Liu Zhen, Chen Shi-Lin, Song Jing-Yuan, Zhang Shou-Jun, Chen Ke-Li
Department of Pharmacy, The 309 Hospital of Chinese People's Liberation Army, Beijing.
Pharmacogn Mag. 2012 Jan;8(29):4-11. doi: 10.4103/0973-1296.93301.
This study aims to determine the candidate markers that can be used as DNA barcode in the Lauraceae family.
Polymerase chain reaction amplification, sequencing efficiency, differential intra- and interspecific divergences, DNA barcoding gap, and identification efficiency were used to evaluate the four different DNA sequences of psbA-trnH, matK, rbcL, and ITS2. We tested the discrimination ability of psbA-trnH in 68 plant samples belonging to 42 species from 11 distinct genera and found that the rate of successful identification with the psbA-trnH was 82.4% at the species level. However, the correct identification of matK and rbcL were only 30.9% and 25.0%, respectively, using BLAST1. The PCR amplification efficiency of the ITS2 region was poor; thus, ITS2 was not included in subsequent experiments. To verify the capacity of the identification of psbA-trnH in more samples, 175 samples belonging to 117 species from the experimental data and from the GenBank database of the Lauraceae family were tested.
Using the BLAST1 method, the identification efficiency were 84.0% and 92.3% at the species and genus level, respectively.
Therefore, psbA-trnH is confirmed as a useful marker for differentiating closely related species within Lauraceae.
本研究旨在确定可作为樟科植物DNA条形码的候选标记。
利用聚合酶链反应扩增、测序效率、种内和种间差异分歧、DNA条形码间隙及鉴定效率，对psbA-trnH、matK、rbcL和ITS2这四种不同的DNA序列进行评估。我们对来自11个不同属的42个物种的68个植物样本测试了psbA-trnH的鉴别能力，发现在物种水平上，psbA-trnH的成功鉴定率为82.4%。然而，使用BLAST1时，matK和rbcL的正确鉴定率分别仅为30.9%和25.0%。ITS2区域的PCR扩增效率较差；因此，ITS2未纳入后续实验。为了在更多样本中验证psbA-trnH的鉴定能力，对来自樟科实验数据和GenBank数据库的117个物种的175个样本进行了测试。
使用BLAST1方法，在物种和属水平上的鉴定效率分别为84.0%和92.3%。
因此，psbA-trnH被确认为区分樟科内近缘物种的有用标记。
