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基于特定分类单元的聚合酶链式反应(PCR)在蝙蝠粪便中鉴定节肢动物猎物的 DNA 条形码。

Taxon-specific PCR for DNA barcoding arthropod prey in bat faeces.

机构信息

School of Biological Sciences, University of Bristol, Woodland Road, Bristol, UK.

出版信息

Mol Ecol Resour. 2011 Mar;11(2):236-44. doi: 10.1111/j.1755-0998.2010.02920.x. Epub 2010 Sep 26.

DOI:10.1111/j.1755-0998.2010.02920.x
PMID:21429129
Abstract

The application of DNA barcoding to dietary studies allows prey taxa to be identified in the absence of morphological evidence and permits a greater resolution of prey identity than is possible through direct examination of faecal material. For insectivorous bats, which typically eat a great diversity of prey and which chew and digest their prey thoroughly, DNA-based approaches to diet analysis may provide the only means of assessing the range and diversity of prey within faeces. Here, we investigated the effectiveness of DNA barcoding in determining the diets of bat species that specialize in eating different taxa of arthropod prey. We designed and tested a novel taxon-specific primer set and examined the performance of short barcode sequences in resolving prey species. We recovered prey DNA from all faecal samples and subsequent cloning and sequencing of PCR products, followed by a comparison of sequences to a reference database, provided species-level identifications for 149/207 (72%) clones. We detected a phylogenetically broad range of prey while completely avoiding detection of nontarget groups. In total, 37 unique prey taxa were identified from 15 faecal samples. A comparison of DNA data with parallel morphological analyses revealed a close correlation between the two methods. However, the sensitivity and taxonomic resolution of the DNA method were far superior. The methodology developed here provides new opportunities for the study of bat diets and will be of great benefit to the conservation of these ecologically important predators.

摘要

DNA 条码技术在饮食研究中的应用允许在没有形态学证据的情况下鉴定猎物分类群,并允许比通过直接检查粪便材料更能确定猎物的身份。对于食虫蝙蝠来说,它们通常吃各种各样的猎物,并且会彻底咀嚼和消化猎物,因此基于 DNA 的饮食分析方法可能是评估粪便中猎物范围和多样性的唯一方法。在这里,我们研究了 DNA 条码技术在确定专门以不同节肢动物猎物为食的蝙蝠物种饮食方面的有效性。我们设计并测试了一种新的分类特异性引物组,并研究了短条码序列在解决猎物物种方面的性能。我们从所有粪便样本中回收了猎物 DNA,并随后对 PCR 产物进行克隆和测序,然后将序列与参考数据库进行比较,为 149/207(72%)个克隆提供了种级别的鉴定。我们检测到了广泛的进化亲缘猎物,同时完全避免了非目标群体的检测。总共从 15 个粪便样本中鉴定出 37 个独特的猎物分类群。DNA 数据与平行形态分析的比较表明两种方法之间存在密切的相关性。然而,DNA 方法的灵敏度和分类分辨率要高得多。这里开发的方法为蝙蝠饮食研究提供了新的机会,并将极大地有益于这些生态重要捕食者的保护。

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