Proceedings of the XLVI Italian Society of Agricultural Genetics - SIGA Annual Congress

Giardini Naxos, Italy - 18/21 September, 2002

ISBN 88-900622-3-1

Poster Abstract - 2.24

DEVELOPMENT OF A TAQMAN ASSAY FOR GENETICALLY MODIFIED DURUM WHEAT QUANTIFICATION

FACCIOLI P.*, FERRARI B.*, FINOCCHIARO F.*, LAMACCHIA C.*, SHEWRY P.**, NAPIER J.**, TERZI V.*

*) Istituto Sperimentale per la Cerealicoltura, Via San Protaso 302, I-29017 Fiorenzuola d’Arda (PC), Italy

**) Department of Agricultural Sciences, University of Bristol, IACR-Long Ashton Research Station, Long Ashton, Bristol BS18 9AF, UK

durum wheat, GMO detection, real time PCR, TaqMan assay

The development of biotechnology in agriculture have put in commerce raw materials and food derived from GM plants and the first normative indicates 1% as the maximum percentage of contamination for ingredients in food. Analytical methods are thus necessary to quantitate the presence of GMO in raw materials, feed and food. Because new transformation events are submitted to the EU council, analytical methods need to be continously up-dated. Real time PCR with TaqMan probes has been shown to be extremely accurate and less labour intensive than quantitative competitive PCR , giving high level of precision and improvement of the range of quantitation. Genetically engineered bread and durum wheat containing gluten with improved processing properties have been obtained using an endosperm specific promoter and transgenes coding for high molecular weight glutenin subunits. In this work, we have developed a qualitative and quantitative assay based on real time PCR using a TaqMan fluorogenic probe for the detection and quantitation of a durum wheat line transformed to express a tobacco gene (rab1) involved in protein trafficking through the secretory system of the cell. Control amplification reactions were carried out using a pair of primers based on the sequence of a low molecular weight glutenin, whereas to track GM durum wheat DNA we construct a real time system based on rab1 gene sequence. The sensitivity of the system was tested on serial dilutions of DNA from transgenic plants.