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R.J. Schnell, J.S. Brown, C.T. Olano, A.W. Meerow, R.J. Campbell and D.N. Kuhn

Mango (Mangifera indica L.) germplasm can be classified by origin with the primary groups being cultivars selected from the centers of diversity for the species, India and Southeast Asia, and those selected in Florida and other tropical and subtropical locations. Accessions have also been classified by horticultural type: cultivars that produce monoembryonic seed vs. cultivars that produce polyembryonic seed. In this study we used 25 microsatellite loci to estimate genetic diversity among 203 unique mangos (M. indica), two M. griffithii Hook. f., and three M. odorata Griff. accessions maintained at the National Germplasm Repository and by Fairchild Tropical Botanic Garden in Miami, Fla. The 25 microsatellite loci had an average of 6.96 alleles per locus and an average polymorphism information content (PIC) value of 0.552 for the M. indica population. The total propagation error in the collection (i.e., plants that had been incorrectly labeled or grafted) was estimated to be 6.13%. When compared by origin, the Florida cultivars were more closely related to Indian than to Southeast Asian cultivars. Unbiased gene diversity (Hnb) of 0.600 and 0.582 was found for Indian and Southeast Asian cultivars, respectively, and both were higher than Hnb among Florida cultivars (0.538). When compared by horticultural type, Hnb was higher among the polyembryonic types (0.596) than in the monoembryonic types (0.571). Parentage analysis of the Florida cultivars was accomplished using a multistage process based on introduction dates of cultivars into Florida and selection dates of Florida cultivars. In total, 64 Florida cultivars were evaluated over four generations. Microsatellite marker evidence suggests that as few as four Indian cultivars, and the land race known as `Turpentine', were involved in the early cultivar selections. Florida may not represent a secondary center of diversity; however, the Florida group is a unique set of cultivars selected under similar conditions offering production stability in a wide range of environments.

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Hilde Nybom, Susan Gardiner and Charles J. Simon

Individual-specific DNA fragment patterns were obtained by hybridization of endonuclease-digested apple (Malus ×domestica Borkh.) DNA with a probe (pAR72) derived from the rDNA spacer region of the `White Angel' crab apple. Fragment detection was carried out with a nonradioactive method, using a horseradish peroxidase-catalyzed luminol oxidation. Paternity could be inferred by comparison of the fragment pattern generated by a seedling with those derived from putative parents.

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Beibei Li, Jianfu Jiang, Xiucai Fan, Ying Zhang, Haisheng Sun, Guohai Zhang and Chonghuai Liu

In this study, we present the molecular characterization of 61 Chinese grape landraces and 33 foreign cultivars by using nine microsatellite DNA markers. A total of 115 distinct alleles were amplified, and the average allele number was 12.78. The average observed and expected heterozygosity values were 0.797 and 0.839, respectively. The effective allele numbers ranged from 5.011 to 8.575. The average polymorphism information content (PIC) was 0.816. Eighty distinct genotypes were detected, and new synonyms and homonyms were found. The clustering dendrogram indicated that 94 Vitis materials could be divided into five major groups, and the cluster analysis showed that part of the Chinese grape landraces had a close relationship with the foreign cultivars. Assessment of the true cultivar identity, and the identification of synonyms and homonyms will be a contribution to improve the grape germplasm management and protect breeders’ intellectual rights.

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Patricia Sweeney, Robert Golembiewski and Karl Danneberger

Random amplified polymorphic DNA (RAPD) markers from leaf tissue extractions are effective for discrimination of turfgrass varieties. The usefulness of RAPD markers for turfgrass variety identification can be enhanced by use of seed rather than leaf tissue for DNA extraction. To determine whether DNA extracted from turfgrass seed was suitable for amplification, DNA was extracted from bulk samples and individual seeds of bermudagrass [Cynodon dactylon (L.) Pers.], chewings fescue (Festuca rubra var. commutata Gaud.), Poa annua L., Poa supina Schrad., creeping bentgrass [Agrostis stolonifera L. var. palustrus (Huds.) Farw.], Kentucky bluegrass (Poa pratensis L.), perennial ryegrass (Lolium perenne L.), and tall fescue (Festuca arundinacea Schreb.). All samples were successfully amplified using an arbitrary primer. Amplification intensity varied among species. With an almost infinite number of arbitrary primers available, it is likely that suitable primers can be found to amplify DNA from most turfgrass species. Amplification of turfgrass seed DNA, whether bulk or individual seed, is possible and should prove more useful than amplification of leaf tissue DNA for discrimination of turfgrass varieties.

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Rogério L. Cansian and Sergio Echeverrigaray

Randomly amplified polymorphic DNA (RAPD) markers were used to discriminate among 16 commercial cultivars of cabbage (Brassica oleracea L. Capitata Group). A set of 18 decamer primers was selected from 100 random sequences and used to characterize cultivars and to evaluate distances. The selected primers produced 105 (54%) polymorphic bands ranging in size from 100 and 2500 base pairs, out of a total of 195 bands, which allowed for discrimination of all cultivars. Similarity indices between cultivars were computed from RAPD data, and ranged from 0.72 to 0.87 with an average of 0.82. Unweighted pair-group method with arithmetic average (UPGMA) cluster analysis revealed two groups, one formed by two cultivars recommended for summer cropping, and the other by 14 cultivars. This large group was additionally divided into two subgroups. RAPD analysis provides a quick and reliable alternative for the identification of cabbage cultivars and for determination of the relationships among them.

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Mike L. Grant, Diana M. Miller and Alastair Culham

Knowledge of the origin of Lavatera L. (tree mallows) cultivars helps to predict their cultural requirements. Eighteen accessions representing 15 cultivars, 14 accessions of 7 species, and 5 accessions of an F1 hybrid between the putative parents of the cultivars were sampled for morphological variation and for randomly amplified polymorphic DNA (RAPD) fingerprint variation. Species-specific molecular markers were identified from the RAPD profiles. Chimeral elements were not distinguishable by RAPD analysis. Principal component analysis identified the majority of the cultivars to be selections of hybrid origin, probably from a narrow genetic base. Two cultivars were derived directly from individual species. The resolving power of RAPD markers and morphology was similar although RAPD data offered greater ability to ascribe parentage while morphology offered optimal discrimination of cultivar selections.

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Maureen C. O'Leary and Thomas H. Boyle

A germplasm collection of 59 Schlumbergera Lemaire clones was assayed for isozymes of aspartate aminotransferase, glucose-6-phosphate isomerase, leucine aminopeptidase, malate dehydrogenase, phosphoglucomutase, shikimate dehydrogenase, and triosephosphate isomerase. The collection included cultivars of holiday cactus [S. truncata (Haworth) Moran and S. ×buckleyi (T. Moore) Tjaden] plus accessions of S. kautskyi (Horobin & McMillan) N.P. Taylor, S. opuntioides (Löfgren & Dusén) D. Hunt, S. orssichiana Barthlott & McMillan, S. russelliana (Hooker) Britton & Rose, S. ×exotica Barthlott & Rauh, and S. ×reginae McMillan & Orssich. Twelve loci with 36 alleles were detected. Percent polymorphic loci, mean number of alleles per locus, and mean heterozygosity were 83, 3.00, and 0.24, respectively, for the entire collection. Forty-one clones (69%) could be distinguished solely on the basis of their isozyme profiles, but the remaining 18 clones shared profiles with one or two other clones. Isozymes proved useful for determining the parentage of some clones and verifying that some progeny were interspecific hybrids. About 28% of the genetic diversity within the entire collection is unique to four Schlumbergera species that have scarcely been exploited for breeding holiday cactus cultivars.

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Lisa J. Rowland, Smriti Mehra, Anik L. Dhanaraj, Elizabeth L. Ogden, Janet P. Slovin and Mark K. Ehlenfeldt

Because randomly amplified polymorphic DNA (RAPD) is the only type of molecular marker that has been used extensively in blueberry (Vaccinium spp.) for mapping and DNA fingerprinting of cultivars, there is a need to develop a new, robust marker system. Expressed sequence tags (ESTs) produced from a cDNA library, derived from RNA from floral buds of cold acclimated plants, were used to develop EST-PCR markers for blueberry. Thirty clones, picked at random from the cDNA library, were single-pass sequenced from the 5' and 3' ends. Thirty PCR primer pairs were designed from the ends of the best quality sequences that were generated and were tested in amplification reactions with genomic DNA from 19 blueberry genotypes, including two wild selections (the original parents of a mapping population), and 17 cultivars. Fifteen of the 30 primer pairs resulted in amplification of polymorphic fragments that were detectable directly after ethidium bromide staining of agarose gels. Several of the monomorphic amplification products were digested with the restriction enzyme AluI and approximately half resulted in polymorphic-sized fragments (cleaved amplified polymorphic sequences or CAPS markers). The polymorphic EST-PCR and CAPS markers developed in this study distinguished all the genotypes indicating that these markers should have general utility for DNA fingerprinting and examination of genetic relationships in blueberry. Similarity values were calculated based on the molecular marker data, and a dendrogram was constructed based on the similarity matrix. Coefficients of coancestry were calculated for each pair of genotypes from complete pedigree information. A fair correlation between similarity coefficients calculated from marker data and coefficients of coancestry was found.

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Kirk W. Pomper, Sheri B. Crabtree, Shawn P. Brown, Snake C. Jones, Tera M. Bonney and Desmond R. Layne

The pawpaw [Asimina triloba (L.) Dunal.] is a tree fruit native to many areas of the southeastern and mid-western United States. Kentucky State University (KSU) is designated as a satellite repository for Asimina for the U.S. Department of Agriculture (USDA), National Plant Germplasm System (NPGS). An assessment of the level of genetic diversity in cultivated pawpaw would assist in development of the future germplasm repository collection strategies for cultivar improvement. The objectives of this study were to identify intersimple sequence repeat (ISSR) markers that segregate in a simple Mendelian fashion and to use these markers to assess genetic diversity in 19 pawpaw cultivars. Leaf samples from the 34 progeny of controlled crosses (1-7-1 × 2-54 and reciprocal) and the parents were collected, DNA was extracted, and subjected to the ISSR methodology using the University of British Columbia microsatellite primer set #9. Seven primers yielded 11 Mendelian markers with either a 3:1 or 1:1 ratio that was confirmed by chi-square analysis. Analysis of genetic diversity using 10 of the ISSR markers from 19 pawpaw cultivars revealed a moderate to high level of genetic diversity, with a percent polymorphic loci P = 80 and an expected heterozygosity He = 0.358. These diversity values are higher than those reported for cultivated pawpaw using isozyme or randomly amplified polymorphic DNA (RAPD) markers, indicating that the ISSR marker methodolgy has a higher level of discrimination in evaluating genetic diversity in pawpaw and/or pawpaw has greater levels of genetic diversity than previously found.

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B.J. Weir, R.G. St. Pierre and R.N. Chibbar

Randomly amplified polymorphic DNA (RAPD) markers were used to distinguish among 16 cultivars of saskatoon (Amelanchier spp.). Eight 9-base, oligonucleotide primers amplified a total of 98 DNA fragments, of which 29 were useful as reproducible polymorphic markers. Twelve cultivars and two pairs of cultivars were uniquely characterized by these 29 markers. Polymorphism was not detected among five sources of the cv. Thiessen, whereas variability was found among seedlings from self-pollinated `Thiessen'. Samples of the cvs. Regent and Parkhill were indistinguishable from one of two sources, suggesting that the cultivars were mislabelled.