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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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