Evaluation of DNA barcodes in discriminating Quercus species from Lam Dong, Vietnam

Nguyen Van Ngoc, Nong Van Duy, Nguyen Thi Minh Phuong, Hoàng Thị Bình
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Abstract

DNA barcoding is a tool for species discrimination and identification which helps overcome the problem of identification that cannot be covered by morphological identification. Quercus is the second biggest genus of Fagaceae in Vietnam as well as Lam Dong province after Lithocarpus. However, the species discrimination study of the Quercus species in Vietnam and Lam Dong province has yet to be well uncovered due to ambiguous species boundaries and the lack of universal molecular markers. In this study, the DNA barcodes were tested to discriminate among the species of the Quercus genus in Lam Dong province. A total of sixteen and two samples of the Quercus and Lithocarpus genus (out-group) were tested using matK, rbcL, and internal transcribed spacer (ITS). Of which, the new sequences in this study were sequenced from six species and one unknown species of Quercus, the rest was retrieved from GenBank. The BLAST, neighbor-joining, and Bayesian methods were employed to examine species discrimination success. The results showed that ITS was an efficient single-locus barcode for Quercus species by yielding the highest rate of universality as well as the best discriminatory and authentication power among the barcodes examined. In addition, the combination of ITS+matK+rbcL achieved the highest species discrimination. Therefore, matK and rbcL should not be used as DNA barcodes for the species identification of Quercus, whereas the combination of three genes that were proposed in this study is the most suitable DNA barcode for identifying Quercus species in Lam Dong province.
DNA条形码在越南林东栎种鉴别中的应用评价
DNA条形码是一种物种鉴别和鉴定的工具,有助于克服形态鉴定无法涵盖的鉴定问题。栎属是越南和林同省的壳斗科第二大属,仅次于石栎属。然而,由于种界不明确和缺乏普遍的分子标记,越南和林同省栎属植物的物种鉴别研究尚未得到很好的开展。本研究利用DNA条形码技术对林东省栎属树种进行了鉴定。采用matK、rbcL和内部转录间隔器(ITS)对外族栎属(Quercus)和石栎属(Lithocarpus)共16份和2份样品进行了鉴定。其中,新序列来源于栎属6个种和1个未知种,其余序列来源于GenBank。采用BLAST、邻域连接和贝叶斯方法对物种识别成功率进行了检验。结果表明,ITS是一种有效的栎属单位点条形码,具有最高的通用性和最佳的鉴别和鉴定能力。ITS+matK+rbcL组合的物种辨识度最高。因此,matK和rbcL不适合作为栎树物种鉴定的DNA条形码,而本研究提出的3个基因组合是最适合用于林东省栎树物种鉴定的DNA条形码。
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