Fernández E., Figueiras M., Benito C.
Departamento de Genética, Facultad de Biología, Universidad Complutense, 28040-Madrid, Spain.
Theor Appl Genet. 2002 Apr;104(5):845-851. doi: 10.1007/s00122-001-0848-2. Epub 2002 Feb 8.
The potential of bulk analyses of RAPD and ISSR-PCR markers for fingerprinting purposes was evaluated using ten RAPD and ten ISSR primers. The phylogenetic relationships of 16 barley cultivars from different countries, and all having a known pedigree, were analysed using 353 PCR markers (125 RAPDs and 228 ISSRs). The band profiles generated were reproducible in spite of the different DNA extractions, PCR techniques, electrophoretic methods and gel scorings used. The RAPD primer S10 and four ISSR primers (811, 820, 835 and 881) were both able to distinguish all cultivars. A strong and quite linear relationship was observed between Resolving Power (Rp) of a primer and its ability to distinguish genotypes. The dendrograms obtained using these two molecular markers are in agreement with their known origin, showing clusters that separate very well the spring/winter and six-rows/two-rows cultivars. Thus, bulk analyses of RAPD and ISSR PCR markers provides a quick, reliable and highly informative system for DNA fingerprinting and also permit to establish genetic relationships which agree with, by other means, known origin of the cultivars.
使用10个随机扩增多态性DNA(RAPD)引物和10个简单序列重复区间(ISSR)引物,评估了RAPD和ISSR-PCR标记进行指纹分析的潜力。利用353个PCR标记(125个RAPD和228个ISSR)分析了来自不同国家、具有已知谱系的16个大麦品种的系统发育关系。尽管使用了不同的DNA提取方法、PCR技术、电泳方法和凝胶评分,但产生的条带图谱具有可重复性。RAPD引物S10和4个ISSR引物(811、820、835和881)都能够区分所有品种。观察到引物的分辨能力(Rp)与其区分基因型的能力之间存在很强且相当线性的关系。使用这两种分子标记获得的聚类图与其已知来源一致,显示出的聚类能够很好地将春/冬性品种和六行/两行品种区分开来。因此,RAPD和ISSR-PCR标记的批量分析为DNA指纹分析提供了一个快速、可靠且信息丰富的系统,还能够建立与品种已知来源相符的遗传关系。