Ferri G, Corradini B, Ferrari F, Santunione A L, Palazzoli F, Alu' M
Dipartimento di Medicina Diagnostica, Clinica e di Sanità Pubblica, Struttura Complessa di Medicina Legale, Università di Modena e Reggio Emilia, Italy.
Dipartimento di Medicina Diagnostica, Clinica e di Sanità Pubblica, Struttura Complessa di Medicina Legale, Università di Modena e Reggio Emilia, Italy.
Forensic Sci Int Genet. 2015 Mar;15:131-6. doi: 10.1016/j.fsigen.2014.10.005. Epub 2014 Oct 16.
The ambitious idea of using a short piece of DNA for large-scale species identification (DNA barcoding) is already a powerful tool for scientists and the application of this standard technique seems promising in a range of fields including forensic genetics. While DNA barcoding enjoyed a remarkable success for animal identification through cytochrome c oxidase I (COI) analysis, the attempts to identify a single barcode for plants remained a vain hope for a longtime. From the beginning, the Consortium for the Barcode of Life (CBOL) showed a lack of agreement on a core plant barcode, reflecting the diversity of viewpoints. Different research groups advocated various markers with divergent set of criteria until the recent publication by the CBOL-Plant Working Group. After a four-year effort, in 2009 the International Team concluded to agree on standard markers promoting a multilocus solution (rbcL and matK), with 70-75% of discrimination to the species level. In 2009 our group firstly proposed the broad application of DNA barcoding principles as a tool for identification of trace botanical evidence through the analysis of two chloroplast loci (trnH-psbA and trnL-trnF) in plant species belonging to local flora. Difficulties and drawbacks that were encountered included a poor coverage of species in specific databases and the lack of authenticated reference sequences for the selected markers. Successful preliminary results were obtained providing an approach to progressively identify unknown plant specimens to a given taxonomic rank, usable by any non-specialist botanist or in case of a shortage of taxonomic expertise. Now we considered mandatory to update and to compare our previous findings with the new selected plastid markers (matK+rbcL), taking into account forensic requirements. Features of all the four loci (the two previously analyzed trnH-psbA+trnL-trnF and matK+rbcL) were compared singly and in multilocus solutions to assess the most suitable combination for forensic botany. Based on obtained results, we recommend the adoption of a two-locus combination with rbcL+trnH-psbA plastid markers, which currently best satisfies forensic needs for botanical species identification.
利用一小段DNA进行大规模物种鉴定（DNA条形码技术）这一宏伟构想，已然成为科学家们手中的有力工具，并且这项标准技术在包括法医遗传学在内的一系列领域的应用前景似乎颇为广阔。虽然通过细胞色素c氧化酶I（COI）分析，DNA条形码技术在动物鉴定方面取得了显著成功，但长期以来，为植物确定单一条形码的尝试一直是徒劳的希望。从一开始，生命条形码联盟（CBOL）在核心植物条形码上就缺乏共识，这反映了观点的多样性。不同的研究小组倡导各种具有不同标准的标记物，直到CBOL植物工作组最近发表相关成果。经过四年努力，国际团队在2009年得出结论，同意采用标准标记物，推广多基因座解决方案（rbcL和matK），在物种水平上的鉴别率为70 - 75%。2009年，我们小组首次提出广泛应用DNA条形码技术原理，通过分析本地植物群中植物物种的两个叶绿体基因座（trnH - psbA和trnL - trnF）来鉴定微量植物证据。遇到的困难和缺点包括特定数据库中物种覆盖不足以及所选标记物缺乏经过验证的参考序列。我们获得了成功的初步结果，提供了一种逐步将未知植物标本鉴定到给定分类等级的方法，可供任何非专业植物学家使用，或在分类专业知识短缺的情况下使用。现在，考虑到法医要求，我们认为必须更新并将我们之前的发现与新选定的质体标记物（matK + rbcL）进行比较。对所有四个基因座（之前分析的两个trnH - psbA + trnL - trnF以及matK + rbcL）的特征进行了单独和多基因座组合比较，以评估最适合法医植物学的组合。基于所得结果，我们建议采用rbcL + trnH - psbA质体标记物的双基因座组合，目前它最能满足植物物种鉴定的法医需求。
