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Genetic diversity in a collection of apple (Malus x domestica Borkh.) cultivars as revealed by RAPD markers
25-35.Views:174A 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:153The 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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RAPD analysis of grapevine hybrids and cultivars
63-66.Views:140Utilization 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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Character association and genetic gain of nine agronomic traits of F1 populations in onion (Allium cepa L.)
50-58.Views:494F1 populations of half diallel cross were considered for this investigation. Phenotypic component of variation (s2P) was higher than genotypic (s2G), interaction ((s2I) and within error (s2w) components of variation. Trait bulb weight showed the highest values for s2P, s2G, s2I and s2w. The noticeable amount of phenotypic, genotypic, interaction and within error covariation was found in the combination of BW×BY, possibly indicating wide scope of selection for this pair of characters. Genotypic correlations were higher than the respective phenotypic correlations. This situation was also marked in the path coefficient analysis. Bulb yield/plot showed highly significant and positive correlation coefficient with other characters both at phenotypic and genotypic levels. When all the nine characters were included in an index, it exhibited the highest genetic gain as percentage. When a combination of two or more characters was studied in a function, the efficacy was higher than that of direct selection for bulb yield. The combination of five, six, seven or eight characters showed higher percentage of expected gain. Due to significantly correlated with BY and having high positive direct effect at phenotypic level characters viz., LL, BW, PH and NLs is considered as primary yield components. Again combinations of these four characters gave the commendable expected genetic gain of 330.7290% may be considered as important selection index for this material.
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Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae)
15-12.Views:267In the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for improving their fungal resistance using genetic engineering techniques. Adventitious shoot formation from leaf pieces of apples studied was achieved using middle leaf segments taken from the youngest leaves from in vitro-grown plants.
Optimum conditions for ‚direct’ shoot organogenesis resulted in high regeneration efficiency of 0%, 95%, 92%, 94% in the studied apples respectively. Putative transgenic shoots could be obtained on MS media with B5 Vitamins, 5.0 mg l-1 BAP, or 2.0 mg l-1 TDZ with 0.2 mg l-1 NAA in the presence of the selection agent “PPT” at 3.0-5.0 mgl-1. Shoot multiplication of transgenic shoots was achieved on: MS + B5 vitamins + 1.0 mg l-1 BAP + 0.3 mg l-1 IBA, 0.2 mg l-1 GA3+1.0 g/l MES+ 30 g/l sucrose + 7.0 g/l Agar, with the selection agent PPT at 5.0 mg l-1 and were subcultured every 4 weeks in order to get sufficient material to confirm transformation of the putative shoots obtained. Six, seven, one and six transgenic clones of the apples studied respectively have been obtained and confirmed by selection on the media containing the selection agent “PPT” and by PCR analysis using the suitable primers in all clones obtained for the presence of the selection” bar gene (447 bp) and the gene-of- interest “g2PS1” (1244 bp), with transformation efficiency of 0.4%, 0.6%, 0.1% and 0.3% respectively. These transgenic clones were multiplied further in vitro in the presence of the selection agent ‘PPT’ and rooted in vitro. Rooted transgenic plantlets were successfully acclimatized and are being kept under-containment conditions according to the biosafety by-law in Syria to evaluate their performance for fungal resistance . -
Experimental results of the effects of Hungarian climatic conditions to German disease-resistant industrial apple varieties
53-56.Views:117In the recent years, several disease-resistant apple varieties appeared through the modern breeding technologies. These varieties can be grown with low usage of pesticides, which mean not only environment friendly fruit growing, but the production costs are also lower. In Eastern-Hungary — it is one of the main apple growing regions — a new apple growing structure started to form by the investment of the German Wink Ltd. — several resistant apple varieties were brought from Germany. 'Resistant' refers genetic resistance that usually transferred from the genome of wild apple species. But the fruit of these apple species is not only resistant to diseases, but its quality is poorer, too. In Germany the Re-apples are grown only for the processing industry. Due to climatic circumstances in Eastern Hungary, the first experiences showed better parameters during laboratory measurement, the fruits have more beautiful view, shape and inner characters than usual industrial apples.
In our paper we discuss the results of sensory (consumer) tests, carried out in Eastern Hungary and in the Budapest-region the data analysis of systematic storing experiments (refraction, flesh firmness, weight loss, etc.) and profile analysis of fresh and stored Re-apples. (In the profile analysis the ProfiSens software [4,5] has been used.)
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Identification of Rabbiteye Blueberry Cultivars (Vaccinium ashei Reade) and Analysis of Genetic Relationships Using Amplified Fragment Length Polymorphism (AFLP)
27-30.Views:109Proper cultivar identification is a requisite for commercial planting and breeding nurseries of cross-pollinated blueberry (Vaccinium ashei Reade) cultivars to insure high crop yields and optimize germplasm maintenance and utilization. Fourteen rabbiteye blueberry cultivars and three non-identified clones were screened with amplified fragment length polymorphism (AFLP) analysis with the aim of developing a fast and reliable identification technique. The selective primer pair applied (M-CTG/ E-ACC), which was previously tested, resulted in a large number of reproducible polymorphic fragments for cultivar identification. After comparison of the AFLP fingerprints, the Jaccard similarity indexes were calculated, and an UPGMA dendrogram was constructed. It was revealed that the three non-identified clones belong to the Tifblue' cultivar. Moreover, AFLP technique proved to be a fast, successful and reliable way in rabbiteye blueberry identification.
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Development of microsatellite markers for Rhodiola rosea
37-42.Views:201Rhodiola rosea L. is an important adaptogen medicinal plant. In this study two new microsatellite markers were developed. The assessment of the genetic diversity of R. rosea has recently started with molecular markers, but only a few species-specific microsatellite markers have been published so far. However the small number of markers allows only a limited insight into the genetic variability of the species therefore the aim of our work was to develop new microsatellite markers for R. rosea with a microsatellite enrichment library technique. Genomic DNA was cleaved with an endonuclease enzyme followed by adaptor ligation and PCR amplification. DNA fragments that contained microsatellites were first isolated using a biotin-streptavidin linkage based magnetic selection and then cloned into plasmids. Out of forty-three sequenced clones three contained microsatellites, in these cases primers were designed for the amplification of the microsatellite repeats. The newly developed primer pairs were tested on individuals from distant R. rosea populations and the variability of the amplified fragments was estimated by fragment-length analysis. The locus RhpB14a was found to be monomorphic while RhpB14b and RhpB13 were polymorphic. As a result of the present study, two novel variable microsatellite loci were identified in the genome of R. rosea.
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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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Genotyping Hungarian apricot cultivars for self-(in)compatibility by isoelectric focusing and PCR analysis
69-72.Views:146Self-incompatibility (SI) in flowering plants is a widespread genetic system that promotes out-crossing. In Prunus species the SI is a gametophytic trait, which is controlled by a single multiallelic locus, termed S-locus. S-alleles codify stylar glycoproteins with ribonuclease activity (S-RNases). Our objective was to assess the S-genotype of some Hungarian apricot varieties by isoelectric focusing of stylar RNases as well as by PCR analysis using cherry consensus primers. Consensus primers amplified one or two bands of various sizes. Primers amplifying the 1st intron gained fragments the size of which ranged from 250 to 500 bp; while those amplifying the 2nd intron resulted in fragments of 800-2000 by length. Our data demonstrated that the first intron of the apricot S-RNase gene is shorter than the second one, which coincides with the structure of cherry S-RNase alleles. `Hargrand' (S1S2) and `Harcoe (S1S4) possessed one common S-RNase isoenzyme. Hungarian 'Orias' apricot cultivars showed different bands compared to the previous cultivars, but they shared completely identical patterns confirming that they possess the same S-genotype. 'Bergeron', `Harmat' and 'Korai zamatos' are characterised by an evidently distinct S-RNase pattern. The self-compatible cultivar (`Bergeron') had one allele, which suggests its correspondence to the Sc. Primers for the 2nd intron was unsuccessful in gaining fragments, which indicates that the 2nd intron in the Sc allele is too long to get any amplification. On the basis of our data, identities and differences were revealed in the S-allele constitution of some economically important Hungarian apricot cultivars at protein and DNA levels.
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Resistance gene Sw-5 of tomato confers resistance to TCSV in Solanum melongena
41-47.Views:137Eggplants transformed with Sw-5 gene, regenerated by organogenesis and somatic embryogenesis, were resistant to the Tomato chlorotic spot virus, while wild plants did present systemic infection. TO plants were selfed and the segregation analysis of T1 and T2 generation indicated the existence of one or more insertion sites. Southern blot analysis confirmed one or two independent insertions in T2 plants. Different lesions associated with the insertion number were observed in TI and T2 plants. T2 plants with two copies displayed faster hypersensitive reactions and characteristic necrotic lesions that contrasted with slower responses and necrotic ring lesions in plants with one copy. These results suggest that the Sw-5 confers resistance to tospovirus in transgenic eggplants and that the resistant phenotype depends on the number of transgene copies.
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Genetic transformation of bean callus via Agrobacterium- mediated DNA transfer
49-53.Views:120Callus cultures were induced from hypocotyl of young bean seedlings. Callus developed and maintenaned on B5 medium supplemented with 2mg/1 2,4-D and 1 mg/1 kinetin. The results demonstrate that A. tumefacins-mediated transformation is a convenient method to obtain transient gene expression in callus of bean. The results have shown that the bean callus co-cultivated with A. tumefaciens can be transformed to get heibicide Finale (glufosinate-ammonium) resistant GUS positive tissues. Southern blot analysis of transformed calli showed integration of gusA marker gene carried by a binary vector. Transformed calli were selected on herbicide containing media. Data of molecular analysis (Southern blotting) confirmed the insertion of gusA gene in the genome of herbicide resistant calli with bar gene. There are three evidences that calli are stable transformants: (1) herbicide resistance, (2) GUS activity which is indicative since the coding region containing an intron, (3) the results of Southern hybridization technique.
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Genetic variation in the rose pathogen Marssonina rosae estimated by RAPD
63-67.Views:156Blackspot, 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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Microsatellite based identification of grapevine cultivars traditional in Hungary and in the Carpathian Basin
71-73.Views:137Grapevine cultivars and clones traditional in Hungary and in the Carpathian Basin are maintained in the genetic collection of the Corvinus University of Budapest. Mostly ancient varieties and clones were genotyped using microsatellite loci. The investigated 6 loci were sufficient to distinguish all cultivars. Some clones could also be separated based mostly on the variable VVS2 loci. For 17 out of the 31 investigated cultivars this is the first report on characterization of the polymorphism of the allele lengths by microsatellite markers on loci: VVS2, VVMD7, VVMD27, vrZAG62.
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The antioxidant capacity of sea buckthorn (Hippophae rhamnoides L.) berries depends on the genotype and harvest time
27-29.Views:246Berries of sea buckthorn (Hippophae rhamnoides L.) are characterized by increasing popularity due to their presumable healtheffects. The aim of this study was to compare the antioxidant capacity and total polyphenolic content in the berries of six Hungarian grown sea buckthorn genotypes and characterize the genetic variability in this trait. The harvest time of sea buckthorn berries affects the antioxidant capacity and total phenolic contents in berries of three popular cultivars of German origin. Berries harvested in October had higher antioxidant capacity compared with those harvested one month later. The extent of the difference was genotype-specific. Our analysis revealed a nearly 3-fold difference between the lowest and highest antioxidant capacities of the 6 tested genotypes with ‘Leikora’ showing the highest ferric reducing antioxidant power and total phenolic content. The TEAC values ranged between 1.76 and 3.13 mmol Trolox/100g fresh weight with Pető 1 and ‘Frugana’ having the highest values. The results presented in this study demonstrated that Hippophae rhamnoides berries possess in vitro antioxidant activity strongly determined by genotype but also influenced by harvest time.
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Production of transgenic bean callus via genetic transformation by DNA-coated tungsten particles
43-47.Views:108Callus cultures were induced from hypocotyl of young bean seedlings. The B5 medium completed with 1 mg/1 KIN and 2mg/1 2,4-D proved the best. Callus developed and maintenaned on B5 medium supplemented with 1mg/1 kinetin and 2mg/I 2,4-D. The B5 medium supplemented with 1mg/1 KIN and 2mg/1 2,4-D induced much more callus than half strength MS medium supplemented with 0.5 or 0.75mg/1 BA and 0.1 mg/1 NAA. The results demonstrate that GeneboosterTM is convenient method to obtain transient gene expression in callus of bean. The results have shown that the bean callus shot by GeneboosterTM can be transformed to get (kanamycin-resistant and stress mannitoltolerant) calli. The presence of mannitol-dehydrogenase gene (mt/) was verified by PCR, showing the integration of mt/ gene carried by two plasmids. Co-transformed calli were selected after bombardment on kanamycin, mannitol and (kanamycin+mannitop-containing media. Data of molecular analysis (PCR) confirmed the insertion of mtl gene in the genome of mannitol-tolerant callus lines.
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Comparative analysis of sour cherry cultivars on their ecological and biological indicators
7-28.Views:329Sour cherries developed in the northern hemisphere, an alloploid hybrid of dwarf sour cherries (Prunus fruticosa) and bird cherries (P. avium), born in the confluence of the two species. However, the ecological and, above all, cold tolerance of the ancestor of cultivated sour cherries is higher than that of wild cherries (De Candolle, 1894; Rehder, 1954; Terpó, 1974; Iezzoni et al., 1991; Faust & Surányi, 1997). The cultivation limits are in the northern hemisphere 38-44. degree. The Carpathian Basin, the Balkans and Asia Minor are considered to be the main birthplaces for sour cherries. The genetic and morphological diversity of sour cherries is greater than that of the basic species (Iezzoni et al. 1991; Faust & Surányi, 1997). In the study, 472 sour cherry cultivars were compared based on 7 relative ecological indicators and 3 biological values. Compared to other Prunus species, we mostly found less variability in sour cherries - not counting their salt tolerance (SB). The partial similarity between open pollination (OP), frost tolerance (FR) and disease resistance (DR) - partly true in terms of varieties, but also reflected the effects of purposeful breeding and selection. The cultivars together - in comparison, showed balance, but in the highlighting, the differences of the 3 cultivar groups became significant. Indeed, the differences between the species of the former Hungarian cultural flora are clearly different (Surányi, 2004), which is also the case when comparing a large number of apricot (Surányi, 2014), plum (Surányi, 2015) and peach (Surányi, 2020) varieties.
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Antioxidant capacity, total phenolics and mineral element contents in fruits of Hungarian sour cherry cultivars
59-64.Views:355Several epidemiological studies revealed that the consumption of antioxidant compounds and the risk of atherosclerosis, increased blood pressure or cancer are inversely proportional. Fruits of sour cherry contain a wide range of antioxidant compounds including melatonin, perillyl alcohol, ellagic acid, several flavonoids, polyphenolics, and anthocyanins. This study was carried out to survey the antioxidant power and mineral element content of seven commercial sour cherry cultivars and three cultivar candidates and to assess the influence of some external conditions on fruits' functional properties. Our analysis revealed nearly 5- and 2-fold differences between the lowest and highest antioxidant capacities and total phenolics content, respectively. Some cultivars (`Kántorjánosi' and ‘Újfehértói fürtös') and cultivar candidates (D, 'Petri' and 'Éva') showed outstanding antioxidant capacity and total phenolic content; in addition, mineral element content in fruits of the ‘Újfehértói fürtös' cultivar was also favourable. Redox parameters of fruits were influenced by the cultivation plot or fruit positions within the canopy in about half of the cultivars tested. Genetic background of cultivars forms the decisive factor in determining fruits' antioxidant capacity, although external factors may have also sizeable modifying effects. Enhanced functional properties of the fruit may also be further increased through breeding programs since considerable variation exists within the tested germplasm.
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Ecological drought resistance and adaptability of apple varieties
113-122.Views:300For adequate yields in apple plantations, during the long growing period of the fruit primordia, one of the decisive factors is water supply. Indicators of stress are valuable signs for the diagnosis of drought and necessity of watering, i.e. planning the irrigation of plantations. The aim of the present study was to find reliable signs of water stress on apple trees and at the same time conclude on the drought tolerance of different varieties. The plantation of apple varieties grafted on various stocks and cultivated according to different systems (irrigated, non irrigated, integrate and biological) has been examined continuously by leaf analysis. Along the period of growing fruits, measurements were made in the field, then the leaf samples were analysed in the laboratory for composition of pigments, carbohydrates and antioxidants, as well as the histology of the tissues checked. Without irrigation, the mean leaf mass and the relative chlorophyll content (SPAD) of the variety Idared on M4 stocks increases beyond the values of 50 SPAD, whereas on M26 stock and integrated system, it declines. In the collection of varieties, grown according to the „integrated” system, during the development of fruits, ‘Gála’ and ‘Remo’ varieties have been affiliated to the “less susceptible” group regarding drought tolerance because of the leaf morphology, structure and content of SPAD and antioxidants. However, ‘Idared’ and ‘Jonagold’ belong to the “susceptible” group. To the same group are ranged the ‘Akane, Red Rome vanWell, Pink Lady’ varieties. Those varieties reacted to a short period of drought by increased production (content) of carbohydrates. Regarding changes of carbohydrate content ‘Greensleeves’ and ‘Ozark Gold’ varieties belong to the moderately water dependent (requiring) group. In the field, SPAD-tests facilitated the measurement of nutrient-uptake and incorporation, which is proved by the tight correlation between the data of SPAD and the increment of leaf weight (r=0.76–r=0.88), however, this depends on the variety too. SPAD is an indicator of water supply and is related with the density of stomata, cannot used for the selection of water-exigent varieties but for (drought) tolerant ones. In integrated culture, the (drought) susceptible varieties display (water-soluble) hydrophilic and lipophilic antioxidants (ACWand ACL resp.) the quantity of which may have some role in drought tolerance.