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实现昆虫群落分子特征描述方法的可比性和标准化的三个步骤。

Three steps towards comparability and standardization among molecular methods for characterizing insect communities.

机构信息

Bioinformatics and Genetics Department, Swedish Museum of Natural History, PO Box 50007, Stockholm, 104 05, Sweden.

Leibniz Institute for the Analysis of Biodiversity Change, Museum Koenig Bonn, 53113, Germany.

出版信息

Philos Trans R Soc Lond B Biol Sci. 2024 Jun 24;379(1904):20230118. doi: 10.1098/rstb.2023.0118. Epub 2024 May 6.

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 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

分子方法目前是最适合用于昆虫监测的技术之一。然而,该领域发展迅速,缺乏关于方法或社区标准的共识。为了在大规模监测中应用基于 DNA 的方法,并在相当的数据中获得洞察力,我们需要易于实施的标准来提高数据可比性。在这里,我们提供了三条关于如何通过宏条形码来改进和协调生物多样性评估和监测工作的建议:(i)我们应该采用合成标准物质的使用,这将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)方法使用多个标记;(iii)我们应该承诺以标准化的方式公布和透明化所有与协议相关的元数据。对于(i),我们提供了一种用于合成细胞色素氧化酶标准物质的即用型配方,这使得可以进行样本间的比较。对于(ii),我们提出了两个用于实施多重 PCR 方法的基因区域,从而实现更全面的群落描述。对于(iii),我们提供了透明和统一的现场、湿实验室和干实验室程序报告指南,这是进行研究之间比较的关键。总的来说,我们认为这三个进展将导致联合质量和校准标准,而不是当前实验室特定的概念验证。本文是主题为“迈向全球昆虫生物多样性监测工具包”的一部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b78/11070264/8856bb6041b2/rstb20230118f01.jpg

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