Crop Genetics Department, Scottish Crop Research Institute (SCRI), DD2 5DA, Invergowrie, Dundee, Scotland.
Theor Appl Genet. 1996 Mar;92(3-4):442-7. doi: 10.1007/BF00223691.
Primers complementary to simple sequence repeats (SSRs) and with variable three-base 'anchors' at their 5' end, were used in PCR analyses to compare pooled DNA samples from various Brassica napus and B. rapa cultivars. Amplification products were resolved on polyacrylamide gels and detected by silver-nitrate staining. The resulting banding patterns were highly repeatable between replicate PCRs. Two of the primers produced polymorphisms at 33 and 23 band positions, respectively, and could each discriminate 16 of the 20 cultivars studied. Combined use of both primers allowed all 20 cultivars to be distinguished. The UPGMA dendrogram, based on the cultivar banding profiles, demonstrated clustering on the basis of winter/spring growth habit, high/low glucosinolate content, and cultivar origin (i.e. the breeder involved). Intracultivar polymorphism was investigated using a minimum of ten individuals for each cultivar and was found to vary considerably between cultivars. It is concluded that anchored SSR-PCR analysis is a highly informative and reproducible method for fingerprinting oilseed rape populations, but that intra-cultivar variation should be investigated before using banding profiles from pooled samples for the identification of individuals.
引物与简单重复序列（SSR）互补，并在其 5'端具有可变的三碱基'锚'，用于 PCR 分析，以比较来自各种油菜和油菜品种的 pooled DNA 样本。扩增产物在聚丙烯酰胺凝胶上进行分离，并通过硝酸银染色检测。重复 PCR 之间的条带模式高度可重复。其中两个引物分别在 33 和 23 个带位置产生多态性，并且每个引物都可以区分研究的 20 个品种中的 16 个。两种引物的组合使用可以区分所有 20 个品种。基于品种带型的 UPGMA 聚类图表明，聚类基于冬/春生长习性、高/低硫代葡萄糖苷含量和品种起源（即涉及的育种者）。使用每个品种至少 10 个个体研究了品种内多态性，发现品种之间差异很大。结论是，锚定 SSR-PCR 分析是一种高度信息丰富且可重复的方法，用于鉴定油菜群体，但在使用 pooled 样本的带型鉴定个体之前，应调查品种内的变异。
