Prabhu R R, Gresshoff P M
Institute of Agriculture, University of Tennessee, Knoxville 37901-1071.
Plant Mol Biol. 1994 Oct;26(1):105-16. doi: 10.1007/BF00039524.
DNA amplification fingerprinting (DAF) using a high primer-to-template ratio and single, very short arbitrary primers, was used to generate amplified fragment length polymorphic markers (AFLP) in soybean (Glycine max (L.) Merr.). The inheritance of AFLPs was studied using a cross between the ancestral Glycine soja PI468.397 and Glycine max (L.) Merr. line nts382, F1 and F2 progeny. The amplification reaction was carried out with soybean genomic DNA and 8 base long oligonucleotide primers. Silver-stained 5% polyacrylamide gels containing 7 M urea detected from 11 to 28 DAF products with primers of varying GC content (ranging from 50 to 100% GC). Depending on their intensity, AFLPs were classified into three classes. DAF profiles were reproducible for different DNA extractions and gels. Forty AFLPs were detected by 26 primers when comparing G. soja and G. max. Most AFLPs were inherited as dominant Mendelian markers in F1 and F2 populations. However, abnormal inheritance occurred with about 25% of polymorphisms. One marker was inherited as a maternal marker, presumably originating from organelle DNA while another showed apparent paternal inheritance. To confirm the nuclear origin and utility of dominant Mendelian markers, three DAF polymorphisms were mapped using a F11 mapping population of recombinant inbred lines from soybean cultivars Minsoy x Noir 1. The study showed that DAF-generated polymorphic markers occur frequently and reliably, that they are inherited as Mendelian dominant loci and that they can be used in genome mapping.
利用高引物与模板比例以及单个非常短的任意引物进行DNA扩增指纹分析（DAF），以在大豆（Glycine max (L.) Merr.）中产生扩增片段长度多态性标记（AFLP）。使用原始大豆Glycine soja PI468.397与Glycine max (L.) Merr.品系nts382的杂交后代F1和F2子代研究AFLP的遗传。扩增反应使用大豆基因组DNA和8个碱基长的寡核苷酸引物进行。含7M尿素的5%银染聚丙烯酰胺凝胶检测到，不同GC含量（50%至100%GC）的引物产生11至28条DAF产物。根据其强度，AFLP被分为三类。DAF图谱对于不同的DNA提取物和凝胶是可重复的。比较G. soja和G. max时，26个引物检测到40个AFLP。在F1和F2群体中，大多数AFLP作为显性孟德尔标记遗传。然而，约25%的多态性出现异常遗传。一个标记作为母本标记遗传，可能源自细胞器DNA，而另一个显示明显的父本遗传。为了确认显性孟德尔标记的核起源和实用性，利用大豆品种Minsoy x Noir 1的重组自交系F11作图群体对三个DAF多态性进行了定位。该研究表明，DAF产生的多态性标记频繁且可靠地出现，它们作为孟德尔显性位点遗传，并且可用于基因组作图。
