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通过下一代测序技术为水黾快速开发微卫星。

Rapid microsatellite development for water striders by next-generation sequencing.

机构信息

Department of Ecology and Evolutionary Biology, University of Toronto, Toronto ON M5S 3B2, Canada.

出版信息

J Hered. 2011 Jan-Feb;102(1):125-9. doi: 10.1093/jhered/esq099. Epub 2010 Sep 1.

Abstract

Water striders have become a model system for studies of sexual conflict and coevolution, but progress is currently limited by a lack of genetic resources. Next-generation sequencing technologies offer the potential for rapid and cost-effective development of molecular markers and hold particular promise for model organisms in ecology for which no reference genome exists. We used Roche 454 sequencing of genomic DNA to identify microsatellite loci for the water strider Gerris incognitus. A modest sequencing volume generated 182,912 reads, of which 30,820 (16.8%) contained microsatellite repeats. We selected 23 loci for primer development, based on criteria that maximized the likelihood of amplifying polymorphic loci, and tested them in G. incognitus and the related species G. buenoi. Of the 16 amplifying loci, 10 yielded reliable amplification and detectable polymorphism, with an average of 6.1 alleles per locus (range: 2-12). These markers should facilitate new avenues of study, including postcopulatory sexual selection, population genetic structure, phylogeography, and sexual coevolution, for a key taxon in studies of mating conflict. The current study demonstrates an effective method for microsatellite development and shows that light sequencing of genomic DNA can provide numerous and highly variable markers.

摘要

水黾已成为研究性冲突和协同进化的模式系统,但目前的进展受到遗传资源缺乏的限制。新一代测序技术为快速和经济有效地开发分子标记提供了可能,对于那些没有参考基因组的生态学模式生物尤其具有前景。我们使用罗氏 454 测序技术对水黾 Gerris incognitus 的基因组 DNA 进行了微卫星标记的鉴定。适度的测序量生成了 182912 条reads,其中 30820(16.8%)条包含微卫星重复序列。我们根据最大限度地增加扩增多态性位点的可能性的标准,选择了 23 个用于引物开发的位点,并在 G. incognitus 和相关物种 G. buenoi 中进行了测试。在 16 个可扩增的位点中,有 10 个产生了可靠的扩增和可检测的多态性,每个位点平均有 6.1 个等位基因(范围:2-12)。这些标记应该为关键分类单元的交配冲突研究提供新的研究途径,包括交配后性选择、种群遗传结构、系统地理学和性协同进化。本研究展示了一种有效的微卫星开发方法,并表明基因组 DNA 的轻度测序可以提供众多高度可变的标记。

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