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为丰富普通荞麦(Fagopyrum esculentum Moench)的序列信息,挖掘有效的SSR标记,基于Hiseq 2000测序平台对普通荞麦的种子转录谱进行测序、分析,利用Misa软件进行SSR位点扫描,采用Primer 5.0引物设计程序对其中的300个SSR位点设计引物,随机挑选40对引物对19个普通荞麦品系进行遗传多样性分析。结果表明:测序共产生20 508 824条高质量的短序列(read),总长度为1 889 111 004bp,通过拼接最终获得54 947条转录本(transcript)和36 133个独立基因(unigene)。在2 226个独立基因中发现了2 666个SSR位点。有20对引物(占50%)能扩增出目标产物,其中,12对有多态性,多态性信息量(PIC)范围为0.10~0.93,平均为0.57,多态性程度高。
In order to enrich the sequence information of Fagopyrum esculentum Moench and find effective SSR markers, the seed transcriptional spectrum of common buckwheat was sequenced and analyzed based on the Hiseq 2000 sequencing platform. Misa software was used to scan the SSR loci. Primer 5.0 primer design The program designed primers for 300 of these SSR loci and randomly selected 40 pairs of primers for 19 common buckwheat lines for genetic diversity analysis. The results showed that 20 508 824 high quality short sequences were generated by sequencing, with a total length of 1 889 111 004 bp. 54 947 transcripts and 36 133 unigene were finally obtained by splicing. A total of 2 666 SSR sites were found in 2 226 independent genes. Twenty pairs of primers (50%) amplified the target product. Among them, 12 pairs were polymorphic. The PIC ranged from 0.10 to 0.93 with an average of 0.57 and high polymorphism.