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大麦(Hordeum vulgare L.)中基于EST的SSR和SNP的开发与比较

Generation and comparison of EST-derived SSRs and SNPs in barley (Hordeum vulgare L.).

作者信息

Kota R, Varshney R K, Thiel T, Dehmer K J, Graner A

机构信息

Institute of Plant Genetics and Crop Plant Research (IPK), Correnstr. 3, 06466 Gatersleben, Germany.

出版信息

Hereditas. 2001;135(2-3):145-51. doi: 10.1111/j.1601-5223.2001.00145.x.

Abstract

The progress of genome sequencing projects of model plants like barley, combined with the recent advances of high throughput assays, has provided a wealth of sequence information. This information is being employed to develop a high density transcript map of barley (Hordeum vulgare L.). To achieve this goal, the available EST database is being used as a resource for the development of novel microsatellite (SSR) and single nucleotide polymorphism (SNP) markers. So far, a total of 692 microsatellites representing different di-, tri- and tetra-nucleotide repeats were identified from a set of 19,000 EST sequences. Non-redundant SSRs have been used for mapping and so far 76 microsatellite loci were mapped. In addition to the 180 SNP primer pairs, which were designed to target specific ESTs, 72 were polymorphic among the seven genotypes examined here. Of these, 60 SNPs have been mapped applying a denaturing HPLC approach. To examine the potential of the EST-derived markers for pedigree studies, EST-derived SSRs (75 loci) and SNPs (72 loci) were used to fingerprint a set of seven genotypes. The results show that although both marker types yielded similar groupings, a larger data set of both SSRs and SNPs is necessary to obtain stable clusters in unrelated germplasm.

摘要

大麦等模式植物基因组测序项目的进展,再加上高通量检测技术的最新进展,已经提供了丰富的序列信息。这些信息正被用于构建大麦(Hordeum vulgare L.)的高密度转录图谱。为实现这一目标,现有的EST数据库被用作开发新型微卫星(SSR)和单核苷酸多态性(SNP)标记的资源。到目前为止,从一组19000条EST序列中总共鉴定出692个代表不同二核苷酸、三核苷酸和四核苷酸重复序列的微卫星。非冗余SSR已用于作图,到目前为止已定位了76个微卫星位点。除了针对特定EST设计的180对SNP引物外,在此检测的7个基因型中有72对具有多态性。其中,60个SNP已采用变性高效液相色谱法进行定位。为了检验EST衍生标记在系谱研究中的潜力,利用EST衍生的SSR(75个位点)和SNP(72个位点)对一组7个基因型进行指纹分析。结果表明,尽管两种标记类型产生了相似的分组,但需要更大的SSR和SNP数据集才能在不相关种质中获得稳定的聚类。

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