基于地黄转录组数据的SNP标记开发与地黄指纹图谱构建

doi:10.13560/j.cnki.biotech.bull.1985.2019-0409

摘要/Abstract

摘要： 地黄（Rehmannia glutinosa）是一种具有较高药用价值和经济价值的植物。准确的品种鉴定对地黄种质管理和育种至关重要,使用SNP分子标记来鉴定地黄种质和构建指纹图谱对地黄分子标记育种提供了新方法。利用地黄转录组数据和SRA数据库中的3个地黄转录组数据比对,寻找候选SNP,用PCR技术扩增和序列分析研究28个地黄品种候选SNP变异。从地黄转录组数据中获得了102 075条Unigenes,其中共有SNP位点35 339个,发生频率为0.51/kb;从中随机选取40个候选SNP位点,设计引物39对,用PCR和序列分析从中筛选出7对好的SNP引物,包含8个多态性好的SNP位点;利用最终筛选出的8个SNP位点构建地黄指纹图谱,可以将17个不同地黄种质区分开,可用于地黄种间和种内品种的鉴定。

关键词: 地黄, 转录组, SNP标记开发, 指纹图谱, 品种鉴定

Abstract: Rehmannia glutinosa possesses well-known medicinal properties and nutritional values. Accurately identifying varieties is essential for germplasm management and breeding. Using the SNP molecular markers to identify R. glutinosa germplasm and to construct fingerprints provides a new method for R. glutinosa molecular marker breeding. Rehmannia transcriptome data was applied and compared with three rehmannia transcriptome data in the SRA database to find candidate SNPs,and PCR technology amplification and sequence analysis were used to study the mutations of candidate SNP of 28 Rehmannia cultivars. Total 102 075 Unigenes were obtained from the transcriptome data of R. glutinosa,including 35 339 SNP loci with a frequency of 0.34. A 40 candidate SNPs were randomly selected,and 39 pairs of primers were designed. Seven pairs of good SNP primers were selected by PCR and sequence analysis,including 8 SNP loci with fine polymorphism. The fingerprint of R. glutinosa was constructed using these 8 SNP loci,which allowed to distinguish 17 different rehmannia germplasm,thus it can be used for the identification of interspecific and intraspecific species of R. glutinosa.

Key words: Rehmannia glutinosa, transcriptome, single nucleotide polymorphism（SNP）markers, fingerprint, variety identification

郭萌萌, 周延清, 段红英, 杨珂, 邵露营. 基于地黄转录组数据的SNP标记开发与地黄指纹图谱构建[J]. 生物技术通报, 2019, 35(11): 224-230.

GUO Meng-meng, ZHOU Yan-qing, DUAN Hong-ying, YANG Ke, SHAO Lu-ying. Development of SNP Marker Based on the Rehmannia glutinosa Transcriptome Database and Construction of DNA Fingerprint in Rehmannia[J]. Biotechnology Bulletin, 2019, 35(11): 224-230.

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