PCR PRIMED WITH MINISATELLITE CORE SEQUENCES YIELDS DNA-FINGERPRINTING PROBES IN WHEAT

Four minisatellite core sequences were used as primers in a polymerase chain reaction (PCR) technique: known as the directed amplification o f minisatellite-region DNA (DAMD), to detect polymorphisms in three pa irs of hexaploid/tetraploid wheat cultivars. In each pair, the tetrapl oid cultivar (genomic formula AABB) was extracted from its correspondi ng hexaploid (genomic formula AABBDD) parent. Reproducible profiles of the amplified products revealed characteristic bands that were presen t only in the hexaploid wheats but not in their extracted tetraploids. Some polymorphisms were observed among the hexaploid cultivars. Twent y-three DAMD-PCR amplified fragments were isolated and screened as mol ecular probes on the genomic DNA of wild wheat species, hexaploid whea t and triticale cultivars, Subsequently, 8 of the fragments were clone d and sequenced, The DAM D-PCR clones revealed various degrees of poly morphism among different wild and cultivated wheats. Two clones yielde d individual-specific DNA fingerprinting patterns which could be used for species differentiation and cultivar identification. The results d emonstrated the use of DAMD-PCR as a tool for the isolation of informa tive molecular probes for DNA fingerprinting in wheal cultivars and sp ecies.