Infection of mature Pinus densiflora with ectomycorrhizal fungi, Tricholoma matsutake

We carried out research work to infect the roots of mature pine (Pinus densiflora) with Tricholoma matsutake for cultivation of the pine mushroom. Four-month-old and one-year-old seedlings and forty-year-old mature pine were treated with α-NAA (α-naphthalene acetic acid) as rooting agent for generating new rootlets. The optimum concentrations of α-NAA for generating new rootlets in four-month-old and one-year-old pine seedlings were 0.5 mg per root, at which numbers of generated new rootlets were approximately 2–3 times higher than in control. The mature pine treated with 1.0 mg of α-NAA per root produced approximately 1.7 times more new rootlets than untreated. Roots in 15 mature pines were treated with α-NAA, and about 79% of the treated roots successfully generated new branching roots. For mycorrhizal synthesis, the new rootlets without contamination were inoculated with mycelia of matsutake cultured in a glass container with sterilized vermiculite substrate. After 4 months, it was identified by ITS specific primer method that about 50% of the analyzed root samples were infected with matsutake. The results showed that the roots of mature pine can be infected by matsutake.