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首页> 美国卫生研究院文献>other >Comparison of RAPD ISSR and AFLP Molecular Markers to Reveal and Classify Orchardgrass (Dactylis glomerata L.) Germplasm Variations
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Comparison of RAPD ISSR and AFLP Molecular Markers to Reveal and Classify Orchardgrass (Dactylis glomerata L.) Germplasm Variations

机译：比较RAPDISSR和AFLP分子标记来揭示和分类果园草（Dactylis glomerata L.）种质变异

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Three different DNA-based techniques, Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Amplified Fragment Length Polymorphism (AFLP) markers, were used for fingerprinting Dactylis glomerata genotypes and for detecting genetic variation between the three different subspecies. In this study, RAPD assays produced 97 bands, of which 40 were polymorphic (41.2%). The ISSR primers amplified 91 bands, and 54 showed polymorphism (59.3%). Finally, the AFLP showed 100 bands, of which 92 were polymorphic (92%). The fragments were scored as present (1) or absent (0), and those readings were entered in a computer file as a binary matrix (one for each marker). Three cluster analyses were performed to express–in the form of dendrograms–the relationships among the genotypes and the genetic variability detected. All DNA-based techniques used were able to amplify all of the genotypes. There were highly significant correlation coefficients between cophenetic matrices based on the genetic distance for the RAPD, ISSR, AFLP, and combined RAPD-ISSR-AFLP data (0.68, 0.78, 0.70, and 0.70, respectively). Two hypotheses were formulated to explain these results; both of them are in agreement with the results obtained using these three types of molecular markers. We conclude that when we study genotypes close related, the analysis of variability could require more than one DNA-based technique; in fact, the genetic variation present in different sources could interfere or combine with the more or less polymorphic ability, as our results showed for RAPD, ISSR and AFLP markers. Our results indicate that AFLP seemed to be the best-suited molecular assay for fingerprinting and assessing genetic relationship among genotypes of Dactylis glomerata.
机译：三种不同的基于DNA的技术，即随机扩增多态性DNA（RAPD），内部简单序列重复（ISSR）和扩增片段长度多态性（AFLP）标记用于指纹鉴定小球藻的基因型并检测三个不同亚种之间的遗传变异。在这项研究中，RAPD分析产生了97条带，其中40条是多态性的（41.2％）。 ISSR引物扩增了91条带，其中54条显示了多态性（59.3％）。最终，AFLP显示了100条带，其中92条是多态的（92％）。将该片段记为存在（1）或不存在（0），并将这些读数作为二进制矩阵输入到计算机文件中（每个标记一个）。进行了三个聚类分析，以树状图的形式表达了基因型与检测到的遗传变异之间的关系。所使用的所有基于DNA的技术都能够扩增所有基因型。基于RAPD，ISSR，AFLP和RAPD-ISSR-AFLP组合数据的遗传距离，在同位矩阵之间存在极高的相关系数（分别为0.68、0.78、0.70和0.70）。提出了两个假设来解释这些结果。它们都与使用这三种类型的分子标记物获得的结果一致。我们得出的结论是，当我们研究紧密相关的基因型时，对变异性的分析可能需要不止一种基于DNA的技术。实际上，正如我们针对RAPD，ISSR和AFLP标记所显示的结果，不同来源中存在的遗传变异可能会干扰或与或多或少的多态性结合。我们的结果表明，AFLP似乎是最适合指纹鉴定和评估小球藻基因型之间遗传关系的分子检测方法。

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