Qi X, Lindhout P
Graduate School of Experimental Plant Sciences, Dept. of Plant Breeding, Wageningen Agricultural University, The Netherlands.
Mol Gen Genet. 1997 Apr 16;254(3):330-6. doi: 10.1007/s004380050423.
To investigate the application of amplified fragment length polymorphism (AFLP) markers in barley, 96 primer combinations were used to generate AFLP patterns with two barley lines, L94 and Vada. With seven primer combinations, only a few intense bands were obtained, probably derived from repeated sequences. With the majority of the remaining 89 primer combinations, on average about 120 amplification products were generated, and the polymorphism rate between the two lines was generally over 18%. Based on the number of amplified products and the polymorphism rate, the 48 best primer combinations were selected and tested on 16 barley lines, again including L94 and Vada. Using a subset of 24 primer combinations 2188 clearly visible bands within the range from 80 to 510 bp were generated; 55% of these showed some degree of polymorphism among the 16 lines. L94 versus Vada showed the highest polymorphism rate (29%) and Proctor versus Nudinka yielded the lowest (12%). The polymorphism rates per primer combination showed little dependence on the barley lines used. Hence the most efficient and informative primer combinations identified for a given pair of lines turned out to be highly efficient when applied to others. Generally, more than 100 common markers (possibly locus specific) among populations or crosses were easily identified by comparing 48 AFLP profiles of the parent lines. The existence of such a large number of markers common to populations will facilitate the merging of molecular marker data and other genetic data into one integrated genetic map of barley.
为了研究扩增片段长度多态性(AFLP)标记在大麦中的应用,使用96对引物组合对两个大麦品系L94和Vada进行AFLP分析。使用7对引物组合时,只获得了少数几条深色条带,可能来自重复序列。使用其余89对引物组合中的大多数时,平均产生约120个扩增产物,两个品系之间的多态性率通常超过18%。根据扩增产物数量和多态性率,选择48对最佳引物组合并在16个大麦品系上进行测试,其中再次包括L94和Vada。使用24对引物组合的子集,产生了2188条80至510 bp范围内清晰可见的条带;其中55%在16个品系中表现出一定程度的多态性。L94与Vada之间的多态性率最高(29%),Proctor与Nudinka之间的多态性率最低(12%)。每对引物组合的多态性率对所用大麦品系的依赖性较小。因此,为给定的一对品系鉴定出的最有效和信息最丰富的引物组合,应用于其他品系时也非常高效。一般来说,通过比较亲本系的48个AFLP图谱,很容易在群体或杂交组合中鉴定出100多个共同标记(可能是位点特异性的)。群体中存在如此大量的共同标记将有助于将分子标记数据和其他遗传数据合并到一个综合的大麦遗传图谱中。