The impact of extraction method and pollen concentration on community composition for pollen metabarcoding

IF 2.7 3区 生物学 Q2 PLANT SCIENCES
Arne Devriese, Gerrit Peeters, Rein Brys, Hans Jacquemyn
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引用次数: 0

Abstract

Premise

Plants and pollinators closely interact with each other to form complex networks of species interactions. Metabarcoding of pollen collections has recently been proposed as an advantageous method for the construction of such networks, but the extent to which diversity and community analyses depend on the extraction method and pollen concentration used remains unclear.

Methods

In this study, we used a dilution series of two pollen mixtures (a mock community and pooled natural pollen loads from bumblebees) to assess the effect of mechanical homogenization and two DNA extraction kits (spin column DNA extraction kit and magnetic bead DNA extraction kit) on the detected pollen richness and community composition.

Results

All species were successfully detected using the three methods, even in the most dilute samples. However, the extraction method had a significant effect on the detected pollen richness and community composition, with simple mechanical homogenization introducing an extraction bias.

Discussion

Our findings suggest that all three methods are effective for detecting plant species in the pollen loads on insects, even in cases of very low pollen loads. However, our results also indicate that extraction methods can have a profound impact on the ability to correctly assess the community composition of the pollen loads on insects. The choice of extraction methodology should therefore be carefully considered to ensure reliable and unbiased results in pollen diversity and community analyses.

Abstract Image

提取方法和花粉浓度对花粉代谢标码群落组成的影响
前言植物与传粉昆虫之间密切互动,形成复杂的物种互动网络。最近有人提出,花粉采集的元条码是构建此类网络的一种有利方法,但多样性和群落分析在多大程度上取决于所使用的提取方法和花粉浓度仍不清楚。方法在这项研究中,我们使用了两种花粉混合物的稀释系列(模拟群落和来自熊蜂的汇集天然花粉量)来评估机械均质和两种 DNA 提取试剂盒(旋柱 DNA 提取试剂盒和磁珠 DNA 提取试剂盒)对检测到的花粉丰富度和群落组成的影响。讨论 我们的研究结果表明,这三种方法都能有效地检测昆虫花粉负载中的植物物种,即使在花粉负载量很低的情况下也是如此。然而,我们的研究结果也表明,提取方法会对正确评估昆虫花粉载荷群落组成的能力产生深远影响。因此,应仔细考虑提取方法的选择,以确保花粉多样性和群落分析结果的可靠性和公正性。
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来源期刊
CiteScore
7.30
自引率
0.00%
发文量
50
审稿时长
12 weeks
期刊介绍: Applications in Plant Sciences (APPS) is a monthly, peer-reviewed, open access journal promoting the rapid dissemination of newly developed, innovative tools and protocols in all areas of the plant sciences, including genetics, structure, function, development, evolution, systematics, and ecology. Given the rapid progress today in technology and its application in the plant sciences, the goal of APPS is to foster communication within the plant science community to advance scientific research. APPS is a publication of the Botanical Society of America, originating in 2009 as the American Journal of Botany''s online-only section, AJB Primer Notes & Protocols in the Plant Sciences. APPS publishes the following types of articles: (1) Protocol Notes describe new methods and technological advancements; (2) Genomic Resources Articles characterize the development and demonstrate the usefulness of newly developed genomic resources, including transcriptomes; (3) Software Notes detail new software applications; (4) Application Articles illustrate the application of a new protocol, method, or software application within the context of a larger study; (5) Review Articles evaluate available techniques, methods, or protocols; (6) Primer Notes report novel genetic markers with evidence of wide applicability.
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