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Yeasts (52, 51, 69, and 04) were received from Biotechnology Center of Karaj and Penicillium expansum Isolates P11 and P12 isolated from Golden Delicious. Isolates were evaluated as a potential biological control agents of apple blue mold caused by P. expansum. Dual culture, cell free metabolites and volatile test were used in vitro assay. All tested of yeast isolates inhibited growth of P. expansum. The inhibition varied among isolates of yeasts and ranged from 19.81% to 40.73%, in dual culture, from 43.16% to 66.44% in volatile metabolite and from 22.16% to 50.23% in cell free metabolite test. Apple fruit wounds were inoculated with 40 μl of yeast cell suspension (107cell/ml) followed 48 h later by P. expansum (105 conidia/ ml). The apples were then incubated at 25°C. Four isolates of Saccharomyces cerevisiae reduced, the decay area from 13.46 to 24.92 cm2 compared to 32.18 cm2 in control after incubation for 14 days at 25°C. At 5°C, the lesion size ranged from 13.58 to 24.68 cm2 for the antagonist treatments compared to 22 cm2 for the control treatments after 32 days. The isolate 69 of S. cervisiae was the most effective isolate at both tempetures in this assay and could be one of important new biological control agents for apple blue mold.
Effect of propionic acid, adipic acid, lactic acid, and salts of these acids, on the growth of Penicillium expansum and the dynamics of patulin formation by this strain, was investigated. Toxin level was assayed by TLC and HPLC methods. Minimal concentration of preservatives inhibiting completely microfungus growth and patulin biosynthesis, was determined. Propionic acid and adipic acid at the concentration of 0.05 mol/dm3, and sodium propionate at the concentration of 0.2 mol/dm3 were found to inhibit completely the gain in weight of Penicillium expansum mycelium and the toxin formation. Calcium lactate at the concentration upwards of 0.25 mol/dm3, was noted to counteract the formation of patulin, and - at 0.5 mol/dm3 - to inhibit completely the fungus growth. Lactic acid was least active of all preservatives under test. At the concentration up. to 0.5 mol/dm3, this agent inhibited the strain growth only by 33%, and patulin biosynthesis - by 85%.
The biocontrol activity of two isolates of Trichoderma virens against blue mould of apple fruits caused by Penicillium expansum and their ability to induce biochemical defense responses in apple tissue were investigated. Apple fruit (Malus domestica) wounds were inoculated with 20μl antagonist suspension (107 conidia/ml) of T. virens and 4 h later with 20 μl of conidial suspension of P. expansum (105 conidia/ml). The apples were then incubated at 20°C for 8 days. Lesion diameters were measured 4 and 8 days after inoculation with the pathogen. Two isolates of T. virens were effective in controlling decay of apple fruits caused by P. expansum. Six days after treatment peroxidase activity increased by more than three-fold in apple treated fruit in combination of antagonist and pathogen, in comparison with in wounded, non-inoculated control tissue. Catalase (CAT) activity increased in inoculated fruits in combination of T. virens and P. expansum in comparison with healthy control at all days and maximum activity level was noted at 6 days after inoculation. The results indicated that T6 and T8 isolates of T. virens caused the increase in β-1,3-glucanase activity that reached maximum levels 4 and 6 days after inoculation with pathogen, respectively. The increase in β-1,3-glucanase activity was triggered by wounding although the level of increase was markedly lower than detected in treated fruits. Phenolic compounds accumulation showed the highest levels 2-4 days after inoculation and then decreased. The ability of T. virens to increase the activity of peroxidase, catalase, β-1,3-glucanase and levels of phenolic compounds may be one of mechanisms responsible for its biocontrol activity.
Metodami chromatografii cienkowarstwowej z detekcją w UV i wysokociśnieniowej chromatografii cieczowej przebadano 50 próbek przetworów pomidorowych na obecność patuliny.
It was found that pathogens of clamp rot in sugar beet cultivated in Belarus are the following fungal species: Fusarium redolens, Fusarium culmorum, Penicillium expansum, Botrytis cinerea, Gliocladium catenulatum, Sclerotinia sclerotiorum, Alternaria tenuis, Phoma betae. G. catenulatum, F. redolens, F. culmorum, P. expansum, S. sclerotiorum proved more aggressive, P. betae, A. tenuis - less aggressive. Experiments in vitro and in vivo have revealed that strain Bacillus subtilis BIM B-439 D - main component of new biopesticide „Betaprotectin” was characterized by maximal antagonistic activity against clamp rot pathogens. According to small-scale experimental data, „Betaprotectin” biological efficiency reached 36.9-39.2%. Technology of manufacturing new biopreparation comprises a procedure of direct regulation of bacterial antagonistic activity under the impact of physical stress factors - application of spore inoculation material exposed to thermal treatment raised antimicrobial activity of B. subtilis BIM B-439 D liquid culture by 1.2-1.4 times. Deleterious effect of pathogens during storage of root crops in large-scale clamps and piles fell 2-fold upon „Betaprotectin” treatment as compared to the control. Spread of clamp rot slowed down likewise. Biological efficiency of biopesticide application ranged from 17.5% to 39.6%, economic efficiency - from 5.1% to 35.8%. Root crop preservation rate reached 94.1%.
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