PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 99pg1

MOLECULAR ANALYSIS OF A POLYMORPHIC DNA REGION TIGHTLY LINKED TO THE SUPERNODULATION (NTS) LOCUS OF SOYBEAN.

Alexander Kolchinsky, Debbie Landau-Ellis, Sieglinde Angermuller, Joanna Deckert, Roel Funke and Peter M. Gresshoff, Plant Molecular Genetics and the Center for Legume Research, The University of Tennessee, Knoxville, TN 37901-1071.


The cloned molecular marker pUTG-132a, known to be tightly linked to the supernodulation (nts) locus of soybean (Landau-Ellis et al, Mol. Gen. Genet. 228:221-226, 1991; Mol. Plant-Microbe Inter., 1992, in press) was restriction mapped and partially sequenced. PCR primers were designed to generate a PCR-based polymorphism between G. max (nts) and G. soja (wild type). PCR amplifications of heterozygotes revealed unexpected bias towards the shorter PCR product. The cause of the RFLP originally used to map the nts locus was a 0.8 kb deletion, removing both an nternal DraI restriction site as well as one of the PCR primer sequences. A genomic library of soybean DNA (cultivar Bragg) in lambda phage was screened for clones homologous to pUTG-132a. The same PCR primers were used to screen the library. The restriction map of lambda UT-a confirms the map deduced from hybridizations with genomic DNA. A large segregating F2 population of an nts x wild type cross was analyzed using a fast PCR test and no recombinants between pUTG-a and the nts locus were found. Together with previous data, this result puts the upper limit of the genetic distance between the nts locus and pUTG-a marker at 0.3 centimorgans. Physical mapping in the three-marker pA-36 cluster, ten centimorgans distant from the nts locus, shows that one centimorgan in the vicinity of these molecular markers equals 550 kb. Therefore, the pUTG-a marker is a valuable starting point for positional cloning of the nts locus.


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