Universal primers and PCR of gut contents to study marine invertebrate diets
Universal primers and PCR of gut contents to study marine invertebrate diets
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DOI：
10.1111/j.1365-294x.2005.02448.x
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发表时间：
2005-03-01
影响因子：
4.9
通讯作者：
Yayanos, AA
中科院分区：
文献类型：
--
作者：
Blankenship, LE;Yayanos, AA
通过肠道内容物分析确定海洋无脊椎动物的饮食是有问题的。许多被食用的生物一旦被部分消化就变得无法识别，而那些具有坚硬残留物的生物（例如硅藻骨骼）可能会使分析产生偏差。在这里，我们适应基于DNA的方法类似于用于微生物多样性调查作为一种新的方法来研究饮食的巨食性（深海端足类Scopelocheirus schellenbergi和Eurythenes gryllus）和微食性（双壳类Lucinoma aequizonata）的饲养员在深海。聚合酶链反应（PCR）结合“通用”引物扩增的线粒体细胞色素c氧化酶I（COI）基因的部分动物摄入的S。schellenbergi和E. gryllus和18S rRNA基因的较小真核生物摄入的L.水杉将扩增的序列与GenBank中的序列相结合，构建摄入微生物的系统发育树。分析表明，S. schellenbergi、黑腹叶蝉E. gryllus和L. aquizonata的饮食比以前认为的要多样化得多，为这些物种的觅食策略提供了新的线索。最后，我们讨论了这种技术的优点和缺点，以及它对其他无脊椎动物的饮食分析的潜在适用性。
Determining the diets of marine invertebrates by gut content analysis is problematic. Many consumed organisms become unrecognizable once partly digested, while those with hard remains (e.g. diatom skeletons) may bias the analysis. Here, we adapt DNA-based methods similar to those used for microbial diversity surveys as a novel approach to study the diets of macrophagous (the deep-sea amphipods Scopelocheirus schellenbergi and Eurythenes gryllus) and microphagous (the bivalve Lucinoma aequizonata) feeders in the deep sea. Polymerase chain reaction (PCR) in conjunction with 'universal' primers amplified portions of the mitochondrial cytochrome c oxidase I (COI) gene for animals ingested by S. schellenbergi and E. gryllus and the 18S rRNA gene for lesser eukaryotes ingested by L. aequizonata. Amplified sequences were combined with sequences from GenBank to construct phylogenetic trees of ingested organisms. Our analyses indicate that S. schellenbergi, E. gryllus and L. aequizonata diets are considerably more diverse than previously thought, casting new light on the foraging strategies of these species. Finally, we discuss the strengths and weaknesses of this technique and its potential applicability to diet analyses of other invertebrates.