Isolation of microsatellite loci and reliable genotyping using noninvasive samples of a critically endangered primate, Trachypithecus leucocephalus.
Isolation of microsatellite loci and reliable genotyping using noninvasive samples of a critically endangered primate, Trachypithecus leucocephalus.
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DOI：
10.1111/1749-4877.12192
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发表时间：
2016-07
影响因子：
3.3
通讯作者：
Weiran Wang;Yu Qiao;Yitao Zheng;Meng Yao
中科院分区：
文献类型：
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作者：
Weiran Wang;Yu Qiao;Yitao Zheng;Meng Yao
遗传信息对于确定保护优先事项和制定保护战略至关重要。目前迫切需要非侵入性的遗传工具来研究野生种群的亚洲colobine猴。这些物种中的大多数面临着生境破坏、种群减少甚至灭绝的威胁，但通常缺乏关于其遗传多样性和种群结构的信息。由于对濒危物种操作的严格规定，以及在其居住的具有挑战性的环境中捕获这些树栖和快速移动的猴子的困难和风险，这些物种的遗传样本和组织收集一直很稀少。这些困难阻碍了通常来自组织或血液的分子遗传标记的发展。在这项研究中，我们提出了一种方法从头微卫星分离和基因分型使用的DNA从非侵入性的起源的一个严重濒危的亚洲colobine，白头叶猴（Trachyphecus leucocephalus）。从头发中分离的基因组DNA被证明是足够的微卫星富集和分离，与类似的分离效率从组织DNA。我们确定了20个多态性微卫星位点，并评估其扩增成功率和基因分型的可靠性与86个现场采集的粪便样本。这些结果表明，该面板的座位可以产生可靠的基因型从粪便样品，并代表了一个有用的工具，非侵入性调查的遗传结构，个体识别和亲缘关系评估在这一高度濒危物种。我们的方法可以应用于其他野生物种的保护遗传学研究，缺乏序列信息和组织样本。
Genetic information can be critical in identifying conservation priorities and developing conservation strategies. There is an urgent need for noninvasive genetic tools to study the wild populations of Asian colobine monkeys. The majority of these species are threatened with habitat destruction, population reduction and even extinction, but generally lack information on their genetic diversity and population structure. Genetic sampling and tissue collection have been scarce in these species owing to strict regulations on manipulation of endangered species, and the difficulties and risks associated with capturing these arboreal and fast-moving monkeys in the challenging environments that they inhabit. These difficulties have hindered the development of molecular genetic markers, which are usually derived from tissues or blood. In this study, we present a method for de novo microsatellite isolation and genotyping using DNA from noninvasive origins of a critically endangered Asian colobine, the white-headed langur (Trachypithecus leucocephalus). Genomic DNA isolated from hair was shown to be sufficient for microsatellite enrichment and isolation, with similar isolation efficiencies as from tissue DNA. We identified and characterized 20 polymorphic microsatellite loci, and evaluated their amplification success and genotyping reliability with 86 field-collected fecal samples. These results show that this panel of loci can produce reliable genotypes from fecal samples, and represent a useful tool for noninvasive investigation of genetic structure, individual identification and kinship assessment in this highly endangered species. Our approach can be applied to conservation genetic studies of other wild species that lack sequence information and tissue samples.