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Abstract: When two strains of Lentinus edodes were cultured in pairs on agar media such as a potato dextrose agar, no clear antagonistic line appeared around confronting margins. We observed that an antagonistic line or a colored zone appeared around the confronting margins when a saturated solution (about 1.496) of guaiacol was poured onto the plate. We examined agar media, reagents for color development and other factors and identified the optimum conditions for strain differentiation. Then we produced 40 strains by crossing which were genetically related to each other in different degrees. Using these strains we tested the effectiveness of the method. We compared the new method (guaiacol method) with the method using the sawdust rice-bran medium and found that the former was more sensitive than the latter. We also compared the guaiacol method with that using esterase isozymes and found in some cases that the former was better.
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