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Jean-Guy Parent and Danièl Pagé

Characterization and identification of 13 red raspberry (Rubus idaeus L.) and two purple raspberry (R. × neglectus Peck) cultivars were obtained by nonradioactive genetic fingerprinting. DNA from leaves was digested with Hae III and Hin f I restriction enzymes and probed with alkaline phosphatase-labeled oligonucleotide. All tested cultivars could be identified by a unique band pattern. No differences were noted within cultivars when the reproducibility of the fingerprints was evaluated by analyzing the effects of age of the raspberry plantation, developmental stage during the growing season, or position of the sampled leaf on stem. These results suggest that simple nonradioactive DNA fingerprinting can be routinely used to identify raspberry cultivars.

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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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Assunta Bertaccini, Robert E. Davis and Ing Ming Lee

A collection of mycoplasma-like organisms (MLOs) was maintained in plant tissues micropropagated in vitro. MLO-infected plants included Chrysanthemum frutescens L. with chyrsanthemum yellows disease, Gladiolus sp. L. with “germ fins,” Hydrangea macrophilla (Thunb.) DC. with virescence, Rubus fruticosus L. with rubus stunt, and periwinkle [Catharanthus roseus (L.) G. Don] singly infected by the following MLOs: Italian periwinkle virescence, chrysanthemum yellows, North American aster yellows, Italian periwinkle stunt, American periwinkle little leaf. Shoots micropropagated in vitro exhibited symptoms of little-leaf and/or abnormal proliferation of axillary shoots resulting in “witches' broom” appearance that resembled symptoms in grafttransmitted greenhouse-grown or naturally infected field-collected plants. These symptoms, typical of infection by MLOs, were not observed in micropropagated healthy shoots of the same plant species, and, compared with the healthy ones, varied with MLO strain and host plant species. Dot hybridizations with a nonradioactive cloned DNA probe provided evidence for the presence of MLOs in propagated tissues through serial subcultures.

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Jean-Guy Parent and Daniele Page

Random amplified polymorphic DNA (RAPD) markers are used in Quebec's certification program to verify the identity of raspberry cultivars. However, sequence characterized amplified region (SCAR) markers, less sensitive to modifications in reaction conditions, could be derived from RAPD markers. Our objective was to evaluate the potential of SCAR markers to replace the RAPD ones. Five RAPD markers obtained with primer OPG06 (length of 520, 700, 825, 1450, and 2000 bp) were cloned in pTZ/PC or pCRII vectors. Extremities of the cloned markers were sequenced by the nonradioactive silver sequence method using pUC/M13 forward and reverse primers. Sequence information was used to make SCAR primers, similar in length to standard PCR primers. Some SCAR primers were elongated RAPD primers, whereas others were from internal regions. Ability of primer pairs and combination of primer pairs to discriminate cultivars of our certification program was compared with their RAPD counterparts as well as with the technical feasibility of both methods.

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Ali Fuat Gokce and Michael J. Havey

Cytoplasmic-genic male sterility (CMS) is used to produce hybrid onion seed. For the most widely used source of CMS in onion, male sterility is conditioned by the interaction of sterile (S) cytoplasm and the homozygous recessive genotype at a single nuclear male-fertility restoration locus (Ms). Maintainer lines used to seed-propagate male-sterile lines possess normal fertile (N) cytoplasm and the homozyous recessive genotype at the Ms locus. Presently, it takes 4 to 8 years to establish if maintainer lines can be extracted from an uncharacterized population or family. We previously developed a PCR marker useful to distinguish N and S cytoplasms of onion. To tag the nuclear male-fertility restoration locus (Ms), we evaluated segregation at Ms over at least three environments. Segregations of AFLPs, RAPDs, and RFLPs revealed molecular markers flanking the Ms locus. We are working to convert these linked molecular markers to nonradioactive PCR-based detection. The organellar and nuclear markers were used to select plants from open-pollinated onion populations and determine if the number of test-crosses required to identify maintaining genotypes.

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Masashi Yamamoto, Takahiro Tomita, Michio Onjo, Kiyotake Ishihata, Tatsuya Kubo, Shigeto Tominaga and Yoshimi Yonemoto

. 1990 A simple and efficient method for identification of hybrids using nonradioactive rDNA as probe Jpn. J. Breeding 40 339 348 Kumar, S. Tamura, K. Nei, M. 2004 MEGA 3: Integrated software for molecular

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Lee Kalcsits, Gregory van der Heijden, Michelle Reid and Katie Mullin

mature trees in the field. 44 Ca is a non-radioactive, stable form of Ca that exists naturally at a concentration of ≈2.086% ( Boulyga, 2010 ). 44 Ca is commercially available as enriched calcium carbonate (CaCO 3 ) comprises any of 42 Ca, 44 Ca, 46

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Qing Xu, Shi-Rong Guo, He Li, Nan-Shan Du, Sheng Shu and Jin Sun

were lower than in the compatible combination and nongrafted plants. The decrease of the F v /F m values in the “D” graft combination could be the result of an increase in the protective nonradioactive energy dissipation, photodamage to PSII centers

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Manuel Di Vecchi Staraz, Roberto Bandinelli, Maurizio Boselli, Patrice This, Jean-Michel Boursiquot, Valérie Laucou, Thierry Lacombe and Didier Varès

. Masi, E. Cresti, M. 2002 Genomic variability in Vitis vinifera L. ‘Sangiovese’ assessed by microsatellite and non-radioactive AFLP test Electron. J. Biotechnol. 5 1 Villifranchi, C.G. 1773 Oenologia