The Method Research on Tuber spp. DNA in Soil
Truffles are ascomata of ectomycorrhizal hypogeous fungi associated with endemic Pinus tabuliformis in Mount Helan National Nature Reserve in Inner Mongolia, China, was investigated f in soil cubes taken from pure P. tabuliformis stands. The objectives of this study were (i) to develop a molecular method to detect mycelia of Tuber spp. in soil and (ii) to test for mycelial distribution around two truffle-bearing P. tabuliformis trees in a truffle orchard. Isolation of total DNA from soil was performed, followed by PCR amplification with Tuber spp.-specific primers and restriction analysis. To address the detection sensitivity level, soil samples were inoculated with known amounts of gleba of Tuber spp. Mycelium was detected primarily within the area defined by the truffle burn and within the top 35 cm of the soil in all directions from the trees.
KeywordsTuber spp Soil mycelium PCR–RFLP Truffle orchard ITS
This work was supported in part by the DMAH-GENCAT, by scholarship 2002FI-00711 to L.M. Suz from the DURSI-GENCAT, and by the research project FMI-REN2002-04068-CO2-01. We are indebted to M. Donate for the open access to his truffle orchard. The authors thank C.R. Fischer for helpful suggestions and English support.
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