Terroir 2020 banner
IVES 9 IVES Conference Series 9 Potential application of indigenous Pichia kluyveri for enhanced wine aroma quality

Potential application of indigenous Pichia kluyveri for enhanced wine aroma quality

Abstract

Aims: In previous work, five indigenous Pichia kluyveri strains, GS1-1, FS-2-7, HS-2-1, C730 and C732, were isolated and selected from spontaneous fermented wines from Ningxia and Gansu. The aims of this study were to 1) evaluate resistance of these strains to environmental stressors that may restrict their growth and the progress of alcoholic fermentation; 2) Investigate their fermentation dynamics; 3) Characterise aroma profiles of Cabernet Sauvignon wines made from mixed cultures of P. kluyveri and Saccharomyces cerevisiae

Methods and Results: Tolerance assays were conducted in YEPD medium to test resistance of each Pichia kluyveri strain to sugar, pH, ethanol, temperature and free SO2. All strains except FS-2-7 were able to tolerate 60% w/v glucose, low pH of 2.0, 16% v/v ethanol, extreme fermentation temperatures (11˚C and 44˚C), and 500 mg/L total SO2. Following this, these strains were inoculated into a synthetic grape juice medium to test their fermentation performance and evaluate basic parameters of the final synthetic wine. Strain HS-2-1 was the first to initiate fermentation, and produced significantly higher amounts of total organic acids and less volatile acids compared to other strains. Thus, strain HS-2-1 was chosen for further characterisation in Cabernet Sauvignon fermentation trials co-fermented with S. cerevisiae NX11424 at different ratios. Viable yeast cell numbers were determined by plate counting. Yeast-derived volatile compounds of the final wine were analysed using head space-solid phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC/MS). Mixed fermentation did not inhibit HS-2-1 growth, and also produced less volatile acid, and significantly more esters and higher alcohols compared to single fermentation by S. cerevisiae. Notably, concentrations of isopentanol, ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl 9-decenoate and ethyl lactate increased in line with the increased proportion of HS-2-1 in the inoculum.

Conclusions: 

This study shows indigenous P. kluyveri HS-2-1 has good resistance to alcoholic fermentation associated common stressors, better fermentation performance, and excellent oenological characteristics when co-fermenting with S. cerevisiae

Significance and Impact of the Study: Chinese wine regions such as Ningxia and Gansu have developed dramatically in recent years. These wine regions are in great need to produce wines with typical regional characteristics. To promote regional typicity, using selected indigenous yeasts could introduce a unique local character or “terroir” during winemaking. Pichia kluyveri widely occurs at earlier stages of spontaneous fermentation, however limited research has been done on its oenological characteristics. This study comprehensively investigated the features of indigenous P. kluyveri strain(s), and highlighted the potential application of strain HS-2-1 in winemaking by co-fermenting with S. cerevisiae for improving the fruity and floral aroma profile of these Chinese wines.

DOI:

Publication date: March 25, 2021

Issue: Terroir 2020

Type: Video

Authors

iao Jiang1, Wenjing Zhang1, Li Feng1, Dongqing Ye1, Yanlin Liu1,2*

1College of Enology, Northwest Agricultural and Forestry University, Yangling, Shaanxi 712100, China
2Shaanxi Engineering Research Center for Viti-viniculture, Yangling, Shaanxi 712100, China

Contact the author

Keywords

Pichia kluyveri, stress tolerance, fermentation, volatile compounds, aroma  

Tags

IVES Conference Series | Terroir 2020

Citation

Related articles…

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.

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Soil quality in Beaujolais vineyard. Importance of pedology and cultural practices

A pedological study was carried out from 2009 to 2017 in Beaujolais vineyard, to improve physical and chemical knowledge of soils. It was completed in 2016 and 2017 by the current study, dealing with microbial aspects, in order to build a reference frame for improved advice in soil management. Microbial biomass was measured on representative plots of the six most common soil types identified in Beaujolais and, for each soil type, on plots with different levels of the main impacting parameters: total organic carbon, pH, cation exchange capacity, extractable copper. A total of 59 soil samples were collected. Confirming the results of various trials carried out in Beaujolais over the past 20 years, the results of the present study showed that the soils were still alive, but exhibited a large variability of biological parameters, which appeared dependant on both pedological and anthropic factors. Therefore, a good interpretation of biological parameters and advice for vine growers must rely on a pedologically-based referential with differentiated main driving factors. For example, the control of pH is of primary importance in granitic soils and in no way organic matter addition can improve soil quality if pH is too low. Conversely, in calcareous soils, biological parameters are more directly affected by direct or indirect (cover crops for example) inputs of organic matter. The use of biological parameters, such as microbial biomass, is of great potential value to improve advice on agro-viticultural practices (soil management, fertilization, liming, etc.), basis of a sustainable wine production on fragile soils.

Effect of the commercial inoculum of arbuscular mycorrhiza in the establishment of a commercial vineyard of the cultivar “Manto negro

The favorable effect of symbiosis with arbuscular mycorrhizal fungi (AMF) has been known and studied since the 60s. Nowadays, many companies took the chance to start promoting and selling commercial inoculants of AMF, in order to be used as biofertilizers and encourage sustainable biological agriculture. However, the positive effect of these commercial biofertilizers on plant growth is not always demonstrated, especially under field conditions. In this study, we used a commercial inoculum on newly planted grapevines of a local cultivar grafted on a common rootstock R110. We followed the physiological status of vines, growth and productivity and functional biodiversity of soil bacteria during the first and second years of 20 inoculated with commercial inoculum bases on Rhizophagus irregularis and Funeliformis mosseaeAMF at field planting time and 20 non-inoculated control plants. All the parameters measured showed a neutral to negative effect on plant growth and production. The inoculated plants always presented lower values of photosynthesis, growth and grape production, although in some cases the differences did not reach statistical significance. On the contrary, the inoculation supposed an increase of the bacterial functional diversity, although the differences were not statistically significant either. Several studies show that the effect of inoculation with AMF is context-dependent. The non-favorable effects are probably due to inoculation ineffectiveness under complex field conditions and/or that, under certain conditions, AMF presence may be a parasitic association. This puts into question the effectiveness of its application in the field. Therefore, it is recommended to only resort to this type of biofertilizer when the cultivation conditions require it (e.g., very low previous microbial diversity, foreseeable stress due to drought, salinity, or lack of nutrients) and not as a general fertilization practice.

Late season canopy management practices to reduce sugar loading and improve color profile of Cabernet-Sauvignon grapes and wines in the high irradiance and hot conditions of California Central Valley

Global warming is accelerating grape ripening, leading to unbalanced wines from fruit with high sugar content but poor aroma and colour development. Reducing the size of the photosynthetic apparatus after veraison has been shown to delay technological ripeness in cool climates, but methods have not been tested in areas with high irradiance and temperature where fruit exposure could have disastrous effects on berry composition. In this Cabernet-Sauvignon trial, we compared the application of an antitranspirant (pinolene), to severe canopy topping and above bunch zone leaf removal, all performed at mid-ripening, with an untouched control. We monitored the vines weekly by measuring stem water potential, gas exchange, fruit zone light exposure. We sampled berries to measure berry weight, total soluble solids, pH, titratable acidity, and the anthocyanin profile. At harvest, we assessed yield components, measured carbon isotope discrimination, rated sunburn on clusters, and produced experimental wines. We submitted harvest samples to metabolomic profiling through PFP-Q Exactive MS/MS and wines to sensory analysis. Application of the antitranspirant significantly reduced stomatal conductance and assimilation rate but did not affect the stem water potential. Inversely, leaf removal and topping increased water potential but did not affect leaf gas exchange. The late topping was the only treatment able to decrease sugar content (up to 2Bx), increase titratable acidity and pH, and improve anthocyanin content because of lower degradation of di-hydroxylated forms. Late leaf removal above the bunch zone increased lightning conditions in the canopy and produced the most significant damage on fruits. Yield components were not affected. This work suggests that late-season canopy management can effectively control ripening speeds and improve grapes and wines. Still, the effect on grape exposure in a critical time must be well balanced to avoid problems with the appropriate technique.