PDF(Pubmed)
Abstract：
Molecular methods are currently some of the best-suited technologies for implementation in insect monitoring. However, the field is developing rapidly and lacks agreement on methodology or community standards. To apply DNA-based methods in large-scale monitoring, and to gain insight across commensurate data, we need easy-to-implement standards that improve data comparability. Here, we provide three recommendations for how to improve and harmonize efforts in biodiversity assessment and monitoring via metabarcoding: (i) we should adopt the use of synthetic spike-ins, which will act as positive controls and internal standards; (ii) we should consider using several markers through a multiplex polymerase chain reaction (PCR) approach; and (iii) we should commit to the publication and transparency of all protocol-associated metadata in a standardized fashion. For (i), we provide a ready-to-use recipe for synthetic cytochrome c oxidase spike-ins, which enable between-sample comparisons. For (ii), we propose two gene regions for the implementation of multiplex PCR approaches, thereby achieving a more comprehensive community description. For (iii), we offer guidelines for transparent and unified reporting of field, wet-laboratory and dry-laboratory procedures, as a key to making comparisons between studies. Together, we feel that these three advances will result in joint quality and calibration standards rather than the current laboratory-specific proof of concepts. This article is part of the theme issue \'Towards a toolkit for global insect biodiversity monitoring\'.
摘要：
分子方法是目前最适合在昆虫监测中实施的一些技术。然而,该领域发展迅速，在方法或社区标准上缺乏共识。为了将基于DNA的方法应用于大规模监测，并获得相应数据的洞察力，我们需要易于实施的标准来提高数据的可比性。这里,我们就如何改善和协调生物多样性评估和监测方面的工作提供了三项建议：（i）我们应该采用合成刺突蛋白，作为阳性对照和内部标准;(ii)我们应该考虑通过多重聚合酶链反应(PCR)方法使用几种标记;和(iii)我们应该致力于以标准化的方式公布和透明所有与方案相关的元数据。对于（i），我们提供了合成细胞色素c氧化酶的现成配方，这可以实现样本之间的比较。对于（ii），我们提出了两个基因区域来实施多重PCR方法，从而实现更全面的社区描述。对于（iii），我们提供透明和统一的现场报告准则，湿实验室和干实验室程序，作为在研究之间进行比较的关键。一起,我们认为，这三个进步将导致质量和校准标准的联合，而不是目前针对实验室的概念证明。本文是“迈向全球昆虫生物多样性监测工具包”主题的一部分。
