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…

Spatial variability of temperature is linked to grape composition variability in the Saint-Emilion winegrowing area

Elevated temperature during the grape maturation period is a major threat for grape quality and thus wine quality. Therefore, characterizing the grape composition response to temperature at a larger scale would represent a crucial step towards adaptation to climate change. In response to changes in temperature, various physiological mechanisms regulate grape composition. Primary and secondary metabolisms are both involved in this response, with well-known effects, for example on anthocyanins, and lesser known effects, for example on aromas or aroma precursors. At the field scale or at the regional scale, however, numerous environmental or plant-specific factors intervene to make the effects of temperature difficult to distinguish from overall variability. In this study, it was attempted to overcome this difficulty by selecting well-characterized situations with differing temperatures.
A long-term study of air temperature variability across several Merlot vineyards in the Saint-Emilion and Pomerol wine producing area found significant temperature differences and gradients at various time scales linked to environmental factors. From this study area, a few sites were selected with similar age, soil and training system conditions, and with repeated and contrasted temperature differences during the maturation period. The average temperature difference during the maturation period was about 2°C between cooler and warmer sites, a difference similar to that expected under future climate change scenarios. In close vicinity to the temperature sensors at each site, grape berries were sampled at different times until full maturity during 2019 and 2020. Also, berries from bunches on either side of the row were analyzed separately, allowing an investigation of bunch exposure effect associated with the coupling of berry temperature and solar radiation. Four replicates of pooled berries for each time – site – bunch exposure combination were obtained and analyzed for biochemical composition. Analyses of variance of the biochemical composition data collected at different sampling times reveal significant effects associated with temperature, site, and bunch azimuth. For instance, anthocyanins in grape skins are clearly influenced by temperature and solar radiation exposure, with up to 30% reduction in warmer conditions.

Adaptation to soil and climate through the choice of plant material

Choosing the rootstock, the scion variety and the training system best suited to the local soil and climate are the key elements for an economically sustainable production of wine. The choice of the rootstock/scion variety best adapted to the characteristics of the soil is essential but, by changing climatic conditions, ongoing climate change disrupts the fine-tuned local equilibrium. Higher temperatures induce shifts in developmental stages, with on the one hand increasing fears of spring frost damages and, on the other hand, ripening during the warmest periods in summer. Expected higher water demand and longer and more frequent drought events are also major concerns. The genetic control of the phenotypes, by genomic information but also by the epigenetic control of gene expression, offers a lot of opportunities for adapting the plant material to the future. For complex traits, genomic selection is also a promising method for predicting phenotypes. However, ecophysiological modelling is necessary to better anticipate the phenotypes in unexplored climatic conditions Genetic approaches applied on parameters of ecophysiological models rather than raw observed data are more than ever the basis for finding, or building, the ideal varieties of the future.

Impact of climate change on the viticultural climate of the Protected Designation of Origin “Jumilla” (SE Spain)

Protected Designation of Origin “Jumilla” (PDO Jumilla) is located in the Spanish provinces of Albacete and Murcia, in the South-eastern part of the Iberian Peninsula, where most of the models predict a severe impact of climate change in next decades. PDO Jumilla covers an area of 247,054 hectares, of which more than 22,000 hectares

Late frost protection in Champagne

Probably one of the most counterintuitive impacts of climate change on vine is the increased frequency of late frost. Champagne, due to its septentrional position is historically and regularly affected by this meteorological hazard. Champagne has therefore developed a strong experience in frost protection with first experiments dating from the end of 19th century. Frost protection can be divided in two parts: passive and active. Passive protection includes all the methods that do not seek to modify the vine’s environment or resistance at the time of frost. The most iconic passive protection in Champagne is the establishment of the individual reserve. This reserve allows to stock a certain quantity of clear wine during a surplus year to compensate a meteorological hazard like frost during the following years. Other common passive methods are the control of planting area (walls, bushes, topography), the choice of grape variety, late pruning, or the impact of grass cover and tillage. Active frost protection is also divided in two parts. Most of the existing techniques tend to modify vine’s environment. Most of the time they provide warmth (candles, heaters, windmills, heating cables…), or stabilise bud’s temperature above a lethal threshold (water sprinkling). The other way to actively fight is to enhance the resistance of buds to frost (elicitors). The Comité Champagne evaluates frost protection methods following three main axes: the efficiency, the profitability, and the environmental impact through a lifecycle assessment. This study will present the results on both passive and active protection following these three axes.

Influence of grapevine rootstock/scion combination on rhizosphere and root endophytic microbiomes

Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.