Search
Search Results
-
Co-transformation of bean callus using high-velocity microprojectiles- mediated DNA transfer
76-78.Views:128We have found that 50 mg/I kanamycin and 0.8 Mo1/1 mannitol concentration was sufficient to kill the control callus of bean (Phaseolus vulgaris L.) and differentiate transgenic from the non-transgenic cells. The GeneBooster particle delivery system was used for the bombardment of bean callus. The kanamycin resistance gene was used as a selectable marker. The test was made by transferring the healthy white callus, subcultured for three months on selective and non-selective medium. After selection on kanamycin containing media, several kanamycin resistant calli had been obtained, survived and grew. After selection on mannitol containing media no drought resistant calli had been obtained. Resistance of the selected calli were verified by their ability to grow repeatedly on selective medium containing 150 mg/I kanamycin. Selective pressure was maintained over a period of 8 months.
-
Bean tissue culture and genetic transformation with Agrobacterium
32-35.Views:134In this paper we report the establishment methods of a rapidly growing callus culture of Phaseolus vulgaris bean as well as the conditions required for a high level of transient gene expression using Agrobacterium-mediated transformation. A vector is containing both the lindan-resistance gene as a selectable marker, and GUS gene as a screenable marker. By using hypocotyl explant and vertical culture on B5 medium supplemented with 1 mg/1 kinetin- and 2,4-D 2 mg/1 and subcultured every 3-4 weeks, we can recommend to get a good and much callus from bean. This will help in introducing foreign DNA into callus cells. One strain of Agrobacterium carrying plasmid as vector for introducing foreign DNA into plant cells was used. At different concentrations of lindan; 3, 4 and 4.5 mg/I, the transformed Maxidor callus survived and grew over a period of 6 month and subcultured every 3-4 weeks, but the control callus died. Callus were assayed for GUS activity to confirm the expression of the GUS gene using the histochemical assay test. The GUS gene was also correctly expressed in callus cultures grown on 4mg/I lindan-selected medium, the typical blue colour in the histochemical assay using the X-gluc as substrate. But the control, non-transformed callus was not able to grow in the presence of lindan, neither showed a positive reaction in the in vitro assays.
-
DNA-based determination of suitable pollinating cultivars for the pear cultivar 'Carola' (Pyrus communis)
15-19.Views:135Pollen-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.
-
The use of SSR markers in family Rosaceae
29-32.Views:144The identification of plant species and study of their genetic relatedness is an important object of plant genetics. The Rosaceae family contains a lot of economically important fruit, ornamental, and wild plant species. The microsatellite markers have been proven to be an efficient tool for description of the genetic relatedness among varieties and species. Their evolutionary conserved regions enable them to differentiate among various accessions. This article intends to show proceeded identification and characterization projects on Rosaceae species by using SSR markers. The article presents sources of already published primer sequences. The use of already published primers can highly reduce the cost and duration of this kind of researches.
-
Attempting Regeneration from Cultured Cotyledons and Plant Regeneration from Cotyledonary Nodes in Common Bean (Phaseolus vulgaris L.)
57-60.Views:162Dry seeds from two cultivars of common bean (Phaseolus vulgaris L.) were germinated on sterile cotton and sterile deionized distilled water. Cotyledonary node tissue of seedlings were cultured on Murashige and Skoog(MS)-based media supplemented with different combination of N6-benzyl-aminopurine (BAP) and indole-3-acetic acid (IAA), and benzyladenine (BA) and a-naphthaleneacetic acid (NAA). The results revealed that the regeneration percent and the average number of buds and shoots per explant were influenced by the type of explants and exogeneously added hormones. Multiple shoot induction on dry bean cotyledonary node that contain 4-5 mm from cotyledons and hypocotyl on a medium containing full concentration of MS inorganic salts supplemented with 0.5mg/1 BA and 0.1mg/1 NAA was feasible and the method can be applied in transformation experiments.
-
Callus induction on standard type Cymbidium cultivars
108-110.Views:135Tissue cultured Cymbidium PLBs (protocormlike body) were used as starting material to induce embryogenic callus which could serve as objects of genetic transformation. We obtained callus using two methods. The first method was culturing the PLB segments for one month in liquid MS medium in the presence of 0.5 mg/1 benzyladenine and 0.05 mg/1 naphtylacetic acid followed by cultivation on the same composition solid medium with 0.5 g/l activated charcoal for an additional month. Callus formation was observed on 30% of the explants. The second way was to propagate the PLB segments on solid MS medium supplemented with 1 mg/1 thidiazuron. In these cultures we also observed callus formation on 20% of the explants.
-
Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple
7-10.Views:357Quince (Cydonia oblongaMill.) is a minor fruit crop, which is primarily used for marmalade, jam and sauce.Very few quince cultivars are known all over the world and in many cases similar names are used for presumably different cultivars. The aim of the present study was to evaluate and characterize the genetic diversity of 36 quince cultivars and selections with SSR markers. Seven out of 8 SSR markers designed from apple sequences could successfully yield amplification also in quince cultivars. Number of alleles per locus ranged from 2 to 3 alleles. These allele numbers are quite low when compared to apple. It is supposed to be the consequence of a genetic bottleneck. In spite of the low allele number per locus, the 36 quince cultivars formed 30 different genotypes. The ratio of homozygosity was low, which might be coupled with the self-(in)compatibility phenotype of quinces. SSR markers proved unable to differentiate putatively closely related cultivars (e.g. ‘Bereczki’ and ‘Bereczki bôtermő’). In general, the level of polymorphism among the tested quince genotypes was much restricted due to the low allele number detected. However, it must be considered that the number of analysed SSR loci is not enough high to estimate the overall heterozygosity of the quince genome. Further experiments are needed and the SSR markers proved to be a reliable and useful tool for such analyses.
-
Microsatellite based identification of grapevine cultivars traditional in Hungary and in the Carpathian Basin
71-73.Views:145Grapevine 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.
-
Preliminary characterization of the self-incompatibility genotypes of European plum (Prunus domestica L.) cultivars
23-26.Views:233European plum is an important fruit crop with complex, hexaploid genome of unknown origin. The characterization of the selfincompatibility (S) locus of 16 European plum cultivars was carried out using the PaConsI-F primer in combination with the EM-PC1consRD primer for the first intron and the EM-PC2consFD and EM-PC3consRD primers for the second intron amplification. Altogether, 18 different alleles were scored indicating high genetic diversity. These alleles were labelled using alphabetical codes from SA to SS. We identified 5 different alleles in 9 cultivars, 4 alleles in 5 cultivars, while 3 alleles were shown in two of the assayed cultivars. A total of 16 different S-genotypes were assigned, and discrimination of all plum cultivars was successful based on their unique S-genotypes. However, further research is required to reliably identify the S-alleles based on their DNA sequence and clarify complete S-genotypes.
-
The effect of different berries in human nutrition
147-151.Views:210The aims of this study were on the one hand to compare of examined compounds according to their importance in nutrition and human-health, and on the other hand we made preexperiments to investigate the relationships among antioxidant capacity and the endogenous substances which contribute the antioxidant status of the plants. The following species were involved in the experiment: raspberry, blackberry, black currant, elderberry and sour cherry. These fruits have potent health-promoting antioxidant power. Glucose, fructose, total phenol, formaldehyde and anthocyanin contents were determined in addition to ferric reducing ability. Our results reflected considerable differences in the measured parameters of the analysed species. In blackberries and elderberries the high antioxidant capacity is coupled with low carbohydrate content. Besides the formerly proven correlations between total phenol content, anthocyanin concentration and antioxidant capacity, these parameters also correlated with the measured formaldehyde concentrations, hereby we can follow the methylation /demethylation status of the plant.
-
Study of different factors of grapevine regeneration systems and genetic transformation
33-36.Views:218The most limitating factor for successful transformation is the absence of high-yielding regeneration protocols. However, the anther-derived embryogenic culture is an optimal technique for genetic transformation and it has been widely applied in many important cultivars, but the necessity of further development of regeneration systems has been proved. We attempted to produce somatic embryos on a wide range of genotypes from various tissues; leaves, petioles, stem segments. We started the examination of grapevine regeneration via organogenesis, succeeded in inducing shoot from the meristematic tissue of the base of bud by testing induction medium contained different concentrations of two types of hormones. To optimize the conditions of the Agrobacterium-mediated transformation, we studied the effectiveness of different Agrobacterium-treatments, the use of antioxidants and the sufficient quantity of kanamycin for selection of transformed cells.
-
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:176In 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.
-
Genotyping Hungarian apricot cultivars for self-(in)compatibility by isoelectric focusing and PCR analysis
69-72.Views:162Self-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.
-
High-velocity microprojectile mediated DNA delivery into Phaseolus vulgaris callus cells
99-102.Views:123We report the method for the establishment of rapidly growing callus cultures of Phaseolus vulgaris and the conditions required for efficient transformation using high velocity microprojectiles and high level of transient gene expression. Using hypocotyl explant and vertical culture on B5 medium with lmg/1 kinetin and 2 mg/1 2,4-D, we can recommend to get a rapidly growing callus from bean which is a good starting material to introduce foreign DNA into bean cells. The GeneBooster particle delivery system was used for the bombardment of bean callus and the Hgm resistance gene (Hgmr) was used as a selectable marker gene. 25mg/I hygromycin (Hgm) concentration was sufficient to kill the control callus. We used the standard physical factors, the appropriate pressure of N2 gas for the bombardment of the callus tissue, the shooting distance and the size of tungsten particles used as microprojectiles. Selective and nonselective tests were made by transferring the healthy green and white calluses, subcultured for 4 months on selective and nonselective medium. Several Hgm resistant calli had been obtained. Selective pressure was maintained over a period of 10 months.
-
Six promising selections from the Hungarian apple breeding program for multiple resistance
23-28.Views:143An apple breeding program has been carried out at the Department of Fruit Science for more than a decade. Several apple selections have been released from the progenies of crosses in 1992 and 1993. Six candidates were submitted for national recognition out of the hybrids examined for more than a decade. The six selections are resistant against all the three most important apple diseases (apple scab, powdery mildew and fire blight). Scab resistance is controlled by the Vf gene originating from the species Malus floribunda 821 and transmitted by cultivar Prima. Heterozygote Vfvf genotype of the six cultivar candidates was proved by molecular genetic examinations of Dept. Genetics and Plant Breeding. Characteristics of these selections from 'Prima' progenies are shown on the base of our own observations.
-
Self-incompatibility alleles in Esatern European and Asian almond (Prunus dulcis) genotypes: a preliminary study
23-26.Views:200Almond [Prunus dulcis (Mill.) D. A. Webb.] as one of the oldest domesticated plants is thought to have originated in central Asia. Gametophytic self-incompatibility of almond is controlled by the highly polymorphic S-locus. The S-locus encodes for an S-ribonuclease (S-RNase) protein in the pistils, which degrades RNA in self-pollen tubes and hence stops their growing. This study was carried out to detect S-RNase allelic variants in Hungarian and Eastern European almond cultivars and Turkish wild growing seedlings, and characterize their S-allele pool. Five new alleles were identifi ed, S31H, S36-S39 in Eastern European local cultivars. The village Bademli and Akdamar island are two distinct places of almond natural occurrence in Turkey. Trees growing wild around Bademli city showed greater genetic diversity than those originated on Akdamar island. Many of the previously described 45 S-RNase alleles have been also detected in these regions. Homology searches revealed that Turkish almonds carried some P. webbii alleles indicating hybridization between the two cultivars and massive introgression events. Our results supply long-awaited information on almond S-allele diversity from regions between the main cultivation centres and the centre of origin of this species; and are discussed from the aspect of methodological developments and evolution of the cultivated almond.
-
Factors affecting apricot fruit antioxidant capacity and mineral element contents
95-99.Views:225Several epidemiological studies revealed that the consumption of antioxidant compounds and the risk of atherosclerosis, increased blood pressure or cancer are inversely proportional. The individual amounts and relative proportions of macro- and micro elements present in food is also of great consideration since these are involved in a wide range of physiological processes including the influence of the redox homeostasis. The antioxidant power and mineral nutrient content of fruits might be affected by several factors including genotype, ripening stage, year-effects or a wide range of environmental conditions. This study was carried out to survey the antioxidant power and mineral element content in fresh fruits of apricot and analyse some genetic and environmental factors that may have important contribution to the inner content of apricot fruits. In addition, the influencing effect of the extraction procedure used for antioxidant analyses was also tested. Our analyses indicate that a considerable fraction of antioxidant capacity is attributable to the hydrophilic antioxidants. The genetic background has crucial importance in determining apricot fruit antioxidant capacity and mineral nutrient content; however, the growing season and the ripening time of fruits may have also important effects.
-
Somatic embriogenesis and regeneration of Vitis sp.
41-45.Views:154Somatic embriogenesis and regeneration of Vitis sp.
-
In vitro regeneration from cotyledons of watermelon
96-98.Views:168Cotyledonary segments of five different genotypes of watermelon were used to induce organogenesis. Five different hormone combinations were applied to enhance the induction of shoot formation on the surface of the segments. The phases of organogenesis were followed with light and scanning electron microscope. Shoots were obtained after four weeks, then the shoots were transferred to hormone free medium for root induction.
This method of regeneration can be applied in transformation experiments. GUS histochemical assay was made to check the expected success of using Agrobacterium for the transformation.
-
Evaluation of fruit quality of old apple cultivars originating from the foot of the Carpathian Mountains, for utilization in breeding and in organic farming
15-24.Views:302In the frame of the apple breeding program at the Department of Fruit Science old cultivars found in the Carpathian basin were involved in examination as well. Beside testing susceptibility to fire blight (Erwinia amylovora of old genotypes (historical, local and regional cultivars) originating from scattered orchards of Visk (foot of the Carpathian Mountains), fruit quality was also evaluated and other open-field observations were made.
According to the results of inner content examinations, 'Szemes alma' and 'Sikulai alma', chosen as gene sources for breeding to resistance against fire blight, had inner content values worthy to mention in some aspects, and competing with those of the control cultivar 'Jonathan'. The quality of 'Pónyik alma', also recommended as a gene source is confirmed by the data of earlier literature. Further three cultivars ('Batul', 'Vilmos renet', 'Zöld sóvári') also proved to be good based on resistance against Erwinia amylovora, the first two cultivars are suitable from the point of view of fruit quality, too.
In our field observations, susceptibility to apple scab and powdery mildew was evaluated. Among cultivars selected from the foot of the Carpathian Mountains on the base of resistance against Erwinia amylovora, the production value of 'Szemes alma' is worsened by its high susceptibility to apple scab. Further selected cultivars ('Sikulai', 'Pónyik alma', 'Batul', 'Vilmos renet') are not too susceptible to fungal diseases and their fruit quality is also suitable, therefore they can be recommended for direct planting in ecological farming or in restoring scattered orchards.
-
Researches concerning the selection of some nectarine varieties as genitors for the breeding program in Romania
44-46.Views:109Researches concerning the selection of some nectarine varieties as genitors for the breeding program in Romania
-
S-locus genotyping on stone fruits in Hungary: a review of the most recent achievements
39-43.Views:208Central Europe can be taken as a geographical and historical connection zone between the western growing countries and Asian gene centres of Prunus tree fruits. The determination of the S-genotype of stone fruit (mainly almond, plum, cherries and apricot) cultivars and landraces has both practical and theoretical significance. Our group has allocated complete S-genotypes for more than 200 cultivars and selections of almond, Japanese plum, sweet cherry and apricot. Among Eastern European almond cultivars, two novel cross-incompatibility groups (CIGs) were identified. S-alleles of a related species were also shown in P. dulcis accessions; a fact seems to be indicative of introgressive hybridization. Our results with Japanese plum clarified and harmonized two different allele nomenclatures and formed a basis for intensive international studies. In apricot, a total of 13 new S-alleles were identified from Eastern European and Asian accessions. Many Turkish and North African cultivars were classified into new CIGs, III–XVII. Results suggest that the mutation rendering apricot self-compatible might have occurred somewhere in south-east of Turkey and we were successful to confirm the presumed Irano-Caucasian origin of North African apricots based on the geographical distribution of S-alleles. In sweet cherry, new alleles have been identified and characterized from Turkish cultivars and selections. In addition, wild sweet cherry and sour cherry S-alleles were also shown indicating a a broader gene pool in Turkey as compared with international cultivars. We also used S-genotype information of Ukrainian sweet cherry cultivars to design crosses in a functional breeding program. Our results exhibit an increased number of S-alleles in tree fruit accessions native to the regions from Eastern Europe to Central Asia, which can be used to develop S-genotyping methods, to assist cultivation and draw inferences for crop evolution.
-
Genetic diversity in a collection of apple (Malus x domestica Borkh.) cultivars as revealed by RAPD markers
25-35.Views:181A 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.
-
Evaluation of different sources of resistance for breeding powdery mildew resistant grapevine varieties
35-40.Views:180The appearance of resistance to powdery mildew was investigated on seedling progenies of interspecific crossings. Hybrids of American and East-Asian wild species and one Vitis vinifera variety (Janjal kara) were used as source of resistance in the crossings. The resistance of foliage and of berries were tested in the field at the time of vintage. The degree of resistance was ranked into six classes.
In American and Asian hybrid derivatives there were few highly resistant progenies, but in the hybrids of Janjal kara variety more than 90 % of the progenies proved to be highly resistant to powdery mildew. According to the distribution of classes of resistance in the populations the resistance of American and Asian sources are mostly influenced by additive gene affects. The resistance of Janjal kara seemed to be mono or digenic.
No close correlation was found between the powdery mildew resistance of the foliage and that of the berries.
-
The Hungarian pear germplasm as source of genetic variability for breeding programmes
7-13.Views:233TheHungarian pear collection (Pyrus communis L.) consists of 423 genotypes distributed over seven genebanks inHungary. This is one of themost extensive collections of native and cultivated pears found in Eastern Europe and includes a wide range of genotypes with small size fruit (referred to as “Miniature pears”). Based on the in situ and ex situ measures taken by governmental and other institutions for fruit tree conservation in Hungary, an overview is given on some activities regarding areas of Pyrus collection and genebanks where pears are collected and grown. Descriptions of traits of miniature pears found in Hungarian genebanks for the interest of genetic characterization and breeding are presented.