Development and validation of rapid environmental DNA (eDNA) detection methods for bog turtle (Glyptemys muhlenbergii)

Bog turtles  Glyptemys muhlenbergii are listed as Species of Greatest Conservation Need (SGCN) for wildlife action plans in every state it occurs and multi-state efforts are underway to better characterize extant populations and prioritize restoration efforts. However, traditional sampling methods can be ineffective due to the turtle’s wetland habitat, small size, and burrowing nature. Molecular methods, such as qPCR, provide the ability to overcome this challenge by effectively quantifying minute amounts of turtle DNA left behind in its environment (eDNA). Developing such methods for bog turtles has proved difficult partly because of the high sequence similarity between bog turtles and closely-related, cohabitating species, most often wood turtles ( Glyptemys insculpta ). Additionally, substrates containing bog turtle eDNA are often rich in organics or other substances that frequently inhibit both DNA extraction and qPCR amplification. Here, we describe the development and validation o...

牟氏水龟（Bog turtle，学名Glyptemys muhlenbergii）在其分布的所有州均被列入野生动物保护行动计划中的最高优先级保护物种（Species of Greatest Conservation Need, SGCN），目前跨州协作项目正有序推进，以更精准地刻画其现存种群特征并优先开展修复工作。然而，受该龟类的湿地栖息环境、小巧体型与穴居习性影响，传统采样方法往往难以奏效。诸如定量聚合酶链式反应（qPCR）在内的分子检测技术，可通过有效量化环境中残留的极微量龟类DNA（环境DNA，eDNA），破解这一难题。但为牟氏水龟开发此类检测方法始终存在挑战，部分原因在于牟氏水龟与其近缘且同域共存的物种（最常见的是木雕水龟，学名Glyptemys insculpta）之间的序列相似性极高。此外，携带牟氏水龟eDNA的基质通常富含有机物或其他干扰物质，往往会抑制DNA提取及qPCR扩增过程。本文详述了该方法的开发与验证工作……