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RAPD analysis of grapevine hybrids and cultivars
63-66.Views:139Utilization of the Randomly Amplified Polymorphic DNA (RAPD) technique as a molecular marker was tested to investigate the relationships between some representative grapevine cultivars and hybrids established at the Department of Genetics and Plant Breeding (CUB), to distinguish clones as well as to characterize various hybrids between species or cultivars and their parents. Vitis vinifera cultivars were easily and successfully distinguished by the RAPD technique and they were grouped according to the traditional taxonomic classification. RAPD patterns of the examined Pinot gris clones proved to be completely identical. Number of generations was reflected by the value of genetic distance of the examined hybrids. Genetic identity of parents and their offsprings was influenced by the selection applied in the process of plant breeding. Parental phenotypic and morphologic characteristics showed high degree of segregation in hybrids, but RAPD analysis revealed that their genetic similarity is considerable. The three Vitis anntrensis clones were properly discriminated from every cultivar and hybrid of Vitis vinifera, i.e. hybrids are much closer to the cultivated grapevine than to V. anzurensis due to the phenotypic selection carried out during the life-cycle of one or two generations.
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Genetic diversity in a collection of apple (Malus x domestica Borkh.) cultivars as revealed by RAPD markers
25-35.Views:173A collection of 151 apple cultivars was investigated with 7 RAPD primers generating 71 informative bands, to evaluate genetic variability and relatedness. All cultivars presumably derived through genetic recombination were distinguished whereas identical DNA profiles indicated that some cultivars had arisen as sports. A cluster analysis and a PCO did not reveal any distinct geographic patterns, but there was a weak tendency for Swedish and foreign cultivars to differentiate.. Many cultivars however clustered together with either one of their parents or with siblings. Overall genetic diversity among the 151 cultivars was estimated with Nei's diversity index (H), 0.269, and with Shannon's index (H'), 0.594. The cultivars were also analysed in six groups, according to time of origination and country of origin, with an average H = 0.262 and H' = 0.546. No major differences in genetic diversity were observed over time or space, although the group with recent, foreign cultivars had the lowest diversity (FL = 0.235, H' = 0.493). Comparison between the entire material and a subset with 94 mandate cultivars chosen for preservation in Sweden, showed similar genetic diversity: HFN.rj1E = 0.268, frE,NTIRE, = 0.593 and HMANDATE = 0.263, WMANDATE = 0.575. No major differences in band frequencies were observed between these two sets, but 5 RAPD bands were missing in the set with mandate cultivars.
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Genetic relatedness among Asian Cotoneaster species investigated with DNA marker analysis
43-46.Views:152The widespread genus Cotoneaster has its centre of diversity in the Himalayas and surrounding areas. Most taxa appear to be polyploid and apomictic, and many of them have become popular ornamentals due to their attractive foliage and berries. One of the taxonomically most critical groups is Section Alpigeni which contains several important ornamental plants. The number of species belonging to this section varies widely between different taxonomic treatises, depending on whether 'splitting' or 'lumping' of species is preferred. Using a rather narrow species definition, we have investigated 13 different species using RAPD analysis. A simple matching (SM) coefficient-based principle coordinate analysis (PCO) was calculated from the RAPD data. Some species were clearly more similar to each other than to other species in the analysis. The levels of similarity did, however, not correspond very well to the lumping together of several taxa under the same species name as performed e.g. in the recent Flora of China. Obviously, the complex hybridogenous origination and, in some cases, still ongoing recombination with sexual species or among the apomictic taxa themselves, produces a genetic variability structure that cannot be properly reflected in a hierarchical taxonomy.
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Severely pollen-limited fruit set in a pear (Pyrus communis) orchard revealed by yield assessments and DNA-based paternity assignment of seedlings
67-74.Views:162In commercial fruit tree orchards, consistently high yields are necessary for a durable economy. The Swedish pear cultivar 'Carola' has been noted for low setting in some orchards, possibly due to insufficient pollination. In this study, fruit set was evaluated in a research orchard where `Carola' had been planted together with four potential pollinators. Total yield and number of fruits was noted during three and four years, respectively. In 2003, seeds were germinated from the harvested `Carola' fruits, and the paternity of three seedlings from 50 trees was determined with RAPD analysis. 'Clapp's Favourite' had sired 39.6% of the seedlings, closely followed by `Seigneur d'Esperen' (30.7%) and 'Clara Frijs' (26.7%) whereas 'Skanskt sockerparon' only sired 1.1% of the seedlings. The remaining 2.3% appeared to have been derived by selfing. Pollen-limited seed set was indicated at surprisingly short distances; accumulated number of fruits on the `Carola' trees was significantly higher when separated by only 2 m from one of the two most efficient pollinators, 'Clapp's Favourite' or 'Seigneur d'Esperen‘, compared to trees 4—l0 in away in the same row. Number of viable seeds per fruit was also higher in fruits from trees immediately adjacent to the pollinators, suggesting an effect of improved pollination success. The importance of very short inter-cultivar distances for efficient pollen transfer became even more clear when comparisons involved the true pollination distances as determined by RAPD; the accumulated yields decreased linearly from 55 kg at a 2 in distance to only 17 kg at 13 m.
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Molecular diversity of Hungarian melon varieties revealed by RAPD markers
11-13.Views:117RAPD markers were used to reveal genetic diversity between nine varieties of Cucumis melo L. and to identify the studied varieties. Of the 60 primers tested 12 primers produced polymorph patterns. A set of 4 primers was sufficient for distinction the nine investigated melon varieties.
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Molecular analysis of strawberry cultivars using RAPD, AP-PCR and STS markers
24-28.Views:121Seventeen strawberry (Fragaria x ananassa Duch.) cultivars representing the national list of Hungary, were subjected to RAPD, AP—PCR and STS analysis. Of the 31 decamer and oligomer primers tested 26 primers produced polymorphic patterns. 45 polymorphic fragments were analysed, ranging between 200-2800 by in size. Based on the data, similarity coefficients (Jaccard index and Simple matching coefficient) were calculated, and dendrograms were constructed using the unweighted pair group method of arithmetic averages (UPGMA). The dendrograms only partly reflect the known pedigree data. Specific RAPD markers were identified for cultivars F5, Pocahontas and Rabunda.
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Genetic variation in the rose pathogen Marssonina rosae estimated by RAPD
63-67.Views:155Blackspot, caused by the fungus Marssonina rosae (Diplocarpon msae), is one of the most devastating and widespread diseases in garden roses, and it has as yet not been fully characterized in molecular terms. In this initial study we used RAPD analysis to investigate the genetic diversity among and within a few geographically diverse groups of single-spore isolates of M. rosae. DNA was extracted from in vitrogrown mycelia of 1 I single spore isolates grown on PDA medium. High levels of polymorphism were detected among the isolates. They clustered into three distinct groups: Group !consisted of isolates from eastern North America plus a European isolate (Germany), Group 2 included isolates from southern Sweden, and Group 3 included the isolates from Manitoba, Canada. The greater similarity of the environmental conditions in eastern North America and Europe as compared to the Canadian prairies suggest that. climate and weather could be key factors in influencing the potential race structure of M. rosae. However, variations among closely situated sites, e.g. southern Sweden, also occurred.
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DNA-based determination of suitable pollinating cultivars for the pear cultivar 'Carola' (Pyrus communis)
15-19.Views:130Pollen-limited fruit set has long been suspected in some relatively low-yielding orchards with the Swedish pear cultivar 'Carola'. Fruit was therefore harvested on 23 'Carola' trees in a commercial pear orchard. The seeds were germinated and five seedlings from each tree were sampled to determine which of the surrounding cultivars had been the most successful pollinators. Leaves of 'Carola', the 7 putative pollinating cultivars and the 115 seedlings were analysed with 6 RAPD primers. By comparison of the band patterns, paternity could be ascertained for 74 seedlings. The by far most successful pollinator was 'Clara Frijs' which had sired approx. half of the seedlings, followed by 'Herzogin Elsa', `Skanskt Sockerpiiron', 'Alexandre Lucas', 'Coloree de Juillet' and 'Doyenne du Cornice'. The latter is the maternal parent of 'Carola', and these two cultivars must therefore share one S-allele and hence can only be semi-compatible. In addition, 6% of the seedlings were in all likelihood derived from selling_ since they showed no bands that did not occur also in 'Carola'. Maximum distance between 'Carola' trees and suitable pollinators should not exceed 15-20 tn. Longer distances may produce a serious dearth of compatible pollen as evidenced by the large percentage of seedlings derived either from selling. (25%) or from long-distance (> 40 m) pollen transfer (25%) when 'Carola' trees were surrounded by non-preferred pollinators.
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New simple and efficient method of DNA isolation from pear leaves rich in polyphenolic compounds
21-24.Views:113This study aimed to establish a new protocol for DNA isolation from the Pyrus genus to get high quality DNA that is suitable for the generation of molecular markers, such as RAPD and AFLP. This method is based on modified CTAB extraction procedure (Aldrich & Cullis, 1993). For isolation of high-quality DNA we used copper (II) acetate treatment that enabled fixation and removing of tannins, present in abundance in Pyrus. DNA yield from this procedure is high. DNA is completely digestible with restriction endonucleases and amplifiable in the PCR, indicating freedom from common contaminating polyphenolic compounds.
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The importance of clonal selection of grapevine and the role of selected clones in production of healthy propagating stocks
15-24.Views:287Genetical alterations and phytosanitary status promote the variability and modify the appearance of vine. Old vine varieties in old vineyards are highly variable and well adapted to selection. Clonal selektion is based on a visual performance: valuable individuals (clones) are picked out according to visible symptoms or characters. The genetical stability of clones is proved by testing the vegetatively propagated progenies on the basis of morphological and molekular (SSR, AFLP, SMPL, RAPD) markes. Authors take great care of the visual phytosanitary selection as part of the clonal selection being the oreliminary step to develop pathogen-free propagation stocks. In Serbia (Vojvodina) the selection breeding has been carried on for several decades resulted in comparative clone trials with home and imported clones of Welsch Riesling, Chardonnay, Pinot gris, White Riesling. Among the clones of home selection SK.54 Welsch Riesling clone is the most valuable. Its clearing from pathogene is being carried on in an interregional IPA programme (HUSRB/0901/214/123) in Kecskemét. In Kecskemét, the centre of the Hungarian Danube vine region 5 vine clones have been registered (Cegléd szépe K.73, Irsai Olivér K.11, Kövidinka K.8, Hárslevelû K.9, Pannónia kincse K.56). Besides them 18 virus-tested clones have also been qualified.Works aiming at their complete exemption are going on in order to obtain clones free of propagation wood-borne diseases.
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Test of the utility of apple retrotransposon insertion patterns for molecular identification of 'Jonathan' somatic mutants
7-10.Views:170Up until today, apple sport mutants proved to be indistinguishable from each other and their progenitors at the molecular level using random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) marker techniques. This is not surprising, since the genomes of these somatic mutants differ only in one or a few small regions that affect economically important characteristics, such as improved fruit colour, size, or flavour. In most cases, these genome differences are probably caused by retrotransposons which are able to convert their RNA transcripts to DNA with reverse transcriptase enzyme prior to reinsertion, but unable to leave the genome and infect other cells. Retrotransposon insertions can alter the expression of other genes and/or the structure of encoded proteins. The sequence-specific amplified polymorphism (S-SAP) technique is capable of revealing the genetic distribution of retrotransposable elements over the whole genome. The present study used this approach to try to characterize and distinguish 'Jonathan' somatic mutants via fingerprinting, which is an unsolved problem.