Abstract: Abstract:The different genetic bands was screened out by molecular markers between parent strain 26 and strain 28,and waw successfully changed into nucleus marker SCAR1 for strain 26 and SCAR2 for strain 28 respectively.With the applicaton conbined by nucleus SCAR markers and protoplast regeneration, hybrid strain 2628 successfully split into two parental strain of protoplasted monocaryons. Simultaneously, two parental strain of protoplasted monocaryons also can be reconstructed into hybrid strain 2628 identified by nucleus SCAR markers and fruiting test. This enriches homothallic fungi genetics and breeding theory and provides a new way of thinking.