Evaluation of Cell Wall Properties of Wine Associated Organisms, Specifically Grapevine (Vitis vinifera), Yeast (Saccharomyces cerevisiae) and Bacteria, in Relation to Berry Ripening, Leaf Development, Abiotic and Biotic Stresses, Fermentation and Wine Properties


Project Number
IWBT-P 09-01

Project title
Evaluation of cell wall properties of wine associated organisms, specifically grapevine (Vitis vinifera), yeast (Saccharomyces cerevisiae) and bacteria, in relation to berry ripening, leaf development, abiotic and biotic stresses, fermentation and wine properties.

Project leader
Moore, J P

Institution
University of Stellenbosch. Faculty of AgriSciences. Institute for Wine Biotechnology

Team members
Moore, J P
Nguema-Ona, E
Young, P R
Stander, C
Potgieter, E
De Beer, A
Tredoux, M
Lashbrooke, J
Bauer, F F
Nieuwoudt, H H
Vivier, M A

Project description
The cell wall of biological organisms (such as plants, yeast or bacteria) constitutes the interface between the external environment and the interior cellular apparatus. All of the major wine-associated organisms (Vitis vinifera, Saccharomyces cerevisiae, and the various wine bacteria) possess cell walls, although, the molecular composition and arrangement differ widely between the specific organisms. The cell wall is of fundamental importance in the vineyard, since grapevine plants undergo changes in wall composition in roots, berries and leaves due to developmental and environmental (e.g. climate and disease) influences. Cell walls play a direct role in textural and colour properties of grape berries during ripening in both the wine- and table grape industries. Wall properties and processes (such as phenolic oxidation commonly referred to as browning reactions) are also known to strongly influence storage potential of berries and post-harvest quality of many fruits (including grapes). Similarly, during the winemaking process numerous wall-derived polymers (commonly polysaccharides, proteins and phenolic polymers) are supplied to the fermenting must from the grape berries as well as yeast and bacteria present. These polymers interact with numerous components present in the must affecting filtration efficiency, aroma compound retention, solution clarity, organoleptic properties (e.g. mouth-feel) and the potential health-improving properties of the wine produced. Clearly, wall derived polymers and processes impart significant desirable and undesirable characteristics, during grape-growing (e.g. berry ripening, browning potential, pathogen susceptibility), grape storage (e.g. texture modification, browning reactions), fermentation (such as yeast and bacterial growth phenotypes) and wine properties (e.g. aroma, mouth-feel, turbidity, health properties).

Despite the obvious importance associated with cell wall properties in the grape and wine industries, very little is known regarding the fundamental scientific processes that relate to these quality parameters (aroma, storage, texture). Accurate, but high-throughput analytical and profiling methods for cell walls are lacking. What is greatly needed is a set of methodologies to study these processes by tracking the careers of grape and wine polymers from the vineyard, through the storage and processing steps, during winemaking, to the shelf and / or glass. Developing relatively rapid, but also highly accurate techniques to dissect and monitor wall processes under defined conditions and controlled field and cellar conditions will be of tremendous value. Understanding the underlying scientifically defined processes (causes) which govern the various quality parameters (effects) discussed previously is an absolute requirement for rational and effective experimentation in the table grape and wine industries.

The aim of this project is to develop cell wall platform analytical technologies and capacity to support the various research themes under study in the table grape- and wine research programmes.

The main project is divided into three sub-projects which integrate with each other through the use of chemometrics and statistical analysis. Subproject 1 is to develop a set of reproducible and highly accurate reference methods for the various wall components to be analysed. Sub-project 1 includes the use of sophisticated analytical techniques such as spectrophotometry, gas chromatography (GC), high performance liquid chromatography (HPLC) and mass spectrometry (MS).

Subproject 2 is to develop rapid non-invasive screening methods using Fourier Transform (FT) Mid or Near Infrared (MIR and NIR) and Nuclear Magnetic Resonance (NMR) spectroscopy. This will allow the application of Chemometric techniques to develop test models with the aid of the reference data generated in Subproject 1.

Subproject 3 will involve the use of non-spectroscopic, semi-rapid but highly specific analytical techniques to analyses wall components. These techniques principally involve the development of glycan arrays using commercial cell wall antibodies and mass profiling using mass spectrometry to analyse wall sub-structures. Each subproject integrates through the use of metric techniques to provide complementary information on the particular cell wall system under study.

Although the main thrust of this project is to develop and evaluate the technologies for cell wall profiling and analysis, it will be integrated with current research projects in grapevine biotechnology. This will provide this project with fully characterised materials to work with and would lead to immediately useful datasets from the analysis of the prototype grapevines.

Poster(s)
Moyo, M, Steyn, J, Nguema-Ona, E E, Moore, J P and Vivier, M A. 2010. Cell wall remodelling in VvPGIP1 overexpressed in tobacco (Nicotiana tabacum) and grapevine (Vitis vinifera) plants. Poster presented at the 4th International Congress of the South African Society for Enology and Viticulture. 28-30 July, Cape Town, South Africa.

Moore, J P, Ngueman-Ona, E E and Vivier, M A. 2009. Tracking the careers of grape and wine polymers. Poster presented at the 32nd National Congress of the South African Society for Enology and Viticulture.12 December, Somerset West, South Africa.

Moore, J P, Nguema-Oma, E E, Farrant, J M and Vivier, M A. 2010. Towards a systems based approach to plant drought and desiccation tolerance – a role for cell wall processes and -omics. Poster presented at the Cost Action F0605 Conference, Plant Abiotic Stress from Signalling to Crop Improvement. 26-27 May, Valencia, Spain.

Zhang, S and Moore, J P. 2011. Evaluating cell wall properties and mannoproteins content in laboratory and industrial wine yeast strains using mid- and near infrared spectroscopy. Poster presented at the International Conference on Near-Infrared Spectroscopy. 13-20 May, Cape Town, South Africa.

Presentation(s)
Nguema-Ona, E E, Moore, J P and Vivier, M A. 2008. Cell wall reinforcement in transgenic tobacco lines expressing Vitis vinifera polygalacturonase inhibiting proteins (PGIPs). Presentation at the Cape Biotechnology Forum. 30 November – 2 December, Somerset West, South Africa.

Nguema-Ona, E E, Moore, J P and Vivier, M A. 2009. The role of polygalacturonase inhibiting proteins (PGIPs) and the cell wall of grapevine in pathogen defence. Paper presented at the 35th Conference of the South African Association of Botanists. 19-29 January, Stellenbosch, South Africa.

Moore, J P, Nguema-Ona, E E and Vivier, M A. 2010. Cell wall-omics: Tracking the careers of grape and wine polymers using biotechnology and systems biology. Presentation at the Cape Biotechnology Forum. 24-26 March, Somerset West, South Africa.

Nguema-Ona, E E, Moore, J P and Vivier, M A. 2010. Cell walls and plant defences: Using microscopical and analytical techniques to elucidate resistance phenotypes. Paper presented at the Cape Biotechnology Forum. 24-26 March, Somerset West, South Africa.

Moore, J P, Nguema-Ona, E E, Ndlovu, T, Divol B, Nieuwoudt, H H, Bauer, F F and Vivier, M A. 2010. Methods to track grape and yeast derived cell wall polymers in wine: Applications for wine biotechnology. Paper presented at the 12th International Cell Wall Meeting. 25-30 July, Porto, Portugal .

Nguema-Ona, E E, Steyn, C, Moore, J P, Alexandersson, E, Jacobson, D and Vivier, M A. 2010. Polygalacturonase-inhibiting proteins (PGIPs), cell wall remodelling and defence: Are there links? Paper presented at the 12th International Cell Wall Meeting. 25-30 July, Porto, Portugal.

Moore, J P, Nguema-Ona, Ndlovu, T, Divol, B, Nieuwoudt, H H, Bauer, F F and Vivier, M A. 2009. Developing cell wall modified and mannoprotein release wine yeast (Saccharomyces cerevisiae) strains for improved wine protectant properties (e.g. haze protection). Paper presented at the 32nd National Congress of the South African Society for Enology and Viticulture. 12 December, Somerset West, South Africa.

Article
Alexandersson, E O, Becker, J v W, Jacobson, D, Nguema-Ona, E, Steyn, C, Denby, K, Vivier, M A. 2011. Constitutive expression of a grapevine polygalacturonase-inhibiting protein affects grape gene expression and cell wall properties in uninfected tobacco, BMC Research Notes, v. 4 (p. 493)

Nguema-Ona, E, Moore, J P, Fagerstrom, A, Willats, W, Hugo, A, Vivier, M A. 2012. Profiling the main cell wall polysaccharides of tobacco leaves using high throughput and fractionation techniques, Carbohydrate Polymers, Mnth Apr v. 88 (3) (p. 939-949)
http://dx.doi.org/10.1016/j.carbpol.2012.01.044

FinalReport.pdf

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