Y-chromosomal testing of brown bears (Ursus arctos): Validation of a multiplex PCR-approach for nine STRs suitable for fecal and hair samples
Y-chromosomal testing of brown bears (Ursus arctos): Validation of a multiplex PCR-approach for nine STRs suitable for fecal and hair samples
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DOI：
10.1016/j.fsigen.2015.07.018
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发表时间：
2015-11-01
影响因子：
3.1
通讯作者：
Eiken, Hans Geir
中科院分区：
文献类型：
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作者：
Aarnes, Siv Grethe;Hagen, Snorre B.;Eiken, Hans Geir
高分辨率Y染色体标记已被应用于人类和其他灵长类动物，以研究种群遗传学、迁徙、社会结构和生殖。Y连锁标记可以直接评估独特的男性遗传谱系的遗传结构和基因流，也可能对野生动物保护和法医学有用，但到目前为止只有少数野生物种可以使用。因此，我们开发了两种多重PCR反应，包括从棕熊(Ursus Arctos)中识别的九个Y-STR标记，并在毛发、粪便和组织样本上进行了测试。对多重聚合酶链式反应方法进行了优化，并对其物种特异性、敏感性和卡顿-峰比进行了分析。九个Y-STR还显示了雄性黑熊和雄性北极熊的特定STR片段，而当使用雌性北极熊或其他12种哺乳动物的雄性DNA时，这九个标记都没有产生任何PCR产物。两次聚合酶链式反应的多重聚合酶链式反应只需0.2 ng的模板即可扩增。来自毛发、粪便扫描和组织的DNA模板的精确度很高，标准偏差小于0.14，中位数口吃比率在0.04到0.63之间。在8个二核苷酸和1个四核苷酸重复标记中，我们在9个标记中的7个标记中检测到了9-25个重复单位的简单重复结构。在9个Y-STR中有8个存在等位基因变异，每个标记有2-9个等位基因，在453个主要来自北欧的雄性棕熊中总共有36个等位基因。我们的结论是，这9个Y-STR的多重PCR方法将提供雄性熊Y染色体的特异性和适用于保护和野生动物法医学样本的证据。(C)2015爱思唯尔爱尔兰有限公司。保留所有权利。
High-resolution Y-chromosomal markers have been applied to humans and other primates to study population genetics, migration, social structures and reproduction. Y-linked markers allow the direct assessment of the genetic structure and gene flow of uniquely male inherited lineages and may also be useful for wildlife conservation and forensics, but have so far been available only for few wild species. Thus, we have developed two multiplex PCR reactions encompassing nine Y-STR markers identified from the brown bear (Ursus arctos) and tested them on hair, fecal and tissue samples. The multiplex PCR approach was optimized and analyzed for species specificity, sensitivity and stutter-peak ratios. The nine Y-STRs also showed specific STR-fragments for male black bears and male polar bears, while none of the nine markers produced any PCR products when using DNA from female bears or males from 12 other mammals. The multiplex PCR approach in two PCR reactions could be amplified with as low as 0.2 ng template input. Precision was high in DNA templates from hairs, fecal scats and tissues, with standard deviations less than 0.14 and median stutter ratios from 0.04 to 0.63. Among the eight di- and one tetra-nucleotide repeat markers, we detected simple repeat structures in seven of the nine markers with 9-25 repeat units. Allelic variation was found for eight of the nine Y-STRs, with 2-9 alleles for each marker and a total of 36 alleles among 453 male brown bears sampled mainly from Northern Europe. We conclude that the multiplex PCR approach with these nine Y-STRs would provide male bear Y-chromosomal specificity and evidence suited for samples from conservation and wildlife forensics. (C) 2015 Elsevier Ireland Ltd. All rights reserved.