Molecular Analysis of Ancient DNA Extracted from 3250-3450 Year-old Plant Seeds Excavated from Tepe Sagz Abad in Iran

To determine ancient seed species, 3250-3450 year-old charred seeds obtained from different Iranian archaeological sites were studied using Scanning Electron Microscopy (SEM) and molecular analysis. SEM analysis of ancient seeds revealed that the surface of the seeds was damaged. Therefore, we could not accurately identify their species. Molecular analysis on ancient specimens was done on different samples obtained from Masjede Kabood (Tabriz), Tepe Rahmat Abad (Pasargad) and Tepe Sagz Abad (Qazvin plain). The specific primer pairs were designed based on a part of the promoter region of the High Molecular Weight (HMW) glutenin gene and a short fragment of the vrs1 gene were verified on samples of modern wheat and barley varieties, respectively. The designated primers failed to amplify ancient DNAs (aDNAs) obtained from Masjede Kabood and Tepe Rahmat Abad, but successfully amplified the aDNA obtained from Tepe Sagz Abad. This finding was expected since the latter seeds had a better morphological preservation in comparison to the former ones. The accuracy of the amplified products was further proved by cloning and sequencing