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Guillaume Besnard, Catherine Breton, Philippe Baradat, Bouchaib Khadari and André Bervillé

One hundred and thirteen olive (Olea europaea L.) accessions were characterized using randomly amplified polymorphic DNA (RAPD) markers. Forty-five polymorphic RAPD markers were obtained enabling us to distinguish 102 different RAPD profiles. The approximate estimation of the probability of obtaining the same RAPD profile for two different trees was between 6.75 × 10-5 and 4.82 × 10-14. A dendrogram was constructed using Ward's minimum variance algorithm based on chi-square distances. This led to a more clear-cut classification of profiles than the classical approach of unweighted pair group method with arithmetic average. Twenty-four clusters of RAPD profiles were shown in Ward's dendrogram. Reliability of the dendrogram structure was checked using variance analysis. RAPD data exhibited an acceptable resolving power for cultivar identification. A combination of three primers was proposed for rapid molecular identification of cultivars in collections and in nurseries.

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Hafid Achtak, Ahmed Oukabli, Mohammed Ater, Sylvain Santoni, Finn Kjellberg and Bouchaib Khadari

combination of two types of markers, codominant (microsatellites) and dominant (ISSR) that may be statistically linked. This study did not seek to optimize molecular techniques for efficient cultivar identification with a suitable cost analysis. In the

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A. Estilai, A. Hashemi and J.G. Waines

Leaf extracts of 500 plants from 47 guayule (Parthenium argentatum Gray) entries including AZ-101, Gila, Cal-3, Cal-6, and Cal-7 germplasms; 12 accessions from Mexico; and a diverse array of diploid, triploid, and tetraploid selections were analyzed for isozyme variation of 17 enzyme systems. Glutamate oxalacetate transaminase (GOT, EC 2.6.1.1), isocitrate dehydrogenase (IDH, EC 1.1.1.42), malate dehydrogenase (MDH, EC 1.1.1.37), phosphoglucoisomerase (PGI, EC 5.3.1.9), shikimate dehydrogenase (SKDH, EC 1.1.1.25), and triosephosphate isomerase (TPI, EC 5.3.1.1) produced sharp and well-resolved bands. With the exception of AZ-101 and Gila, intra- and inter-accession polymorphisms were present for the above enzymes. Plants of AZ-101 and Gila showed identical banding patterns for every enzyme, supporting the view that these two germplasms may be the apomictic progenies of a single selection. Isozyme variations within entries indicated that most of the available guayule germplasms and selections are heterogeneous. Differences between entries suggested that isozymes may provide useful markers for cultivar identification.

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Gayle M. Volk and Adam D. Henk

important historic cultivars that are not currently available in NPGS. The existence of a DNA fingerprinting database aids in cultivar identification and also serves as collection management tool in the identification of duplicates or possibly mislabeled

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M. Hubbard, J. Kelly, S. Rajapakse, A. Abbott and R. Ballard

We have identified cloned rose DNA fragments that detect restriction fragment length polymorphisms (RFLP) in rose (Rosa ×hybrida) cultivars. RFLP can be used as genetic markers for identification, certification, and patent protection. By comparing RFLP patterns for each of six probes, we have been able to characterize eight cultivars. These results confirm that RFLP analyses are useful for rose cultivar identification and may provide a means for protecting patent rights to new cultivars.

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Elizabeth J. Parks and James W. Moyer

Fingerprinting using molecular markers is a highly effective method of cultivar identification that is a powerful aid to traditional methods based on morphology. Amplified fragment length polymorphism (AFLP) is a robust and reliable method for generating molecular markers that has been used to evaluate many crops for a variety of applications. In this study, AFLP was used to develop and validate robust genetic fingerprints for poinsettia (Euphorbia pulcherrima Willd. ex Klotzch) cultivars. Polymorphism selection was completed to facilitate the identification of useful polymorphisms and minimize future fingerprinting costs and time. Poinsettia is a highly variable crop subject to genetic drift and variable cultivars. Validation of polymorphisms to remove those associated with intracultivar variation improved the reliability of the fingerprinting. The result was a poinsettia AFLP database that defines the genetic fingerprints of 104 cultivars. Cluster analysis illustrated differentiation of most poinsettia cultivars tested. Selection of a subset of AFLP polymorphisms resulted in clustering of cultivars according to known origin and breeding program. This method has applications not only for cultivar identification for cultivar protection, and maintenance of cultivar uniformity, but also has the potential application of developing markers for important traits.

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D.P. Zhang, M. Ghislain, A. Golmirzaie and J.C. Cervantes

Detecting inter- and intra-varietal variation is essential for the management of a plant germplasm bank. The sensitivity and efficiency of randomly amplified polymorphic DNA (RAPD) for cultivar identification and somaclonal mutation in sweetpotato were evaluated. RAPD demonstrated a highly significant inter-varietal variation. Every one of the 23 tested cultivars can be identified with a RAPD profile generated by a single primer. Suspected duplicates that are morphologically indistinguishable can be unambiguously verified with a combination of three decamers. No intra-varietal variation was found using RAPD. Clones of `Jewel' and `Beauregard' collected from different sources all have the same RAPD profiles. Moreover, with 150 markers, the transgenic `Chogoku' sweetpotato cannot be differentiated from its untransformed counterparts, even though the transgenic plant shows significant morphological changes. These results demonstrate that RAPD is a sensitive and efficient tool for identifying cultivar duplicates, but it is not efficient for detecting intra-clonal variation or somaclonal mutation in sweetpotato.

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Gerald S. Dangl, Keith Woeste, Mallikarjuna K. Aradhya, Anne Koehmstedt, Chuck Simon, Daniel Potter, Charles A. Leslie and Gale McGranahan

One hundred and forty-seven primer pairs originally designed to amplify microsatellites, also known as simple sequence repeats (SSR), in black walnut (Juglans nigra L.) were screened for utility in persian walnut (J. regia L.). Based on scorability and number of informative polymorphisms, the best 14 loci were selected to analyze a diverse group of 47 persian walnut accessions and one J. hindsii (Jepson) Jepson ex R.E. Sm × J. regia hybrid (Paradox) rootstock. Among the 48 accessions, there were 44 unique multi-locus profiles; the accessions with identical profiles appeared to be synonyms. The pairwise genetic distance based on proportion of shared alleles was calculated for all accessions and a UPGMA (unweighted pair group method with arithmetic mean) dendrogram constructed. The results agree well with what is known about the pedigree and/or origins of the genotypes. The SSR markers distinguished pairs of closely related cultivars and should be able to uniquely characterize all walnut cultivars with the exception of budsports. They provide a more powerful and reliable system for the molecular characterization of walnut germplasm than those previously tested. These markers have numerous applications for the walnut industry, including cultivar identification, verification of pedigrees for cultivar and rootstock breeding programs, paternity analysis, and understanding the genetic diversity of germplasm collections.

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Kang Hee Cho, Seo Jun Park, Su Jin Kim, Se Hee Kim, Han Chan Lee, Mi Young Kim and Jae An Chun

fragment length polymorphisms, single sequence repeats (SSRs), and inter simple sequence repeats (ISSRs) for DNA fingerprinting. The choice of a DNA marker depends on the scale and purpose of cultivar identification; several different DNA marker types have

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Viji Sitther, Dapeng Zhang, Sadanand A. Dhekney, Donna L. Harris, Anand K. Yadav and William R. Okie

. Discussion SSR markers in Prunus species have been widely used for cultivar identification, genetic mapping, and phylogenetic analyses ( Aranzana et al., 2003 ; Testolin et al., 2000 ). One of the major criteria in cultivar identification using molecular