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Saad B. Javed, Abdulrahman A. Alatar, Mohammad Anis, and Mohamed A. El-Sheikh

in a greenhouse (RH, 50% ± 20%) for 4 weeks before field transfer. Molecular characterization. Genomic DNA was extracted from 10 tissue culture-derived plantlets and the donor plant using the cetyl-methylammonium bromide method, as described by Doyle

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

germplasm management in an Italian olive collection J. Amer. Soc. Hort. Sci. 131 352 359 Muzzalupo, I. Lombardo, N. Salimonti, A. Perri, E. 2008a Molecular characterization of Italian olive cultivars by

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Joseph C. Kuhl, Kelly Zarka, Joseph Coombs, William W. Kirk, and David S. Douches

-DNA where the RB primers anneal is designated by “PCR Product.” Molecular characterization. DNA extractions were conducted using a 2× CTAB procedure ( Doyle and Doyle, 1987 ). Detection of transgene insertions used 30 ng of total genomic DNA

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Rose Palumbo, Wai-Foong Hong, Guo-Liang Wang, Jinguo Hu, Richard Craig, James Locke, Charles Krause, and David Tay

genetic similarity of the accessions through molecular characterization by target region amplified polymorphism (TRAP) markers ( Hu and Vick, 2003 ). TRAP is a technique that combines the AT- and GC-rich primers of SRAP (sequence-related amplification

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Salih Kafkas, Sezai Ercişli, Yıldız Doğan, Yaşar Ertürk, Ayhan Haznedar, and Remzi Sekban

germplasm: Comparison of AFLP, ISSR and RAPD markers J. Amer. Soc. Hort. Sci. 131 522 529 Kafkas, S. Ozgen, M. Dogan, Y. Ozcan, B. Ercisli, S. Serce, S. 2008 Molecular characterization

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Kelly A. Zarka, Ria Greyling, Inge Gazendam, Dean Olefse, Kimberly Felcher, Gurling Bothma, Johan Brink, Hector Quemada, and David S. Douches

cry1Ia1 vector was developed and transformed into the potato cultivar Spunta. The public release of any GM crop is a highly regulated event and requires the rigorous molecular characterization of the transgenic event, detailed data on protein safety

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June Liu, Zhimin Yang, Weiling Li, Jingjin Yu, and Bingru Huang

physiological data indicated that the improved cold tolerance through in vitro selection was associated with the maintenance of chlorophyll accumulation and cell membrane stability as well as the suppression of membrane lipid peroxidation. The molecular

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María Ferriol, Belén Picó, and Fernando Nuez

Cucurbita maxima Duch. is one of the most morphologically variable cultivated species. The Center for Conservation and Breeding of the Agricultural Diversity (COMAV) holds a diverse germplasm collection of the Cucurbita genus, with more than 300 landraces of this species. Morphological and molecular characterization are needed to facilitate farmer and breeder use of this collection. With this aim, the morphological variation of a collection of 120 C. maxima accessions was evaluated. The majority of these accessions originated from Spain, which has acted as a bridge since the 16th century for spreading squash morphotypes between the Americas and Europe. South American landraces (the center of origin of this species) were also included. Eight morphological types were established based on this characterization and previous intraspecific classifications. A subset of these accessions, selected from these classification and passport data, was employed for molecular characterization. Two marker types were used; sequence related amplified polymorphism (SRAP), which preferentially amplifies open reading frames (ORF), and amplified fragment length polymorphism (AFLP). In the main, SRAP marker analysis grouped accessions in accordance to their type of use (agronomic traits) and AFLP marker analysis grouped accessions as to their geographical origin. AFLP marker analysis detected a greater genetic variability among American than among Spanish accessions. This is likely due to a genetic bottleneck that may have occurred during the introduction of squash into Europe. The disparity of the results obtained with the two markers may be related to the different genome coverage which is characteristic of each particular marker type and/or to its efficiency in sampling variation in a population.

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Laura L. Benson, Warren F. Lamboy, and Richard H. Zimmerman

The U.S. National Plant Germplasm System (NPGS) currently holds 36 separate accessions of the `Yichang' clone of Malus hupehensis (Pamp.) Rehd. The `Yichang' clone originally entered the United States in 1908 as seed collected for the Arnold Arboretum by E.H. Wilson near Yichang, Hubei Province, China. The original description of M. hupehensis omits fruit characters, and botanists frequently augment these omissions with descriptions of the `Yichang' clone. Apomixis occurs in Malus, including M. hupehensis, and is strongly associated with elevated ploidy levels. Simple sequence repeats (SSRs) were used to characterize 65 accessions of M. hupehensis. To check for polyploidy, a set of M. hupehensis accessions was evaluated with flow cytometry. The simple sequence repeat phenotypes and ploidy information revealed the `Yichang' clone under various accession names in arboreta. It was neither known nor suspected that the U.S. National Plant Germplasm System held many duplicate accessions of the `Yichang' clone prior to their molecular characterization. Germplasm conservation decisions for Malus species can benefit from an increased knowledge of the genetic variation or lack thereof in naturalized populations and ex situ collections.

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Laura L. Benson, Warren F. Lamboy, and Richard H. Zimmerman

Simple sequence repeats (SSRs) are highly polymorphic regions of DNA that can be used for the molecular characterization of apple (Malus) germplasm. SSR markers are sufficiently variable to distinguish between individual plants in wild Malus species. In this study, accessions of Malus hupehensis were screened for fragment length variation in PCR amplified simple sequence repeat regions of DNA. The fragment length phenotype produced by five SSR primer pairs showed no variation between two lineages of M. hupehensis collected in the Changjiang (Yangtse) River valley. One lineage was collected by E.H. Wilson in 1908 near the city of Ichang, Hubei Province. The second lineage was collected by cooperators at China's Southwest Agricultural University (SWAU) in 1997 near the city of Chongqing (Chungking). M. hupehensis Plant Introduction No. 588760 from the National Plant Germplasm System lacks provenance, but displays a fragment length phenotype identical to both the Wilson and SWAU lineages. The spread of a clone may be aided by asexual reproduction through seed, which is not uncommon in polyploid apples. Two seedlings each of 15 maternal trees from the SWAU lineage were assayed for ploidy level by flow cytometry. The DNA content per nucleus for all SWAU progeny fell within the range for triploids, 2.19 to 2.68 pg DNA/nucleus. It appears that plant explorers in China separated by almost 90 years have succeeded in sampling a single clonal lineage of M. hupehensis.