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…

Modeling island and coastal vineyards potential in the context of climate change

Climate change impacts regional and local climates, which in turn affects the world’s wine regions. In the short term, these modifications rises issues about maintaining quality and style of wine, and in a longer term about the suitability of grape varieties and the sustainability of traditional wine regions. Thus, adaptation to climate change represents a major challenge for viticulture. In this context, island and coastal vineyards could become coveted areas due to their specific climatic conditions. In regions subject to warming, the proximity of the sea can moderate extremes temperatures, which could be an advantage for wine. However, coastal and island areas are particular prized spaces and subject to multiple pressures that make the establishment or extension of viticulture complex.
In this perspective, it seems relevant to assess the potentialities of coastal and island areas for viticulture. This contribution will present a spatial optimization model that tends to characterize most suitable agroclimatic patterns in historical or emerging vineyards according to different scenarios. Thanks to an in-depth bibliography a global inventory of coastal and insular vineyards on a worldwide scale has been realized. Relevant criteria have been identified to describe the specificities of these vineyards. They are used as input data in the optimization process, which will optimize some objectives and spatial aspects. According to a predefined scenario, the objectives are set in three main categories associated with climatic characteristics, vineyards characteristics and management strategies. At the end of this optimization process, a series of maps presents the different spatial configurations that maximize the scenario objectives.

Soil, vine, climate change – what is observed – what is expected

To evaluate the current and future impact of climate change on Viticulture requires an integrated view on a complex interacting system within the soil-plant-atmospheric continuum under continuous change. Aside of the globally observed increase in temperature in basically all viticulture regions for at least four decades, we observe several clear trends at the regional level in the ratio of precipitation to potential evapotranspiration. Additionally the recently published 6th assessment report of the IPCC (The physical science basis) shows case-dependent further expected shifts in climate patterns which will have substantial impacts on the way we will conduct viticulture in the decades to come.
Looking beyond climate developments, we observe rising temperatures in the upper soil layers which will have an impact on the distribution of microbial populations, the decay rate of organic matter or the storage capacity for carbon, thus affecting the emission of greenhouse gases (GHGs) and the viscosity of water in the soil-plant pathway, altering the transport of water. If the upper soil layers dry out faster due to less rainfall and/or increased evapotranspiration driven by higher temperatures, the spectral reflection properties of bare soil change and the transport of latent heat into the fruiting zone is increased putting a higher temperature load on the fruit. Interactions between micro-organisms in the rhizosphere and the grapevine root system are poorly understood but respond to environmental factors (such as increased soil temperatures) and the plant material (rootstock for instance), respectively the cultivation system (for example bio-organic versus conventional). This adds to an extremely complex system to manage in terms of increased resilience, adaptation to and even mitigation of climate change. Nevertheless, taken as a whole, effects on the individual expressions of wines with a given origin, seem highly likely to become more apparent.

The use of rootstock as a lever in the face of climate change and dieback of vineyard

As viticulture faces challenges such as climate change or vineyard dieback, the choice of the variety and rootstock becomes more and more crucial. To study rootstock levers in the Bordeaux region, a parcel of Cabernet Sauvignon (CS) was planted with four rootstocks in 2014. Twenty repetitions of each of the following four rootstocks were set up: 101-14 MGt, Nemadex AB, 420A MGt and Gravesac. The number of bunches, yields and pruning weights of the vine shoots were measured individually on 240 vines from 2017 to 2021. Since 2020, nitrogen status assessed by assimilable nitrogen level, hydric status assessed by δ13C and berry maturity were measured on 80 samples taken from 20 repetitions of the four rootstocks. A lower yield was measured for CS grafted onto Nemadex AB due to the lower number of bunches and the lower weight of berries. The differences between the other three rootstocks are small, but CS grafted onto 420A MGt was the most productive. The CS grafted onto Nemadex AB had the lowest pruning weight while 101-14 MGt had the highest. In 2020, δ13C showed a more moderate water stress with 101-14 MGt and 420A MGt than with Nemadex AB. Surprisingly, the Gravesac was under more stress than the 101-14 MGt. The nitrogen status in the berries was better for Nemadex AB but this was perhaps due to the significantly lower weight of the berries.Rootstock 101-14 MGt attained the highest accumulation of sugars in the berries while 420A MGt allows to preserve higher acidity. The parcel is still young which may explain some of the results. These measures must therefore be continued over the next several years to fully assess the effects of these rootstocks on the development of the vines and the quality of the production under new climatic conditions.

Grapevine sugar concentration model in the Douro Superior, Portugal

Increasingly warm and dry climate conditions are challenging the viticulture and winemaking sector. Digital technologies and crop modelling bear the promise to provide practical answers to those challenges. As viticultural activities strongly depend on harvest date, its early prediction is particularly important, since the success of winemaking practices largely depends upon this key event, which should be based on an accurate and advanced plan of the annual cycle. Herein, we demonstrate the creation of modelling tools to assess grape ripeness, through sugar concentration monitoring. The study area, the Portuguese Côa valley wine region, represents an important terroir in the “Douro Superior” subregion. Two varieties (cv. Touriga Nacional and Touriga Franca) grown in five locations across the Côa Region were considered. Sugar accumulation in grapes, with concentrations between 170 and 230 g l-1, was used from 2014 to 2020 as an indicator of technological maturity conditioned by meteorological factors. The climatic time series were retrieved from the EU Copernicus Service, while sugar data were collected by a non-profit organization, ADVID, and by Sogrape, a leading wine company. The software for calibrating and validating this model framework was the Phenology Modeling Platform (PMP), version 5.5, using Sigmoid and growing degree-day (GDD) models for predictions. The performance was assessed through two metrics: Roots Mean Square Error (RMSE) and efficiency coefficient (EFF), while validation was undertaken using leave-one-out cross-validation. Our findings demonstrate that sugar content is mainly dependent on temperature and air humidity. The models achieved a performance of 0.65

What are the optimal ranges and thresholds for berry solar radiation for flavonoid biosynthesis?

In wine grape production, canopy management practices are applied to control the source-sink balance and improve the cluster microclimate to enhance berry composition. The aim of this study was to identify the optimal ranges of berry solar radiation exposure (exposure) for upregulation of flavonoid biosynthesis and thresholds for their degradation, to evaluate how canopy management practices such as leaf removal, shoot thinning, and a combination of both affect the grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) yield components, berry composition, and flavonoid profile under context of climate change. First experiment assessed changes in the grape flavonoid content driven by four degrees of exposure. In the second experiment, individual grape berries subjected to different exposures were collected from two cultivars (Cabernet Sauvignon and Petit Verdot). The third experiment consisted of an experiment with three canopy management treatments (i) LR (removal of 5 to 6 basal leaves), (ii) ST (thinned to 24 shoots per vine), and (iii) LRST (a combination of LR and ST) and an untreated control (UNT). Berry composition, flavonoid content and profiles, and 3-isobutyl 2-methoxypyrazine were monitored during berry ripening. Although increasing canopy porosity through canopy management practices can be helpful for other purposes, this may not be the case of flavonoid compounds when a certain proportion of kaempferol was achieved. Our results revealed different sensitivities to degradation within the flavonoid groups, flavonols being the only monitored group that was upregulated by solar radiation. Within different canopy management practices, the main effects were due to the ST. Under environmental conditions given in this trial, ST and LRST hastened fruit maturity; however, a clear improvement of the flavonoid compounds (i.e., greater anthocyanin) was not observed at harvest. Methoxypyrazine berry content decreased with canopy management practices studied. Although some berry traits were improved (i.e. 2.5° Brix increase in berry total soluble solids) due to canopy management practices (ST), this resulted in a four-fold increase in labor operations cost, two-fold decrease in yield with a 10-fold increase in anthocyanin production cost per hectare that should be assessed together as the climate continues to get hot.