Plant Genetics and Ecosystem Management, Pacific Northwest Research Station, USDA Forest Service, 3200 SW Jefferson Way, Corvallis, Oregon 97331, USA.
Am J Bot. 2011 Nov;98(11):e323-5. doi: 10.3732/ajb.1100317. Epub 2011 Oct 19.
Microsatellite primers were developed for the Pacific Northwest conifer, Chamaecyparis lawsoniana (Cupressaceae), to enhance efficiencies in disease-resistance breeding and germplasm screening for gene conservation of this rare species.
Using multiplexed massively parallel Illumina sequencing, we identified over 300000 microsatellite-containing sequences from 2 million paired-end microreads. After stringent filtering and primer evaluation, we selected 11 primer pairs and used these to screen variation in four populations of C. lawsoniana. Loci show between three and 10 repeats per locus, with an average of eight. Screening of these markers in the North American relative Callitropsis nootkatensis demonstrated limited marker transferability, but these markers could have utility in Asian species of Chamaecyparis.
These microsatellite primers show high polymorphism and should provide a high level of individual discrimination for paternity analysis in defined pedigrees, and routine screening of wild variation in Chamaecyparis lawsoniana.
为了提高太平洋西北地区针叶树柏木(柏科)抗病性育种和遗传资源筛选的效率,促进该稀有物种的基因保护,我们为其开发了微卫星引物。
利用多重大规模平行 Illumina 测序,我们从 200 万个配对末端微读数中鉴定出超过 300000 个包含微卫星的序列。经过严格的过滤和引物评估,我们选择了 11 对引物,并使用这些引物来筛选柏木的四个种群的变异。每个位点的重复数在 3 到 10 个之间,平均为 8 个。在北美亲缘种北美柏木中对这些标记进行筛选表明标记转移的局限性,但这些标记可能在柏木属的亚洲种中有一定的应用价值。
这些微卫星引物显示出较高的多态性,应该为定义的家系中的亲子分析提供高水平的个体鉴别能力,并为柏木属的野生变异进行常规筛选。