Abstract：
The use of genetic data for timber species and population assignment is a powerful tool for combating the illegal timber trade, but the challenges of extracting DNA from timber have prevented the routine use of genetics as a supply chain management tool. To overcome these challenges, we explored the feasibility of focused ultrasound extraction (FUSE) for rapid DNA release from timber. Using high-pressure ultrasound pulses, FUSE generates a cavitation bubble cloud that disintegrates samples into acellular debris, resulting in the mechanical release of DNA. In this work, FUSE was applied to white oak (Quercus alba) timber shavings to test the feasibility of using FUSE for timber DNA extraction for the first time. Results showed that FUSE processing disintegrated the tissue samples and released significant quantities of DNA. After five minutes of tissue processing DNA quantities of 0.21 ± 0.02 ng/mg, 0.99 ± 0.32 ng/mg, and 0.14 ± 0.01 ng/mg, were released from medium, coarse, and combination shaving groups, respectively. Amplification and sequencing of regions within the matK and rbcL chloroplast genes confirmed that the quality of DNA prepared with FUSE was suitable for PCR and short-read sequencing applications. Overall, these results show that FUSE can serve as a DNA sample preparation method capable of releasing high-quality DNA from timber in a fraction of the time required by conventional extraction methods. Based on the improved efficiency of DNA release with FUSE, ongoing work aims to develop this technology into portable systems that can be used to rapidly prepare timber samples for genetic species identification.
摘要：
利用木材树种和种群分配的遗传数据是打击非法木材贸易的有力工具,但是从木材中提取DNA的挑战阻碍了遗传学作为供应链管理工具的常规使用。为了克服这些挑战，我们探索了聚焦超声提取（FUSE）从木材中快速释放DNA的可行性。使用高压超声波脉冲，保险丝产生空化气泡云，将样品分解成无细胞碎片，导致DNA的机械释放。在这项工作中,将FUSE应用于白橡木（Quercusalba）木材刨花，以首次测试使用FUSE提取木材DNA的可行性。结果表明，FUSE处理分解了组织样品并释放了大量的DNA。经过五分钟的组织处理DNA量为0.21±0.02ng/mg，0.99±0.32ng/mg，和0.14±0.01ng/mg，从培养基中释放出来，粗糙，和组合剃须组，分别。matK和rbcL叶绿体基因内区域的扩增和测序证实，用FUSE制备的DNA质量适合PCR和短读取测序应用。总的来说,这些结果表明，FUSE可以作为DNA样品制备方法，能够在常规提取方法所需的一小部分时间内从木材中释放出高质量的DNA。基于FUSE的DNA释放效率的提高，正在进行的工作旨在将这项技术开发到便携式系统中，该系统可用于快速制备木材样品以进行遗传物种鉴定。
