Theoretical and Applied Genetics

, Volume 101, Issue 5, pp 747–755

Two large-insert soybean genomic libraries constructed in a binary vector: applications in chromosome walking and genome wide physical mapping

  • K. Meksem
  • K. Zobrist
  • E. Ruben
  • D. Hyten
  • T. Quanzhou
  • H-B. Zhang
  • D. A. Lightfoot

DOI: 10.1007/s001220051540

Cite this article as:
Meksem, K., Zobrist, K., Ruben, E. et al. Theor Appl Genet (2000) 101: 747. doi:10.1007/s001220051540

Abstract 

Large DNA insert libraries in binary T-DNA vectors can assist in the isolation of the gene(s) underlying 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.

Key words Bacterial artificial chromosomeSoybeanPhysical mappingTargeted microsatellitesPositional cloningDisease resistanceGenomicsCyst nematode

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • K. Meksem
    • 1
  • K. Zobrist
    • 1
  • E. Ruben
    • 1
  • D. Hyten
    • 1
  • T. Quanzhou
    • 2
  • H-B. Zhang
    • 2
  • D. A. Lightfoot
    • 1
  1. 1.Department of Plant Soil and General Agriculture, Southern Illinois University at Carbondale, Room 176. Carbondale, IL 62901–4415, USA Fax: +618 453 7457 e-mail: http://www.coalab.siu.edu/genome/meksemk@siu.eduUS
  2. 2.Department of Soil and Crop Sciences, Crop Biotechnology Center, Texas A & M University, College Station, TX 77843, USAUS