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Abstract: Botrytis cinerea Pers., the fungal plant pathogen and the causal agent of gray mould diseases in grapevine, is vastly responsible for substantial economic losses in table and wine grapes production worldwide by negatively affecting plant growth and causing the reduction of grape and wine quality. The conventional approach for pathogen control has been up to date based on synthetic fungicides with good effectiveness against pathogens but a negative impact on the environment. The growing level of harmful residues in the environment and some also detected in wines have led the European Union and many winemakers to limit the application of synthetic fungicides to earlier season. However, with a high risk of disease also late in the season, the need for other solutions is clear. Consequently, more and more research is focused on finding potential alternatives in the form of effective biological control agents. Although there are several reports of yeast’s biocontrol activity, they are up to date still poorly commercialized for such purposes. As the yeasts represent an important part of the grape microflora, competing with other microorganisms (including pathogens) for nutrients and space, we decided to examine the potential of autochthonous yeasts as "green" alternatives in fighting against phytopathogens such as B. cinerea in viticulture. With this aim, we tested the biocontrol activity of 119 different indigenous yeasts belonging to 30 different species of 17 genera against filamentous fungus B. cinerea, the causal agent of grey mould or botrytis bunch rot in grape. The yeasts were screened for putative multidimensional modes of action such as antifungal volatiles (VOC), in vitro inhibition of fungal mycelial growth, competition for nutrients, hydrolytic enzyme activities, and yeast tolerance to fungicides like copper, iprodione and cyprodinil/fludioxonil combination. With a qualitative detection of the hydrolysis activity by using screening methods based on solid medium with chitin or βD-glucosides as substrates, we found that many tested yeasts were capable of producing lytic enzymes with the ability to degrade the cell wall of phytopathogenic fungi and are potentially also able to produce VOCs via hydrolysis of grape glycosides as a result of β-glucosidase presence. Furthermore, we observed the capability of tested yeast to inhibit fungal mycelia growth on plate and assimilation of a wide variety of carbon sources; however, no siderophore producers were detected. In general, the yeasts under observation were tolerant to the tested fungicides. Their fungicide resistance can indeed be regarded as a beneficial trait for potential biofungicide agent (PBA) candidates due to open possibilities of applications and combinations within low input pest management strategies in the vineyard. Finally, a field experiment in Pinot noir and Pinot gris vineyards was designed to study different combinations of optimized canopy microclimate manipulation (CMM) techniques and potential biocontrol agent (PBA) application. In experimental conditions, the ability of PBA’s to maintain appropriate population density for disease prevention was observed. In addition, the grape and wine quality parameters were analysed to observe the possible impact of implemented biocontrol yeast on final products. The biocontrol yeast Pichia guilliermondii ZIM 624 was selected and applied in experimental vineyards based on yeast testing results. We were able to detect and confirm PBA yeast’s suitable density on grapes until harvest. In the case of early defoliation for both varieties lower grape compactness was observed together with lower yield/ plant, regardless of PBA yeast/ no yeast application. Among grape basic quality parameters, the optimized techniques showed a positive effect on sugar content. Still, unexpectedly, in the treatments with biocontrol yeasts some trends toward higher acidity were noticed in Pinot gris.
Keywords: sustainable viticulture, Botrytis cinerea, gray mould, yeasts, biocontrol, canopy microclimate manipulation, grapevine metabolite, dissertations
Published in RUNG: 07.07.2022; Views: 1771; Downloads: 132
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Abstract: Different analytical methods were tested and optimized for the determination of four biogenic amines (BA) histamine, putrescine, cadaverine and tyramine produced by grape-associated lactic acid bacteria (LAB). Autochthonous LAB were isolated from ‘Refošk’ grapes belonging to Slovenian-Italian Karst region as they represent a potential pool of beneficial LAB starter bacteria for improving the typicality and quality of wine. Six isolated strains were screened by multiplex PCR, of which four strains were positive for BA-forming genes (MKBT-49, MKBT-282, MKBT-568, MKBT-570). The production of BA was evaluated by thin-layer chromatography (TLC), high-performance liquid chromatography coupled to diode array detection (HPLC-DAD) and enzymatic method. The HPLC-DAD results showed that strain MKBT-49 (tyrdc+) had high tyramine production (386.6 ± 0.14 mg/L), which was confirmed by TLC. The ability to produce putrescine was confirmed in strain MKBT-282 by PCR, HPLC-DAD (16.4 ± 1.72 mg/L), and TLC. Histamine-producing ability was detected in strain MKBT-570, with a concentration below the limit of detection of the HPLC-DAD (<0.2 mg/L), while the other two methods were not sensitive enough for confirmation. This study shows that the production of BA can detected in native LAB and that relatively simple method such as TLC can be used effectively for the initial screening.
Keywords: grapes, wine-like matrices, biogenic amines, autochthonous lactic acid bacteria, analytical methods
Published in RUNG: 03.01.2022; Views: 1673; Downloads: 7
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Abstract: A novel method was developed for the preparation of porous hematite (α-Fe2O3) thin films. First, a solution containing iron precursor was spin-coated on fluorine-doped tin oxide substrate and later short heat-treated at 750 °C. The prepared α-Fe2O3 thin films were applied as dual-function catalyst in photoelectrochemical (PEC) water oxidation and textile dye degradation studies. For the first time, α-Fe2O3 thin films were used in efficient PEC degradation of a textile dye (Basic Blue 41 – B41) using in-situ generated reactive chlorine species. In comparison with photocatalytic and electrocatalytic approaches, the PEC technique allows faster degradation of B41 dye at an applied bias potential of 1.5 V versus reversible hydrogen electrode and under visible light illumination. In the presence of Cl− using the PEC approach the degradation of B41 reaches 99.8%. High-performance liquid chromatography coupled with UV–VIS system confirmed the degradation of B41 dye using PEC. Gas-chromatography coupled to mass spectrometry was used to study the by-products obtained during PEC degradation. Chemical oxygen demand analyses confirmed that the mineralization level of B41 is in the order of 68%. The α-Fe2O3 films developed in this study give a higher level of PEC degradation efficiency compared to other iron oxide-based systems.
Keywords: thin films, photoelectrocatalysis, kinetics, visible light, degradation, textile dye
Published in RUNG: 10.05.2021; Views: 2296; Downloads: 12
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Keywords: samoevalvacijsko poročilo
Published in RUNG: 23.12.2020; Views: 3312; Downloads: 0
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Abstract: In this work high performance liquid chromatography coupled to thermal lens spectrometry (HPLC-TLS) was applied for monitoring of vinylphenolic pyranoanthocyanins formation during the fermentation of Pinot Noir wines. Vinylphenolic pyranoanthocyanins are wine pigments, present in low concentrations, but very important for wine colour stability. Fermentation process was conducted with four different yeast strains, used as starters, either in sequential fermentation of non-Saccharomyces with S. cerevisiae yeast or as single fermentation with S. cerevisiae yeasts in order to test the applicability of developed method for monitoring of selected compounds in real wine fermentation experiments. The developed HPLC-TLS method showed higher sensitivity compared to HPLC coupled to diode array detection (DAD) technique for particular wine colour compounds. Obtained limits of detection (LODs), were 6- and 22-times lower in comparison to HPLC–DAD in gradient and isocratic elution mode, respectively, whereas limits of quantification (LOQs) 5 and 18-times lower. Lower LODs enabled earlier observation of vinylphenolic pyranoanthocyanins formation during fermentation (already at day 7) in the case of HPLC-TLS method in gradient mode, while by using HPLC–DAD in gradient elution mode the formation of vinylphenolic pyranoanthocyanins was noticed only after 12 days of fermentation.
Keywords: Thermal lens spectrometry (TLS), High performance liquid chromatography (HPLC), Pyranoanthocyanins, Wine, Yeasts
Published in RUNG: 18.06.2020; Views: 3262; Downloads: 0
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