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

Published online by Cambridge University Press:  22 February 2007

G. Gyulai ,
M. Humphreys ,
R. Lagler ,
Z. Szabo ,
Z. Toth ,
A. Bittsanszky ,
F. Gyulai  and
L. Heszky
G. Gyulai*
Affiliation:
St. Stephanus University Genetics and Pland Breeding and the Hungarian Academy of Sciences–St. Stephanus University Research Group for Molecular Plant Breeding, Godollo, H-2103, Hungary Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, SY23 3EB, UK
M. Humphreys
Affiliation:
Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, SY23 3EB, UK
R. Lagler
Affiliation:
St. Stephanus University Genetics and Pland Breeding and the Hungarian Academy of Sciences–St. Stephanus University Research Group for Molecular Plant Breeding, Godollo, H-2103, Hungary
Z. Szabo
Affiliation:
St. Stephanus University Genetics and Pland Breeding and the Hungarian Academy of Sciences–St. Stephanus University Research Group for Molecular Plant Breeding, Godollo, H-2103, Hungary
Z. Toth
Affiliation:
St. Stephanus University Genetics and Pland Breeding and the Hungarian Academy of Sciences–St. Stephanus University Research Group for Molecular Plant Breeding, Godollo, H-2103, Hungary
A. Bittsanszky
Affiliation:
St. Stephanus University Genetics and Pland Breeding and the Hungarian Academy of Sciences–St. Stephanus University Research Group for Molecular Plant Breeding, Godollo, H-2103, Hungary
F. Gyulai
Affiliation:
Institute of Agrobotany, Tapioszele, H-2766, Hungary
L. Heszky
Affiliation:
St. Stephanus University Genetics and Pland Breeding and the Hungarian Academy of Sciences–St. Stephanus University Research Group for Molecular Plant Breeding, Godollo, H-2103, Hungary
*
*Correspondence: Email: [email protected]
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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–MseCAA was found to be the most effective selective-primer 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 (MboI), 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.

Keywords

ancient DNAexcavated seedsPanicum miliaceum
Type
Research Article
Information
Seed Science Research , Volume 16 , Issue 3 , September 2006 , pp. 179 - 191
Copyright
Copyright © Cambridge University Press 2006

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