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通过对非侵入性粪便和降解样本进行测序实现数千样本的基因分型(GT-seq):用于持续监测加拿大北极熊种群的新方法。

Genotyping-in-thousands by sequencing (GT-seq) of noninvasive faecal and degraded samples: A new panel to enable ongoing monitoring of Canadian polar bear populations.

作者信息

Hayward Kristen M, Clemente-Carvalho Rute B G, Jensen Evelyn L, de Groot Peter V C, Branigan Marsha, Dyck Markus, Tschritter Christina, Sun Zhengxin, Lougheed Stephen C

机构信息

Department of Biology, Queen's University, Kingston, Ontario, Canada.

Hakai Institute, Campbell River, British Columbia, Canada.

出版信息

Mol Ecol Resour. 2022 Jul;22(5):1906-1918. doi: 10.1111/1755-0998.13583. Epub 2022 Feb 8.

Abstract

Genetic monitoring using noninvasive samples provides a complement or alternative to traditional population monitoring methods. However, next-generation sequencing approaches to monitoring typically require high quality DNA and the use of noninvasive samples (e.g., scat) is often challenged by poor DNA quality and contamination by nontarget species. One promising solution is a highly multiplexed sequencing approach called genotyping-in-thousands by sequencing (GT-seq), which can enable cost-efficient genomics-based monitoring for populations based on noninvasively collected samples. Here, we develop and validate a GT-seq panel of 324 single nucleotide polymorphisms (SNPs) optimized for genotyping of polar bears based on DNA from noninvasively collected faecal samples. We demonstrate (1) successful GT-seq genotyping of DNA from a range of sample sources, including successful genotyping (>50% loci) of 62.9% of noninvasively collected faecal samples determined to contain polar bear DNA; and (2) that we can reliably differentiate individuals, ascertain sex, assess relatedness, and resolve population structure of Canadian polar bear subpopulations based on a GT-seq panel of 324 SNPs. Our GT-seq data reveal spatial-genetic patterns similar to previous polar bear studies but at lesser cost per sample and through use of noninvasively collected samples, indicating the potential of this approach for population monitoring. This GT-seq panel provides the foundation for a noninvasive toolkit for polar bear monitoring and can contribute to community-based programmes - a framework which may serve as a model for wildlife conservation and management for species worldwide.

摘要

使用非侵入性样本进行基因监测为传统种群监测方法提供了补充或替代方案。然而,下一代测序监测方法通常需要高质量的DNA,而使用非侵入性样本(如粪便)往往因DNA质量差和非目标物种污染而面临挑战。一种有前景的解决方案是一种名为测序数千分型(GT-seq)的高度多重测序方法,它能够基于非侵入性收集的样本,实现对种群进行经济高效的基于基因组学的监测。在此,我们开发并验证了一个包含324个单核苷酸多态性(SNP)的GT-seq面板,该面板针对基于非侵入性收集的粪便样本中的北极熊DNA进行基因分型进行了优化。我们证明了:(1)来自一系列样本来源的DNA成功进行了GT-seq基因分型,包括对62.9%经确定含有北极熊DNA的非侵入性收集粪便样本成功进行基因分型(>50%位点);以及(2)我们能够基于一个包含324个SNP的GT-seq面板可靠地区分个体、确定性别、评估亲缘关系并解析加拿大北极熊亚种群的种群结构。我们的GT-seq数据揭示了与以往北极熊研究相似的空间遗传模式,但每个样本成本更低且通过使用非侵入性收集的样本,表明了这种方法在种群监测方面的潜力。这个GT-seq面板为北极熊监测的非侵入性工具包奠定了基础,并可为基于社区的项目做出贡献——这一框架可作为全球物种野生动物保护和管理的典范。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f260/9305793/6991463c2eb1/MEN-22-1906-g001.jpg

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