Skip to main content
ARS Home » Midwest Area » Columbia, Missouri » Plant Genetics Research » Research » Publications at this Location » Publication #65404
Title: REPETITIVE, GENOME-SPECIFIC PROBES IN WHEAT (TRITICUM AESTIVUM L. EM THELL)AMPLIFIED WITH MINISATELLITE CORE SEQUENCES

Author
item SOMERS, DARYL - UNIVERSITY OF MISSOURI
item ZHOU, Z - UNIVERSITY OF MISSOURI
item BEBELI, P - ATHENS AGRI UNIVERSITY
item Gustafson, J

Submitted to: Journal of Theoretical and Applied Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/15/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary: DNA fingerprinting has been utilized in analyzing humans for years, and recently, we succeeded in isolating and characterizing a DNA fingerprinting marker from cultivated rice. However, we were unable to obtain DNA fingerprinting sequences from wheat using the standard techniques. An experiment was designed to see if we could isolate and characterize DNA fingerprinting markers from wheat using a newly developed molecular technique involving the direct amplification of the DNA which would save years of time and expense. We succeeded in using the technique in isolating new DNA fingerprinting markers from wheat for potential use in marker-mediated wheat cultivar development and germplasm evaluation. This new technique has been adapted for use in wheat. It will allow for the isolation of sequences that are specific to each of the genomes that make up hexaploid wheat. Now scientists can use DNA fingerprinting to speed up gene manipulation and backcross breeding within the individual genomes of wheat. It will also allow for the identification of a wheat variety using a single DNA probe instead of using several probes thereby speeding up the process of germplasm identification as well as making this process less expensive.

Technical Abstract: The detection and analysis of DNA polymorphism in crops is an essential component of marker assisted selection and cultivar identification in plant breeding. We have explored the direct amplification of minisatellite DNA by polymerase chain reaction (DAMD-PCR) as a means for generating DNA probes that are useful for detecting DNA polymorphism and DNA fingerprinting in wheat. This technique was facilitated by high stringenc PCR with known plant and animal minisatellite core sequences as primers on wheat genomic DNA. The DAMD-PCR profiles from Triticum aesitvum, T. durum, T. monococcum, T. speltoides and T. tauschii, showed a high degree of polymorphism, and the various genomes could be identified. Several types of probes were generated following cloning of the DAMD-PCR products, with the most frequent type being polymorphic probes specific to the various genomes of wheat. In addition, polymorphic, single locus and moderately dispersed probes were isolated that may have potential for DNA fingerprinting. Our experiments were limited primarily to diploid wheats and the results indicated that DAMD-PCR may isolate genome-specific probes from wild diploid wheat species that could be used to monitor genome introgression into hexaploid wheat.