Seed remains of common millet from the 4th (Mongolia) and 15th (Hungary) centuries: AFLP, SSR and mtDNA sequence recoveries

Abstract

Seed remains of common millet (Panicum miliaceum L.) were excavated from sites of AD 4th-century Darhan (Mongolia), and AD 15th-century Budapest (Hungary). Because the 15th-century medieval grains looked so intact, a germination test was carried out under aseptic conditions, which resulted in swelling of the grains but no cell proliferation or germination. Ancient DNA (aDNA) was extracted from the aseptic grains; analysed for amplified fragment length polymorphisms (AFLP), simple sequence repeats (SSR) and mitochondrial DNA (mtDNA); and compared with the modern millet cultivar 'Topaz'. AFLP analysis revealed that extensive DNA degradation had occurred in the 4th-century ancient millet, resulting in only 2 (1.2%) AFLP fragments (98.8% degradation) amplified by MseCAA -EcoAGT, compared to the 15th-century medieval millet, with 158 (40%) fragments (60% degradation), and modern millet cultivar 'Topaz' with 264 fragments (100%). EcoAGT-Mse-CAA was found to be the most effective selectiveprimer combination for the analysis of medieval and modern millet. Eight AFLP fragments were sequenced after re-amplification and cloning. Microsatellite (SSR) analysis at the nuclear gln4, sh1, rps28 and rps15 loci revealed one SNP (single nucleotide polymorphism) at the 29th position (A ! G) of rps28 locus, compared to modern millet. An mtDNA fragment (Mbo I), amplified at the 18S -5S ribosomal DNA (rDNA) locus in the medieval millet, showed no molecular changes compared to modern millet. The results underline the significance of aDNA extraction and analysis of excavated seeds for comparative analysis and molecular reconstruction of ancient and extinct plant genotypes.

Ancient plant DNA in archaeobotany

“…This aDNA damage results in absent bands (=allelic dropouts) or in artificial bands, produced by the presence of co-extracted contaminating modern or archaeological plant material or by polyploidy or gene duplication. Nevertheless, some reports have been published (Siles et al 2000;Suh et al 2000;Iniguez et al 2003;Gyulai et al 2006).…”

Agro-morphological Characterization of Proso millet Germplasm for Utilization

“…The rapid and widespread adoption of proso millet during its early domestication can be attributed to its short growing duration and resilience to biotic and abiotic stress [6,7]. Additionally, proso millet is rich in protein [8], and recent studies have reported the presence of high levels of antioxidants and secondary metabolites with health benefits [9,10].…”

The Cucurbits of Mediterranean Antiquity: Identification of Taxa from Ancient Images and Descriptions

“…Archaeobotanical records include findings of several seeds purportedly of cucumber, but it is extremely difficult, even for experts, to differentiate between the seeds of C. sativus and C. melo (Bates and Robinson, 1995). Possibly, an identification of the species of these seeds could be accomplished by analysis of ancient DNA (Gyulai et al, 2006). In this survey of Mediterranean iconography and verbal sources of Roman times, we have found no hard evidence of the presence of cucumbers.…”