LIB/Zoological Research Museum Alexander Koenig (ZFMK), Centre for Biodiversity Monitoring, Bonn, Germany.
Mol Ecol Resour. 2023 Jan;23(1):92-105. doi: 10.1111/1755-0998.13694. Epub 2022 Aug 18.
DNA metabarcoding is routinely used for biodiversity assessment, in particular targeting highly diverse groups for which limited taxonomic expertise is available. Various protocols are currently in use, although standardization is key to its application in large-scale monitoring. DNA metabarcoding of arthropod bulk samples can be conducted either destructively from sample tissue, or nondestructively from sample fixative or lysis buffer. Nondestructive methods are highly desirable for the preservation of sample integrity but have yet to be experimentally evaluated in detail. Here, we compare diversity estimates from 14 size-sorted Malaise trap samples processed consecutively with three nondestructive approaches (one using fixative ethanol and two using lysis buffers) and one destructive approach (using homogenized tissue). Extraction from commercial lysis buffer yielded comparable species richness and high overlap in species composition to the ground tissue extracts. A significantly divergent community was detected from preservative ethanol-based DNA extraction. No consistent trend in species richness was found with increasing incubation time in lysis buffer. These results indicate that nondestructive DNA extraction from incubation in lysis buffer could provide a comparable alternative to destructive approaches with the added advantage of preserving the specimens for postmetabarcoding taxonomic work but at a higher cost per sample.
DNA 代谢组学通常用于生物多样性评估,特别是针对那些具有有限分类学专业知识的高度多样化群体。目前有各种不同的方法,但标准化是其在大规模监测中应用的关键。对节肢动物混合样本进行 DNA 代谢组学分析,可以从样本组织进行破坏性分析,也可以从样本固定液或裂解缓冲液进行非破坏性分析。非破坏性方法非常适合保存样本完整性,但尚未进行详细的实验评估。在这里,我们比较了 14 个大小排序的马勒氏陷阱样本的连续处理结果,其中包括三种非破坏性方法(一种使用固定液乙醇,两种使用裂解缓冲液)和一种破坏性方法(使用均质组织)。从商业裂解缓冲液中提取的 DNA 与地面组织提取物的物种丰富度和物种组成高度重叠。从基于保存剂乙醇的 DNA 提取中检测到一个明显不同的群落。在裂解缓冲液中孵育时间的增加与物种丰富度没有一致的趋势。这些结果表明,从裂解缓冲液的孵育中进行非破坏性 DNA 提取可以为破坏性方法提供一个可比较的替代方案,其额外的优势是可以为后续代谢组学分类工作保存标本,但每个样本的成本更高。