Development of Taxon-Specific Sequences of Common Wheat for the Detection of Genetically Modified Wheat

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• 作者：Mayu Iida ; Satomi Yamashiro ; Hirohito Yamakawa ; Katsuyuki Hayakawa ; Hideo Kuribara ; Takashi Kodama ; Satoshi Furui ; Hiroshi Akiyama ; Tamio Maitani ; and Akihiro Hino
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2005
• 出版时间：August 10, 2005
• 年：2005
• 卷：53
• 期：16
• 页码：6294 - 6300
• 全文大小：370K
• 年卷期：v.53,no.16(August 10, 2005)
• ISSN：1520-5118

文摘

Qualitative and quantitative Polymerase Chain Reaction (PCR) systems aimed at the specific detectionand quantification of common wheat DNA are described. Many countries have issued regulations tolabel foods that include genetically modified organisms (GMOs). PCR technology is widely recognizedas a reliable and useful technique for the qualitative and quantitative detection of GMOs. Detectionmethods are needed to amplify a target GM gene, and the amplified results should be comparedwith those of the corresponding taxon-specific reference gene to obtain reliable results. This paperdescribes the development of a specific DNA sequence in the waxy-D1 gene for common wheat(Triticum aestivum L.) and the design of a specific primer pair and TaqMan probe on the waxy-D1gene for PCR analysis. The primers amplified a product (Wx012) of 102 bp. It is indicated that theWx012 DNA sequence is specific to common wheat, showing homogeneity in qualitative PCR resultsand very similar quantification accuracy along 19 distantly related common wheat varieties. In Southernblot and real-time PCR analyses, this sequence showed either a single or a low number of copygenes. In addition, by qualitative and quantitative PCR using wx012 primers and a wx012-T probe,the limits of detection of the common wheat genome were found to be about 15 copies, and thereproducibility was reliable. In consequence, the PCR system using wx012 primers and wx012-Tprobe is considered to be suitable for use as a common wheat-specific taxon-specific reference genein DNA analyses, including GMO tests.Keywords: Triticum aestivum L.; common wheat; genetically modified; GMO detection; real-time PCR;endogenous reference gene; waxy; granule-bound starch synthase; quantitative analysis