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Salih Kafkas and Rafael Perl-Treves

Phylogenetic relationships among nine species in the genus Pistacia were studied by randomly amplified polymorphic DNA (RAPD) analysis. The following species were included: P. atlantica, P. terebinthus, P. eurycarpa, P. vera, P. integerrima, P. mexicana, P. palaestina, P. lentiscus, and P. khinjuk. Genomic DNA was extracted from leaf tissue and RAPD analysis was performed using 20 primers. A total of 242 fragments were generated and 228 bands were polymorphic at the inter-specific level. Subjecting these data to phylogenetic analysis yielded a shortest cladogram that is 338 steps long, featuring two main groups. P. vera, P. khinjuk, P. eurycarpa, P. atlantica, and P. integerrima were included in one group, while P. terebinthus, P. palaestina, P. mexicana, and P. lentiscus formed the second group. The first group included species with single-trunked and big trees, whereas the species included in the second group mostly grow as shrubs or small trees. The cladogram showed that the closest pairs of species were P. terebinthus and P. palaestina, P. eurycarpa and P. atlantica, P. vera and P. khinjuk, and P. mexicana and P. lentiscus. We suggest that P. palaestina is in fact a variety of P. terebinthus in view of the small genetic distance between them. This study also showed that P. eurycarpa (syn. P. atlantica var. kurdica) is a distinct species from P. atlantica, rather than a variety within the same species.

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Jason D. Zurn, Katie A. Carter, Melinda H. Yin, Margaret Worthington, John R. Clark, Chad E. Finn, and Nahla Bassil

and hybridize readily with nearby plants ( Clark and Finn, 2011 ). DNA fingerprinting sets have become a useful tool to identify cultivars, validate pedigrees, study population diversity, and protect breeders’ rights ( Laurentin, 2009 ; Peace, 2017

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Paul A. Wiersma, Deniz Erogul, and Shawkat Ali

DNA fingerprinting of highly related individuals. Although SSR markers can be used to discriminate sweet cherry cultivars as previously suggested, finding markers that differentiate closely related cultivars, such as the Lapins and ‘Sweetheart, groups

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Barbara S. Gilmore, Nahla V. Bassil, Danny L. Barney, Brian J. Knaus, and Kim E. Hummer

yellow cedar [ Callitropsis nootkatensis ( Jennings et al., 2011 )] and mile-a-minute weed [ Mikania micrantha ( Yan et al., 2011 )]. The objective of this study was to develop SSR markers from short-read DNA sequences and use them to fingerprint the

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A. Vainstein and H. Ben-Meir

Mini- and microsatellite probes were hybridized to DNA of 24 rose (Rosa×hybrida) genotypes. The resultant DNA fingerprints were shown to be genotype-specific, thereby enabling cultivar identification at the DNA level. Restriction enzyme Dra I yielded the most informative band patterns. Full-sib family analysis of DNA fingerprints revealed 32 parental-specific bands out of the 128 observed in the parents. These bands were revealed cumulatively by phage (M13), human (33.6), and oligonucleotide (GACA)4 probes. Only one pair of these loci was found to be allelic, and no linked pairs were detected in the progeny analyzed. The probability of two offspring from this cross having identical DNA fingerprints was calculated to be 2 × 10-8.

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R.N. Trigiano, B.H. Ownley, A.N. Trigiano, J. Coley, K.D. Gwinn, and J.K. Moulton

tools in plant biotechnology, which includes genomics and proteomics, are gel electrophoresis, polymerase chain reaction (PCR), and DNA fingerprinting. To incorporate these techniques into middle and high school and college curricula, content instruction

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Mark Hubbard, John Kelly, Albert Abbott, and Robert Ballard

To protect plant patents, rose breeders would benefit from a reliable and sensitive method for differentiating cultivars at the genetic level. Rcombinant DNA technologies are being employed to characterize individual DNA structure of numerous rose cultivars. Restriction fragment length polymorphisms (RFLPs) are being studied to develop a characteristic pattern, or fingerprint for each cultivar. DNA from various cultivars is restriction enzyme digested and the fragments separated by agarose gel electrophoresis. The gel is Southern blotted and hybridized with probes from the rose DNA library to yield RFLPs. RFLPs are being located and will eventually result in a characteristic fingerprint for each cultivar.

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Naomi R. Smith and Robert N. Trigiano

Flowering dogwood (Cornus florida L.) is an important tree of forests and urban landscapes in the eastern United States. Currently, there are over 100 cultivars of flowering dogwood commercially available. An identification process based on genotype would be of use to researchers, breeders, and nurserymen, as many cultivars are similar phenotypically. Molecular markers offer a promising way of definitively identifying flowering dogwood cultivars. Amplified fragment length polymorphism (AFLP) is a technique that can be used to generate DNA fingerprints. DNA was isolated from leaves of 17 common cultivars of dogwood and AFLP fingerprints were generated by a Beckman Coulter CEQ™ 8000. Fingerprints were converted to binary data and verified manually. Two drafts of a cultivar identification key were generated based on the corrected, verified binary data and cultivar-specific peaks. Six primer combinations were used to construct all keys and were tested with seven unknown dogwood cultivar samples. Six unknown samples were correctly identified using the keys. Only one unknown, `Cherokee Brave', was unidentifiable with any key. In all cases, some intracultivar variation was observed. A similarity index was calculated and visualized with a tree of genetic relatedness using NTSYSpc. Intracultivar variation was observed in the similarity index as well. This database for cultivar-specific molecular markers will serve as a starting point to which other cultivars can be added and also can be used in breeding applications, patent application and other projects, such as mapping the C. florida genome.

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Victor Luk and John Carlson

DNA fingerprinting is a potentially powerful molecular genetic technique that can be used to distinguish subtle differences in genome structure among closely related genotypes, such as many horticultural varieties. A DNA fingerprinting project is currently in progress at the Univ. of British Columbia (UBC) Biotechnology Laboratory to produce a set of DNA markers and an easy, reliable, and legally recognized analysis protocol that will enable the UBC Botanical Garden Plant Introduction Scheme (PISBG) to unambiguously identify any of their released varieties, even in dormant or juvenile form, wherever it is being propagated or sold. High-quality genomic DNA was isolated from the leaf samples of six PISBG species (Anagallis monellii, Artemesia stelleriana, Clematis, Genista pilosa, Microbiota decussata, and Penstemon fruticosa) using a modified CTAB DNA isolation protocol, and further purified by cesium chloride/ethidium bromide gradient. Samples of these genomic DNA preparations (10 ng) were then amplified by a 45-cycle polymerase chain reaction (PCR) protocol using 1.5-μm 10-nucleotide primers of arbitrary nucleotide sequence that amplify a variety of sites distributed across the genome. Following the amplification, PCR products [random amplified polymorphic DNA (RAPD) markers] were separated by agarose gel electrophoresis and visualized by ethidium bromide staining. More than 70% of the 51 primers tested so far generated distinctive banding patterns (2–11 bands) with DNA samples from each species. Subtle changes in the genome or differences between genotypes can be detected by screening a series of such primers against DNA samples from the genotypes in question. Once a RAPD primer has been identified that consistently generates a different banding pattern between genotypes, it can be used as an identification tool for discriminating between those genotypes at any time in the future.

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Ashish K. Pathak, Sudhir P. Singh, and Rakesh Tuli

fingerprint and classify lychee cultivars using isozymes ( Aradhya et al., 1995 ; Degani et al., 1995 ) and DNA polymorphism using random amplified polymorphic DNA [RAPD ( Anuntalabhochai et al., 2002 ; Chundet et al., 2007 ; Kumar et al., 2010