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Carbohydrate utilization of Erwinia amylovora in vitro
31-34.Views:188Nectar is a multi-component aqueous solution that promotes bacterial multiplication. The concentration of nectar in plant flowers is not stable since it is under the influence of environmental conditions, especially free moisture and relative humidity. Experiments were conducted with "artificial nectar" and directed along two lines: (1) determination of the optimal concentrations of carbohydrates for the growth of E. amylovora development (2) consumption of different carbohydrates besides basic sugars.
Solutions of "artificial nectar" were prepared in different compositions by changing the dominance of basic sugars (fructose — glucose —sucrose) in proportions of 2:1:1, 1:2:1, 1:1:2 and between concentrations of 10-0.6% (diluted with Basal minimum broth) in order to determine optimal conditions for the development of E. amylovora.
At a basic sugar concentration of 10% bacterial multiplication started and continued until I log degree (from 106 to 107 cfu/ml). At concentrations of 5% and 2,5 % cells developed with nearly the same kinetics (from 106 to 8x107 cfu/ml and from 106 to 9x107 cfu/ml, respectively). Multiplication was more pronounced and nearly the same at concentrations of 1.2 % and 0.6 % (from106 to 2x108 cfu/ml). At a basic sugar concentration 30% total sugars bacterial multiplication did not occur, while at 20 % it was negligible, not measurable photometrically.
At minimal concentrations of F, G, S (between 1-0.1 %) bacterial cells were still able to multiply, producing organic acids from sugars.
Our study showed that E. amylovora requires only a small amount of sugars (0.1%) for multiplication (acid production) while high concentrations inhibit multiplication. There was a negative correlation between sugar content and cell density. The optimal range of sugar concentration was at about 1%.
Effect of "less frequent carbohydrates" to E. amylovora multiplication was also determined using the API 50 CH strip. We could provide information on utilization of 39 carbohydrates by the bacterium at different categories as follows: Not utilized-, Slowly and weakly utilized-, Slowly and completely utilized-, Quickly and completely utilized carbohydrates. We suppose that carbohydrates that belong to the latter two groups could play an important role as nectar components in promoting E. amylovora multiplication in the blossoms of pome fruit trees.
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Reactions of different plant organs of pear cultivars to Erwinia amylovora infection
17-21.Views:151Research project has been initiated in 1999 with the aim of evaluating the degree of susceptibility/resistance of pear cultivars grown in Hungary to fire blight disease caused by Erwinia amylovora. The recently selected promising cultivars were also examined. Inoculation experiments were conducted in controlled greenhouse conditions because of quarantine regulations in Hungary. Following the disease process, development of symptoms of plant organs (shoots, flower parts, fruits) was observed. Suspension of two E. amylovora strains (Ea 21, Ea 23) isolated from pear was used in a mixture (5x108 cells x m1-1) for the inoculation. Twenty-six pear cultivars were examined and grouped into four categories: low susceptibility, moderately susceptible, susceptible and very susceptible. Most of the cultivars were susceptible or very susceptible while some promising 'Eldorado', 'Harrow Delight' and `Hosui' showed low susceptibility.
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Fire blight (Erwinia amylovora) susceptibility of old Hungarian apple cultivars
35-38.Views:179The aim of the Hungarian apple breeding program started in the Department of Fruit Science was to find resistant apple cultivars against major diseases (scab, powdery mildew, fire blight). The outbreak of fire blight (Erwinia amylovora) in 1996 motivated us to search new resistant sources principally from old traditional apple cultivars. First of all, cultivars have been gathered since 1997 from Carpathia (Visk) and evaluated between 2001 and 2003. In this recent study evaluation of resistance of old Hungarian genotypes to fire blight collected from the English National Fruit Collection (Brogdale, Faversham) is presented.
13 old Hungarian apple cultivars in 2002, and 38 genotypes in 2005 have been evaluated. We used `Idared' and 'Jonathan M41' as susceptible controls and 'Liberty' and 'Remo' as resistant ones. Shoots of two-year-old potted plants were inoculated with a mixture of virulent E. amylovora isolates (Ea2, Ea60, Ea67) at a concentration of 5 x 108 cells/ml. Resistance of apple cultivars was evaluated weekly, four times after inoculation by disease severity of symptoms. Numbers of bacterial colonies in 1 cm length shoot were determined in the fourth week after infection.
8 cultivars in 2002 and 9 cultivars in 2005 displayed notable resistance to fire blight based on one-year data. Based on the coincident data of both years, out of the cultivars collected also from Carpathia ‘Pónyik', and `Sikulai' were found to be resistant and gene sources additional old Hungarian valuable apple genotypes could be selected: `Szabadkai szercsika' and `Tordai piros !davit'. The cultivar `Szemes alma' originated from Visk has proved to be recurrently resistant.
The number of bacterial cells in shoots of the investigated cultivars correlated with the severity of symptoms. With this method, which was introduced by us earlier, we can screen cultivars displaying weak visible symptoms, which cannot be proposed as a source of resistance because of their latent infection.
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Testing the virulence of some Hungarian Erwinia amylovora strains on in vitro cultured apple rootstocks
52-55.Views:206A useful method was improved to test and to evaluate the susceptibility of plants to fire blight and the virulence of E. amylovora strains. Six Hungarian strains from different host plants were tested on in vitro cultured apple rootstocks. Disease rating was used for the characterization of the process of disease development. The different strains had different capacity to cause disease, mainly in the first period of incubation. There were significant differences between the virulence of the strains.
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Susceptibility of some traditional pear cultivars of Hungarian and foreign origin to the pathogenic bacterium Erwinia amylovora
41-45.Views:158Fire blight, a disease caused by the bacterium Erwinia amylovora (Burrill) Winslow et al., has been causing serious damage in Hungarian pear plantations since 1996. A prospective control measure could be the use of resistant cultivars. For that purpose ten pear cultivars have been tested under laboratory and greenhouse conditions for resistance to Erwinia amylovora strains collected in Hungary. Six of these cultivars are Hungarian ones of unknown origin, while four are traditional old varieties. Resistant cultivars should serve as germplasm for future breeding. Inoculations were made with a mixture of different pear isolates of the bacteria collected from various growing regions of Hungary (Ea 21, 23), at a density of 5x108 cells/ml. Susceptibility/resistance has been assessed on the basis of intensity of blight symptoms observed on shoots, flower parts and fruits. Cultivars were assigned to three susceptibility groups (low, moderate and high). Complete resistance was not found among the cultivars tested. The highest level of resistance was found in cultivar 'Kieffer', while the other cultivars displayed either moderate or high susceptibility to infection.
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Knot formation by Pseudomonas syringae subsp. savastanoi on the in vitro shoots of Sorbus redliana
59-62.Views:183Two strains of Pseudomonas syringae subsp. savastanoi were isolated from Forsythia sp. and Nerium oleander in Hungary in 1997. The effects of growth regulators produced by the bacteria were studied in different experiments. The strains were co-cultured with Sorbus redliana in vitro shoots without being in contact with the plant on solid media. Further culture filtrates in different concentrations were added to the culture medium. The growth regulators presented in the agar caused knot formation on the shoots and on the leaves in both kinds of culture. There were significant differences in the cultural and physiological characters, auxin and cytokinin activity of the strains of different origin.
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Effect of Erwinia amylovora infection on peroxidase enzyme activity in resistant apple cultivars
37-39.Views:159Two apple cultivars that display enhanced resistance to fire blight (causal agent: Erwinia amylovora) were selected. The aim of the present study was to characterize the peroxidase (POD) enzyme activity of `Szemes alma' (a historical cultivar) and MR-03, (a Hungarian multiresistant hybrid of 'Prima') and compare them to susceptible 'Jonathan M 40' and resistant 'Remo' controls. Peroxides enzyme activity during E. amylovora infections was investigated in artificially infected apple shoots. Increases in enzyme activities were observed in a `Jonathan M40' and in 'Remo', MR-03, `Szemes alma' cultivars. There was a consistent relationship between total enzyme activity and fire blight disease severity. High activity of the peroxidase was positively correlated with the degree of resistance to fire blight. A general hypothesis that POD activity is related to fire blight susceptibility/resistance is supported by our results.
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The effect of modified bacterial virulence to host-pathogen relationship (Phaseolus vulgaris L. Pseudomonas savastanoi pv. phaseolicola)
53-56.Views:221The Pseudomonas savastanoi pv. phaseolicola is one of the most expressive biogen stressors of the bean (Phaseolus vulgaris L.) in Hungary. The chemical and agrotechnological defence is inefficient, so breeding is the only workable way. The conventional cultivars are susceptible to PS while most of the new industrial varieties have genetic resistance to the pathogen. The genetic background of resistance is, however, a complex system in the bean. Leaf resistance is a monogenic system, but this gene is not expressed in juvenile stage of the host. The pathogen species can be divided into different races. After inoculation with virulent strains, typical symptoms appeared on the leaves. To understand the details of host-pathogen relationships, there were carried out experiments using bacterial strains with altered virulence. Six transposon mutants of the PS were tested. Our main objective was to test these modified bacterial strains on bean cultivars of known genetic background. First we analysed the symptoms, and then the correlation between the symptoms and the multiplication of mutant bacteria. Three cultivars (Cherokee, Inka and Főnix) were tested.
The infection by the virulent PS isolate produced typical symptoms on the three cultivars tested. Mutant bacteria (except strain 756) did not cause any significant symptoms on the hosts. The mutant 756 induced visible symptoms on the cultivars Cherokee and Inka. On Cherokee there were small watersoaked lesions, and HR (hypersensitivity reaction) was detected on Inka, but this was restricted to some cells only (mikro HR). The rate of multiplication of the wild type strain was much higher than the multiplication of the mutants. Bacteria were detected in the cotyledons and primordial leaf, but there is not any substantial number of bacteria in leaves, except for strains 757, 1212 and 1213. The rate of multiplication of strain 756 was intermediate. These, and other experiments can help to understand the genetic background of resistance and the host-pathogen relationship in the Pseudomonas-bean pathosystem.
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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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First selections of the Hungarian apple breeding program for multiple resistance
9-13.Views:169The aim of the first Hungarian apple breeding program for multiple resistance started in the beginning of the nineties is to widen Hungarian apple assortment by good quality, resistant apple cultivars with excellent productivity and ecological capability to the most important fruit growing areas in Hungary. In the first years of seedling production we made early selection for susceptibility to apple scab in greenhouse. Alter this, field observation of susceptibility to powdery mildew, scab and canker and a yearly negative selection was carried on. From 1997, fruit quality was evaluated as well, and from 2001 the resistance of shoots to Erwinia ainylovora (Burrill/Winslow et al.) was examined using inoculations in greenhouse conditions. From the progenies of crosses in 1992 and 1993, six candidates were announced to national recognition out of hybrids examined for more than a decade. Descriptions of these selections from 'Prima' progenies and the most important data of their resistance, growing habit, morphological characteristics and fruit quality are shown in this article.