701 Chloroplast DNA Diversity in Portuguese Grape Cultivars I.D. Castro and O. Pinto-Carnide Dpto de Genética e Biotecnologia Centro de Genómica e Biotecnologia da Universidade de Trás-os-Montes e Alto Douro Instituto de Biotecnologia e Bioengenharia (CGB-UTAD/IBB) Vila Real Portugal J.M. Ortiz and J.P. Martín Dpto de Biología Vegetal Escuela Técnica Superior de Ingenieros Agrónomos Universidad Politécnica de Madrid Ciudad Universitaria s/n Madrid Spain Keywords: Vitis vinifera L., cpSSR, PCR-RFLP, chloroplast variability, grapevine domestication, autochthonous Portuguese grapevines Abstract Chloroplast genome (cpDNA) analysis is an excellent tool in phylogenetic and phylogeographic studies. Investigations about grapevine domestication are being carried out based on chloroplast microsatellites (cpSSR) variation in samples of Vitis vinifera ssp. sylvestris and sativa from the whole area of the species’ distribution (from the Iberian Peninsula to the Middle East). Polymorphisms detected suggest the existence of at least two important origins for the cultivated germplasm, one in the Near East and another in the western Mediterranean region. The objective of this study was to assess the diversity in the chloroplast genome of a group of Portuguese cultivars, with the purpose to know their regional genetic patterns as well as to infer their putative origin. A total of 38 cultivars, representative of cultivated grapevine material in the north of Portugal, and three international cultivars (outgroups) were studied using three cpSSR loci and analysing several fragments of the large single copy (LSC) region of the cpDNA by cleaved amplified polymorphic sequences (CAPS) or PCR-RFLP methodology. The three cpSSR loci were polymorphic, two different alleles were found in each locus. Allele variants of the three loci combined in a total of three different haplotypes. The most frequent haplotype (70.7%) was the same previously reported as the most frequent in the Iberian Peninsula and western Europe. One of the haplotypes was unique in ‘Rabigato’. This haplotype was previous- ly proposed to be an ancestral haplotype. Inference about the progenitors of the cultivar ‘Touriga Franca’ is done. This cultivar could be the progeny of ‘Touriga Nacional’ and ‘Marufo’. The cpSSR analysis revealed that ‘Touriga Nacional’ could have been the male progenitor. Ten fragments of the LSC region from Vitis vinifera cpDNA were amplified and digested with two restriction enzymes (HinfI and TaqI), according to CAPS methodology. No polymorphisms were found in the 41 cultivars, but this analysis will be expanded to the rest of the LSC region. INTRODUCTION Portugal, compared with other western countries, such as Spain, France and Italy, with a long history in grape cultivation has the highest density of grape landrace cultivars (Martins, 2009). Chloroplast genome analysis can be an excellent tool helping in tracking the course of grapevine material dispersion to ascertain its phylogenetic origin. Chloroplast DNA markers have been used in comparative analysis involving wild and cultivated grapevines and in parent/offspring studies due to its conserved nature and to maternal inheritance. In particular, chloroplast microsatellites (cpSSRs), have been widely used in characterization of cultivated and wild grapevine genotypes, either from all areas of the species’ cultivation (Arroyo-Garcia et al., 2006) and in more limited studies relative to central and southern Portugal (Cunha et al., 2009), northern Italy (Rossoni et al., 2003) or northern central Spain (Santana et al., 2010) cultivars. Genetic analysis by using cpSSRs combined with PCR-RFLP markers would be informative Proc. XXVIII th IHC – III rd IS on Plant Genetic Resources Ed.: K.E. Hummer Acta Hort. 918, ISHS 2011