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Deborah Dean, Phillip A. Wadl, Xinwang Wang, William E. Klingeman, Bonnie H. Ownley, Timothy A. Rinehart, Brian E. Scheffler and Robert N. Trigiano

hypervariabilty and are easily detected using polymerase chain reaction (PCR) and two unique primers ( Powell et al., 1996 ). Microsatellite markers are ideal for identification and genetic fingerprinting of plants because of the high levels of polymorphism they

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Imen Rekik, Amelia Salimonti, Naziha Grati Kamoun, Innocenzo Muzzalupo, Oliver Lepais, Sophie Gerber, Enzo Perri and Ahmed Rebai

). In this article, we used 10 microsatellite markers to study 20 Tunisian olive cultivars of major commercial interest. The objective of the present study was the first to assess the potential of SSR markers to differentiate a number of Tunisian olive

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Fuad Gasi, Kenan Kanlić, Belma Kalamujić Stroil, Naris Pojskić, Åsmund Asdal, Morten Rasmussen, Clive Kaiser and Mekjell Meland

, compared with traditional morphological (e.g., pomological) characterizations, in revealing mislabeled plant accession ( Nybom and Weising, 2010 ). Microsatellite markers, also known as simple sequence repeats (SSRs), have shown great promise as a tool for

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Rohollah Karimi, Ahmad Ershadi, Kourosh Vahdati and Keith Woeste

study, seven populations of J. regia were characterized using 11 highly polymorphic microsatellite markers. The average number of alleles per locus was 5.73, much higher than 1.3 and 3.9 detected in J. regia with RAPDs ( Nicese et al., 1998 ) and

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Michael Dossett, Jill M. Bushakra, Barbara Gilmore, Carol A. Koch, Chaim Kempler, Chad E. Finn and Nahla V. Bassil

., 2010 ) was then used to filter redundant sequences (90% or greater similarity) to a single unique “clustered” read. Microsatellite marker discovery and screening. After read clustering, primers were designed for all of the SSRs using the Genome Database

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Fuad Gasi, Silvio Simon, Naris Pojskic, Mirsad Kurtovic, Ivan Pejic, Mekjell Meland and Clive Kaiser

these genetic resources ( Hokansson et al., 1998 ). Examining ex situ collections with microsatellite markers has so far been performed in a number of studies ( Garkava-Gustavsson et al., 2008 ; Gharghani et al., 2009 ; Guarino et al., 2006 ; Guilford

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Hua Wang, Dong Pei, Rui-sheng Gu and Bao-qing Wang

.K. Koehmstedt, A. Simon, C. 2005 Characterization of 14 microsatellite markers for genetic analysis and cultivar identification of walnut J. Amer. Soc. Hort. Sci. 130 348 354 Derory, J. Mariette, S

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Innocenzo Muzzalupo, Nicola Lombardo, Aldo Musacchio, Maria Elena Noce, Giuseppe Pellegrino, Enzo Perri and Ashif Sajjad

Genetic diversity studies using microsatelite analysis were carried out in a set of 39 accessions of Olea europaea L., corresponding to the majority of the regional autochthon germplasm in Apulia. Samples of olive leaves were harvested from plants growing in the olive germplasm collection of the Consiglio per la Ricerca e Sperimentazione in Agricoltura (C.R.A.) - Istituto Sperimentale per l'Olivicoltura at Rende in Cosenza Italy. Herein, we evaluated the extent to which microsatellite analysis using electrophoresis was capable of identifying traditional olive cultivars. In addition, the DNA sequence of all amplicons was determined and the number of repeat units was established for each sample. Using five loci, electrophoretic analysis identified 24 genotype profiles, while DNA sequence analysis detected 28 different genotype profiles, identifying 54% of cultivars. The remaining 46% were composed of seven different accession groups containing genetically indistinguishable cultivars, which are presumably synonyms. This study demonstrates the utility of microsatellite markers for management of olive germplasm and points out the high level of polymorphisms in microsatellite repeats when coupled with DNA sequence analysis. The establishment of genetic relationships among cultivars in the Apulian germplasm collection allows for the construction of a molecular database that can be used to establish the genetic relationships between known and unknown cultivars.

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Peter Boches, Lisa J. Rowland, Kim Hummer and Nahla V. Bassil

Microsatellite markers for blueberry (Vaccinium L.) were created from a preexisting blueberry expressed sequence tag (EST) library of 1305 sequences and a microsatellite-enriched genomic library of 136 clones.

Microsatellite primers for 65 EST-containing simple sequence repeats (SSRs) and 29 genomic SSR were initially tested for amplification and polymorphism on agarose gels. Potential usefulness of these SSRs for estimating species relationships in the genus was assessed through cross-species transference of 45 SSR loci and cluster analysis using genetic distance values from five highly polymorphic EST-SSR loci. Cross-species amplification for 45 SSR loci ranged from 17% to 100%, and was 83% on average in nine sections. Cluster analysis of 59 Vaccinium species based on genetic distance measures obtained from 5 EST-SSR loci supported the concept of V. elliotii Chapm. as a genetically distinct diploid highbush species and indicated that V. ashei Reade is of hybrid origin. Twenty EST-SSR and 10 genomic microsatellite loci were used to determine genetic diversity in 72 tetraploid V. corymbosum L. accessions consisting mostly of common cultivars. Unique fingerprints were obtained for all accessions analyzed. Genetic relationships, based on microsatellites, corresponded well with known pedigree information. Most modern cultivars clustered closely together, but southern highbush and northern highbush cultivars were sufficiently differentiated to form distinct clusters. Future use of microsatellites in Vaccinium will help resolve species relationships in the genus, estimate genetic diversity in the National Clonal Germplasm Repository (NCGR) collection, and confirm the identity of clonal germplasm accessions.

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LJ Grauke, Maria Azucena Mendoza-Herrera, Carol Loopstra and Tommy E. Thompson

Microsatellite or Simple Sequence Repeat (SSR) markers are being developed in ongoing research in the USDA ARS Pecan Breeding Program. These co-dominant markers provide a powerful tool for the verification of parentage. To confirm their utility, SSR profiles were used to confirm the parentage of 19 of the 25 controlled crosses released by the breeding program. Questions were raised concerning the parentage of some crosses thought to be known. When the genotype of the maternal parent is known, the paternal genotype necessary to have produced the progeny can be determined. A SAS program was written to query a database that includes 288 pecan accessions to find appropriate paternal genotypes given a maternal pattern. If neither parent is known, all possible parental combinations can be derived based on the progeny. Putative parents can be qualified on the basis of genotype as well as other evidence, such as nut morphology, dates of origin, locations of origin, and dichogamy. Using these techniques, putative parents are suggested for the historic cultivars `Riverside' and `Western'. Although the probabilities for a particular genotypic pattern can be determined based on allele frequencies within the population, assigning numeric probabilities to other evidence is more challenging. Meticulous records are necessary to establish the linkage between an inventory of an accession and its historic origin, thereby placing putative parents in combination at the proper place and appropriate time. Records of USDA–ARS National Plant Germplasm System, as exemplified by logbooks and vouchers of the McKay Collection of the National Arboretum, provide evidence for confident molecular genetic verification of cultivar identity and parentage, increasing the value of the living accessions in the NPGS.