IDENTIFICATION OF MARKERS LINKED TO DISEASE-RESISTANCE GENES BY BULKED SEGREGANT ANALYSIS - A RAPID METHOD TO DETECT MARKERS IN SPECIFIC GENOMIC REGIONS BY USING SEGREGATING POPULATIONS
IDENTIFICATION OF MARKERS LINKED TO DISEASE-RESISTANCE GENES BY BULKED SEGREGANT ANALYSIS - A RAPID METHOD TO DETECT MARKERS IN SPECIFIC GENOMIC REGIONS BY USING SEGREGATING POPULATIONS
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DOI：
10.1073/pnas.88.21.9828
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发表时间：
1991-11-01
影响因子：
11.1
通讯作者：
KESSELI, RV
中科院分区：
文献类型：
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作者：
MICHELMORE, RW;PARAN, I;KESSELI, RV
我们开发了一种快速鉴定与任何特定基因或基因组区域相关的标记的方法，即批量分离分析。从来自单次杂交的分离群体产生两个混合的DNA样品。每个池或批量包含特定性状或基因组区域相同但所有非连锁区域任意的个体。因此，这两个批量在选定区域中具有遗传差异，但在所有其他区域似乎是杂合的。这两个批量可以针对任何基因组区域和从任何分离群体制备。使用限制性片段长度多态性探针或随机扩增多态性DNA引物筛选原液的差异。我们使用批量分离分析来鉴定生菜中与霜霉病抗性基因相关的三个随机扩增多态性DNA标记。我们发现，标记可以可靠地确定在一个25厘摩的窗口上的任何一方的目标基因座。与使用近等基因系来鉴定基因组特定区域中的标记相比，混合分离分析具有几个优点。通过多轮混合分离分析，遗传步移将是可能的;每对新的混合物将在前一轮分析中鉴定的基因座处不同。这种方法将在那些可能自交的物种和那些必须远系繁殖的物种中得到广泛应用。
We developed bulked segregant analysis as a method for rapidly identifying markers linked to any specific gene or genomic region. Two bulked DNA samples are generated from a segregating population from a single cross. Each pool, or bulk, contains individuals that are identical for a particular trait or genomic region but arbitrary at all unlinked regions. The two bulks are therefore genetically dissimilar in the selected region but seemingly heterozygous at all other regions. The two bulks can be made for any genomic region and from any segregating population. The bulks are screened for differences using restriction fragment length polymorphism probes or random amplified polymorphic DNA primers. We have used bulked segregant analysis to identify three random amplified polymorphic DNA markers in lettuce linked to a gene for resistance to downy mildew. We showed that markers can be reliably identified in a 25-centimorgan window on either side of the targeted locus. Bulked segregant analysis has several advantages over the use of near-isogenic lines to identify markers in specific regions of the genome. Genetic walking will be possible by multiple rounds of bulked segregation analysis; each new pair of bulks will differ at a locus identified in the previous round of analysis. This approach will have widespread application both in those species where selfing is possible and in those that are obligatorily outbreeding.