18S rRNA V9 metabarcoding for diet characterization: a critical evaluation with two sympatric zooplanktivorous fish species.
18S rRNA V9 metabarcoding for diet characterization: a critical evaluation with two sympatric zooplanktivorous fish species.
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DOI：
10.1002/ece3.1986
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发表时间：
2016-03
影响因子：
2.6
通讯作者：
Estonba A
中科院分区：
文献类型：
--
作者：
Albaina A;Aguirre M;Abad D;Santos M;Estonba A
使用MiSeq配对末端（PE; 2 × 150 bp）技术探索了18 S rRNA V9元条形码方法用于饮食评估的潜力。为了评价该方法在降解/消化DNA时的性能，分析了来自比斯开湾的两种食动物性鱼类欧洲沙丁鱼（Sardina pilchardus）和欧洲鲱鱼（Sprattus sprattus）的饲料。18 S V9元条形码的分类分辨率和定量潜力，首先在硅片和模拟和现场浮游生物样品进行评估。我们的方法能够以可靠的方式区分参考数据库中的物种，只要在18 S V9区域中存在至少一个可变位置。此外，它成功地区分了两种鱼类之间的饮食，包括沙丁鱼之间的栖息地和昼夜差异，克服了传统视觉饮食分析方法的一些局限性。18 S V9元条形码编码方法的高灵敏度和半定量性质得到了目视显微镜和基于qPCR的结果的支持。这种分子方法为食物网分析提供了一种替代的成本和时间有效的工具。
The potential of the 18S rRNA V9 metabarcoding approach for diet assessment was explored using MiSeq paired‐end (PE; 2 × 150 bp) technology. To critically evaluate the method′s performance with degraded/digested DNA, the diets of two zooplanktivorous fish species from the Bay of Biscay, European sardine (Sardina pilchardus) and European sprat (Sprattus sprattus), were analysed. The taxonomic resolution and quantitative potential of the 18S V9 metabarcoding was first assessed both in silico and with mock and field plankton samples. Our method was capable of discriminating species within the reference database in a reliable way providing there was at least one variable position in the 18S V9 region. Furthermore, it successfully discriminated diet between both fish species, including habitat and diel differences among sardines, overcoming some of the limitations of traditional visual‐based diet analysis methods. The high sensitivity and semi‐quantitative nature of the 18S V9 metabarcoding approach was supported by both visual microscopy and qPCR‐based results. This molecular approach provides an alternative cost and time effective tool for food‐web analysis.