Mehlenbacher S A, Brown R N, Davis J W, Chen H, Bassil N V, Smith D C, Kubisiak T L
Department of Horticulture, Oregon State University, 4017 ALS Building, Corvallis, OR 97331, USA.
Theor Appl Genet. 2004 Feb;108(4):651-6. doi: 10.1007/s00122-003-1476-9. Epub 2003 Oct 21.
A total of 1,110 decamer primers were screened for RAPD markers linked to a dominant allele in hazelnut ( Corylus avellana) that confers resistance to eastern filbert blight caused by Anisogramma anomala. Twenty RAPD markers linked in coupling, and five markers linked in repulsion, were found. A seedling population was used to construct a linkage map of the region flanking the resistance locus. The map spans 46.6 cM, with 14 markers on one side of the resistance locus and eight on the other side. Eleven markers showed less than 3% recombination with resistance, including three that showed no recombination. Seven of these 11 markers are sufficiently robust to allow their use in marker-assisted selection. These include AA12(850) which shows no recombination, and six markers on one side of the resistance locus: 173(500), 152(800), 122(825), 275(1130), H19(650) and O16(1250). Marker 268(580), which flanks the resistance locus on the other side, is also suitable for use in marker-assisted selection, but shows 5.8% recombination with resistance. Other markers are less suitable for marker-assisted selection because of sensitivity to changes in primer or MgCl(2) concentration, or the long time required for electrophoresis to separate bands of similar size. The 16 markers closest to the resistance locus were cloned and sequenced. The W07(365) marker, which showed no recombination with the resistance locus but is difficult to score, includes a CT microsatellite repeat. The sequence information will allow the design of SCAR primers and eventual map-based cloning of the resistance allele.
共筛选了1110个十聚体引物,用于寻找与榛子(欧洲榛)中一个显性等位基因连锁的随机扩增多态性DNA(RAPD)标记,该等位基因赋予对由奇异盘二孢菌引起的东部榛子枯萎病的抗性。发现了20个呈耦合连锁的RAPD标记和5个呈互斥连锁的标记。利用一个实生苗群体构建了抗性基因座侧翼区域的连锁图谱。该图谱跨度为46.6厘摩,抗性基因座一侧有14个标记,另一侧有8个标记。11个标记与抗性的重组率低于3%,其中3个没有重组。这11个标记中有7个足够稳定,可用于标记辅助选择。其中包括不发生重组的AA12(850),以及抗性基因座一侧的6个标记:173(500)、152(800)、122(825)、275(1130)、H19(650)和O16(1250)。位于抗性基因座另一侧的标记268(580)也适用于标记辅助选择,但与抗性的重组率为5.8%。其他标记由于对引物或MgCl₂浓度变化敏感,或电泳分离相似大小条带所需时间长,不太适合用于标记辅助选择。对最接近抗性基因座的16个标记进行了克隆和测序。与抗性基因座不发生重组但难以计分的W07(365)标记包含一个CT微卫星重复序列。序列信息将有助于设计序列特异性扩增区域(SCAR)引物,并最终基于图谱克隆抗性等位基因。