Identification of Uvaria sp by barcoding coupled with high-resolution melting analysis (Bar-HRM).
Identification of Uvaria sp by barcoding coupled with high-resolution melting analysis (Bar-HRM).
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DOI：
10.4238/gmr.15017405
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发表时间：
2016-01
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通讯作者：
M. Osathanunkul;P. Madesis;S. Ounjai;Kitti Pumiputavon;R. Somboonchai;P. Lithanatudom;T. Chaowasku;J. Wipasa;C. Suwannapoom
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作者：
M. Osathanunkul;P. Madesis;S. Ounjai;Kitti Pumiputavon;R. Somboonchai;P. Lithanatudom;T. Chaowasku;J. Wipasa;C. Suwannapoom
DNA条形码是大约十年前开发的，它依赖于基因组的短而标准化的区域来识别植物和动物物种。这种方法不仅可以用来鉴定已知的物种，也可以用来发现新的物种。许多序列存储在世界各地的在线数据库中。在物种鉴定中节省成本和时间（通过省略测序步骤）的方法之一是使用可用的条形码数据来设计优化的引物以用于进一步分析，例如高分辨率解链分析（HRM）。本研究的目的是确定混合方法Bar-HRM（DNA条形码结合HRM）的有效性，以确定具有相似外部形态特征的物种，而不是进行传统的分类鉴定，需要标本的主要部分（叶，花，果实）。用于检测的标本是那些无法在种属水平上鉴别的标本，通过形态学鉴别表明可能是长柄紫玉盘（Uvaria longipes）或紫玉盘（Uvaria wrayias）。在Bar-HRM中使用来自叶绿体区域（matK、psbA-trnH、rbcL和trnL）的引物对。从psbA-trnH引物获得的结果是足够好的，以帮助鉴定标本，而其余的不是。Bar-HRM分析被证明是一种快速、经济的植物物种鉴定方法。
DNA barcoding, which was developed about a decade ago, relies on short, standardized regions of the genome to identify plant and animal species. This method can be used to not only identify known species but also to discover novel ones. Numerous sequences are stored in online databases worldwide. One of the ways to save cost and time (by omitting the sequencing step) in species identification is to use available barcode data to design optimized primers for further analysis, such as high-resolution melting analysis (HRM). This study aimed to determine the effectiveness of the hybrid method Bar-HRM (DNA barcoding combined with HRM) to identify species that share similar external morphological features, rather than conduct traditional taxonomic identification that require major parts (leaf, flower, fruit) of the specimens. The specimens used for testing were those, which could not be identified at the species level and could either be Uvaria longipes or Uvaria wrayias, indicated by morphological identification. Primer pairs derived from chloroplast regions (matK, psbA-trnH, rbcL, and trnL) were used in the Bar-HRM. The results obtained from psbA-trnH primers were good enough to help in identifying the specimen while the rest were not. Bar-HRM analysis was proven to be a fast and cost-effective method for plant species identification.