University of Connecticut, Ecology and Evolutionary Biology, 75 N. Eagleville Road, Storrs, Connecticut 06269 USA.
Am J Bot. 2012 Mar;99(3):e88-90. doi: 10.3732/ajb.1100392. Epub 2012 Feb 16.
New microsatellite primers were developed for testing genetic differentiation within Nothoceros aenigmaticus and their potential use in other Nothoceros species. The microsatellites are designed to investigate partitioning of genetic variation in a taxon with a peculiar sex allopatry in the southern Appalachian Mountains and relationships with conspecific sexual populations from Mexico.
We used two methods for microsatellite development: an enriched library and second-generation shotgun sequence reads. From these two methods, a total of nine primer pairs were selected and tested on 89 southern Appalachian N. aenigmaticus accessions, nine Mexican accessions, and 16 N. vincentianus accessions. Three mitochondrial loci were recovered from the enriched library method and six loci from 454 shotgun sequencing: three were from the chloroplast and three from the nucleus. The primers amplified repeats with two to 20 alleles per locus.
New microsatellite primers were developed for testing genetic differentiation within N. aenigmaticus and potentially for use in other Nothoceros species. We present one of the first reports of highly polymorphic mitochondrial microsatellites in plants.
为了测试 Nothoceros aenigmaticus 内的遗传分化,开发了新的微卫星引物,并探讨了其在其他 Nothoceros 物种中的潜在用途。这些微卫星设计用于研究在南部阿巴拉契亚山脉中具有奇特性别异地分布的分类群中遗传变异的分配,以及与来自墨西哥的同种类有性种群的关系。
我们使用了两种微卫星开发方法:富集文库和第二代鸟枪法测序。从这两种方法中,总共选择了 9 对引物,并在 89 个来自南部阿巴拉契亚山脉的 N. aenigmaticus 样本、9 个来自墨西哥的样本和 16 个 N. vincentianus 样本中进行了测试。从富集文库方法中回收了 3 个线粒体基因座,从 454 鸟枪法测序中获得了 6 个基因座:3 个来自叶绿体,3 个来自细胞核。引物扩增的重复序列每个基因座有 2 到 20 个等位基因。
开发了新的微卫星引物,用于测试 N. aenigmaticus 内的遗传分化,并可能用于其他 Nothoceros 物种。我们首次报道了植物中高度多态性的线粒体微卫星。