Pisum &amp; Ervilia Tetovac: Made in Early Iron Age Leskovac, Part two: Extraction of the ancient DNA from charred seeds from the site of Hissar in South Serbia

Zivko Jovanovic,Svetlana Radovic,Vuk Djordjevic,Aleksandar Mikic,Aleksandra Nikolic,Nemanja Stanisavljevic,Aleksandar Medovic

doi:10.5937/ratpov1101227j

Abstract

The extracts were prepared from the samples of 3,200-year-old charred pea and bitter vetch seeds from the site of Hissar near Leskovac, South Serbia, using two different DNA extraction procedures. We used CTAB method with some modification and obtained low quantity of ancient DNA in comparison with the second method used - commercial available kit. After the extraction, a whole genome amplification using Phi29 DNA polymerase was performed. The amplified DNAs were used for PCR reaction using primers for 26S rDNA gene, which is located on the nuclear genome. The single band corresponding to 26S rDNA fragment from modern relatives was obtained. We conclude that DNA from charred pea and vetch seed can be extracted and used for further archaeobotanical analysis at the molecular level.

Highlights

After death DNA is degraded by various biotic and abiotic processes, resulting in fragmentation and modification of original sequence information (Schlumbaum et al 2008)
We described the methods for Ancient DNA (aDNA) isolation from pea (Pisum sativum), one of the first crops cultivated by man, and from bitter vetch (Vicia ervilia), two charred pulse crop storaged of the fortified hill settlement Hissar
There are data about aDNA isolation from ancient plants, this is the first time that aDNA from ancient pea and bitter vetch seed was isolated

Summary

Introduction

After death DNA is degraded by various biotic and abiotic processes, resulting in fragmentation and modification of original sequence information (Schlumbaum et al 2008). It has emerged that cold, dry and low oxygen environments are beneficial for DNA survival; for this reason, freshly excavated material is best stored cold or frozen (Burger et al 1999, Smith et al 2001, Pruvost et al 2007). These DNA fragments still contain information which may help us to understand agricultural or vegetation history. Some of them are the proof of authenticity, contamination with other ancient and/or modern DNA and reproducibility of obtained results (Schlumbaum et al 2008)

Objectives

Methods

Results

Conclusion