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Carbohydrate utilization of Erwinia amylovora in vitro
31-34.Views:187Nectar 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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The effect of different carbohydrates on the multiplication of Hosta cultivars
115-117.Views:159The effect of seven concentrations of two carbohydrate sources were compared to determine the best source and the most suitable source and concentration for micropropagation of some Hosta cultivars: H. 'Gold Haze', H. 'Gold Drop' and H. 'Dew Drop'. 0, 5, 10, 20, 30, 40 and 50 g/1 sucrose or glucose were added to a MS basic medium supplemented with 3 mg/1 kinetin and 0.1 mg/1 IAA. For 'Gold Haze' 40 g/1 sucrose proved to be the best source and concentration, the proliferation ratio was 15 shoots per explant. Thirty g/1 sucrose concentration was the optimum for 'Gold Drop', the proliferation rate was 14.6 shoots per explant. In 'Dew Drop,' the best results were obtained with 30 g/1 sucrose but 40 g/l sucrose gave good results too. Both cultivars rooted well on these media. On glucose containing media, very low propagation rates were found in all concentrations and all examined cultivars.
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Physiological and biochemical evolution of peach leaf buds during dormancy course under two contrasted temperature patterns
15-19.Views:120Budbreak anomalies in temperate fruit trees grown under mild conditions have often been described. However, only few authors approached the physiological evolution of leaf buds all along the dormancy period according to the temperature pattern. The aim of this study was to characterize the evolution of peach leaf bud dormancy through some physiological and biochemical parameters under temperate winter conditions and under total cold deprivation after the endodormancy onset. Two treatments were applied in peach trees cv. Redhaven: (i) Regular Chilling Amounts — RCA and (ii) Total Chilling Deprivation — TCD. Buds were sampled periodically from different parts of the stem (terminal, medium and basal ones). We recorded the evolution of: carbohydrate concentrations (glucose, fructose, sucrose, sorbitol and starch), respiration rate, water contents and energy metabolism (ATP and ADP ratio). The dynamics of these parameters were compared and correlated with dormancy evolution ("one node cuttings" test) and budbreak patterns in plank:. The endodormancy intensity of terminal buds was significantly lower than those of median and basal buds in early October. Under RCA treatment, this gradient faded and the bud endodormancy release was completed at the same time in all positions along the stem. Thereafter, the "cuttings" test indicated that terminal buds grew slightly faster than median and basal buds, and, consistently, budbreak in planta started with the terminals buds, followed by the medians and then by the basal ones. The carbohydrate contents showed a transitory change only when the buds began to grow after the endodormancy was released under RCA. Respiration, water content and ATP/ADP changed dynamics only under RCA and only after the end of the endodormancy (their respective changes were very parallel). The dynamics of none of the tested parameters could be related with the endodormancy dynamics, but respiration, water content and ATP/ADP could be consistent markers of the actual bud growth before bud break (in this respect, ATP/ADP could not show differences between the terminal and axillary buds while respiration and water content could).
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Production of transgenic carnation with a heterologous 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase bifunctional enzyme cDNA
75-79.Views:145Transgenic carnations were produced with a modified mammalian bifunctional enzyme cDNA coding 6-phosphofructo-2- kinaseffructose 2,6-bisphosphatase. Relative activity of this enzyme determines the fructose 2,6-bisphosphate (fru 2,6-P2) cytosolic concentration. This metabolite — as a signal molecule — is one of the carbohydrate metabolism regulators. The regenerated Dianthus chinensis and Dianthus caryophyllus shoots were selected on MS basal medium containing 150 mg/1 kanamycin. Transgene integration was proven by PCR analysis with cDNA specific primers followed by Southern hybridization of DNA isolated from selected green shoots, which survived on kanamycin containing medium, so 3 D. chinensis and 20 D. caryophyllus transgenic plants were produced. Transgene expression were examined by RT-PCR. Transformed and control plants were potted in glasshouse to evaluate the effect of modified fru 2,6-P2 on development, growth and carbohydrate metabolism.
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The effect of different berries in human nutrition
147-151.Views:225The 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.