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检测小麦品种中的 DNA 序列多态性。

Detection of DNA sequence polymorphisms among wheat varieties.

机构信息

Department of Agronomy, Purdue University, 47907, West Lafayette, IN, USA.

出版信息

Theor Appl Genet. 1992 Aug;84(5-6):573-8. doi: 10.1007/BF00224154.

Abstract

A DNA marker detection strategy that allows the rapid, efficient resolution of high levels of polymorphism among closely related lines of common wheat (Triticum aestivum) has been developed to circumvent the apparent lack of restriction fragment length polymorphism in many important self-pollinated crop species. The technique of randomly amplified polymorphic DNA (RAPD) was combined with a denaturing gradient gel electrophoresis system (DGGE) to explore DNA sequence polymorphisms among different genotypes of wheat. Of the 65 primer combinations used for the polymerase chain reaction (PCR) amplifications, over 38% of them produced readily detectable and reproducible DNA polymorphisms between a spring wheat line, SO852, and a winter wheat variety, 'Clark'. A high level of polymorphism was observed among a number of commercial varieties and breeding lines of wheat. This procedure was also used to detect polymorphisms in a recombinant inbred population to test the feasibility of its application in genome mapping. This DNA polymorphism detection system provides an opportunity for pedigree analysis and fingerprinting of developed wheat lines as well as construction of a high density genetic map of wheat. Without the need for (32)P and sophisticated DNA extraction procedures, this approach should make it feasible to utilize marker-based selection in a plant breeding program.

摘要

已经开发出一种 DNA 标记检测策略,该策略允许快速、有效地解决普通小麦(Triticum aestivum)近缘系之间高水平的多态性,以规避许多重要自花授粉作物物种中明显缺乏限制性片段长度多态性的问题。随机扩增多态性 DNA(RAPD)技术与变性梯度凝胶电泳系统(DGGE)相结合,探索了不同小麦基因型之间的 DNA 序列多态性。在用于聚合酶链反应(PCR)扩增的 65 个引物组合中,超过 38%的组合在春小麦系 SO852 和冬小麦品种“Clark”之间产生了可轻易检测和重现的 DNA 多态性。许多商业品种和小麦育种种质中观察到高水平的多态性。该程序还用于检测重组自交系群体中的多态性,以检验其在基因组图谱构建中的应用可行性。该 DNA 多态性检测系统为开发的小麦系进行系谱分析和指纹图谱提供了机会,也为小麦高密度遗传图谱的构建提供了机会。由于不需要(32)P 和复杂的 DNA 提取程序,这种方法应该可以在植物育种计划中实现基于标记的选择。

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