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ARS Home » Midwest Area » Columbia, Missouri » Plant Genetics Research » Research » Publications at this Location » Publication #71810
Title: PCR PRIMED WITH MINISATELLITE CORE SEQUENCES YIELDS DNA FINGERPRINTING PROBES IN WHEAT

Author
item BEBELI, P - ATHENS AGRI UNIVERSITY
item ZHOU, Z - UNIV OF MISSOURI
item SOMERS, D - UNIV OF MISSOURI
item GUSTAFSON, J

Submitted to: Journal of Theoretical and Applied Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/3/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary: DNA fingerprinting has been utilized in analyzing humans for years. It has most often been associated with uses in forensic medicine and court cases for individual identification, but has also been used in population studies. Over the past several years we succeeded in isolating and characterizing DNA fingerprinting markers from cultivated rice. These markers have proven valuable in evaluating rice germplasm and for potential use in cultivar development. An experiment was designed to see if we could isolate DNA fingerprinting markers from wheat in a much accelerated fashion that would save years of time and expense. We succeeded in utilizing a newly developed molecular technique to isolate new DNA fingerprinting markers for use in DNA fingerprinting wheat germplasm. Utilization of a single molecular marker to fingerprint wheat will greatly simplify cultivar identification and enhance study of the wheat species with differing numbers of chromosomes.

Technical Abstract: Four minisatellite core sequences were used as primers in a polymerase chain reaction (PCR), known as the directed amplification of minisatellite-region DNA (DAMD), to detect polymorphisms in three pairs of hexaploid/tetraploid wheat cultivars. In each pair, the tetraploid cultivar (genomic formula AABB) was extracted from its corresponding hexaploid (genomic formula AABBDD) parent. Reproducible profiles of the amplified products revealed characteristic bands of the same molecular size that were present only in the haxaploid wheats, but not in their extracted tetraploids. Some polymorphisms were observed among the hexaploid cultivars. Twenty-three DAMD-PCR amplified fragments were isolated and screened as molecular probes on the genomic DNA of wild wheat species, hexaploid wheat, and triticale cultivars. Subsequently, eight of the fragments were cloned and sequenced. The DAMD-PCR clones revealed various degrees of polymorphism among different wild and cultivated wheats. Two clones yielded individual-specific DNA fingerprinting patterns which could be used for species differentiation and cultivar identification. Based on the fingerprint data, a dendrogram was constructed depicting genetic relationships among wild species, and the cultivated wheats and triticales. The results demonstrated the use of DAMD-PCR as a tool for the isolation of informative molecular probes for DNA fingerprinting in wheat cultivars and species.