摘要
以12个灰树花(Grifola frondosa)菌株及已公布的灰树花基因组为基础,采用生物信息学和实验验证相结合的方法,设计192对SSR引物研究灰树花基因组SSR类型及分布.结果表明:总计检索到5种SSR类型的666个位点,其中,三核苷酸重复的SSR位点最多(52.70％),是灰树花SSR的主要类型;二核苷酸SSR位点占比为32.43％;四、五、六核苷酸重复类型较多,但数量较少.由验证实验得到15对SSR引物,共检测到55个等位基因位点,15对SSR引物的平均Shannon's信息指数、观测杂合度、期望杂合度和多态性信息含量分别为0.817、0.506、0.442和0.397.基于15对SSR引物进行灰树花DNA指纹编码构建,获得12个灰树花菌株唯一的30位数分子指纹数据库编码,可对每一个灰树花菌株进行精准鉴定.
Abstract
Based on 12 Grifola frondosa strains and the published genomic sequence of G.frondosa,192 pairs of SSR primers were designed to study the types and distribution of SSR markers in the G.frondosa,genome and the SSR primers were validated by PCR.The results showed that a total of 666 SSR loci of five SSR types were retrieved.Among them,trinucleotide repeats are the most abundant in G.frondosa,accounting for 52.70％,and the proportion of dinucleotide SSR loci is 32.43％.Repeats of four,five and six bases have a high diversity of repeat motifs,but their quantity was low.Fifteen pairs of polymorphic primers were selected from the validation test,and a total of 55 allele loci were detected.The average Shannon's information index,observed heterozygosity,expected heterozygosity and polymorphism information content of the 15 pairs of SSR primers were 0.817,0.506,0.442 and 0.397,respectively.Using the 15 pairs of SSR primers,the DNA fingerprint coding of G.frondosa was constructed,and the unique 30-digit molecular fingerprint database coding for the 12 G.frondosa strains was obtained,facilitating accurate identification of G.frondosa strains.
维普
万方数据