| A MIS A search revealed simple sequence repeats (SSRs) based on the genomes of Agaricus bisporus. Among, single nucleotide type (52.9%) is the most abundant, followed by tri-(34.3%), di-(11.4%). The most abundant mono-nucleotide, di-nuclotide and tri-nucleotide repeats were A/Tã€AG/CTã€AAC/GTT. A total of112primer pairs were successfully designed by Primer5.0, of which97primers produced bands and25primer pairs showed polymorphism in8A.bisporus genotypes. Further study showed that67alleles were detected with the average of2.68. The average of Nei’s genetic diversity index (H) was0.5023. The genetic similarity coefficient ranged from0.3363to0.8641with a average of0.5385. The UPGMA cluster analysis based on SSR data showed that these parental lines could be classified into two groups at the threshold of0.46, which was some related to their pedigree analysis.19primer pairs followed the law of segregation in single spore isolates population. Based on testing of different combinations and concentration of lysozymes, osmotic stabilizer and different methods of mycelium incubation, an efficient system of protoplast preparation and regeneration in A.bisporus was established in present paper. It was proved that after the mycelium being digested with1.5%lywallzyme in0.6M KC1for3-4h at25℃, the protoplast yield reached106/ml, and that after the purified protoplasts being incubated in PDMA medium with0.6M sucrose as osmotic stabilizer for about10days at25℃, the protoplast would be regenerated. Baased on SSR markers,4homokaryotic protoplasts were identified from12protoplast strains with a rate of1/3. At the end of the paper, we make a bioinformatics comparsions of the SNPs and Indels in the genomes of A.bisporus and A.burnettii.This result indicated that molecular markers will be more usable in map structure, analysis of genetic polymorphism in A.bisporus. It also provided the genetic basis for the breeding of new varieties of A.bisporus or the collection and management of germplasm resources. |
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