Wenzl Peter, Carling Jason, Kudrna David, Jaccoud Damian, Huttner Eric, Kleinhofs Andris, Kilian Andrzej
Center for the Application of Molecular Biology to International Agriculture, G.P.O. Box 3200, Canberra, ACT 2601, Australia.
Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9915-20. doi: 10.1073/pnas.0401076101. Epub 2004 Jun 10.
Diversity Arrays Technology (DArT) can detect and type DNA variation at several hundred genomic loci in parallel without relying on sequence information. Here we show that it can be effectively applied to genetic mapping and diversity analyses of barley, a species with a 5,000-Mbp genome. We tested several complexity reduction methods and selected two that generated the most polymorphic genomic representations. Arrays containing individual fragments from these representations generated DArT fingerprints with a genotype call rate of 98.0% and a scoring reproducibility of at least 99.8%. The fingerprints grouped barley lines according to known genetic relationships. To validate the Mendelian behavior of DArT markers, we constructed a genetic map for a cross between cultivars Steptoe and Morex. Nearly all polymorphic array features could be incorporated into one of seven linkage groups (98.8%). The resulting map comprised approximately 385 unique DArT markers and spanned 1,137 centimorgans. A comparison with the restriction fragment length polymorphism-based framework map indicated that the quality of the DArT map was equivalent, if not superior, to that of the framework map. These results highlight the potential of DArT as a generic technique for genome profiling in the context of molecular breeding and genomics.
多样性阵列技术(DArT)能够在不依赖序列信息的情况下,同时检测数百个基因组位点的DNA变异并进行分型。在此我们表明,它可有效地应用于大麦的遗传图谱构建和多样性分析,大麦是一种基因组大小为5000兆碱基对的物种。我们测试了几种降低复杂性的方法,并选择了两种能产生最多多态性基因组表现形式的方法。包含这些表现形式中单个片段的阵列产生了DArT指纹图谱,其基因型检出率为98.0%,评分重现性至少为99.8%。这些指纹图谱根据已知的遗传关系对大麦品系进行了分组。为了验证DArT标记的孟德尔遗传行为,我们构建了栽培品种Steptoe和Morex杂交的遗传图谱。几乎所有多态性阵列特征都可归入七个连锁群之一(98.8%)。所得图谱包含约385个独特的DArT标记,跨度为1137厘摩。与基于限制性片段长度多态性的框架图谱比较表明,DArT图谱的质量即便不优于框架图谱,也与之相当。这些结果凸显了DArT作为分子育种和基因组学背景下基因组分析通用技术的潜力。