Devriese Arne, Peeters Gerrit, Brys Rein, Jacquemyn Hans
Department of Biology, Plant Conservation and Population Biology KU Leuven Leuven B-3001 Belgium.
Research Institute for Forest and Nature Gaverstraat 4 Geraardsbergen B-9500 Belgium.
Appl Plant Sci. 2024 Aug 6;12(5):e11601. doi: 10.1002/aps3.11601. eCollection 2024 Sep-Oct.
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.
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.
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.
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.
植物和传粉者相互密切作用，形成复杂的物种相互作用网络。花粉收集的代谢条形码分析最近被提议作为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依赖于所使用的提取方法和花粉浓度仍不清楚。
在本研究中，我们使用了两种花粉混合物（一个模拟群落和来自熊蜂的混合天然花粉负载）的稀释系列，以评估机械匀浆和两种DNA提取试剂盒（旋转柱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。
使用这三种方法均成功检测到了所有物种，即使在最稀释的样本中也是如此。然而，提取方法对检测到的花粉丰富度和群落组成有显著影响，简单的机械匀浆会引入提取偏差。
我们的研究结果表明，所有这三种方法对于检测昆虫身上花粉负载中的植物物种都是有效的，即使在花粉负载非常低的情况下也是如此。然而，我们的结果还表明，提取方法可能会对正确评估昆虫身上花粉负载的群落组成的能力产生深远影响。因此，在花粉多样性和群落分析中，应仔细考虑提取方法的选择，以确保获得可靠且无偏差的结果。
