Laury-Ann Dumoulin, Marion Chevrinais, Richard St-Louis, Geneviève J. Parent
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引用次数: 0
摘要
天然基质会影响环境 DNA(eDNA)的检测。基质浸润化合物对聚合酶链反应(PCR)步骤的影响是大多数抑制研究的重点。在典型的实验室工作流程(即 DNA 提取和 PCR 步骤)中,影响 eDNA 检测的因素大多不为人知。在此,我们评估了海洋沉积物中的四种金属离子(Ca2+、Fe3+、Mn2+、Cu2+)对提取和 PCR 检测步骤中 DNA 可检测性的影响。将单一金属离子和外源 DNA 添加到经过化学处理以去除抑制剂的海洋沉积物中。我们的研究结果表明,表层海洋沉积物中天然富集的钙、铁和锰离子会完全阻碍DNA的检测。而加入基质中的铜离子则可将 DNA 检测率提高 7.7%。我们还观察到钙离子和铁离子对 DNA 检测能力的双峰抑制作用,这表明提取和 PCR 步骤都会受到影响。我们的发现凸显了 eDNA 检测的新局限性。我们还讨论了如何优化适用于多种基质的 eDNA 检测方案。
Metal ions limit or enhance environmental DNA detectability in marine sediments
Natural matrices affect environmental DNA (eDNA) detections. Effects of matrix-eluted compounds on the polymerase chain reaction (PCR) step have been the focus of most inhibition studies. Factors affecting eDNA detections in a typical laboratory workflow, i.e., DNA extraction and PCR steps, are mostly unknown. Here, we assessed the effect of four metal ions (Ca2+, Fe3+, Mn2+, Cu2+) present in marine sediments on DNA detectability for both the extraction and the PCR detection steps. A single metal ion and exogenous DNA were added to marine sediments treated chemically to remove inhibitors. Our results showed that natural concentrations of calcium, iron, and manganese ions in surface marine sediments can impede completely DNA detections. Alternatively, copper ions added to the matrix increased DNA detectability by 7.7%. We also observed bimodal inhibitory effects of calcium and iron ions on DNA detectability, suggesting that the extraction and the PCR steps are both affected. Our findings highlight new limitations of eDNA detections. Avenues to optimize eDNA detection protocols applicable to multiple matrices are discussed.