Recovery, identification and relationships by microsatellite analysis of ancient grapevine cultivars from Castilla-La Mancha: the largest wine growing region in the world

A total of 374 grapevine accessions collected during prospecting works in different wine regions of Castilla-La Mancha in the period 2004–2011, in addition to 55 accessions belonging to national and international cultivars, conserved in the grapevine variety collection of Castilla-La Mancha were studied. For genetic characterization of all accessions, 26 nuclear and 5 chloroplast microsatellite markers were analyzed, yielding 158 different genetic profiles. Of the profiles, 44 were recorded as “new genotypes”, not previously described in the literature consulted. Some of them belonged to traditional cultivars cited as being present in the region in the early twentieth century, such as Churriago, Mizancho, Tortozona Tinta, Maquias and Zurieles and whose head was lost, while others were nameless cultivars hidden in ancient vineyards and unknown to science. The 26 microsatellite loci used reflected a high discrimination power (99.998 %) and a low probability that two randomly chosen individuals had identical genotypes, the probability of identity (1.02 × 10−22) was comparable to those reported for other grapevine germplasms. Four chlorotypes, A, B, C and D, were defined among the studied cultivars, most of the cultivars (53 %) carried chlorotype A, followed by chlorotypes D (28 %), C (11 %) and B (7 %). Moreover, the 158 single genotypes were analyzed for possible parent-offspring relationships. A total of nine putative parentages not previously published were obtained, and another seven were confirmed.