Department of Zoology and Animal Biology, University of Geneva, CH-1211 Geneva 4, Switzerland.
J Eukaryot Microbiol. 2010 Mar-Apr;57(2):197-205. doi: 10.1111/j.1550-7408.2009.00468.x. Epub 2010 Jan 22.
Ribosomal DNA (rDNA) sequences have been shown to be very useful for identification of microbial eukaryotes. Usually, complete or long partial sequences of the rDNA genes are analysed. However, the development of new massive sequencing technologies producing a large amount of relatively short sequences raises the question about the minimum length of rDNA fragments necessary for species distinction in environmental sampling. To answer this question, we compared six variable regions of the small subunit (SSU) rDNA of foraminifera, known to have rapidly evolving ribosomal genes. For each region, we analysed (1) the sequence divergence between and within foraminiferal morphospecies, (2) the intraspecific polymorphism, and (3) the ability of each region to recognize the phylotypes inferred from analysis of a longer fragment. Our results show that although the variable regions differ considerably between taxonomic groups, most of them perform very well as species identifiers. Taking into account different analyses, the expansion segment of Helix 37 appears to be the best candidate for barcoding foraminifera. We propose that this relatively short region, averaging 50-60 nt in length, could be an ideal barcode for identification of foraminifera in environmental samples using massive sequencing approach.
核糖体 DNA(rDNA)序列已被证明非常有助于鉴定微生物真核生物。通常,分析 rDNA 基因的完整或长部分序列。然而,新的大规模测序技术的发展产生了大量相对较短的序列,这就提出了关于环境采样中区分物种所需的 rDNA 片段最小长度的问题。为了回答这个问题,我们比较了有孔虫小亚基(SSU)rDNA 的六个可变区,已知这些基因的核糖体进化迅速。对于每个区域,我们分析了(1)有孔虫形态种之间和内部的序列差异,(2)种内多态性,以及(3)每个区域识别从更长片段分析推断出的系统发育型的能力。我们的结果表明,尽管不同的分类群之间的可变区有很大差异,但它们中的大多数作为物种鉴定者表现得非常好。考虑到不同的分析,Helix 37 的扩展片段似乎是有孔虫条形码的最佳候选者。我们建议,这个相对较短的区域,平均长度为 50-60nt,可以成为使用大规模测序方法识别环境样本中有孔虫的理想条形码。