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芝麻(Sesamum indicum L.)物种中微卫星标记(SSR)的开发和特征描述。

Development and characterization of microsatellite markers (SSR) in Sesamum (Sesamum indicum L.) species.

机构信息

Institute of Biotechnology, A.N.G.R.A.U, Rajendranagar, Hyderabad 500 030, India.

出版信息

Appl Biochem Biotechnol. 2012 Nov;168(6):1594-607. doi: 10.1007/s12010-012-9881-7. Epub 2012 Sep 13.

Abstract

Microsatellites, also known as simple sequence repeats (SSRs), are the class of repetitive DNA sequences present throughout the genome of many plant and animal species. Recent advances in molecular genetics had been the introduction of microsatellite markers to investigate the genetic structuring of natural plant populations. We have employed an enrichment strategy for microsatellite isolation by using multi-enzymes digestion, microsatellite oligoprobes, and streptavidin magnetic beads in Sesamum (Sesamum indicum L.). More than 200 SSR motifs were detected (SSR motifs ≥2 repeat units or 6 bp); 80 % of the clones contained SSR motifs. When regarding SSRs with four or more repeat units and a minimum length of 10 bp, 132 of them showed repeats. Eighteen SSR markers were initially characterized for optimum annealing temperature using a gradient PCR technique. Among the 18 SSR markers characterized, five were found to be polymorphic and used to analyze 60 Sesamum germplasm accessions. The maximum number of alleles detected was four with a single primer and the least number of two alleles with three primers with an average PIC value of 0.77. SSRs are a valuable tool for estimating genetic diversity and analyzing the evolutionary and historical development of cultivars at the genomic level in sesame breeding programs.

摘要

微卫星,也被称为简单重复序列(SSR),是存在于许多植物和动物物种基因组中的一类重复 DNA 序列。分子遗传学的最新进展是将微卫星标记引入到对自然植物群体的遗传结构的研究中。我们采用了一种富集策略来分离微卫星,该策略使用了多酶消化、微卫星寡探针和链霉亲和素磁珠在芝麻(Sesamum indicum L.)中进行。检测到了 200 多个 SSR 基序(SSR 基序≥2 个重复单元或 6bp);80%的克隆包含 SSR 基序。当考虑到具有四个或更多重复单元且最小长度为 10bp 的 SSR 时,其中 132 个显示出重复。最初使用梯度 PCR 技术对 18 个 SSR 标记进行了最佳退火温度的特征描述。在所鉴定的 18 个 SSR 标记中,有 5 个表现出多态性,并用于分析 60 份芝麻种质资源。用单个引物检测到的等位基因数量最多为 4 个,用 3 个引物检测到的等位基因数量最少为 2 个,平均 PIC 值为 0.77。SSR 是在芝麻育种计划中从基因组水平评估遗传多样性和分析品种进化和历史发展的有价值的工具。

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