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Journal of Plant Diseases and Protection

, Volume 111, Issue 1, pp 30–38 | Cite as

Development of Pcr-based markers closely linked to rym5

  • Bettina Pellio
  • W. Friedt
  • A. Graner
  • F. Ordon
Article

Abstract

In the present study, two new dominant randomly amplified polymorphic Dna (Rapd) fragments and two amplified fragment length polymorphism (Aflp) markers linked to the recessive resistance gene rym5 which is effective against BaMMV, BaYMV, BaYMV-2 were identified. For initial primer screening, bulked segregant analysis was employed on resistant and susceptible doubled haploid (Dh) barley lines. By screening 1,200 decamer primers and 256 EcoRI/MseI Aflp primer combinations, new markers were developed which are co-segregating or very tightly linked (1.3 cM ) to rym5. As demonstrated in this study one of the most powerful and efficient ways to develop new markers is the use of the Aflp-technique. However, the availability of closely linked markers that allow a reliable and easy to handle detection of respective alleles is of special interest in marker-based selection procedures. Therefore, Rapd-marker Op-Af18h971 has to be considered being well suited for this approach and further experiments were conducted to convert it into a more specific marker. By elongating the respective primer at the 3’-end with selective nucleotides C and G, respectively, the specificity of the banding pattern was improved. Because of its close linkage to rym5, this marker offers the opportunity for marker-assisted selection in practical barley breeding programmes.

Key words

Hordeum vulgare barley yellow mosaic virus disease (BaMMV, BaYMV, BaYMV-2) Rapds Aflps molecular markers marker-assisted selection 

Entwicklung von Pcr-gestutzten Markern für das Resistenzgen rym5

Zusammenfassung

In der vorliegenden Untersuchung konnten zwei neue dominante Randomly Amplified Polymorphic Dna (Rapd) Marker sowie zwei Amplified Fragment Length Polymorphism (Aflp) Marker identifiziert werden, die mit dem rezessiven Resistenzgen rym5 gekoppelt sind, welches Resistenz gegen BaMMV, BaYMV und BaYMV-2 bedingt. Mit Hilfe der ‘bulked segregant analysis’ wurden zunächst Primer auf Polymorphismen zwischen dem resistenten und anfälligen bulk (DNA-Ramsch) getestet. Durch die Untersuchung von 1200 Decamer-Primern und 256 EcoRI/MseI Aflp Primer-Kombinati- onen konnten Marker identifiziert werden, welche mit dem Gen cosegregieren bzw. sehr eng gekoppelt sind (1,3 cM). Im Rah men der Untersuchungen konnte die Leistungsfahigkeit und Effizienz der Aflp-Methode demonstriert werden. Von besonderem Interesse in markergestützten Selektions- verfahren ist jedoch die Verfugbarkeit von eng gekoppelten Markern, die eine zuverlässige sowie insbesondere eine schnelle und einfach zu handhabende Detektion erlauben. Zu diesem Zweck wurde der Rapd-Marker Op-Af18h971 in einen spezifischeren Marker umgewandelt. Eine Erhöhung der Spezifitat des Primers wurde durch selektive Basenverlangerung am 3’-Ende mit C bzw. G erreicht. Aufgrund der engen Kopplung mit rym5 steht damit der praktischen Gerstenzüchtung ein weiterer Marker zur Verfügung, der eine effektive markergestutzte Selektion erlaubt.

Stichwörter

Hordeum vulgare Gelbmosaikvirose (BaMMV, BaYMV, BaYMV-2) Rapds Aflps molekulare Marker markergestützte Selektion 

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Copyright information

© Deutsche Phythomedizinische Gesellschaft 2004

Authors and Affiliations

  • Bettina Pellio
    • 1
  • W. Friedt
    • 1
  • A. Graner
    • 2
  • F. Ordon
    • 1
  1. 1.Institute of Crop Science and Plant Breeding IJustus-Liebig-UniversityGiessenGermany
  2. 2.Institute of Plant Genetics and Crop Plant Research (Ipk)GaterslebenGermany

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