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Power and limitations of the chloroplast trnL (UAA) intron for plant DNA barcoding

机译:叶绿体trnL(UAA)内含子用于植物DNA条形码的功能和局限性

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摘要

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.
机译:DNA条形码应使用标准化的DNA区域作为标签,以提供快速,准确和可自动化的物种识别。根据GenBank中可用的序列和本研究产生的序列,我们评估了整个叶绿体trnL(UAA)内含子(254–767 bp)和该内含子的较短片段(P6环的10–143 bp)的分辨能力)用高度保守的引物扩增。整个trnL内含子用于DNA条形码的主要限制仍然是其相对较低的分辨率(明确鉴定了来自GenBank的67.3%的物种)。 P6回路的分辨率较低(确定为19.5%),但仍高于现有替代系统的分辨率。在特定情况下,例如来自单一生态系统的物种或通常食用的植物,分辨率更高。尽管分辨率较低,但整个trnL内含子及其P6环仍具有许多优势:引物高度保守,扩增系统非常耐用。当使用来自加工食品或多年冻土样品的高度降解的DNA时,P6环甚至可以被扩增,并且有可能被广泛用于食品工业,法医学,基于粪便的饮食分析以及古代DNA研究中。

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