAGCGGG-3'), generating a single amplicon of 440 bp. The molecular detection of
T. melanosporum by the methods presented here will allow the rapid and accurate detection of mycorrhization of trees, both
under nursery and field conditions. This technology will also provide more security to farmers by controlling the quality of
the mycorrhized trees they will plant and also by following the mycorrhization status of established orchards.
