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…

Ecophysiological performance of Vitis rootstocks under water stress

The use of rootstocks tolerant to soil water deficit is an interesting strategy to cope with limited water availability. Currently, several nurseries are breeding new genotypes, but the physiological basis of its responses under water stress are largely unknown. To this end, an ecophysiological assessment of the conventional 110-Richter (110R) and SO4, and the new M1 and M4 rootstocks was carried out in potted ungrafted plants. During one season, these Vitis genotypes were grown under greenhouse conditions and subjected to two water regimes, well-watered and water deficit. Water potentials of plants under water deficit down to < -1.4 MPa, and net photosynthesis (AN) <5 μmol m-2 s-1 did not cause leaf oxidative stress damage compared to well-watered conditions in any of the genotypes. The antioxidant capacity was sufficient to neutralize the mild oxidative stress suffered. Under both treatments, gravimetric differences in daily water use were observed among genotypes, leading to differences in the biomass of root, shoot and leaf. Under well-watered conditions, SO4 and 110R were the most vigorous and M1 and M4 the least. However, under water stress, SO4 exhibited the greatest reduction in biomass while M4 showed the lowest. Remarkably, under these conditions, SO4 reached the least negative stem water potential (Ψstem), while M1 reduced stomatal conductance (gs) and AN the most. In addition, SO4 and M1 genotypes also showed the highest and lowest hydraulic conductance values, respectively. Our results suggest that there are differences in water use regulation among genotypes, not only attributed to differences in stomatal regulation or intrinsic water use efficiency at the leaf level. Therefore, because no differences in canopy-to-root ratio were achieved, it is hypothesized that xylem vessel anatomical differences may be driving the reported differences among rootstocks performance. Results demonstrate that each Vitis rootstock differs in its ecophysiological responses under water stress.

Leaf vine content in nutrients and trace elements in La Mancha (Spain) soils: influence of the rootstock

The use of rootstock of American origin has been the classic method of fighting against Phylloxera for more than 100 years. For this reason, it is interesting to establish if different rootstock modifies nutrient composition as well as trace elements content that could be important for determining the traceability of the vine products. A survey of four classic rootstocks (110-Richter, SO4, FERCAL and 1103-Paulsen) and four new ones (M1, M2, M3 and M4) provided by Agromillora Iberia. S.L.U., all of them grafted with the Tempranillo variety, has been carried out during 2019. The eight rootstocks were planted in pots of 500 cc, on three soils with very different characteristics from Castilla-La Mancha (Spain). In the month of July, the leaves were collected and dried in a forced air oven for seven days at 40ºC. Then, the samples were prepared for the analysis determination, carried out by X-Ray fluorescence spectrometry. The results obtained showed that in the case of content in mineral elements in leaf, separated by soil type, we can report the importance of few elements such as Si, Fe, Pb and, especially, Sr. The rootstock does not influence the composition of the vine leaf for the studied elements that are the most important in determining the geochemical footprint of the soil. The influence of the soil can be discriminated according to some elements such as Fe, Pb, Si and, especially, Sr.

The interplay between grape ripening and weather anomalies – A modeling exercise

Current climate change is increasing inter- and intra-annual variability in atmospheric conditions leading to grapevine phenological shifts as well altered grape ripening and composition at ripeness. This study aims to (i) detect weather anomalies within a long-term time series, (ii) model grape ripening revealing altered traits in time to target specific ripeness thresholds for four Vitis vinifera cultivars, and (iii) establish empirical relationships between ripening and weather anomalies with forecasting purposes. The Day of the Year (DOY) to reach specific grape ripeness targets was determined from time series of sugar concentrations, total acidity and pH collected from a private company in the period 2009-2021 in North-Eastern Italy. Non-linear models for the DOY to reach the specified ripeness thresholds were assessed for model efficiency (EF) and error of prediction (RMSE) in four grapevine cultivars (Merlot, Cabernet Sauvignon, Glera and Garganega). For each vintage and cultivar, advances or delays in DOY to target specified ripeness thresholds were assessed with respect to the average ripening dynamics. Long-term meteorological series monitored at ground weather station by means of hourly air temperature and rainfall data were analyzed. Climate statistics were obtained and for each time period (month, bimester, quarter and year) weather anomalies were identified. A linear regression analysis was performed to assess a possible correlation that may exist between ripening and weather anomalies. For each cultivar, ripeness advances or delays expressed in number of days to target the specific ripening threshold were assessed in relation to registered weather anomalies and the specific reference time period in the vintage. Precipitation of the warmest month and spring quarter are key to understanding the effect of climate change on sugar ripeness. Minimum temperatures of May-June bimester and maximum temperatures of spring quarter best correlate with altered total acidity evolution and pH increment during the ripening process, respectively.

Diagnosis of soil quality and evaluation of the impact of viticultural practices on soil biodiversity in a vineyard in southwestern France

Viticulture is facing two major changes – climate change and agroecological transition. In both cases, soil quality is seen as a lever to move towards a more sustainable viticulture. However, soil biological quality is little considered in the implementation of viticultural practices. Gascogn’Innov (2017-2022) is an Operational Group funded by the European Innovation Partnership for Agriculture. As such, it brings together winegrowers from the south-west of France, scientists, advisors and technicians, around a project focused on viticultural soil biological functioning and the design of technical routes more respectful toward soil heritage. To achieve this, the project aims to acquire references on the impact of viticultural practices on soil biology from a dynamic way, and to test a methodology to integrate information provided by the soil bioindicators to manage farming systems. A set of indicators of soil biological quality are evaluated in the project: microorganisms (bacteria and fungi abundance and diversity), fauna (abundance and diversity of nematodes and earthworms), physico-chemical characteristics, soil structure assessment and degradation rate of organic matter. Based on a network of 13 plots that have been subject to an initial diagnosis in 2017, several agronomical practices to restore soil fertility are experimented to redesign the cropping system (for instance plant cover, organic matter inputs, reduction of herbicides, mineral fertilizers). System redesign was made in collaboration by winegrowers and an interdisciplinary group of experts (agronomists, biologists). Several indicators are measured on vine and soil at each vintage to assess vine health and productivity. At the end of the project (2021), a final diagnosis was carried out. Gascogn’Innov allowed to create a regional database on the quality of wine-growing soils, which permitted to evaluate the effect of practices according to soil types. Especially, decreasing the intensity of tillage and increasing the duration and diversity of grass coverage tends to increase the abundance of all the organisms studied. This project confirmed the value of soil biological quality indicators to drive the sustainability of practices, but also highlighted the key-role of expertise, in both agronomy and soil biology, to help winegrowers understand and appropriate their soil quality diagnoses.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).