biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 45:337-345, 2002 | DOI: 10.1023/A:1016253214182

Development and Characterization of Microsatellite Markers from Chromosome 1-Specific DNA Libraries of Vicia Faba

D. Požárková1, A. Koblížková1, B. Román2, A.M. Torres2, S. Lucretti3, M. Lysák4, J. Doležel4, J. Macas5
1 Institute of Plant Molecular Biology, ASCR, České Budějovice, Czech Republic
2 Departamento de Mejora y Agronomía, C.I.F.A., Córdoba, Spain
3 Biotechnology and Agriculture Division, ENEA C.R. Casaccia, Rome, Italy
4 Institute of Experimental Botany, ASCR, Olomouc, Czech Republic
5 Institute of Plant Moelcular Biology, ASCR, České Budějovice, Czech Republic

An integrated approach has been developed for targeted retrieval of microsatellite markers from selected regions of the field bean (Vicia faba L.) genome. The procedure is based on a combination of advanced physical and genetic mapping techniques and includes the following steps: 1) flow-sorting of metaphase chromosomes, 2) construction of microsatellite-enriched chromosome-specific DNA libraries, 3) isolation of polymorphic microsatellite sequences from the libraries, 4) testing chromosome specificity of the microsatellites using polymerase chain reaction with purified fractions of individual chromosome types, and 5) integration of chromosome-specific markers into a genetic map. Several strategies for isolation of microsatellite clones were tested, including direct screening of non-enriched libraries with single or mixed probes and screening of the libraries after one or two rounds of enrichment. Finally, the usefulness of this approach was demonstrated by the retrieval of novel markers from a selected portion of the largest field been chromosome (No. 1).

Keywords: field bean; flow-sorting; genetic map; physical mapping; plant chromosomes
Subjects: chromosome; field bean, microsatellite markers; flow-sorting; genetic map; markers; microsatellite markers; polymorphism, proteins; proteins, polymorphism; Vicia faba

Published: September 1, 2002  Show citation

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Požárková, D., Koblížková, A., Román, B., Torres, A.M., Lucretti, S., Lysák, M., Doležel, J., & Macas, J. (2002). Development and Characterization of Microsatellite Markers from Chromosome 1-Specific DNA Libraries of Vicia Faba. Biologia plantarum, 45(3), 337-345. doi: 10.1023/A:1016253214182
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