Department of Infectious Diseases and Pathology, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611, USA.
Infect Genet Evol. 2012 Dec;12(8):1968-74. doi: 10.1016/j.meegid.2012.08.020. Epub 2012 Sep 13.
Recent advances in high throughput molecular techniques have allowed the development of cost- and time-effective libraries of molecular markers, such as microsatellites, for population genetic studies in non-model species. The American crow, Corvus brachyrhynchos, is recognized to be one of the species that has been most negatively affected by the emergence of West Nile virus (WNV) in North America in 1999. Genetic monitoring of the process of a declining population after the introduction of an infectious disease can provide insights into the demographic and evolutionary impact of a pathogen in a natural host population over time. In this study, shotgun pyrosequencing and validation of previously published cross-species markers were the approaches used to identify and develop a set of 32 polymorphic loci for the C. brachyrhynchos. Since the American crow is morphologically similar to the sympatric species Fish crow (Corvus ossifragus), we also designed a real-time PCR protocol to rapidly differentiate these two species using a set of primers and probes that can discriminate a section of the COI gene at the mitochondrial DNA. These new markers together with a faster method for species verification will allow further detailed studies to characterize and compare genetic diversity of historic and contemporary C. brachyrhynchos populations.
高通量分子技术的最新进展使得能够开发成本效益高、耗时短的分子标记物文库，如微卫星，用于非模式物种的群体遗传学研究。美洲乌鸦（Corvus brachyrhynchos）被认为是 1999 年在北美出现西尼罗河病毒（WNV）后受负面影响最大的物种之一。在引入传染病后对下降种群的过程进行遗传监测，可以深入了解病原体在自然宿主种群中的人口和进化影响。在这项研究中，使用 shotgun 焦磷酸测序和验证先前发表的跨物种标记物的方法，确定并开发了一组 32 个多态性位点用于 C. brachyrhynchos。由于美洲乌鸦在形态上与同域分布的鱼鸦（Corvus ossifragus）相似，我们还设计了一种实时 PCR 协议，使用一组可以区分线粒体 DNA 中 COI 基因部分的引物和探针，快速区分这两个物种。这些新标记物与一种更快的物种验证方法相结合，将允许进一步详细研究以表征和比较历史和当代 C. brachyrhynchos 种群的遗传多样性。
