Establishment of effective gene disruption tools for gene functional analysis in basidiomycetous fungi.

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To establish effective gene disruption tool for edible basidiomyceteous fungi(mushrooms), gene disruption vector system and resistant marker gene recycling system were constructed in Lentinula edodes. First, a gene disruption vector system was constructed by using DelsGate method, and then the loxP sequences were inserted the vector construction system. Unfortunately, no gene disrupted mutant was obtained yet. The pcc1 gene that is a negative regulator in monokaryotic strain, and the ku80 that is involved in non-homologous DNA joint were cloned in L. edodes. If the gene could disrupted, the strain can form fruiting body without mating, and the strain have high efficiency for homologous recombination. To test Cre-loxP system in basidiomyceteous fungi, faster than L. edodes, loxP sequences were inserted gene disruption cassette for C. cinerea. Marker recycling system will be test in C. cinerea in the near future.