Macrowine 2021
IVES 9 IVES Conference Series 9 Ochratoxin a degradation by Botrytis cinerea laccase: effect of oenological factors and redox mediators

Ochratoxin a degradation by Botrytis cinerea laccase: effect of oenological factors and redox mediators

Abstract

AIM: This study evaluates the effect of different oenological factors and natural mediators on the degradation of Ochratoxin A (OTA) using Botrytis cinerea laccase. Because of its risk to human health, different detoxification techniques have been developed in various kinds of foodstuffs. The use of fungal or bacterial laccases is a biological method to decrease the OTA concentration [1, 2]. Laccases can oxidize a wide range of substrates, some of which cannot be oxidized directly by these enzymes and require the use of redox mediators [3]. Due to this, several natural mediators present in wine and different SO2 and ethanol concentrations were tested in the current work.

METHODS: The ability of laccase to degrade OTA was studied by incubation of the enzyme in acetate buffer pH 4.0 and model wine, with OTA and mediators at 28 ºC during 24 h. To determine the impact of SO2 and ethanol on the OTA degradation caused by laccase, different concentrations of SO2 (10, 20 and 30 mg/L) and ethanol (5, 10 and 15% v/v) were used. The quantification of this mycotoxin was carried out in a HPLC-QTOF-MS system.

RESULTS: Under these conditions, OTA cannot be oxidized directly by laccase from Botrytis cinerea and the use of redox mediators is required. Among natural mediators tested, (-)-epicatechin and (+)-catechin were the phenolic compounds with higher impact on the biodegradation of this mycotoxin, achieving a decrease of OTA concentration over 50%. The degradation of OTA was completely inhibed by 30 mg of SO2/L,  while 20 mg of SO2/L reduced lacasse activity by a half and 10 mg of SO2/L hardly caused any effect on the biodegradation of this mycotoxin. A concentration of 15% of ethanol led to a 50% reduction in the activity of laccase over OTA. 

CONCLUSIONS

These preliminary results may be a first step in finding biological alternative strategies to eliminate undesirable substances such as mycotoxins (OTA) present in wine.

DOI:

Publication date: September 14, 2021

Issue: Macrowine 2021

Type: Article

Authors

osé Pérez-Navarro

Regional Institute for Applied Scientific Research (IRICA), University of Castilla-La Mancha, Av. Camilo José Cela 10, 13071 Ciudad Real, Spain. Higher Technical School of Agronomic Engineering, University of Castilla-La Mancha, Ronda de Calatrava 7, 13071 Ciudad Real, Spain.,Tania, PANIAGUA MARTÍNEZ, Faculty of Chemical Sciences and Technologies, University of Castilla-La Mancha, Av. Camilo José Cela, 10, 13071 Ciudad Real, Spain. Pol, GIMÉNEZ, Faculty of Oenology, University of Rovira I Virgili, C/Marcel.li Domingo s/n, 43007 Tarragona, Spain. Joan Miquel, CANALS, Faculty of Oenology, University of Rovira I Virgili, C/Marcel.li Domingo s/n, 43007 Tarragona, Spain. Fernando, ZAMORA, Faculty of Oenology, University of Rovira I Virgili, C/Marcel.li Domingo s/n, 43007 Tarragona, Spain. Sergio, GÓMEZ-ALONSO, Regional Institute for Applied Scientific Research (IRICA), University of Castilla-La Mancha, Av. Camilo José Cela 10, 13071 Ciudad Real, Spain. Faculty of Chemical Sciences and Technologies, University of Castilla-La Mancha, Av. Camilo José Cela, 10, 13071 Ciudad Real, Spain.

Contact the author

Keywords

mycotoxin, enzyme, biodetoxification, fungi, SO2, ethanol

Citation

Related articles…

Water deficit differentially impacts the performances and the accumulation of grape metabolites of new varieties tolerant to fungi

The use of resistant varieties is a long-term but promising solution to reduce chemical input in viticulture. Several important breeding programs in Europe and abroad are now releasing a range of new hybrids performing well regarding fungi susceptibility and producing good quality wines. Unfortunately, insufficient attention is paid by the breeders to the adaptation of these varieties to climatic changes, notably to the increased climatic demand and water deficit (WD). Thus, prior to the adoption of such varieties by the wine industry in Mediterranean regions, there is a need to consider their suitability to WD. This study aimed to characterize the different drought-strategies adopted by 6 new resistant varieties selected by INRAE in comparison to Syrah. To allow the assessment of long-term impacts of WD, field-grown vines were exposed to contrasted WD from 2018 to 2021 under a semi-arid Mediterranean climate. A gradient of WD was applied in the field and controlled through plant measurements at the single plant level. Grape development was non-destructively monitored to determine the arrest of berry phloem unloading. The impacts of WD on berry composition, including water, primary metabolites (sugars, organic acids), secondary metabolites (anthocyanins, thiols precursors) and main cations contents, were assessed at this specific stage. Results showed different varietal responses during the year and inter-annual acclimation in terms of plant water use efficiency, biomass accumulation, as well as yield components and berry composition. WD differentially reduced the accumulation of primary metabolites at plant and berry levels, but it little changed their concentrations in the fruits at the ripe stage. Moreover, WD differentially impacted the accumulation of secondary metabolites and major cations between the varieties. In the talk, we’ll present the main results regarding the WD impacts on fruit metabolites and enlarge the reflection about the practical assessment of the grapevine acclimation to WD.

Impact of long term agroecological and conventional practices on subsurface soil microbiota in Macabeu and Xarel·lo vineyards

There is a growing trend on the transition from conventional to agroecological management of vineyards. However, the impact of practices, such as reduced-tillage, organic fertilization and cover crops, is not well-understood regarding the soil microbial diversity, and its relationship with the soil physicochemical properties in the subsurface depth near the rooting zone. Soil bacterial diversity is an important contributor towards plant health, productivity and response to environmental stresses. A field experiment was conducted by sampling subsurface soil bacterial community (NGS and qPCR) near to the root zone of Macabeu and Xarel·lo vineyards, located at the Penedes. 3 organic (ECO) and 3 conventional (CON) vineyards, with more than 10 years of respective management were sampled (n=5 each plot). ECO practices did not affect bacterial and fungal abundance but increased significantly the ammonium oxidizing bacteria and alpha-diversity (Inv.Simpson). Interestingly beta-diversity was significantly affected by the management strategy. ANOSIM-tests revealed a significative effect of the management (ecological vs conventional) and plot, on the soil microbial structure (ASV abundance). Main phyla depicted were Proteobacteria, Actinobacteria and Acidobacteria, whose relative abundances were not affected by the management. EdgeR assay revealed a significant increase of Cyanobacteria and decrease of Gemmatimonadetes and Firmicutes phyla in ECO. Interestingly, the grapevine variety was not correlated with the soil microbial community structure. Mantel-test revealed an important correlation (Spearman) of some physicochemical parameters with the soil microbiota structure, in order of importance: texture, EC, pH Ca/Mg, Mg/P, K+, Mg2+, Ca2+, SO42-, and OM. N-NH4 and NTK, which were higher in the ECO managed soils, did not correlated significantly with the soil microbiome population. The results revealed the importance of combining a deep physicochemical characterization of each replicate with the microbial diversity assessment to gain better insights on the relationship between soil microbiome and vineyard management.

Spatial determination of areas in the Western Balkans region favorable for organic production

In problematic conditions for production of grapes and wine caused by the COVID-19 pandemic and the resulting occurrence of wine surpluses, producers are increasingly turning to the innovative viticulture and winemaking of products that are more appealing to the market and the consumers. On the other hand, consumption of the food safety or organic products, and therefore of organic grapes and wine, is increasingly common in the world, in particular in Europe. The Regional Rural Development Standing Working Group (SWG RRD), as a regional intergovernmental organization gathers actors in the viticulture and winemaking sector from states and territories of the Western Balkans (South-East Europe) in the Expert Working Group for Wine, with the aim of improving viticulture and winemaking in this region through joint activities. In accordance with the aforementioned, the SWG RRD is working on advancing organic production of grapes and wine, and on recognition of specificities of the terroir of wine-growing areas in Western Balkans. In addition, as part of the project “Facilitation of Exchange and Advice on Wine Regulations in Western Balkan Countries” helmed by the German Federal Ministry of Food and Agriculture, in addition to harmonization of relevant legislation with EU regulations, efforts are being invested towards recognition of organic wines. Within activities and project implemented by this organization, expert analyses and scientific research of the terroir of Western Balkans were carried out, and some of the results are presented in this paper.

Current climate change in the Oplenac wine-growing district (Serbia)

Serbian autochthonous vine varieties Smederevka (for white wines) and Prokupac (for rosé and red wines) are the primary representatives of typical characteristics of wines and terroir of numerous wine-growing areas in Serbia. In the past, these varieties were the leading vine varieties, however, as the result of globalization of winemaking and the trend of consumption of wines from widely prevalent vine varieties, they were replaced by introduced international varieties. Smederevka and Prokupac vine varieties are characterized by later time of grape ripening, and relative sensitivity to low temperatures. Climate conditions can be a restrictive factor for production of high-quality grapes and wine and for the spatial spreading of these varieties in hilly continental wine-growing areas.
This paper focuses on the spatial analysis of changes of main climate parameters, in particular, analysis of viticultural bioclimatic indices that were determined for the purposes of viticulture zoning of wine-growing areas in the period 1961-2010, and those same parameters determined for the current, that is, referential climate period (1988-2017). Results of the research, that is, analysis of climate changes indicate that the majority of examined climate parameters in the Oplenac wine-growing district improved from the perspective of Smederevka and Prokupac vine varieties. These studies of climate conditions indicate that changes of analyzed climate parameters, that is, bioclimatic indices will be favorable for cultivation of varieties with later grape ripening times and those more sensitive to low temperatures, such as the autochthonous vine varieties Smederevka and Prokupac, therefore, it is recommended to producers to more actively plant vineyards with these varieties in the territory of the Oplenac wine-growing district.

Mechanisms involved in the heating of the environment by the aerodynamic action of a wind machine to protect a vineyard against spring frost

One of the main consequences of global warming is the rise of the mean temperature. Thus, the heat summation by the plants begins sooner in the early spring, and by cumulating growing degree-days, phenological development tends to happen earlier. However, spring frost is still a recurrent phenomenon causing serious damages to buds and therefore, threatening the harvests of the winegrowers. The wind machine is a solution to protect fruit crops against spring frost that is increasingly used. It is composed of a 10-m mast with a blowing fan at its peak. By tapping into the strength of the nocturnal thermal inversion, it sweeps the crop by propelling warm air above to the ground. Thus, stratification is momentarily suppressed. Furthermore, the continuous action of the machine, alone or in synergy, or the addition of a heater allow the bud to be bathed in a warmer environment. Also, the punctual action of the tower’s warm gust reaches the bud directly at each rotation period. All these actions allow the bud to continuously warm up, but with different intensities and over a different period. Although there is evidence of the effectiveness of the wind machines, the thermal transfers involved in those mechanisms raise questions about their true nature. Field measurements based on ultrasonic anemometers and fast responding thermocouples complemented by laboratory measurements on a reduced scale model allow to characterize both the airflow produced by the wind machine and the local temperature in its vicinity. Those experiments were realized in the vineyard of Quincy, in the framework of the SICTAG project. In the future paper, we will detail the aeraulic characterization of the wind machine and the thermal effects resulting from it and we will focus on how the wind machine warms up the local atmosphere and enables to reduce the freezing risk.