To be able to breed for durable disease resistance it is important to use genetically diverse resistance sources. As a prerequisite to select suitable parents for crosses, it is necessary to evaluate and analyse the available germplasm. The aim of the present study is to: 1) Investigate how the diversity within barley (Hordeum) is structured, and how big a part of the diversity is included in Hordeum vulgare, for instance modern varieties, landraces or exotic varieties, versus Hordeum bulbosum and wild barley species. 2) Elucidate the phylogeny of barley using microsatellite DNA fingerprints; does it correlate with traditional phylogeny and/or with pedigree information? 3) Study the frequency of different microsatellite alleles in barley.
Three sets of materials comprising approximately 180 different barley lines from various barley growing regions will be analysed. Wild species have been provided by the nordic gene bank, H.bulbosum were provided by The Abed Foundation, and some modern European and exotic varieties is included as well to give a reasonable coverage of the whole genus.
Genomic DNA is extracted from green leaves using the CTAB method, and used as template in a PCR to amplify selected microsatellites. Microsatellites are thus used as molecular markers to produce a DNA fingerprint for each accession . A number of published microsatellite primers have been tested and the most polymorphic chosen to give a unique fingerprint for each accession. Following the PCR the product is run on a polyacrylamide gel, and the DNA is visualized by silverstaining, or by using a labeled primer in the PCR and following running the gel in an ABI sequencer where the DNA is detected through fluorescence.
Data is analysed using "Statistica", "Sigma Plot" and "microsat".
Author: phd@dina.kvl.dk. Updated: 5 April 2000