Ultrastructural study of archaeological Vitis vinifera L. seeds using rapid-freeze fixation and substitution

The ultrastructure of Vitis vinifera seeds from different archaeological sites was studied. Preservation status differed between sites. Preliminary investigations of grape seeds from Poggio Bacherina (Chianciano Terme, Siena) and Miranduolo (Chiusdino, Siena) showed collapsed or charred tegument, making this material suitable for morphometric studies only. Rapid-freeze fixation and substitution of grape seeds from Shahr-I Sokhta in Iran and via De’ Castellani in Florence revealed well preserved tegument suitable for chemical and cytochemical analysis. Energy dispersive X-ray microanalysis was used to determine chemical composition. Cytochemical analysis based on fluorescent staining with DAPI suggested the presence of cytoplasm residues.