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为了开发杏鲍菇分子标记,利用前期高通量测序技术获得的杏鲍菇转录组数据,采用生物信息学软件Trinity拼接,然后通过MISA软件检索并分析杏鲍菇转录组简单重复序列(Simple sequence repeats,SSR)位点.共获得45 833条unigene,在其中的3 256条unigene中检测到3 811条符合软件参数的SSR序列,发生频率为8.31%.SSR位点平均长度为15 bp,平均分布频率是1/15.91 kb.在杏鲍菇转录组的SSRs中,单核苷酸与三核苷酸重复基元为主要重复类型,分别占总SSRs的45.24%和35.53%.SSR基元的重复次数为5-24次,其中5次或10次重复是发生频率最高的,SSR重复基元共有85个类型,其中A/T是最常见的重复基元,比例为38.97%,其次为AG/CT,比例为8.24%.对3 811个SSR位点设计出3 644对SSR引物,随机选择20对引物进行PCR扩增,其中9对在3个杏鲍菇品种中表现出多态性.本研究表明杏鲍菇转录组SSR位点多态性丰富,具有较高的应用潜力.
In order to develop the molecular marker of Pleurotus eryngii, the transcriptome data of Pleurotus eryngii obtained by pre-high-throughput sequencing was stitched using Trinity bioinformatics software. The sequence of Simple transcripts of Pleurotus eryngii was searched and analyzed by MISA software. repeats and SSR loci.A total of 45 833 unigene were found, of which 3 811 SSR sequences with software parameters were detected in 3 256 unigene, and the frequency of occurrence was 8.31% .The average length of SSR loci was 15 bp, the average The frequency of distribution was 1 / 15.91 kb. Among the SSRs of the Pleurotus eryngii transcriptome, the mononucleotide and trinucleotide repeat motifs were the main repeat types, accounting for 45.24% and 35.53% of the total SSRs, respectively The number of repeats was 5-24 times. Among them, 5 or 10 repeats were the most frequently occurring. There were 85 types of SSR repeat motifs, of which A / T was the most common repeat motif with a ratio of 38.97%, followed by AG / CT with a ratio of 8.24%, 3 644 pairs of SSR primers were designed for 3 811 SSR loci and 20 pairs of primers were randomly selected for PCR amplification. Nine of them showed polymorphism in three Pleurotus eryngii varieties. This study shows that Pleurotus eryngii transcriptome SSR polymorphism rich, with high potential applications.