陆地植物的DNA条形码方法。
DNA barcoding methods for land plants.
作者信息
Fazekas Aron J, Kuzmina Maria L, Newmaster Steven G, Hollingsworth Peter M
机构信息
Department of Integrative Biology, University of Guelph, Guelph, ON, Canada.
出版信息
Methods Mol Biol. 2012;858:223-52. doi: 10.1007/978-1-61779-591-6_11.
DNA barcoding in the land plants presents a number of challenges compared to DNA barcoding in many animal clades. The CO1 animal DNA barcode is not effective for plants. Plant species hybridize frequently, and there are many cases of recent speciation via mechanisms, such as polyploidy and breeding system transitions. Additionally, there are many life-history trait combinations, which combine to reduce the likelihood of a small number of markers effectively tracking plant species boundaries. Recent results, however, from the two chosen core plant DNA barcode regions rbcL and matK plus two supplementary regions trnH-psbA and internal transcribed spacer (ITS) (or ITS2) have demonstrated reasonable levels of species discrimination in both floristic and taxonomically focused studies. We describe sampling techniques, extraction protocols, and PCR methods for each of these two core and two supplementary plant DNA barcode regions, with extensive notes supporting their implementation for both low- and high-throughput facilities.
与许多动物类群的DNA条形码相比，陆地植物的DNA条形码存在一些挑战。动物的细胞色素氧化酶亚基1（CO1）DNA条形码对植物无效。植物物种频繁杂交，并且存在许多通过多倍体和繁殖系统转变等机制近期形成物种的情况。此外，有许多生活史性状组合，这些组合共同降低了少数标记有效追踪植物物种界限的可能性。然而，最近从两个选定的核心植物DNA条形码区域——叶绿体大亚基核糖体RNA基因（rbcL）和成熟酶K基因（matK），加上两个补充区域——转运RNA基因（trnH）-光系统Ⅱ蛋白D1基因（psbA）和内部转录间隔区（ITS）（或ITS2）得到的结果表明，在植物区系研究和分类学重点研究中，这些区域都具有合理水平的物种鉴别能力。我们描述了这两个核心和两个补充植物DNA条形码区域各自的采样技术、提取方案和聚合酶链反应（PCR）方法，并给出了大量注释，以支持它们在低通量和高通量设施中的应用。
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