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High-velocity microprojectile mediated DNA delivery into Phaseolus vulgaris callus cells
99-102.Views:129We 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.
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Six promising selections from the Hungarian apple breeding program for multiple resistance
23-28.Views:168An 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.
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Self-incompatibility alleles in Esatern European and Asian almond (Prunus dulcis) genotypes: a preliminary study
23-26.Views:211Almond [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.
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Factors affecting apricot fruit antioxidant capacity and mineral element contents
95-99.Views:235Several 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.
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Somatic embriogenesis and regeneration of Vitis sp.
41-45.Views:157Somatic embriogenesis and regeneration of Vitis sp.
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In vitro regeneration from cotyledons of watermelon
96-98.Views:172Cotyledonary 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.
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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:307In 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.
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Researches concerning the selection of some nectarine varieties as genitors for the breeding program in Romania
44-46.Views:114Researches concerning the selection of some nectarine varieties as genitors for the breeding program in Romania
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Molecular characterization of apricot (Prunus armeniaca L.) cultivars using cross species SSR amplification with peach primers
53-57.Views:241Apricot takes an important place in Hungarian fruit production. Considering morphological characteristics of apricots it was concluded that the genetics background of European cultivars is very limited. Molecular markers and their use for genotyping have revolutionized the identification of cultivars. In a classic apricot breeding program, it is important to be able to establish unique DNA profiles of selections to identify them unambiguously and to determine their genetic relationship. Presently SSR is far the most frequently performed technique for genetic diversity studies. In this study there were used peach and apricot primer pairs from four different sources in order to examine microsatellite polymorphism among cultivars and investigate relationships among them. The possibility of cross species amplification among different Prunus species using SSR primers allowed us to use primers developed in peach to study genetic diversity in apricot. In this work, 90% of the primers used were able to amplify SSRs in apricot and more than half of them were polymorphic. With the 10 primer pairs utilized were proven to be sufficient to set unique fingerprint for several cultivars studied. The obtained dendrogram classified of the 45 cultivars included in this study into two major groups and several subgroups.
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Fire blight (Erwinia amylovora) resistance in apple varieties associated with molecular markers
53-57.Views:262The invasive bacterial disease fire blight, caused by Erwinia amylovora has the potential to destroy fruit tree orchards all over Europe. Effective plant protection methods are lacking in many countries, highlighting the increasing importance placed on identification of germplasm with heritable disease resistance. Recent l y. a promising QTL (quantitative trait locus) was identified on linkage group 7 in the apple cultivar 'Fiesta· which is derived from ·cox's Orange Pippin' . I n the present study, 144 Swedish and foreign apple cultivars were analysed with the SCAR markers AE I 0-375 and GE-8019. which flank-. this QTL. Twenty-nine of the analysed cultivars had both markers. 78 had either AE I 0-375 or GE-8019, and 37 cultivars did not carry an) of the two markers. Seventeen cultivars. 7 with both markers and I 0 not having either of the two markers, were then inocoluted with the bacterium in a 4uaran1i ne greenhouse test. Cultivars carrying both DNA markers were significantly less susceptible than cultivars lacking the markers, P<0.001. Cultivars that were most resistant had both markers and had 'Cox· in their pedigree. Unrelated cultivars with the markers may still lack the QTL.
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Examinations of 600-year-old seeds by means of archaeobotanical and genetical methods
79-80.Views:162About 600-year-old plant seeds were discovered in a well of a mediaeval cellar in the course of an excavation in Budapest. After the archaeobotanic purification seed of 16 species were found in large quantities. Seeds preserved in the best state were selected from each group. The existence of endosperm was analysed in these subfossils, which turned to be successful mainly in the case of grapes (Vitis vinifera) and cornels (Cornus mas). Seeds of these two species contained the most endosperm and remains of the embryo. DNA was extracted with the help of DNEasy Plant Mini Kit and analysed by RAPD-PCR method. The amplification of DNA extracted from cornel seeds resulted in detecting a 1500 by fragment, which makes the comparison of these samples with present-day cornels possible.
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Co-transformation of bean callus using high-velocity microprojectiles- mediated DNA transfer
76-78.Views:133We 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.
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Bean tissue culture and genetic transformation with Agrobacterium
32-35.Views:154In 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.
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DNA-based determination of suitable pollinating cultivars for the pear cultivar 'Carola' (Pyrus communis)
15-19.Views:141Pollen-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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The use of SSR markers in family Rosaceae
29-32.Views:179The 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.
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Attempting Regeneration from Cultured Cotyledons and Plant Regeneration from Cotyledonary Nodes in Common Bean (Phaseolus vulgaris L.)
57-60.Views:167Dry 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.
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Callus induction on standard type Cymbidium cultivars
108-110.Views:166Tissue 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.
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Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple
7-10.Views:386Quince (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.
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Microsatellite based identification of grapevine cultivars traditional in Hungary and in the Carpathian Basin
71-73.Views:149Grapevine 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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Preliminary characterization of the self-incompatibility genotypes of European plum (Prunus domestica L.) cultivars
23-26.Views:238European 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.
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The effect of different berries in human nutrition
147-151.Views:227The 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.
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Historical apple cultivars that display high level of resistance to fire blight
19-23Views:142Following the first outbreak of fire blight caused by the bacterium Erwinia amylovora (Burrill) Winslow et al. in Hungary, we have started research with the aim to screen domestic gene sources, mostly historical Hungarian apple cultivars, for disease resistance as part of an apple breeding program for multiple resistance. The present study was conducted with the aim to choose the most tolerant historical apple cultivars among 25 selected cultivars by screening their behavior towards fire blight under controlled conditions. Six cultivars were shown to be the most disease tolerant: `Batul', 'London pepin', Nemes Sóvári Alma', `Sikulai', `Szemes alma', Wilmos renet'. We evaluated these cultivars by investigating both morphological- characteristics under original environmental circumstances and fruit quality. The cultivars had a remarkable degree of fire blight resistance compared to the control cultivars. These were not competitive with the commercially grown 'Jonathan M40' during cultivar tests but on the basis of certain characteristics they could serve as genetic sources for breeding new varieties.
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Determination of the cold tolerance off sour cherry cultivars with frost treatments in climatic chamber
49-54.Views:268Nowadays, sour cherry buds can be seriously damaged by spring and winter frosts. Unlike other fruit species threatened by high frost damage, sour cherry cultivars have not been assessed for frost tolerance. The aim of .our survey was to establish the relative cold tolerance of the Hungarian cultivars after treatment in a climatic chamber, and to optimize the methodology formerly elaborated for the frost treatment of apricot. Fourteen cultivars of Hungarian sour cherry (Prunus cerasus) were used in the experiments, which spanned the winters of 2005/2006 and 2006/2007. Our data were used to rank cultivars in two groups according to their levels of cold resistance. We also recommend critical temperatures and treatment times for the testing of sour cherry cultivar resistance to cold in climatic chambers.
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Pomology of gene resources bearing resistance to Erwinia amylovora
29-33.Views:139The ecological farming and an increasing demand for healthy fruits free from chemical residuals necessitated the reevaluation of the existing cultivars and breeding suitable cultivars for the new requirements. We have gathered the old genotypes known in the Carpathian Basin, and we have selected the most suitable ones according to fruit quality and resistance to diseases in the last fifteen years. The main aim was the selection of resistant genotypes against fire blight and powdery mildew as gene sources for our breeding program. On the other hand, we consider the best quality traditional cultivars as extensive landscape elements (wayside tree rows or extensive squash producing plantations).
During the pathological evaluation 'Szemes alma', 'Pónyik', 'Sikulai', 'Tordai piros 'cavil' and 'Szabadkai szercsika' cultivars proved to be good resistance sources (due to their outstanding resistance to fire blight). The latter four cultivars are not susceptible even to fungal diseases, and their quality is also acceptable so those are commendable for ecological farming or for renewing the sparse orchards. According to fruit quality and low susceptibility to diseases 'Batul' and 'Vilmos renet' can also be offered for extensive organic production.
We have made original or renewed pomological descriptions of the selected cultivars, and we also publish their characters according to the terms of UPOV TG/14/8 based on to our evaluations.
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Reactions of some cucumber (Cucumis sativus L.) lines and hybrids to zucchini yellow mosaic virus (ZYMV) and selection of tolerant breeding lines
66-68.Views:173In the past years zucchini yellow mosaic virus (ZYMV) has been appeared as a new pathogen of cucurbitaceous plants in Hungary. It caused severe disease outbreaks on the fields of pickling cucumber hybrids which are highly tolerant to cucumber mosaic virus (CMV). Preliminary inoculation tests have showed that all of the important pickling cucumber hybrids produced in Hungary were susceptible to ZYMV-P. However, a selection of Chinese Long and the breeding line I- KS 10C proved to be highly tolerant. The Fl plants of the crosses between the tolerant lines and the susceptible genotype H I were susceptible to ZYMV-P. F2 populations segregated for susceptible and tolerant individuals at a ratio of 3:1. The results strongly suggest that the resistance to ZYMV in our sources is controlled by a single recessive gene. Inoculations of the ZYMV and CMV susceptible cultivar Budai csemege and the CMV tolerant hybrid Perez Fl with the complex of ZYMV+CMV resulted extremely severe symptoms (strong mosaic and necrotic spotting of the leaves) on both cultivars. On the Chinese Long line, which is tolerant to ZYMV and CMV, respectively, the complex of the two viruses caused mild symptoms. The results show a synergistic pathological effect of ZYMV and CMV on differrent cucumber genotypes. More detailed studies on the interactions among the plant genotypes, viruses and virus strains are needed to develop cucumber hybrids that are highly resistant to the ZYMV+CMV complex.