PCR-based sensitive detection of the edible fungus Boletus edulis from rDNA ITS sequences

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Abstract

Identification of commercially important fungi, such as the valuable edible fungus Boletus edulis can be difficult considering visual or metabolic approaches. Based on phylogenetic analysis of the rDNA ITS sequence, a pair of specific primers was designed for differentiating B. edulis from other mushrooms by PCR. PCR was performed with total DNA as a template at an annealing temperature between 56-60ºC. Positive amplicons were obtained from B. edulis with all DNA templates from fruit bodies and cultured mycelium, but not from other fungal species at an annealing temperature of 60ºC. The result indicated that B. edulis could be clearly distinguished from other fungi by PCR, and there were no misidentifications under the reaction conditions used. The primers were also successfully employed to identify various tissues of B. edulis.

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Soil fungal communities in a Castanea sativa (chestnut) forest producing large quantities of Boletus edulis sensu lato (porcini): where is the mycelium of porcini?

Ursula Peintner, Alessandra Zambonelli, P Jennifer Klotz … (2007)

A study was conducted in a Castanea sativa forest that produces large quantities of the edible mushroom porcini (Boletus edulis sensu lato). The primary aim was to study porcini mycelia in the soil, and to determine if there were any possible ecological and functional interactions with other dominant soil fungi. Three different approaches were used: collection and morphological identification of fruiting bodies, morphological and molecular identification of ectomycorrhizae by rDNA-ITS sequence analyses and molecular identification of the soil mycelia by ITS clone libraries. Soil samples were taken directly under basidiomes of Boletus edulis, Boletus aestivalis, Boletus aereus and Boletus pinophilus. Thirty-nine ectomycorrhizal fungi were identified on root tips whereas 40 fungal species were found in the soil using the cloning technique. The overlap between above- and below-ground fungal communities was very low. Boletus mycelia, compared with other soil fungi, were rare and with scattered distribution, whereas their fruiting bodies dominated the above-ground fungal community. Only B. aestivalis ectomycorrhizae were relatively abundant and detected as mycelia in the soil. No specific fungus-fungus association was found. Factors triggering formation of mycorrhizae and fructification of porcini appear to be too complex to be simply explained on the basis of the amount of fungal mycelia in the soil.