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Agaricus bisporus mushroom cultivation is one of China's major, ranked third, is China's traditional export products. With bisporus industry's rapid development, the production of the urgent needs of new varieties of Agaricus bisporus replace existing varieties have been used for many years. Breeding work looking for new hybrid strain of early and rapid identification of forecasting methods and means to effectively reduce the fruiting part of the workload and reduce the breeding costs and improve breeding efficiency, become hot and breeders to explore key topics. In recent years, with the development of molecular biology, a variety of molecular markers are also beginning to open new hybrid strains used in the identification of, and improve the breeding efficiency. This study is the use of single spore hybridization breeding of new varieties of Agaricus bisporus, the parent strain F56 using bisporus strains, As2796 and Zhenong the 1st strain. Obtained with strains 41 nuclear bodies, paired crosses 420, get 11 hybrid heterokaryon strains. In this study, ISSR markers for 11 hybrid strains and the parent strain analysis. Test screened out 10 clear amplified bands, polymorphic ISSR primers amplified a total of 40 polymorphic bands, clearly distinct, using cluster analysis software NTSYSpc2.1 biological construct phylogenetic trees . Cluster analysis showed that 14 strains tested genetic similarity coefficients ranged from 0.6700 to 1.000, the similarity coefficient 0.8140, the 14 strains were clustered into four categories. AG95 and F56 similarity coefficient is 1, proved to be not new hybrid strains. ISSR molecular marker results indicate that in addition to the remaining 10 strains AG95 are outside the new hybrid strains, new hybrid strains with the parent strain genetic difference. The hybrid strains fruiting trials, AG97 proved to be an excellent hybrid strains. In this study, a single hybrid strains AG97 spore spore separation and tissue separation, screening of 40 single spore isolates the F2 generation and the two organizations isolates. The 43 strains with the parental As2796 and F56 for genetic analysis, including ISSR molecular markers, colony morphology and growth rate determination and fruiting tests. ISSR molecular marker results showed that 45 strains of genetic similarity coefficient ranged from 0.4250 ~ 1.0000. AG97 and tissue isolates AG97-1 and AG97-3 is similar to the largest coefficient of 1.0000, indicating that the organization can better maintain the separation of hybrid strains of genetic traits. F2 generation Isolates genetic similarity coefficient ranged from 0.5250 ~ 0.9000, indicating that a single spore isolates at the DNA level has a relatively significant genetic variation, showing rich genetic diversity. Fruiting test results show that the yield was higher than hybrid strains AG97 single spore strains AG975, AG9721, AG9722, AG9718 and AG9717; Isolates fruiting body morphology and AG97 no significant differences. These preliminary results show that the hybrid F2 generation from a single spore isolates, the ability to filter to the excellent quality and yield are relatively new strain. In this study, the F2 strain AG9710 hybrid strains of single spore isolates, screened 10 single spore isolates the F3 generation. The F3 generation strains AG9710, AG97 and parental As2796 and F56 for ISSR markers. The results showed that 14 strains tested genetic similarity coefficient ranged from 0.5897 ~ 0.9231. F3 generation big gap between the genetic, while F3 generation and hybrid strains AG97 and the parental strain genetic gap increases. Not only can the use of ISSR markers bisporus strains reveal genetic relationships among, but also for excellent hybrid strains to predict and determine the basis for providing screening, but also for the use of ISSR molecular marker assisted conventional crossbreeding method provides a new way.
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