Osathanunkul Maslin, Ounjai Sarawut, Osathanunkul Rossarin, Madesis Panagiotis
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand.
Center of Excellence in Bioresources for Agriculture, Industry and Medicine, Chiang Mai University, Chiang Mai, Thailand.
PLoS One. 2017 Oct 11;12(10):e0186283. doi: 10.1371/journal.pone.0186283. eCollection 2017.
It is long believed that some spices may help protect against certain chronic conditions. Spices are usually parts of plants that have been powdered into small pieces. Have you ever wondered what the curry powder in your dish is made of? The aim of this work was to develop an appropriate DNA-based method for assessment of spice identity. Selecting the best marker for species recognition in the Zingiberaceae family. Six DNA regions were investigated in silico, including ITS, matK, rbcL, rpoC, trnH-psbA and trnL. Then, only four regions (ITS, matK, rbcL and trnH-psbA) were included in the simulated HRM (High-resolution Melting) analysis as the results from previous analysis showed that rpoC and trnL may not be suitable to be used to identify Zingiberaceae species in HRM analysis based on both the percentage of nucleotide variation and GC content. Simulated HRM analysis was performed to test the feasibility of Bar-HRM. We found that ITS2 is the most effective region to be used for identification of the studied species and thus was used in laboratory HRM analysis. All seven tested Zingiberaceae plants were then able to be distinguished using the ITS2 primers in laboratory HRM. Most importantly the melting curves gained from fresh and dried tissue overlapped, which is a crucial outcome for the applicability of the analysis. The method could be used in an authentication test for dried products. In the authentication test, only one of seven store-sold Zingiberaceae products that were tested contained the species listed on their labels, while we found substitution/contamination of the tested purchased products in the rest.
长期以来,人们一直认为某些香料可能有助于预防某些慢性疾病。香料通常是植物的一部分,已被磨成小碎片。你有没有想过你盘子里的咖喱粉是由什么制成的?这项工作的目的是开发一种合适的基于DNA的方法来评估香料的身份。在姜科植物中选择用于物种识别的最佳标记。通过计算机模拟研究了六个DNA区域,包括ITS、matK、rbcL、rpoC、trnH-psbA和trnL。然后,只有四个区域(ITS、matK、rbcL和trnH-psbA)被纳入模拟的高分辨率熔解(HRM)分析,因为先前的分析结果表明,基于核苷酸变异百分比和GC含量,rpoC和trnL可能不适用于在HRM分析中鉴定姜科植物物种。进行了模拟HRM分析以测试Bar-HRM的可行性。我们发现ITS2是用于鉴定所研究物种的最有效区域,因此被用于实验室HRM分析。然后,在实验室HRM中使用ITS2引物能够区分所有七种测试的姜科植物。最重要的是,从新鲜组织和干燥组织获得的熔解曲线重叠,这对于该分析的适用性来说是一个关键结果。该方法可用于干燥产品的鉴定测试。在鉴定测试中,测试的七种市售姜科产品中只有一种含有其标签上列出的物种,而我们发现其余购买的测试产品存在替代/污染情况。