基于大球盖菇基因组的SSR分子标记开发及指纹数据库应用

Development of SSR molecular markers and application of fingerprint database based on whole genome of Stropharia rugosoannulata

以13个大球盖菇菌株及其已公布的基因组为试材,采用生物信息学和试验验证相结合的方法,研究了大球盖菇基因组上的SSR类型及分布,设计了100对SSR引物开展试验验证,以期为大球盖菇的种质资源鉴定、品种选育、分子标记辅助育种等研究提供参考依据。结果表明:总计检索到6种SSR类型的2 124条序列,其中主要以三核苷酸类型为主,共计704,占SSR总数的33.15%;五、六碱基重复类型较多,但总和只占全部SSR数量的14.59%。由验证试验获得16对引物,其中引物DQGSSR020、DQGSSR031、DQGSSR077的多态性较高、特异性好、无杂峰、荧光亮度高、区分度好。同时,基于16对SSR引物进行大球盖菇DNA指纹编码构建,获得了13个大球盖菇菌株的32位数分子指纹数据库编码。

Based on experimental validation and bioinformatics methods,SSR analysis result were exploited from the genomic of Stropharia rugosoannulata and 13 strains.In the meantime,100 SSR were selected for primer design and experimental verification.In order to provide a theoretical basis for the germplasm resources identification,variety selection and molecular marker-assisted breeding of S.rugosoannulata.The results showed that a total of 2 124 sequences of 6 SSR types were retrieved.The main type of trinucleotide(P3) was 704,accounting for 33.15% of the total SSR number.Pentanucleotide and hexanucleotide repeat types were more,the total number accounted for 14.59% of the total SSR number.16 pairs of primers were obtained by verification test,three primers with high pair property,repeatability and specificity were obtained from the validation experiment,which were DQGSSR020,DQGSSR031 and DQGSSR077.At the same time,the DNA fingerprint coding of 16 pairs of primers was constructed and the 32-digit molecular fingerprint database codes of 13 S.rugosoannulata strains were obtained.