Abstract
Analysis of DNA from cultivated mushrooms is difficult for growers who are not familiar with molecular biology techniques. We developed a simple method for distinguishing commercial strains of shiitake mushrooms (Lentinula edodes) using the FTA filter card. Three kinds of shiitake samples were examined: colony grown on an agar plate, mycelium grown on an agar slant, and a fruiting body. Each sample was pressed directly on an FTA card. A portion of the xylanase (xyl) and glucoamylase (gla1) genes was amplified by PCR from 1.2-mm discs cut from the FTA card. After digesting the PCR products with restriction enzymes MspI and HaeIII, the restriction fragment length polymorphism (RFLP) results of two commercial strains were compared. FTA discs originating from three samples of shiitake were successfully amplified by PCR. Two commercial strains were easily distinguished by RFLP of xly and gla1 on an agarose gel. The described method does not require purifying DNA from samples, and no equipment other than the apparatus for PCR and agarose gel electrophoresis is necessary. The method would be widely applicable for mushroom growers as a simple method for typing strains.
