The project addresses the traceability of DNA and metabolites as instrumental to reconstruct the cultivar origin and composition of high quality olive oils and to identify eventual frauds. This objective will be addressed with a complementary new approach based on combination of genomic, metabolomic, bioinformatic and socio-economical methodologies.

The workplan will be articulated in five main steps following an initial feasibility study.

 

 

The aims of the study are: application and comparison of DNA extraction procedures to olive oils at different stages of the production chain; analysis of the amount of DNA extracted and evaluation of its integrity in terms of fragment sizes; applicability of these DNAs to PCR, RealTime PCR, exploitation for molecular markers and microarray technologies. These techniques will be utilised to assess DNA presence in olive products along the production chain, to assess DNA degradation at different stages of oil production and processing and to identify and quantify DNA in oil samples.

 

 

Collection and database of information will concern up to a maximum of 500 cultivars, based on those growing in EU and elsewhere in the Mediterranean basin as well as the relative information about areas of cultivation for each cultivar. The cultivars present in the World Olive Germplasm Bank which includes about 350 cultivars from all the Mediterranean Countries will be considered as basic information. A different database will collect information regarding oil production, with priority to cultivars belonging to PDO or RDO, mixing of varieties, fluxes of import/export through the Mediterranean basin. A list of high quality olive oils and their variety composition will also be prepared.

 

 

Application of genomic techniques to DNA requires the availability of descriptors of genetic diversity allowing the recognition of the cultivar. All available information will be collected in a database and new additional markers will be designed and tested on DNA extracted from olive plants. Molecular markers will be developed as microsatellites, RAPDs, AFLPs, retrotransposon-based, ITS sequences, SNPs. All informative markers will be converted into Sequence Characterised Amplified Regions (SCARs) for PCR analysis.
Metabolomics requires analytical methods such as HPLC and ES-MS/MS. Different cultivars and olive oils provided by Partners across Europe will be analysed for similarities and differences in composition. During the project partners will be involved in ring tests and inter-laboratory exchanges to ascertain the reliability of metabolomic descriptors, and to evaluate the entity of the fluctuations which can be caused by environments, processing and production techniques. Only the more reliable  among descriptors will be chosen for application in other WPs.

 

 

Quantitative PCR analysis with RealTime technology will be developed for implementing quantitative diagnostic tests for detecting adulteration in oil. The use of DNA microarray technology will allow multiple screening for the presence of DNA sequences in oil samples. Peptide nucleic acids (PNAs) will be used to increase the specificity of molecular tools resolution. All previously identified diagnostic markers will be adapted to application in these platforms prior to forensic testing.

 

 

The techniques and the molecular descriptors of metabolic and genetic diversity developed will be finally applied to forensic analysis of olive oil samples, derived from producers or obtained from reconstruction experiments. The aim is to test the applicability of the traceability techniques to identification of the cultivars in pure or mixed oils, discrimination of EU and non-EU cultivars, definition of low quality oils. At the end of the project, the tool kit for forensic analysis will include two separate components: the molecular tools and the statistical tools for data handling and analysis. The barcode for ID card will transfer all molecular data achieved, in information of simple interpretation to the consumers: origin, composition, quality of cultivars.

 

 

Contacts with regulators, industrialists and consumers associations will be established to ensure an extended audience for the results and to facilitate the broader dissemination of them toward industry. Efforts will be made in the EU and outside to promote traceability and the policy of EU in this field at scientific level.

 

 

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