Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species.
Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species.
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DOI：
10.1371/journal.pone.0138888
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
de Boer H
中科院分区：
文献类型：
--
作者：
Osathanunkul M;Suwannapoom C;Ounjai S;Rora JA;Madesis P;de Boer H
DNA条形码结合高分辨解链（Bar-HRM）是一种基于DNA解离动力学的物种鉴别方法。这项工作的目的是评估不同的引物组，来自选定的DNA区域，在巴豆（大戟科），最大的植物属之一，超过1,200种的物种的Bar-HRM分析的适用性。从四个质体区域，matK，rbcL，rpoC，和trnL，和核核糖体标记ITS 1的七个引物对进行了评估（matK，rbcL 1，rbcL 2，rbcL 3，rpoC，trnL和ITS 1）。来自ITS 1区域的引物对是用于鉴定测试物种的单一最有效区域，而rbcL 1引物对给出了最低分辨率。观察到ITS 1条形码是所有评估的区域和引物中总体上用于物种区分的最有用的DNA条形码区域。我们的酒吧HRM结果在这里也提供了进一步的支持的假设，即序列和碱基组成影响DNA双链体的稳定性。
DNA barcoding coupled high resolution melting (Bar-HRM) is an emerging method for species discrimination based on DNA dissociation kinetics. The aim of this work was to evaluate the suitability of different primer sets, derived from selected DNA regions, for Bar-HRM analysis of species in Croton (Euphorbiaceae), one of the largest genera of plants with over 1,200 species. Seven primer pairs were evaluated (matK, rbcL1, rbcL2, rbcL3, rpoC, trnL and ITS1) from four plastid regions, matK, rbcL, rpoC, and trnL, and the nuclear ribosomal marker ITS1. The primer pair derived from the ITS1 region was the single most effective region for the identification of the tested species, whereas the rbcL1 primer pair gave the lowest resolution. It was observed that the ITS1 barcode was the most useful DNA barcoding region overall for species discrimination out of all of the regions and primers assessed. Our Bar-HRM results here also provide further support for the hypothesis that both sequence and base composition affect DNA duplex stability.