DNA AMPLIFICATION FINGERPRINTING AND HYBRIDIZATION ANALYSIS OF CENTIPEDEGRASS

DNA amplification fingerprinting (DAF) with arbitrary oligonucleotide primers provides germplasm analyses for cultivar identification and pr otection of proprietary rights, and constitutes an effective tool in m arker-assisted breeding and in the study of genetic variation within n atural and domesticated populations. In this study, DAF was used to in fer genetic relationships in centipedegrass [Eremochloa ophiuroides (M unro) Hack.]. An optimization of reaction components defined amplifica tion conditions that reproducibly fingerprinted centipedegrass and oth er turfgrass species. Cultivars Tennessee Hardy, Tennessee Tuff, Oklaw n, Centennial, and Tifton common were analyzed with 14 octamer primers . Four primers produced polymorphic patterns, two of which differentia ted all cultivars. Phylogenetic analysis using parisomy (PAW) distinct ly separated cold-tolerant Tennessee Hardy. Numerous monomorphic bands were obtained, suggesting that the centipedegrass cultivars are close ly related and are from a common origin. Several DAF bands were isolat ed from silver-stained polyacrylamide gels for use as hybridization pr obes. A monomorphic 200-base pair (bp) band that hybridized to a singl e amplification product detected eight homologous regions in genomic c entipedegrass DNA. When hybridized to DAF patterns obtained from other turfgrass species, such as bermudagrass [Cynodon dactylon (L) Pers. x Cynodon transvaalensis Burtt-Davyl, rough bluegrass (Poa trivialis L. ) and Zoysia (Zoysia japonica Steud.), the 200-bp fragment, and a 175- bp fragment generated with a different primer, resulted in smearing pa tterns. While these products are only present within one size amplicon in centipedegrass fingerprints, the smearing indicated that the putat ive sequence was present in multiple copies throughout the amplificati on profile, was abundant in the heterologous grass genomes, and may ha ve represented a family of repetitive DNA sequences. Our results indic ated that DAF can generate molecular markers that can be used reliably for DNA fingerprinting of important centipedegrass cultivars. This to ol would be useful in determining genetic relationships and diversity, and in establishing possible origins of cultivars.