GiESCO 2019 banner
IVES 9 IVES Conference Series 9 GiESCO 9 GiESCO 2019 9 Hexose efflux from the peeled grape berry

Hexose efflux from the peeled grape berry

Abstract

Context and purpose of the study ‐ After the onset of grape berry ripening, phloem unloading follows an apoplasmic route into the mesocarp tissue. In the apoplast, most of the unloaded sucrose is cleaved by cell wall invertases, and imported into the cells as glucose and fructose. Alternatively, sucrose can be imported directly from the apoplast and cleaved into glucose and fructose, either in the cytoplasm or vacuoles. In low‐sucrose cultivars, such as Shiraz, glucose and fructose are the dominant sugars in vacuoles. Transport of sugars across the plasma membrane and tonoplast is a complex process, not fully understood. Some of the elements of the sugar transport mechanism may work in a reverse mode. The purpose of this study was to indirectly observe the nature of the transport mechanism by creating conditions inducing hexose efflux from a peeled berry.

Material and methods ‐ Potted plants of cv. Shirazwere grown in a glass‐house (25/16°C), from the end of anthesis onward. The experimental method was derived from the “berry‐cup” technique: a peeled berry, still attached to the plant, was immersed in a MES buffer (2‐(N‐morpholino)ethanesulfonic acid, pH 5,5)) solution that was collected every 30 minutes over a 3 hour period. The experiment was repeated weekly during the ripening phase. Additionally, during the period of intensive sugar accumulation (one to three weeks after veraison), three treatments were imposed: (i) a comparison of sugar unloading from berries detached or attached to the vine, (ii) the addition of the membrane‐ impermeant sulfhydryl‐specific cytotoxin p‐chloromercuribenzenesulfonic (PCMBS, 1mM) to the buffer solution, (iii) exposing the berry to cold (10°C), room temperature (27°C) or warm (40°C) buffer. Collected samples were analyzed for glucose and fructose concentration.

Results ‐ During five weeks of ripening, the rate of hexose (mg of glucose+fructose per g of berry fresh weight) efflux from the peeled berry into the buffer solution increased.There was no difference in efflux rate between attached or detached berries, however efllux rates were temperature dependent. The non‐penetrating enzyme inhibitor, PCMBS, depressed glucose and fructose efflux at the first sampling date, but not two weeks later. The inhibitory effect of PCMBS on fructose efflux was different from glucose, however for both hexoses the reversible nature of PCMBS was confirmed. During ripening, the glucose to fructose ratio within the collected buffer was significantly lower than in the grape juice, and had the opposite trend. These results lead us to the conclusion that the origin of the collected hexoses was vacuolar, and that the hexose efflux mechanism is differently sensitive to PCMBS at the two stages of ripening. 

DOI:

Publication date: June 19, 2020

Issue: GiESCO 2019

Type: Article

Authors

Predrag BOŽOVIĆ (1,3), Suzy ROGIERS (2,3), Alain DELOIRE (4)

(1) University of Novi Sad,Faculty of Agriculture, Serbia
(2) New South Wales Department of Primary Industries, Wagga Wagga, NSW, Australia
(3)National Wine and Grape Industry Centre, Charles Sturt University, Wagga Wagga, Australia
(4)University of Montpellier, SupAgro, Department of Biology-Ecology, France

Contact the author

Keywords

Grapevine, Sugar transport, Glucose, Fructose, Efflux, PCMBS

Tags

GiESCO 2019 | IVES Conference Series

Citation

Related articles…

The rootstock, the neglected player in the scion transpiration even during the night

Water is the main limiting factor for yield in viticulture. Improving drought adaptation in viticulture will be an increasingly important issue under climate change. Genetic variability of water deficit responses in grapevine partly results from the rootstocks, making them an attractive and relevant mean to achieve adaptation without changing the scion genotype. The objective of this work was to characterize the rootstock effect on the diurnal regulation of scion transpiration. A large panel of 55 commercial genotypes were grafted onto Cabernet Sauvignon. Three biological repetitions per genotype were analyzed. Potted plants were phenotyped on a greenhouse balance platform capable of assessing real-time water use and maintaining a targeted water deficit intensity. After a 10 days well-watered baseline period, an increasing water deficit was applied for 10 days, followed by a stable water deficit stress for 7 days. Pruning weight, root and aerial dry weight and transpiration were recorded and the experiment was repeated during two years. Transpiration efficiency (ratio between aerial biomass and transpiration) was calculated and δ13C was measured in leaves for the baseline and stable water deficit periods. A large genetic variability was observed within the panel. The rootstock had a significant impact on nocturnal transpiration which was also strongly and positively correlated with maximum daytime transpiration. The correlations with growth and water use efficiency related traits will be discussed. Transpiration data were also related with VPD and soil water content demonstrating the influence of environmental conditions on transpiration. These results highlighted the role of the rootstock in modulating water deficit responses and give insights for rootstock breeding programs aimed at identifying drought tolerant rootstocks. It was also helpful to better define the mechanisms on which the drought tolerance in grapevine rootstocks is based on.

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

Organic recycled mulches in sustainable viticulture: assessment of spontaneous plants communities and weed coverage

In recent years, developing more efficient and sustainable viticulture management has been essential due to the impact of climate change in semiarid regions. For this reason, the use of recycled organic mulching (ROM) in the vineyard has become an interesting strategy to cope with water stress, isolated soil from extreme temperatures and improving soil humidity, control the presence of weeds and therefore reduce the inputs of herbicides and improve soil fertility. This work aimed to analyse the effect of three different organic mulches [straw (S), grape pruning debris (GPD) and spent mushroom compost (SMC)] and two traditional soil management techniques [herbicide (H) and interrow (IN)] on weed coverage and the spontaneous plant communities’ presence. Data sampling was collected throughout the vine vegetative cycle of 2021 in La Rioja, Spain. The different soil management techniques had a clear effect on weed coverage and his development during the vine vegetative cycle. SMC and H were the treatments with the highest and the lowest coverage percentage, respectively. IN had a delayed weed emergence at the beginning of the vine vegetative cycle, but finally it reached maximum values nearby SMC. GPD and S had similar effects on weed emergence, reaching 25-30% of the maximum coverage values. A total of 29 herbaceous species were identified during the vegetative cycle, some of them very isolated and occasional. Principal component analysis (PCAs) showed a good association between spontaneous species and treatments, furthermore, specific species-treatment associations were found. Moreover, three clear groups of herbaceous communities were identified by cluster analysis. This study provides interesting information about the effect of different alternative soil management on herbaceous plant coverage and weed species communities which could contribute to making more sustainable viticulture.

Climate change impacts: a multi-stress issue

With the aim of producing premium wines, it is admitted that moderate environmental stresses may contribute to the accumulation of compounds of interest in grapes. However the ongoing climate change, with the appearance of more limiting conditions of production is a major concern for the wine industry economic. Will it be possible to maintain the vineyards in place, to preserve the current grape varieties and how should we anticipate the adaptation measures to ensure the sustainability of vineyards? In this context, the question of the responses and adaptation of grapevine to abiotic stresses becomes a major scientific issue to tackle. An abiotic stress can be defined as the effect of a specific factor of the physico-chemical environment of the plants (temperature, availability of water and minerals, light, etc.) which reduces growth, and for a crop such as the vine, the yield, the composition of the fruits and the sustainability of the plants. Water stress is in many minds, but a systemic vision is essential for at least two reasons. The first reason is that in natural environments, a single factor is rarely limiting, and plants have to deal with a combination of constraints, as for example heat and drought, both in time and at a given time. The second reason is that plants, including grapevine, have central mechanisms of stress responses, as redox regulatory pathways, that play an important role in adaptation and survival. Here we will review the most recent studies dealing with this issue to provide a better understanding of the grapevine responses to a combination of environmental constraints and of the underlying regulatory pathways, which may be very helpful to design more adapted solutions to cope with climate change.

Adaptability of grapevines to climate change: characterization of phenology and sugar accumulation of 50 varieties, under hot climate conditions

Climate is the major factor influencing the dynamics of the vegetative cycle and can determine the timing of phenological periods. Knowledge of the phenology of varieties, their chronological duration, and thermal requirements, allows not only for the better management of interventions in the vineyard, but also to predict the varieties’ behaviour in a scenario of climate change, giving the wine producer the possibility of selecting the grape varieties that are best adapted to the climatic conditions of a certain terroir. In 2014, Symington Family Estates, Vinhos, established two grape variety libraries in two different places with distinctive climate conditions (Douro Superior, and Cima Corgo), with the commitment of contributing to a deeper agronomic and oenological understanding of some grape varieties, in hot climate conditions. In these research vineyards are represented local varieties that are important in the regional and national viticulture, but also others that have over time been forgotten — as well as five international reference cultivars. From 2017 to 2021, phenological observations have been made three times a week, following a defined protocol, to determine the average dates of budbreak, flowering and veraison. With the climate data of each location, the thermal requirements of each variety and the chronological duration of each phase have been calculated. During maturation, berry samples have been gathered weekly to study the dynamics of sugar accumulation, between other parameters. The data was analysed applying phenological and sugar accumulation models available in literature. The results obtained show significant differences between the varieties over several parameters, from the chronological duration and thermal requirements to complete the various stages of development, to the differences between the two locations, confirming the influence of the climate on phenology and the stages of maturation, in these specific conditions.