Taberlet Pierre, Coissac Eric, Pompanon François, Gielly Ludovic, Miquel Christian, Valentini Alice, Vermat Thierry, Corthier Gérard, Brochmann Christian, Willerslev Eske
Laboratoire d'Ecologie Alpine, CNRS UMR 5553, Université Joseph Fourier, BP 53, 38041 Grenoble Cedex 9, France.
Nucleic Acids Res. 2007;35(3):e14. doi: 10.1093/nar/gkl938. Epub 2006 Dec 14.
DNA barcoding should provide rapid, accurate and automatable species identifications by using a standardized DNA region as a tag. Based on sequences available in GenBank and sequences produced for this study, we evaluated the resolution power of the whole chloroplast trnL (UAA) intron (254-767 bp) and of a shorter fragment of this intron (the P6 loop, 10-143 bp) amplified with highly conserved primers. The main limitation of the whole trnL intron for DNA barcoding remains its relatively low resolution (67.3% of the species from GenBank unambiguously identified). The resolution of the P6 loop is lower (19.5% identified) but remains higher than those of existing alternative systems. The resolution is much higher in specific contexts such as species originating from a single ecosystem, or commonly eaten plants. Despite the relatively low resolution, the whole trnL intron and its P6 loop have many advantages: the primers are highly conserved, and the amplification system is very robust. The P6 loop can even be amplified when using highly degraded DNA from processed food or from permafrost samples, and has the potential to be extensively used in food industry, in forensic science, in diet analyses based on feces and in ancient DNA studies.
DNA条形码技术应通过使用标准化的DNA区域作为标签,来提供快速、准确且可自动化的物种鉴定。基于GenBank中可用的序列以及本研究产生的序列,我们评估了整个叶绿体trnL(UAA)内含子(254 - 767 bp)以及用高度保守引物扩增的该内含子较短片段(P6环,10 - 143 bp)的分辨能力。整个trnL内含子用于DNA条形码技术的主要局限性仍然是其相对较低的分辨率(GenBank中67.3%的物种能够明确鉴定)。P6环的分辨率更低(19.5%能够鉴定),但仍高于现有的其他系统。在诸如源自单一生态系统的物种或常见食用植物等特定情况下,分辨率要高得多。尽管分辨率相对较低,但整个trnL内含子及其P6环具有许多优点:引物高度保守,扩增系统非常稳健。即使使用来自加工食品或永久冻土样本的高度降解DNA,P6环也能被扩增,并且有潜力在食品工业、法医学、基于粪便的饮食分析以及古DNA研究中广泛应用。