Chase Mark W, Salamin Nicolas, Wilkinson Mike, Dunwell James M, Kesanakurthi Rao Prasad, Haider Nadia, Savolainen Vincent
Jodrell Laboratory, Royal Botanic Gardens, Kew Richmond, Surrey TW9 3DS, UK.
Philos Trans R Soc Lond B Biol Sci. 2005 Oct 29;360(1462):1889-95. doi: 10.1098/rstb.2005.1720.
Land plants have had the reputation of being problematic for DNA barcoding for two general reasons: (i) the standard DNA regions used in algae, animals and fungi have exceedingly low levels of variability and (ii) the typically used land plant plastid phylogenetic markers (e.g. rbcL, trnL-F, etc.) appear to have too little variation. However, no one has assessed how well current phylogenetic resources might work in the context of identification (versus phylogeny reconstruction). In this paper, we make such an assessment, particularly with two of the markers commonly sequenced in land plant phylogenetic studies, plastid rbcL and internal transcribed spacers of the large subunits of nuclear ribosomal DNA (ITS), and find that both of these DNA regions perform well even though the data currently available in GenBank/EBI were not produced to be used as barcodes and BLAST searches are not an ideal tool for this purpose. These results bode well for the use of even more variable regions of plastid DNA (such as, for example, psbA-trnH) as barcodes, once they have been widely sequenced. In the short term, efforts to bring land plant barcoding up to the standards being used now in other organisms should make swift progress. There are two categories of DNA barcode users, scientists in fields other than taxonomy and taxonomists. For the former, the use of mitochondrial and plastid DNA, the two most easily assessed genomes, is at least in the short term a useful tool that permits them to get on with their studies, which depend on knowing roughly which species or species groups they are dealing with, but these same DNA regions have important drawbacks for use in taxonomic studies (i.e. studies designed to elucidate species limits). For these purposes, DNA markers from uniparentally (usually maternally) inherited genomes can only provide half of the story required to improve taxonomic standards being used in DNA barcoding. In the long term, we will need to develop more sophisticated barcoding tools, which would be multiple, low-copy nuclear markers with sufficient genetic variability and PCR-reliability; these would permit the detection of hybrids and permit researchers to identify the 'genetic gaps' that are useful in assessing species limits.
陆地植物因两个普遍原因而在DNA条形码技术应用方面存在问题:(i)藻类、动物和真菌中使用的标准DNA区域变异性极低;(ii)典型的陆地植物质体系统发育标记(如rbcL、trnL-F等)变异似乎过少。然而,没有人评估过当前的系统发育资源在鉴定(相对于系统发育重建)背景下的效果如何。在本文中,我们进行了这样的评估,特别是针对陆地植物系统发育研究中常用的两个标记,质体rbcL和核糖体DNA大亚基的内部转录间隔区(ITS),发现尽管GenBank/EBI中目前可用的数据并非用于条形码目的而产生,且BLAST搜索也不是用于此目的的理想工具,但这两个DNA区域表现良好。这些结果对于一旦广泛测序后使用质体DNA的更多可变区域(例如psbA-trnH)作为条形码来说是个好兆头。短期内,使陆地植物条形码技术达到目前其他生物所使用标准的努力应会迅速取得进展。DNA条形码用户有两类,即分类学以外领域的科学家和分类学家。对于前者,使用线粒体和质体DNA这两个最易评估的基因组,至少在短期内是一种有用工具,能让他们继续开展研究,这些研究依赖于大致了解所处理的是哪些物种或物种组,但同样这些DNA区域在分类学研究(即旨在阐明物种界限的研究)中存在重要缺陷。出于这些目的,来自单亲(通常是母系)遗传基因组的DNA标记只能提供提高DNA条形码技术所使用分类标准所需信息的一半。从长远来看,我们需要开发更复杂的条形码工具,即多个具有足够遗传变异性和PCR可靠性的低拷贝核标记;这些标记将允许检测杂种,并使研究人员能够识别有助于评估物种界限且有用的“遗传间隙”。