Agronomy and Horticulture Department

 

Date of this Version

10-2000

Citation

Theoretical and Applied Genetics 101:5–6 (October 2000), pp. 747–755

doi: 10.1007/s001220051540

Comments

Copyright © 2000 Springer-Verlag. Used by permission.

Abstract

Large DNA insert libraries in binary T-DNA vectors can assist in the isolation of the gene(s) under-lying a quantitative trait locus (QTL). Binary vectors facilitate the transfer of large-insert DNA fragments containing a QTL from E. coli to Agrobacterium sp. and then to plants. We constructed two soybean large-insert libraries from cv. Forrest in the pCLD04541 (V41) binary vector after partial digestion of genomic high-molecular-weight DNA with BamHI or HindIII. The libraries contain 76,800 clones with an average insert size of 125 kb, and therefore represent 9.5-fold haploid genome equivalents. Colony hybridization using a chloroplast-specific probe infers that the libraries contain less than 0.5% clones of chloroplast DNA origin. These two libraries have provided clones for physical mapping of the soybean genome and for isolation of a number of disease resistance genes. One microsatellite marker was identified from the clone that hybridized to the Bng122 RFLP probe. The sequence-tagged site was used for genetic mapping and marker-assisted selection for genes underlying resistance to the soybean cyst nematode and sudden death syndrome.

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