Plant-pollinator interactions over time: Pollen metabarcoding from bees in a historic collection
Plant-pollinator interactions over time: Pollen metabarcoding from bees in a historic collection
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DOI：
10.1111/eva.12707
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发表时间：
2019-02-01
影响因子：
4.1
通讯作者：
Willows-Munro, Sandi
中科院分区：
文献类型：
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作者：
Gous, Annemarie;Swanevelder, Dirk Z. H.;Willows-Munro, Sandi
授粉是农业粮食生产和生态系统维护的关键组成部分，植物与传粉媒介的相互作用是生态和进化研究的重要研究主题。自然历史收藏提供了对在不同空间和时间尺度收集的样本的独特访问。对这些收集品中授粉媒介体内捕获的花粉的植物起源进行鉴定，可以深入了解历史植物群落和授粉媒介偏好的花卉类群。在这项研究中，花粉是从南非国家昆虫保藏中心的Megachile venusta Smith 蜜蜂中采集的，时间跨度为93 年。使用下一代测序方法（MiSeq，Illumina）对混合花粉样品中的三个条形码区域、内部转录间隔区 1 和 2（ITS1 和 ITS2）以及核酮糖 1,5-二磷酸羧化酶 (rbcL) 进行测序。将测序读数与通过从 GenBank 提取序列和分类数据生成的序列参考数据库进行比较。 ITS1 和 ITS2 在所有（或大多数）样本中成功扩增，而 rbcL 的表现不一致。样本年龄对测序成功没有影响。使用 ITS2 进行植物分类比使用 ITS1 条形码数据提供更多信息。这项研究还强调了对综合参考数据库的需求，因为参考数据库中有限的本地植物序列表示导致更高级别的分类单元分类得到更自信的解释。结果表明，早在 1914 年收集的历史蜜蜂标本中昆虫携带的小花粉样本可用于获取花粉元条形码。混合来源花粉样本的 DNA 元条形码提供了一种更快、更准确的确定花粉来源的方法，而不需要专家孢粉学家。使用历史收藏品直接从传粉媒介采集花粉样本为这些收藏品提供了额外的价值。从历史收集中采集花粉样本可以为研究面对全球气候变化时植物群落结构或传粉媒介花卉选择的变化提供空间和时间尺度。
Pollination is a key component in agricultural food production and ecosystem maintenance, with plant-pollinator interactions an important research theme in ecological and evolutionary studies. Natural history collections provide unique access to samples collected at different spatial and temporal scales. Identification of the plant origins of pollen trapped on the bodies of pollinators in these collections provides insight into historic plant communities and pollinators' preferred floral taxa. In this study, pollen was sampled from Megachile venusta Smith bees from the National Collection of Insects, South Africa, spanning 93 years. Three barcode regions, the internal transcribed spacer 1 and 2 (ITS1 and ITS2) and ribulose-1,5-biphosphate carboxylase (rbcL), were sequenced from mixed pollen samples using a next-generation sequencing approach (MiSeq, Illumina). Sequenced reads were compared to sequence reference databases that were generated by extracting sequence and taxonomic data from GenBank. ITS1 and ITS2 were amplified successfully across all (or most) samples, while rbcL performed inconsistently. Age of sample had no impact on sequencing success. Plant classification was more informative using ITS2 than ITS1 barcode data. This study also highlights the need for comprehensive reference databases as limited local plant sequence representation in reference databases resulted in higher-level taxon classifications being more confidently interpreted. The results showed that small, insect-carried pollen samples from historic bee specimens collected from as early as 1914 can be used to obtain pollen metabarcodes. DNA metabarcoding of mixed origin pollen samples provided a faster, more accurate method of determining pollen provenance, without the need for expert palynologists. The use of historic collections to sample pollen directly from pollinators provided additional value to these collections. Sampling pollen from historic collections can potentially provide the spatial and temporal scales for investigations into changes in plant community structure or pollinator floral choice in the face of global climate change.