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IJUP08 - Universidade do Porto

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Genetic characterization by SSR of six Olea europaea regional<br />

varieties for authenticity evaluation of olive oils<br />

J. Lopes 1 , R. Coelho 2 , M. A. Faria 1 , E. Nunes 2 and M.B.P.P Oliveira 1<br />

1 REQUIMTE, Serviço de Bromatologia, Faculty of Pharmacy, University of <strong>Porto</strong>, Rua Aníbal<br />

Cunha, 164, 4099-030 <strong>Porto</strong>, Portugal.<br />

2 CIBIO, Faculdade de Ciências da <strong>Universidade</strong> <strong>do</strong> <strong>Porto</strong>, Campus Agrário de Vairão, Rua <strong>do</strong><br />

Monte-Castro, Vairão, 4480-661 Vila <strong>do</strong> Conde<br />

Olive cultivation plays an important role in rural economy of certain regions as Trás-os-<br />

Montes. The importance attributed to this cultivar is expressed in the number of regional<br />

varieties that constitutes local germplasm collections. In order to assist germplasm<br />

management and to develop markers for olive oil varietal authentication genotyping the<br />

referred varieties is of crucial importance. Several metho<strong>do</strong>logies have been used to<br />

genotype and evaluate the relationships among cultivars of olive tree, including isozymes,<br />

RFLP, RAPD, ISSR, SSR and SNPs. SSR are the markers of choice since they are highly<br />

polymorphic, co-<strong>do</strong>minant and relatively easy to use.<br />

In this work we have studied the six most relevant regional varieties (Cobrançosa, Verdeal<br />

Transmontana, Madural, Santulhana, Re<strong>do</strong>ndal e Negrinha) at the molecular level using 15<br />

selected SSR loci [1-3]. Genomic ADN was obtained from silica gel dried leaves using<br />

both a CTAB classical method and a commercially available extraction kit.<br />

Results include the construction of a database for further olive oil authenticity studies and<br />

the evaluation of the genetic relationships amongst the varieties. The chosen markers<br />

permitted the complete discrimination of the studied varieties with a probability of identity<br />

of 1,52×10 -5 . Genetic relationships amongst the varieties are graphically represented in Fig.<br />

1.<br />

Linkage Distance<br />

88<br />

78<br />

68<br />

58<br />

48<br />

38<br />

28<br />

18<br />

REDONDAL<br />

COBRANÇOSA<br />

SANTULHANA<br />

NEGRINHA VERDEAL TRANSMONTANA MADURAL<br />

Fig. 1. Dendrogram showing the genetic relations<br />

of the 6 studied varieties<br />

[1] Sefc, K.M., Lopes, M.S., Men<strong>do</strong>nça, D., Santos, M.R., Macha<strong>do</strong>, M.L.C., Macha<strong>do</strong>, A.C.<br />

(2000), Identification of microsatellite loci in olive (Olea europaea) and their characterization in<br />

Italian and Iberian olive trees ,Molecular Ecology, 9, 1171-1193.<br />

[2] Cipriani, G., Marrazzo, M.T., Marconi, R., Cimato, A., Testolin, R. (2002), Microsatellite<br />

markers isolated in olive (Olea europaea L.) are suitable for individual fingerprinting and reveal<br />

polymorphism within ancient cultivars ,Theor Appl Genet., 104, 223-228.<br />

[3] Rallo, P., Dora<strong>do</strong>, G., Martín, A. (2000), Development of simple sequence repeats (SSRs) in<br />

olive tree (Olea europaea L.), Theor. Appl. Genet., 101, 984-989.<br />

208

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