Abstract: 【Objective】 To identify Lentinula edodes(Berk.) Pegler hybrid progenies from dikaryon-monokaryon mating using SCAR molecular marker, so as to provide technical support to molecular-assisted breeding.【Method】Special bands used to mark two nucleus of the donor parental strain YX7 were obtained through ISSR analysis. SCAR primers were designed accor-ding to the sequences of the special bands, 40 hybrid strains from the mating with strain YX7 as donor and spore monokaryons of strain 808 as receptor were identified through SCAR-PCR. Conventional methods of microscopy observation and antagonistic test were adopted to check and verify the results of SCAR identification.【Result】 The specific bands, a and b, that marked the two karyotypes in the nucleus of donor strain YX7 were separately obtained, and the primer PA and PB for SCAR identification that were accordingly designed had specific amplification effect on the two different nucleus of strain YX7 respectively. The SCAR-PCR identification showed that 27 of 40 strains were heterozygote, 19 possessed the a-band marker karyotype, and 8 possessed the b-band karyotype. 13 strains were non-heterozygote, and 11 of them were strain YX7 with both a-band and b-band karyotype, and no a-band or b-band karyotype was detected in other 2 of them, indicating they were the spore monokaryons of strain 808. The results got through conventional method were consistent with SCAR marker identification. Compared to the conventional method, SCAR marker identification was more convenient, effective and had more stable and reliable results. The heterozygotes could be further divided into two groups by karyotype identification.【Conclusion】 SCAR marker identification is an effective, comprehensive and accurate technique for recognizing the heterozygote and non-heterozygote from dikaryon-monokaryon mating of L. edodes strain, and can be applied to hybrid molecular assisted breeding and genetic analysis of L. edodes.