Grapevine carpological remains revealed the existence of a Neolithic domesticated Vitis vinifera L. specimen containing ancient DNA partially preserved in modern ecotypes

• Taxonomic identification of Neolithic carpological remains.
• Morphological discrimination of wild and cultivated Vitis vinifera seeds.
• Sequencing of Barcoding genes starting from grapevine ancient DNA.
• Ancient microsatellite allelic variants preserved in modern ecotypes.
• Domestication and distribution of the grapevine in Italy.

Thirty-four carpological remains, found in the Neolithic layers of Grotta della Serratura (Southern Italy), were subjected to morphological analysis in order to identify their botanical origin. The phenotype of these samples and the measurement of morphometrical indices clearly revealed that they were Vitis vinifera ssp. sativa seeds, suggesting the existence of this plant species in that specific context. Molecular investigations detected the presence of ancient DNA inside the pips. The principal plastidial Barcoding genes were amplified, sequenced and aligned with the accessions of GenBank nucleotide database, to confirm the taxonomic identity of the specimens and to individuate all the genetic changes that occurred in these selected regions of the grapevine genome during the evolution, since Neolithic until today. Then, ten microsatellite loci were successfully typified, starting from the ancient DNA. The genetic profile was compared to the CRA-VIT molecular database of Conegliano and to the Italian Vitis Database. The results showed a high conservation rate of the antique allelic variants in modern grapevine accessions and the existence of possible evolutionary relationships with current Vitis vinifera ecotypes. All these data improved the knowledge about grapevine cultivation, diffusion and use in Southern Italy by Neolithic human civilization. Moreover, they also gave new information for reconstruction and interpretation of past natural environments.