Dissertations / Theses on the topic 'Oenology and viticulture'

Thesis (MScAgric (Viticulture and Oenology))--Stellenbosch University, 2008.
The production of good quality wine is essential to ensure competitiveness on an international level. Wine quality is usually evaluated for the visual, olfactory and taste characteristics of that specific wine. The winemaking process starts with the grapes in the vineyard followed by oenological practises in the winery until the final wine is bottled. Factors that could influence wine quality include the grape quality from which the wine is made and different techniques used during wine production. Other factors include the presence as well as the interaction between microorganisms found in the grape juice and wine, and the biochemical effect these microorganisms have on certain chemical compounds in the wine. The different microorganisms found in grape juice and wine can either have a negative or positive contribution to the final quality of the wine. During certain stages of the winemaking process the growth and metabolic activity of certain microorganisms is a necessity to produce good wine. During other stages the presence of certain microorganisms can lead to the development of compounds that is regarded as off-flavours and therefore lead to unpalatable wines of low quality. Yeast strains that naturally present on the grapes and in the winery can also contribute to the final quality of the wine. Brettanomyces yeasts are part of the natural flora of winemaking and can drastically influence the aroma characters of a wine through the production of volatile phenols. The general aroma descriptions of volatile phenols include "smoky", "spicy", "barnyard", "animal" and "medicinal". Although some wine drinkers believe that these characters can add to the complexity of a wine, high levels of volatile phenols is mostly regarded as off-flavours and mask the natural fruity flavours of a wine. With this study we wanted to generate a better understanding of the effect of different winemaking practises on the production of volatile phenols by B. bruxellensis. We evaluated the difference in volatile phenol production when B. bruxellensis was introduced before or after alcoholic fermentation. We have shown that B. bruxellensis could grow and produce volatile phenols during alcoholic fermentation. Results obtained also showed that commercial wine yeast strains could produce the vinyl derivatives that serve as precursors for Brettanomyces yeast to produce the ethyl derivatives. The commercial yeast strains differed in their ability to produce vinyl derivatives. Different malolactic fermentation scenarios were evaluated, namely spontaneous versus inoculated, and with or without yeast lees. Results showed that spontaneous malolactic fermentation had higher volatile phenol levels in the wine than inoculated malolactic fermentation. The treatment with lees reduced the level of volatile phenols, probably due to absorption by yeast cells. The presence of the phenyl acrylic decarboxylase (PAD1) gene and the production of volatile phenols by S. cerevisiae commercial yeast strains were evaluated in Shiraz grape juice and in synthetic grape juice. The results indicated that the yeast strains differ in their ability to produce 4-vinylphenol and 4-vinylguaiacol. All the yeast strains tested had the PAD1 gene. We also evaluated the presence of the phenolic acid decarboxylase (padA) gene and the ability of different lactic acid bacteria strains to produce volatile phenols in synthetic wine media. Although some of these strains tested positive for the phenolic acid decarboxylase gene most of them only produced very low levels of volatile phenols. This study made a valuable contribution on the knowledge about the effect of Brettanomyces yeast on the volatile phenol content of red wines during different stages of the winemaking process and when applying different winemaking practices. It also showed the effect between Brettanomyces yeast and other wine microorganisms and the possible influence it could have on the final quality of wine. Research such as this can therefore aid the winemaker in making certain decisions when trying to manage Brettanomyces yeast spoilage of wines.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Because of long-term climate changes, apparently associated with higher temperatures and fewer rainfall events, factors such as water-use efficiency and site selection for new cultivars are a matter of increasing importance for viticulture. Within this context, the root system is expected to play a key role. Its relevance to grapevine functioning is due to the numerous functions in which it is involved. In the light of this, the development of the root system is highly relevant to the viticulturist because of the fact that grapevine growth and functioning are dependent on the development of the root system. Differences can, therefore, be expected in terms of berry ripening on single grapevines of the same scion for situations with differing development of root systems, despite being grafted on the same rootstock. Root growth is influenced by several factors, among the ecological aspects. Soil parameters have a predominant influence on root growth and distribution but also annual root production can be altered by canopy manipulation. Due to the importance of root growth to the aboveground development of the vine, it is critical to gain understanding of the relationship between soil factors and root growth and distribution, and the central role that the subterranean environment plays in the concept of terroir. This study aimed to investigate the effect of selected soil physical and chemical parameters on root growth and distribution and to investigate whether having very different canopies influences root growth. In order to achieve these goals, two experiments were conducted; the first was performed in two commercial Sauvignon blanc vineyards each grafted onto Richter 110, non-irrigated, with two treatments: undisturbed lateral growth and complete lateral removal. The second study included the analysis of eight commercial Sauvignon blanc vineyards grafted onto Richter 99 and Richter 110 located in the Stellenbosch Wine of Origin District. Measurements of physical and chemical soil parameters, root growth and distribution, canopy growth and functioning, vine water status and berry composition were performed. The edaphic factors appeared to be one of the most important parameters that affected root development by changing soil water availability and possibly causing physical or chemical limitations on root growth. From the results of this study, it is clear that severe water stress and a pH (KCl) lower than 4.5 play a key role in the limitation of root growth. Due to the fact that most of the soils from the Stellenbosch Wine of Origin District, especially the subsoils, are acidic, this is a factor to consider before planting. On the other hand, the combination of favourable edaphic conditions, such as a subsoil pH of higher than 5.0, light- to mediumtextured subsoil and moderate water stress, allow increased growth of thin roots. However, the effect of canopy management on root growth cannot be discounted due to its importance in the variation of carbohydrate demand by competing sinks. This study showed that lateral removal done from when the berries are at pea size results in an increase in the number of thin roots (0.5-2.0 mm). The secondary leaf area represents at least the same leaf area as the primary leaf area in all the vineyards evaluated, which reveals the relative importance of the laterals in the total leaf area of the vine and the potential importance in terms of microclimate and leaf area available for photosynthesis. Studies of root growth should take the vineyard canopy architecture into account.
AFRIKAANSE OPSOMMING: As gevolg van langtermyn klimaatsveranderinge wat toegeskryf kan word aan die voorkoms van hoër temperature en laer reënval, is faktore soos effektiwiteit van waterverbruik en liggingseleksie vir nuwe kultivars van kardinale belang vir wingerdkunde. Binne hierdie konteks, speel die wortelsisteem ‘n belangrike rol. Die belangrikheid hiervan vir wingerdfunksionering kan toegeskryf word aan die talle funksies waarby dit betrokke is. Die ontwikkeling van die wortelsisteem is dus hoogs relevant vir die wingerdkundige, omdat wingerdgroei en funksionering afhanklik is van die ontwikkeling van die wortelsisteem. Verskille kan daarom dus verwag word in terme van korrelrypwording op ‘n enkele wingerdstok van dieselfde onderstok vir gevalle met verskillende ontwikkeling van die wortelsisteem, ten spyte daarvan dat dit op dieselfde onderstok geënt is. Wortelgroei word, onder ekologiese aspekte, deur verskillende faktore beïnvloed. Grondfaktore het meerendeels ‘n predominante invloed op wortelgroei en -verspreiding, terwyl jaarlikse wortelproduksie deur lowermanipulasie beïnvloed kan word. Weens die belangrikheid van wortelgroei vir die bogrondse ontwikkeling van die wingerd, is dit krities om kennis op te doen oor die verhouding tussen grondfaktore en wortelgroei en –verspreiding, asook die sentrale rol wat die subterreinomgewing op die terroir-konsep speel. Die studie was daarop gemik om die invloed van geselekteerde fisiese en chemiese parameters van grond op wortelgroei en -verspreiding vas te stel, en ook te ondersoek of verskillende lowers wortelgroei sal beïnvloed. Om laasgenoemde doelwitte te bereik, is twee eksperimente uitgevoer. Die eerste is uitgevoer in ‘n kommersïele Sauvignon blanc-wingerd wat geënt is op Richter 110, sonder besproeïng en met twee behandelings, naamlik onversteurde sêkondere lootgroei en volledige sêkondere lootverwydering. Die tweede studie het die analise van agt kommersïele Sauvignon blancwingerde geënt op Richter 99 en Richter 110 in die Stellenbosch Wyn van Oorsprong Distrik. Metings van fisiese en chemiese grondfaktore, wortelgroei en -verspreiding, lowergroei en -funksionering, plantwaterstatus en korrelsamestelling is uitgevoer. Dit blyk dat edafiese faktore een van die belangrikste parameters is wat wortelontwikkeling beïnvloed deur beskikbaarheid van grondwater te verander, en wat moontlik fisiese en chemiese beperkings op wortelgroei kan veroorsaak. Uit die resultate van die studie is dit duidelik dat intense waterspanning en ‘n pH (KCl) laer as 4.5 ‘n belangrike rol in die beperking van wortelgroei speel. Aangesien die meeste van die grondsoorte in die Stellenbosch Wyn van Oorsprong Distrik, veral al die subgronde, suur is, is dit ‘n faktor wat in oorweging geneem moet word voor aanplantings. Die kombinasie van gunstige edafiese toestande, soos ‘n subgrond met ‘n pH hoër as 5.0, ‘n lig tot medium tekstuur en matige waterspanning, sal dus aanleiding gee tot ‘n toename in die groei van dun wortels. Die effek van lowerbestuur op wortelgroei kan egter nie buite rekening gelaat word nie weens die belangrikheid daarvan in die variasie van koolhidraataanvraag deur kompeterende vraagpunte. Hierdie studie toon dat, indien sêkondere lootverwydering tydens ertjiekorrelgrootte toegepas is, dit aanleiding gee tot ‘n toename in die dun wortels (0.5 tot 2.0 mm). Die sêkondere blaaroppervlakte verteenwoordig minstens dieselfde blaaroppervlakte as die primêre blaaroppervlakte in al die wingerde wat ondersoek is, wat dui op die belangrikheid van sêkondere lote in die totale blaaroppervlakte van die wingerd en die potensiële belangrikheid daarvan in terme van mikroklimaat en blaaroppervlakte wat vir fotosintese beskikbaar is. Studies van wortelgroei moet lowerargitektuur in ag neem.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2006.
The quality of wine is influenced by numerous factors of which the grapes, winemaking techniques and effective quality control are to name a few. The use of new techniques should be cost effective and always have a positive influence on the wine quality. The addition of commercial tannins to wine is a fairly new technique and the effects of these additions at the concentrations prescribed have not been investigated in detail. The commercial tannins can be added to wine for different reasons, which may include: stabilisation of colour, increasing the aging potential, to modify aromas, promote precipitation of proteins, limit the effect of laccase activity, substrate for micro-oxygenation, to act as a redox buffer and structural and mouth feel modification. The reason for the addition determines the type of commercial tannin that is used, the timing of the addition and the dosage used.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Previous research has shown that carotenoids are precursors of C13-norisoprenoid aroma compounds in wine. C13-norisoprenoids have low threshold values in wine with the most prominent C13-norisoprenoids being β-damascanone and β-ionone which contribute honey and floral like aroma to wine. Chlorophyll and its derivates have also been detected in wine with potential to be precursors to aroma compounds. Apart from the contribution of these pigments to wine aroma and quality they are vital role players in photosynthesis and are widely found in plants and plant products. The main functions of these pigments in plants are light collection and light-protection. Research has shown that environmental conditions, climate, light exposure of bunches and soil water deficit influence the carotenoid content of grape berries. Furthermore the concentration of carotenoids and chlorophylls has also been shown to differ between cultivars. No research in this regard has been done on Merlot grape berries. With this in mind, the aim of this study was to evaluate the effect of vigour and soil water content on the evolution of carotenoids and chlorophylls through ripening of grape berries from the cv. Merlot. However, when looking at methods to analyse carotenoids and chlorophylls in berry tissue, especially lyophilised tissue, there were no readily available methods. Thus, an extraction method to identify and quantify the carotenoid and chlorophyll profile of lyophilised tissue from unripe (green) to ripe (red) Merlot grape berries was needed. In this study the RPHPLC method of Taylor et al. (2006) for carotenoids and the extraction method of Mendes-Pinto et al. (2004) were adapted to analyse both carotenoids and chlorophylls in lyophilised grape tissue. The RP-HPLC method baseline separated all the carotenoids and chlorophylls and their derivatives. Recovery of standards from mock extractions was high, indicating that the extraction procedure was acceptable. However, extraction recovery tested in the matrix of the grape tissue showed less promising results due to the high acid content of grape tissue. Violaxanthin, neoxanthin and the chlorophylls were especially sensitive to low pH conditions which facilitated their degradation. The degradation products of these compounds under acidic conditions were identified as pheophytin a, b, chlorophillide a, pyropheophytin b, cisviolaxanthin, cis-neoxanthin, neochrome, mutatoxanthin and luteoxanthin. There is a possibility that some degradation products were already present in the tissue due to lyophilisation (since the water in the berry was then removed and the acid concentrated). More work is needed to investigate the effect of lyophilisation and storage on the composition of grape tissue of different maturity. The extraction method for grape berry tissue at different ripening stages should also be optimised further to effectively neutralise tissue acidity, without compromising the extraction of carotenoids significantly, in especially green berry tissue. The question as to whether cisisomers and chlorophyll degradation products are naturally present in grape berries or are formed during sampling and processing remains unanswered in the current study. This study confirmed that in general carotenoids and chlorophylls decrease on a per berry (μg/berry) and concentration (μg/g) basis from veraison to harvest. Furthermore, this study was inconclusive in showing that vigour differences have an effect on the rate of synthesis/degradation of carotenoids, chlorophyll and some other ripening parameters, namely malic acid, total glucose and fructose, total tannin and total anthocyanin, from pre-veraison (pea size) to harvest. Additionally, no significant effect of soil water content on carotenoids, chlorophylls and ripeness parameters was found in this study, most likely due the fact that high soil water capacity was found in lower soil layers which may have prevented significant differences in grapevine water status. Experimental plots selected for vigour differences based on normalised difference vegetation index (NDVI) images, pruning mass and soil water measurements by means of a neutron probe, showed significant differences in soil water content in only the first 30 cm of the soil for the ripening seasons studied. Predawn plant water potential measurements, however, indicated that none of the experimental vines experienced severe water stress which was previously shown to effect carotenoid content of grapes. The carotenoid 5,8-epoxy--carotene was quantified for the first time in grapes and represents a significant amount of the total carotenoids present at harvest. All the carotenoids and chlorophylls except -carotene appeared to be sensitive to seasonal variation in climatic conditions. Lutein and β-carotene were found to be the most abundant carotenoids present in Merlot grape berries together with chlorophyll a for both seasons studied. The values of these carotenoids also correlated well with previous research. However, chlorophyll a was found in much larger quantities in Merlot berries compared to reported data. This is possibly because in this study the chlorophyll degradation products were included in the calculation of chlorophyll a. Multivariate analysis showed promising preliminary prediction models (with correlation values of above 0.8 for both seasons analysed) for the prediction of the concentration of ripeness parameters (glucose, fructose, malic acid, total tannins and anthocyanins) with carotenoid and chlorophyll content. This result highlights the opportunity for the development of a rapid non-destructive method to measure carotenoids and chlorophylls in berries which in turn can predict optimal ripeness. Furthermore, since carotenoids are the precursors to C13- norisoprenoid aroma compounds in wine a preview of the potential contribution of these aromas to wine might be evaluated. Further research is necessary to investigate the possibility of building and validating such models.
AFRIKAANSE OPSOMMING: Vorige navorsing het getoon dat karotenoïede die voorlopers is van C13-norisoprenoïed aromaverbindings in wyn. C13-norisoprenoïede het lae drempelwaardes in wyn, met β- damassenoon en β-jonoon as die prominentste C13-norisoprenoïede wat ‘n bydrae tot die heuning en blomagtige aroma van die wyn maak. Chlorofil en sy derivate is ook reeds in wyn bespeur, met die potensiaal om voorlopers van aromaverbindings te wees. Buiten die bydrae van hierdie pigmente tot wynaroma en -kwaliteit is hulle ook belangrike rolspelers in fotosintese en kom hulle wydverspreid in plante en plantprodukte voor. Die vernaamste funksies van hierdie pigmente in plante is om lig te versamel en om as beskerming teen lig op te tree. Navorsing het getoon dat omgewingstoestande, klimaat, ligblootstelling van die trosse en grondwatertekorte die karotenoïedinhoud van druiwekorrels beïnvloed. Verder is ook getoon dat die konsentrasie van karotenoïede en chlorofille tussen kultivars verskil. Geen navorsing is al in hierdie opsig op Merlot-druiwekorrels gedoen nie. Met hierdie aspek in gedagte was die doelwit van hierdie studie om die effek van groeikrag en grondwaterinhoud op die evolusie van karotenoïede en chlorofille tydens die rypwording van druiwekorrels van die cv. Merlot te evalueer. Wanneer mens egter kyk na die metodes waarvolgens die karotenoïede en chlorofille in korrelweefsel geanaliseer word, is daar geen geredelik beskikbare metodes nie. ‘n Ekstraksiemetode om die karotenoïed- en chlorofilprofiel van geliofiliseerde weefsel van onryp (groen) tot ryp (rooi) Merlot-bessies te identifiseer en kwantifiseer was dus nodig. In hierdie studie is die RP-HPLC metode van Taylor et al. (2006) vir karotenoïede en die ekstraksiemetode van Mendes-Pinto et al. (2004) aangepas om beide karotenoïede en chlorofille in geliofiliseerde druiweweefsel te analiseer. Die basislyn van die RP-HPLC metode het all karotenoïede en chlorofille en hul derivate geskei. Herwinning van die standaarde vanaf skynekstraksies was hoog, wat aandui dat die ekstraksieprosedure aanvaarbaar was. Ekstraksieherwinning wat in die matriks van die druiweweefsel getoets is, het egter minder belowende resultate getoon as gevolg van die hoë suurinhoud van die druifweefsel. Violaxantien, neoxantien en die chlorofille was veral sensitief vir toestande van lae pH, wat hulle afbreking gefasiliteer het. Die afbrekingsprodukte van hierdie verbindings onder suurtoestande is geïdentifiseer as feofitien a en b, chlorofillied a, pirofeofitien b, cis-violaxantien, cis-neoxantien, neochroom, mutatoxantien en luteoxantien. Daar is ‘n moontlikheid dat sommige afbreekprodukte reeds in die weefsel teenwoordig was as gevolg van liofilisering (aangesien die water in die korrel reeds verwyder was en die suur gekonsentreerd was). Meer werk is nodig om die effek van liofilisering en berging op die samestelling van druifweefsel van verskillende rypheid te bepaal. Die ekstraksiemetode vir druifkorrelweefsel op verskillende stadia van rypwording moet ook verder geoptimaliseer word om weefselsuurheid doeltreffend te neutraliseer, sonder om die ekstraksie van karotenoïede noemenswaardig te kompromitteer, veral in groen korrelweefsel. Die vraag of cis-isomere en chlorofil afbreekprodukte natuurlik in die druifkorrels teenwoordig is en of hulle tydens monsterneming en prosessering gevorm word, kon nie in hierdie studie beantwoord word nie. Hierdie studie het bevestig dat karotenoïede en chlorofille oor die algemeen op ‘n korrel (μg/korrel) en konsentrasie (μg/g) basis afneem vanaf deurslaan tot oes. Hierdie studie het nie daarin geslaag om te toon dat groeikragverskille vanaf voor-deurslaan (ertjiekorrelgrootte) tot oes ‘n effek het op die tempo van sintese/afbreking van karotenoïede, chlorofil en ander rypwordingsparameters nie, naamlik op appelsuur, totale glukose en fruktose, totale tannien en totale antosianien. Daar is ook in hierdie studie geen noemenswaardige effek van grondwaterinhoud op karotenoïede, chlorofille en rypheidsparameters gevind nie, heel moontlik as gevolg van die feit dat hoë grondwaterkapasiteit in die laer grondlae gevind is, wat betekenisvolle verskille in wingerdwaterstatus kon verhoed het. Eksperimentele persele wat gekies is vir groeikragverskille op grond van genormaliseerde verskil plantegroei indeks (NDVI) beelde, snoeimassa en grondwatermetings met ‘n neutronvogmeter het net in die eerste 30 cm van die grond noemenswaardige verskille in grondwaterinhoud getoon vir die rypwordingseisoene wat bestudeer is. Voor-sonopkoms plantwaterpotensiaalmetings het egter aangedui dat geen van die eksperimentele wingerdstokke ernstige waterstres ervaar het nie. Sulke stres is voorheen aangedui om ‘n effek op die karotenoïedinhoud van druiwe te hê. Die karotenoïed 5,8-epoksi--karoteen is vir die eerste keer in druiwe gekwantifiseer en verteenwoordig ‘n noemenswaardige hoeveelheid van die totale karotenoïede wat met oes teenwoordig is. Al die karotenoïede en chlorofille behalwe -karoteen blyk sensitief vir seisoenale verskille in klimaatstoestande te wees. Luteïen en β-karoteen was die volopste karotenoïede in die Merlot-druifkorrels, tesame met chlorofil a, vir beide seisoene wat bestudeer is. Die waardes van hierdie karotenoïede was ook goed gekorreleer met vorige navorsing. Chlorofil a is egter in baie groter hoeveelhede in Merlot-korrels gevind in vergelyking met dít wat in die data gerapporteer is. Die rede hiervoor is moontlik dat die chlorofil-afbreekprodukte in hierdie studie in die berekening van chlorofil a ingesluit is. Meerveranderlikeontleding het belowende voorlopige voorspellingsmodelle getoon (met korrelasiewaardes van meer as 0.8 vir beide die seisoene wat geanaliseer is) vir die voorspelling van die konsentrasie van rypheidsparameters (glukose, fruktose, appelsuur, totale tanniene en antosianiene) met karotenoïed- en chlorofilinhoud. Hierdie resultaat beklemtoon die geleentheid vir die ontwikkeling van ‘n vinnige, nie-destruktiewe metode om karotenoïede en chlorofille in korrels te meet, wat op sy beurt optimate rypheid kan voorspel. Aangesien karotenoïede die voorlopers van C13-norisoprenoïed aromaverbindings in wyn is, kan ‘n voorskou van die potensiële bydrae van hierdie aromas tot wyn moontlik verder evalueer word. Verdere navorsing is nodig om die moontlikheid van die bou en geldigheidsbepaling van sulke modelle te ondersoek.

MScAgric
ENGLISH ABSTRACT: The Little Karoo region of South Africa stretches from Montagu in the west, through Barrydale on the Langeberg Mountain, towards Ladismith, Calitzdorp, Oudtshoorn and De Rust in the east, with the Swartberg mountain range in the north. The Wine of Origin district of Calitzdorp is a small, demarcated area around Calitzdorp in the Little Karoo, surrounded by the Rooiberg, Swartberg and Kleinberg mountains. With a mean February temperature (MFT) of 23.7ºC and a low annual rainfall of 233 mm, the district of Calitzdorp has a similar climate to that of the Douro Demarcated Region (DDR). The MFT is comparable to the DDR mean July temperature, and it seems that the Douro Superior sub-region to the east of the DDR has a mean July temperature of higher than 25°C. In the Cima Corgo sub-region (in the centre of the DDR), and the Baixo Cargo sub-region, the mean July temperatures are ±25°C and ±22°C respectively. Annual rainfall in the DDR is much higher, with Baixo Cargo recording 1 018 mm, Cima Corgo recording 658 mm and Douro Superior in the east recording only 437 mm. Touriga Naçional is one of the highest quality Portuguese red grape varieties. It produces high-quality port-style wine as well as table wines. Excellent quality Touriga Naçional wines have a dark black/purple colour, good extract, high, elegant tannin content and intense aromas, with typical plum, raisin, wild fruit, mulberry, “fynbos” and cherry aromas. The most suitable terroir for Touriga Naçional in the DDR has been found to be on sites that restrain the natural vigour of the grapevine. Soils with moderate to low water-holding capacity, in association with low rainfall, result in water deficits during the growing season and are considered optimal to restrict growth vigour. A steep, northern middle slope is ideal in the southern hemisphere for high temperatures and sunlight interception. Warm temperatures (25 to 30 °C) during the day and cooler temperatures during the night are optimal for photosynthesis and colour development. In order to study factors affecting the quality of Touriga Naçional in Calitzdorp, two Vitis vinifera L. cv. Touriga Naçional commercial vineyards in the Calitzdorp district were selected. Each vineyard was divided into two separate management blocks based on their empirically determined quality of production. Two crop-reduction treatments, the standard 50% crop reduction (which was considered to be the control) and a further less drastic treatment of 25% crop reduction, were applied. Significant differences were found in viticultural performance between the two adjacent Touriga Naçional management blocks in each vineyard, especially with respect to vigour. The upper management blocks, which provided grapes for reserve-quality port-style wines, experienced a higher water deficit due to the moderate soil water-holding capacity and higher temperatures in comparison to the lower sites. The higher water deficits had a restraining effect on the Touriga Naçional vines, and therefore the upper sites had lower vigour, which contributed to better quality of both the wine and port-style wine, and this could be recognised sensorially. However, it was not reflected in the chemical analytical results. Crop load also appeared to have an effect on the Touriga Naçional grapevines, but this appeared to be dependent on the management block. The 50% crop reduction had a significant positive effect on the sensory analyses, but did not significantly affect the chemical analyses. Calitzdorp terroir has a similar effect on Touriga Naçional compared to the DDR terroir, and that is why Calitzdorp can produce good table and port-style wines from Touriga Naçional.
AFRIKAANSE OPSOMMING: Die Klein Karoo-streek in Suid Afrika strek vanaf Montagu in die weste, deur Barrydale teen die Langeberg, na Ladismith, Calitzdorp, Oudtshoorn en De Rust in die ooste, met die Swartberg in die noorde. Die distrik van Calitzdorp is ‘n klein area rondom Calitzdorp, in die Klein Karoo, wat deur die Rooiberg, Swartberg en Kleinberg omring is. Calitzdorp het ‘n gemiddelde Februarie-temperatuur (GFT) van 23.7°C en ‘n lae jaarlikse reënval van 223 mm, wat soortgelyk is aan die klimaat van die Douro Vallei in Portugal. Die Douro Vallei se gemiddelde Julie-temperatuur (GJT) in vergelyking met die GFT van Calitzdorp is hoër, met temperature van meer as 25°C in die substreek Douro Superior. Vir die substreke Cima Cargo en Baixo Cargo is die GJT ±25°C en ±22°C onderskeidelik. Die jaarlikse reënval is ook hoër by Baixo Cargo, met 1 018 mm, Cima Cargo met 658 mm en Douro Superior met slegs 437 mm. Touriga Naçional is een van die beste Portugese rooi kultivars wat hoëkwaliteit tafel- en portwyne produseer. ‘n Tipiese hoëkwaliteit Touriga Naçional-wyn het ‘n swartpers kleur, hoë ekstrak, hoë elegante tanniene en intense aromatiese geure wat tipiese pruim, rosyne, wilde vrugte, moerbei, fynbos en kersie aromas insluit. Die geskikste terroir vir Touriga Naçional is op swak gronde wat die natuurlike groeikrag van die wingerdstok strem. Gronde met matige tot lae grondwaterhouvermoë tesame met lae reënval veroorsaak ‘n waterstremming in die wingerdstok gedurende die groeiseisoen en word as optimaal beskou omdat dit beheersde groei veroorsaak. In die suidelike halfrond word relatief steil, noordelike middelhange as ideaal beskou vir hoë temperature en maksimale sonligonderskepping. Gepaardgaande hiermee is die interne dreinasie verantwoordelik vir vinniger uitdroging van die grond. Hoë temperature (25 tot 30°C) gedurende die dag en koue nagte is optimaal vir fotosintese en kleurontwikkeling. Twee Vitis vinifera L. cv. Touriga Naçional kommersiële wingerde in die Calitzdorp-distrik is geselekteer en in twee afsonderlike bewerkingsblokke verdeel, gebaseer op kwaliteitsverskille. In elke blok was die verdeling van so ‘n aard dat daar ‘n hoërliggende helfte en ‘n laerliggende helfte was. Twee trosverminderingsbehandelings, nl. 50% (kontrole) en 25%, gebaseer op trosgetalle, is toegepas. By elkeen van die wingerde was daar betekenisvolle groeiverskille tussen die twee aangrensende helftes. Die boonste helftes (of gedeeltes) het minder gegroei a.g.v. ‘n hoër waterstremming sowel as hoër temperatuur as die laer helftes. Dit het geblyk om ‘n positiewe invloed op die kwaliteit van beide die tafel- en portwyne uit te oefen. Troslading het ook ‘n effek op die Touriga Naçional-wingerde gehad, maar dit blyk of dit blok-afhanklik is. Die 50% trosverminderingsbehandeling het ‘n beduidende positiewe verskil in die sensoriese analises gemaak, maar nie ‘n beduidende verskil in die chemiese analises van die wyne nie. Calitzdorp se terroir het ‘n soortgelyke effek op Touriga Naçional as dié van die DDR terroir en daarom kan Calitzdorp soortgelyke goeie tafel- en portwyne van Touriga Naçional produseer.

Thesis (MScAgric) -- University of Stellenbosch, 2001.
ENGLISH ABSTRACT: There is an increasing demand by the consumer for knowledge and understanding of the origin of each wine produced. This origin is directly linked to the interaction between the environment and grapevine, and therefore to the terroir. A terroir can be defined as a complex of natural factors being expressed through the final product and must therefore be studied in two steps, namely, the identification of relatively homogenous natural terroir units followed by their ecophysiological characterisation. The aim of this study was to characterise the Bottelaryberg-Simonsberg-Helderberg wine growing area according to existing digital information and to identify natural terroir units. The study area is situated to the southwest of Stellenbosch and covers an area of approximately 25 000 ha. Topography is a static feature of the landscape and affects the sunlight interception by a slope, exposure of a site to winds and drainage of soil water and air. It forms an important component of the terroir concept and has a strong interaction with the environmental components of climate and soil. The study area is bordered by mountains and bisected by a river valley resulting in a large variation in aspect and altitude affecting both spatial and temporal temperature variability. There is no doubt as to the important effect of climate on wine character and quality. A number of indices can be used to describe the regional climate. The study area has a Mediterranean climate with notable spatial variation of all climatic parameters due to its complex topography and proximity to the ocean. The effect of geology on wme character is less clear but appears to act through its contribution to the physical properties of the soil. In the Bottelaryberg-Simonsberg- Helderberg study area the geology is complex due to the high degree of tectonic movement and mixing of parent material. In situ weathering of rocks is seldom the only source of soil formation. Soil has a number of contributing factors affecting wine character and quality, inter alia, soil colour, temperature, chemical composition, depth and texture (affecting the water supplying properties of the soil). It is the last two that appear to have the most significant effect. Soil depth affects the buffer capacity of the soil to temper climatic extremes and the water supply has a well-recorded effect on vine growth and functioning. There is a high degree of soil variation in the Bottelaryberg-Simonsberg-Helderberg study area that is difficult to represent in soil associations. A pattern of soil distribution, however, can be noticed in relation to landscape variation. Terrain morphological units, altitude and aspect were used as pnmary keys for the identification of natural terroir units. Broad soil categories and geological information were included at a secondary level. This resulted in 195 units. These natural terroir units will form the basis for future ecophysiological characterisation in order to determine possible future cultivar distribution as well as the terroir effect on wine character.
AFRIKAANSE OPSOMMING: Daar is toenemende aanvraag vanaf die verbruiker vir kennis en begrip van die oorsprong van elke wyn wat geproduseer word. Die oorsprong is direk aan die interaksie tussen die omgewing en wingerd gekoppel, en daarom ook die terroir. Terroir kan omskryf word as die kompleks van natuurlike faktore wat in die finale produk uitdrukking vind en studies daarvan moet dus in twee stappe plaasvind, d.i. die identifisering van relatiewe homogene natuurlike terroir eenhede gevolg deur die ekofisiologiese karakterisering daarvan. Die doel van die studie was om die Bottelaryberg-Simonsberg-Helderberg wynproduserende gebied te karakteriseer, met behulp van bestaande versyferde data en om natuurlike terroir eenhede te identifiseer. Topografie is 'n vaste landskapskenmerk en beïnvloed sonligonderskepping deur hellings, blootstelling van 'n ligging aan wind en die dreinerin~ van grondwater en beweging van koue lug. Dit is 'n belangrike deel van die terroir konsep en het sterk interaksie met omgewingsfaktore van grond en klimaat. Die studiegebied word begrens deur berge en is deursny deur 'n riviervallei, met groot variasie in helling en aspek. Beide hiervan beïnvloed die ruimtelike- en daaglik-variasies in daaglikse maksimum temperatuur. Die belangrike effek van klimaat op wynkarakter en kwaliteit is duidelik soos weerspieël deur die beskikbaarheid van 'n aantal makroklimaatsindekse. Alle klimaatsparameters word deur topografie beïnvloed. Temperatuur en relatiewe humiditeit word ook deur afstand vanaf die see beïnvloed. Die studie area het 'n Meditereense klimaat, met merkwaardige ruimtelike variasie van alle klimaatsparameters. Die effek van geologie op wynkarakter is minder duidelik, maar het waarskynlik 'n effek op die fisiese eienskappe van 'n grond. Die geologie in die Bottelaryberg-Simonsberg- Helderberg studiegebied is kompleks, as gevolg van die hoë graad van tektoniese beweging en vermenging van moedermateriaal. In situ vorming van gronde uit gesteentes vind selde plaas: Grond het 'n groot aantal bydraende faktore, wat wynkarakter en -kwaliteit kon beïnvloed, onder andere, grondkleur, -temperatuur, -chemiese samestelling, -diepte en -tekstuur (wat die waterleweringsvermoë van die grond bepaal). Dit is veral laasgenoemde twee faktore wat blykbaar die mees opvallendste effek openbaar. Gronddiepte bepaal die buffer kapasiteit van die grond om klimaatsuiterstes te kan weerstaan en die waterleweringsvermoë het 'n alombekende effek op wingerdgroei en funksionering. Daar is 'n hoë graad van grondvariasie in die Bottelaryberg-Simonsberg-Helderberg studiegebied wat moeilik is om met grondassosiasies weer te gee. Daar is wel 'n patroon van grond distribusie wat verband hou met landskapvariasie. Terrein morfologiese eenhede, hoogte en aspek is gebruik as primêre sleutels vir die indentifisering van natuurlike terroir eenhede. Globale grondkategorieë en geologiese inligting is op 'n sekondêre vlak gebruik. Dit het 195 eenhede tot gevolg. Die natuurlike terroir eenhede sal die basis vorm vir toekomstige ekofisiologiese karakterisering, om die terroir effek op wynkarakter te kan bepaal.

Thesis (PhD(Agric))--Stellenbosch University, 2012.
ENGLISH ABSTRACT: A lack of defects is required for successful table grape marketing, which pre-suppose optimal vine performance, berry development and post-harvest quality. The supply of mineral nutrients affects vine development, physiology and berry quality. Despite a vast amount of research conducted over decades, there remain many unresolved issues regarding table grape vine nutrition to ensure optimal table grape quality and shelve-life. Unjustified fertilisation practices often include excessive applications of nitrogen (N), potassium (K) and calcium (Ca). A four-year field trial was therefore conducted on a sandy soil in the Paarl district of South Africa, using grafted on Ramsey, and trained to a gable trellis system. Nitrogen, potassium and calcium were applied, singular or in combination, at rates up to 300% the calculated annual nutritional requirement. The effect of these excessive applications on table grape performance under typical South African cultivation conditions was investigated for Vitis vinifera L. cv. Prime Seedless, a very early seedless table cultivar that is produced with minimum berry diameter of 18mm, with special reference to 1) vegetative growth, 2) expression of grapevine nutrient availability through foliar analyses, 3) berry nutrient accumulation patterns of this early cultivar, 4) manipulation of berry nutrient content through soil and bunch directed applications and 5) the effect of berry nutrient content on its quality. No definite vegetative growth responses (expressed as shoot length, leaf surface area and shoot mass) and leaf chlorophyll content differences were obtained for all the treatments. These results were obtained in a vineyard on a sandy soil where excessive N fertilisation caused a reduction of soil pH to detrimentally low levels and where the excessive N, K and Ca applications reduced mutual concentrations and that of Mg, in the soil. A lack of stimulation in vegetative growth may therefore be ascribed to the combined negative effect of these excessive applications on soil pH and vine nutrition. Although the N content of petioles was higher for treatments where N was applied, consistent significant increases in petiole N with N fertilisation were not observed. Petiole N concentration showed a decreasing trend throughout the season. Petiole K concentrations were significantly increased by the K fertilisation at all phenological stages. None of the K fertilisation treatments, however, succeeded to raise petiole K concentrations above the accepted maximum norms and petiole K concentration at a specific sampling stage varied significantly between the four seasons. A general decrease in petiole K concentration was found for all seasons. Calcium fertilisation did not increase soil Ca content, resulting in a lack of differences in petiole Ca concentrations between treatments. An increase in petiole Ca concentration towards harvest was obtained. Correlations between petiole nutrient concentration and berry mineral content at harvest were poor. The only way of knowing the mineral content of berries would seem to be by measuring it directly instead of deducing it from the results of leaf or petiole analyses. The dynamics of berry growth impacted on berry nutrient concentration. Early rapid berry growth, predominantly due to cell division and cell growth, was associated with the most rapid decreases in N, P and Ca concentration. Due to mobility of K and Mg in the plant, that exceeds other nutrients, the decrease in concentration of these two mineral elements was not as pronounced as that of the others. Nutrient accumulation was most rapid during the pre-véraison period, but only Ca showed a definite termination during the early ripening period. The continued inflow of N, P, K and Mg, albeit at slower rates immediately after véraison, should be taken into consideration when fertilisation is applied. As a table grape, total accumulation of each nutrient in Prime Seedless berries also far exceeded that of other cultivars studied thus far. A particular difference is that the berry flesh:skin ratio is much higher than that of previously studied cultivars, leading to higher levels of nutrient accumulation in the flesh. Slightly larger berry size was obtained for N applications and is ascribed to slight increases in early vegetative growth, allowing a better response to GA3 treatments. The use of GA3 for berry enlargement is also considered the reason why K fertilisation, resulting in increased berry K levels, did not affect berry size, as is often found for wine grapes. Higher available NO3 - in the soil on account of excessive N applications resulted in higher levels of berry N, despite sub-optimal soil pH regimes that were created by these treatments. Berry K concentration and content were increased by K fertilisation. Rapid vine K uptake and translocation to the berries seem to negate the reduced vine nutritional status as observed in petioles for situations of over-fertilisation with N. Berry Ca levels were not increased by Ca fertilisation or by bunch applied Ca. The rapid rates of berry growth, together with low rates of berry Ca uptake and Ca uptake that terminates at the onset of ripening, are assumed to be the main reasons for this result. Low levels of decay as well as a lack of consistently increased decay were obtained for N containing treatments. Nitrogen levels in the berries above which their susceptibility to fungal infection is increased, should be established. Information on specific N compounds that may lead to more susceptibility is required. Potentially increased berry browning on account of high rates of K fertilisation needs to be further investigated; indications that this may occur were observed. Neither soil applied Ca nor bunch applied Ca improved berry quality, although Ca treatments seemed to reduce decay during the only season that significant differences were obtained. The negative effect of excessive fertilisation on soil chemistry of sandy soils has again been highlighted by this study. This annuls the fertilisation, leading to inefficient fertilisation and a lack of the desired responses. As indicator of vine nutrient availability, petiole analysis, was proven unreliable and should be evaluated in parallel with soil analyses, taking seasonal variation into consideration. The danger of being only guided by published norms for leaf nutrient concentrations when establishing fertilisation practices has again been highlighted by this study. This research indicated that for a very early cultivar like Prime Seedless, nutrient accumulation dynamics can already start to change during the pre-véraison period in some seasons. This is due to different edaphic and climatic conditions as well as berry size, which leads to much higher flesh:skin ratios. Future research on table grapes would need to develop an understanding of the various factors and dynamics that determine berry nutrient concentration and accumulation of early ripening, large berry sized, seedless table grape cultivars.
AFRIKAANSE OPSOMMING: Suksesvolle bemarking van tafeldruiwe is ten nouste afhanklik van die beskikbaarheid van druiwe sonder defekte, wat ‘n direkte verband met optimale wingerdprestasie, korrelontwikkeling en na-oes kwaliteit inhou. Voorsiening van minerale voedingstowwe beïnvloed die stok se groei, fisiologie en korrelgehalte. Ten spyte van ‘n oorweldigende hoeveelheid navorsing wat oor dekades reeds gedoen is, is daar steeds onopgeloste kwessies aangaande bemesting van tafeldruiwe vir optimale druifgehalte en houvermoë. Die gevolg is onoordeelkundige bemestingspraktyke wat o.a. aanleiding gee tot oorbemesting met stikstof (N), kalium (K) en kalsium (Ca). ‘n Vier-jaar-lange veldproef is gevolglik op ‘n sandgrond in die Paarl distrik (Suid-Afrika) onderneem deur gebruik te maak van Vitis vinifera L. cv. Prime Seedless geënt op Ramsey en op ‘n dubbel-gewel prieelstelsel opgelei is. Stikstof, K en Ca is alleen, of in kombinasie, toegedien teen hoeveelhede gelykstaande aan 300% van die wingerd se jaarlikse behoefte. Die effek van hierdie oormatige toedienings op tafeldruif prestasie onder Suid-Afrikaanse verbouingstoestande is ondersoek, met spesiale verwysing na 1) vegetatiewe groei, 2) uitdrukking van voedingstofbeskikbaarheid deur blaarontledings, 3) die voedingstof akkumulasie patrone van korrels van hierdie vroeë kultivar, 4) manipulasie van korrel voedingstofinhoud deur grond en trosgerigte toedienings en 5) die effek van korrel voedingstofinhoud op kwaliteit. Die doel van die proef was om bemestinspraktyke van Prime Seedless, ‘n baie vroeë pitlose tafeldruifkultivar met ‘n minimum korrelgrootte van 18 mm, te verfyn. Deur die akkumulasie patrone van die druiwe uit te klaar is daar ook ondersoek ingestel of oestyd en na-oes gehalte deur oormatige toediening van voedingstowwe affekteer word. Geen duidelike verskille betreffende vegetatiewe groeireaksies (uitgedruk as lootlengte, blaaroppervlaktes en lootmassas) asook verskille in blaar chlorofilinhoud is vir die behandelings verkry nie. Hierdie resultate is verkry in ‘n wingerd op ‘n sandgrond, waar oormatige N-bemesting aanleiding gegee het tot grond pH verlagings tot die peil van nadelige vlakke. Verder het die oormatige N, K en Ca toedienings wederkerige verlagings in konsentrasies, asook op dié van Mg, in die grond teweeggebring. Die tekort aan vegetatiewe groeiresponse op die behandelings kon dus toegeskryf word aan ‘n gekombineerde effek van die oormatige toedienings op grond pH en voedingstofbalanse. Hoewel die N-inhoud van bladstele hoër was vir behandelings wat N toediening ingesluit het, was daar nie konstante toenames in die vlakke verkry nie. Bladskyf N-konsentrasie het afgeneem deur die loop van die groeiseisoen. Vir alle fenologiese stadiums was bladskyf Kkonsentrasies betekenisvol verhoog deur K-bemesting. Nie een van die Kbemestingsbehandelings het egter daarin geslaag om bladskyf K inhoud vir enige monstertyd bo die algemeen aanvaarde maksimum norms te lig nie. Verder het bladskyf K inhoud by ‘n spesifieke fenologiese stadium ook betekenisvol tussen seisoene verskil. Die K-inhoud van bladskywe het afgeneem met verloop van die seisoen. Kalsiumbemesting het nie die grond se Ca inhoud deurgans verhoog nie, wat dus die tekort aan verskille in Ca konsentrasies tussen die behandelings verklaar. ‘n Toename in Ca konsentrasie en korrel Ca inhoud is vanaf set tot oes waargeneem. Swak korrelasies tussen bladskywe se voedingstofinhoude en korrels se voedingstofinhoude is verkry. Die enigste manier waarop korrels se voedingstofinhoude dus afgelei kan word, blyk te wees deur direkte bepaling daarvan. Voedingstofinhoude van korrels is deur groeipatrone daarvan beïnvloed. Vroeë korrelgroei, hoofsaaklik a.g.v. seldeling en selgroei, het met die vinnigste afnametempo van N, P en Ca gepaard gegaan. As gevolg van die hoër beweeglikheid van K en Mg in die plant in vergelyking met ander voedingstowwe, was die afname in konsentrasie van hierdie twee elemente nie so groot soos vir die ander nie. Voedingstofakkumulasie was die vinnigste in die periode voor deurslaan. Slegs Ca het ‘n beeïndiging van opname aan die einde van hierdie periode getoon. Die voortgesette opname van N, P, K en Mg, alhoewel stadiger kort na deurslaan, moet in ag geneem word wanneer bemesting toegedien word. Vir hierdie kultivar het die totale opname van elke bemestingstof dié van die ander kultivars wat tot hede bestudeer is, ver oorskry. ‘n Spesifieke verskil is ‘n baie hoër vleis:dop verhouding as wat vir ander kultivars verkry is. Dit gee aanleiding tot baie hoër vlakke van voedingstofakkumulasie in die vleis. Effens groter korrelgroottes is verkry waar N toedienings gemaak is. Dit word toegeskryf aan klein toenames in vroeë vegetatiewe groei, wat dus beter reaksie op GA3 behandelings tot gevolg gehad het. Die gebruik van GA3 vir korrelvergroting word ook beskou as die rede waarom K-bemesting, wat tot hoër vlakke van K in die korrels aanleiding gegee het, nie korrelgrootte, soos by wyndruiwe, bevorder het nie. Hoër NO3 - in die grond (water), na aanleiding van N toedienings, het aanleiding gegee tot hoer vlakke van N in die korrels. Dit het plaasgevind ten spyte van sub-optimale grond pH wat deur die oormatige N toedienings veroorsaak is. Korrel K konsentrasie en -inhoud is deur K-bemesting verhoog. Vinnige opname en translokasie van K na die korrels het ook geblyk die rede te wees waarom die verlaagde voedingstatus van die stokke a.g.v. oorbemesting met N nie die korrels se K inhoud geaffekteer het nie. Die vinnige groeitempo van die korrels, tesame met lae vlakke van Ca opname, asook korrels se Ca opname wat tydens rypwording ophou, word as die redes vir die tekorte aan behandelingseffekte beskou. Lae vlakke van bederf, asook ‘n tekort aan betroubare tendense dat bederf deur Nbemesting verhoog word, is verkry. Daar moet vasgestel word of daar N vlakke in die korrels is waarbo hul vatbaarheid vir swaminfeksies verhoog word, en of daar spesifieke N verbindings is wat die korrels meer vatbaar maak vir bederf. Indikasies dat K-bemesting interne verbruiningsvlakke verhoog het, regverdig verdere ondersoek. Korrelkwaliteit is nie deur grond- of trosgerigte toedienings bevoordeel nie. Die negatiewe effek van oormatige bemesting op die chemiese samestelling van sandgronde is weer deur hierdie navorsing uitgelig. Dit lei tot oneffektiewe bemesting en ‘n tekort aan die verlangde effekte. Blaarontledings blyk onbetroubaar te wees as aanduiding van voedingstof beskikbaarheid. Dit moet evalueer word saam met grondontledings en ook seisoenale variasie in ag neem. Die gevaar om slegs deur gepubliseerde norme gelei te word wanneer bemestingspraktyke bepaal word, is weer deur hierdie navorsing uitgelig. Voorst is daar in hierdie navorsing gevind dat voedingstof akkumulasiepatrone van ‘n baie vroeë kultivar soos Prime Seedless alreeds voor deurslaan begin verander a.g.v. omgewingstoestande en korrelgroei wat tot ‘n veel hoër vleis:dop verhouding aanleiding gee. Toekomstige navorsing op tafeldruiwe behoort die faktore en dinamika wat voedingstofkonsentrasie en -akkumulasie in korrels van vroeë, groot korrel, pitlose tafeldruifkultivars beïnvloed verder te ondersoek.

Thesis (MScAgric)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: Due to the extraordinary drought resistance of the grapevine, viticulture without irrigation in the winter rainfall coastal areas of South Africa is a feasible and commonly used practice. Wine quality is largely determined by the quality of the grapes from which it is made. Grapevine physiology is affected both directly and indirectly by water stress, which may vary according to soil type and prevailing atmospheric conditions. The water status of the grapevine can affect grape composition profoundly, either directly or indirectly, in either a positive or negative way, depending on the degree as well as the duration of water stress. There are three important factors involved in the development of water stress, namely the transpiration rate, the rate of water movement from the soil to the roots, and the relationship of soil water potential to leaf water potential. All three these factors are affected by atmospheric and/or soil conditions. In warm winelands such as South Africa (Western Cape), with a mediterranean climate which is characterised by a hot, dry summer period, the most important characteristic of soil is its ability to supply sufficient water to the grapevine during the entire growing season. Leaf water potential (Ψl) has gained wide acceptance as a fundamental measure of grapevine water status, and has been widely applied in viticultural research. Shortly before dawn, Ψl approaches equilibrium with soil water potential and reaches a maximum daily value. The study formed an integral part of a comprehensive, multi-disciplinary research project (ARC Infruitec-Nietvoorbij Project No. WW13/01) on the effect of soil and climate on wine quality, which commenced in 1993 and will be completed in 2004. This study was conducted during the 2002/03 growing season in two Sauvignon blanc vineyards situated at Helshoogte and Papegaaiberg, both in the Stellenbosch district, approximately nine kilometres apart. Two experiment plots, representing contrasting soil types in terms of soil water regime, were selected in each vineyard. At Helshoogte the two soils represented the Tukulu and Hutton forms, and the soils at Papegaaiberg were of the Avalon and Tukulu forms. The aim of this study was to determine the effect of atmospheric conditions and soil water status on the level of water stress in the grapevines for each soil at each locality, as well as the effect of grapevine water stress on yield and wine quality. This was done by determining and comparing the soil water status, soil water holding capacity of the soils and the evapotranspiration of the grapevines on the two different soils, at each of the two localities differing in mesoclimate and topography. The atmospheric conditions at the two localities during the 2002/03 season were also determined and compared to the long-term average atmospheric conditions, and the level of water stress of grapevines on each soil at each locality was measured. During the 2002/03 growing season, atmospheric conditions were relatively warm and dry in comparison to the long-term averages of previous seasons. These conditions accentuated the effects of certain soil properties that may not come forward during wetter, normal seasons. The usually wet Tukulu soil at Helshoogte was drier than expected during the 2002/03 season compared to the Hutton soil. Due to more vigorous growth on the Tukulu soil, grapevines extracted more soil water early in the season, leading to a low soil water matric potential and more water stress in the grapevines. Due to the higher vigour, resulting in more canopy shading, and more water stress, the dominant aroma in wines from the Tukulu soil was fresh vegetative. The Hutton soil maintained consistency with regards to both yield and wine quality compared to previous seasons. On the other hand the Tukulu soil supported a higher yield, but with lower than normal wine quality. The Avalon soil at Papegaaiberg maintained the highest soil water potential towards the end of the season, probably due to capillary supplementation from the sub-soil. Grapevines on the Tukulu soil at Papegaaiberg experienced much higher water stress than ones on the other three soils, especially during the later part of the season. This could be ascribed to a combination of factors, the most important being the severe soil compaction at a shallow depth, seriously limiting rooting depth and root distribution, which is detrimental to grapevine performance. Both the soil water status and atmospheric conditions played important roles in determining the amount of water stress that the grapevines experienced at different stages. The air temperature and vapour pressure deficit throughout the season were consistently lower at Helshoogte, the cooler terroir, compared to Papegaaiberg, the warmer terroir. At flowering, Ψl was lower for grapevines at Helshoogte than at Papegaaiberg, showing that diurnal grapevine water status was primarily controlled by soil water content. The difference in grapevine water status between the two terroirs gradually diminished until it was reversed during the post harvest period when Ψl in grapevines at Papegaaiberg tended to be lower compared to those at Helshoogte. The relatively low pre-dawn Ψl at Helshoogte indicated that the grapevines were subjected to excessive water stress resulting from the low soil water content. However, grapevines at Helshoogte did not suffer material water stress (i.e. Ψl < -1.20 MPa) during the warmest part of the day, suggesting that partial stomatal closure prevented the development of excessive water stress in the grapevines. This suggests that low pre-dawn Ψl values do not necessarily imply that grapevines will experience more water stress over the warmer part of the day, or visa versa. This does not rule out the possibility that side-effects of partial stomatal closure, such as reduced photosynthesis, can have negative effects on grapevine functioning in general. These results also suggest that measurement of diurnal Ψl cycles at various phenological stages is required to understand and quantify terroir effects on grapevine water status.
AFRIKAANSE OPSOMMING: Danksy die droogteweerstand van die wingerdstok is die verbouing van wingerde sonder besproeiing ‘n praktiese en algemene verskynsel in die winterreënval-areas van Suid-Afrika. Wynkwaliteit word grootliks bepaal deur die kwaliteit van die druiwe waarvan dit gemaak word. Wingerdfisiologie word direk en indirek beïnvloed deur waterstres, wat kan varieer volgens die grondtipe en die heersende atmosferiese toestande. Die waterstatus van die wingerdstok beïnvloed druifsamestelling, direk of indirek, en positief of negatief, afhangend van die graad en tydsduur van die waterstres. Daar is drie belangrike faktore betrokke by die ontwikkeling van waterstres, naamlik die transpirasietempo, die tempo van waterbeweging vanaf die grond na die wortels, en die verhouding tussen die grondwatermatrikspotensiaal tot blaarwaterpotensiaal. Al drie die faktore word beïnvloed deur die atmosferiese en/of grondtoestande. In warm wynboulande soos Suid-Afrika (Weskaap), met ‘n meditereense klimaat wat gekarakteriseer word deur ‘n warm, droë somerperiode, is die belangrikste eienskap van grond die vermoë om voldoende water aan die wingerdstok te verskaf gedurende die hele seisoen. Blaarwaterpotensiaal (Ψl) het wye aanvaarding bekom as die fundamentele meting van wingerstokwaterstatus, en word wyd toegepas in wingerdkundige navorsing. Kort voor sonsopkoms, nader Ψl ‘n ewewig met die grondwatermatrikspotensiaal en bereik ‘n maksimum daaglikse waarde. Die studie vorm ‘n integrale deel van ‘n omvattende, multi-dissiplinêre navorsingsprojek (ARC Infruitec-Nietvoorbij Projek No. WW13/01) op die effek van grond en klimaat op wynkwaliteit, wat in 1993 in aanvang geneem het en in 2004 afgehandel sal word. Hierdie studie is uitgevoer gedurende die 2002/03 seisoen in twee Sauvignon blanc wingerde geleë by Helshoogte en Papegaaiberg, beide in die Stellenbosch distrik, ongeveer nege kilometer van mekaar. Twee eksperimentele persele, elkeen verteenwoordigend van kontrasterende grondtipes in terme van grondwaterregime, is geselekteer in elke wingerd. By Helshoogte word die twee gronde verteenwoordig deur die Tukulu en Hutton grondvorms, en die gronde by Papegaaiberg is van die Avalon en Tukulu vorms. Die doel van die studie was om die effek van atmosferiese toestande en grondwaterstatus op die wingerdstok se waterstatus vir elke grond by die twee lokaliteite te bepaal, sowel as die effek van die wingerdstok se waterstatus op die opbrengs en wynkwaliteit. Dit is gedoen deur die grondwaterstatus, die grondwaterhouvermoë, sowel as die evapotranspirasie van die wingerdstokke op die twee verskillende gronde by elk van die twee lokaliteite, wat verskil in mesoklimaat en topografie, te bepaal en vergelyk. Die atmosferiese toestande by die twee lokaliteite gedurende die 2002/03 seisoen is ook bepaal en vergelyk met die langtermyn gemiddelde atmosferiese toestande. Die vlakke van waterstres in wingerdstokke op elke grond by elke lokaliteit is ook gemeet. Gedurende die 2002/03 groeiseisoen, was die atmosferiese toestande relatief warm en droog in vergelyking met die langtermyn gemiddeldes van vorige seisoene. Hierdie kondisies aksentueer die effek van sekere grondeienskappe wat nie noodwendig na vore kom gedurende normale, natter seisoene nie. Die gewoonlike nat Tukulu grond by Helshoogte was droër as verwag gedurende 2002/03 in vergelyking met die Hutton grond. As gevolg van sterker groekrag op die Tukulu grond, het wingerdstokke meer grondwater onttrek vroeg in die seisoen, wat gelei het tot ‘n lae grondwatermatrikspotensiaal en meer waterstres in die wingerdstokke. Die sterker groeikrag het meer beskaduwing van die lower asook meer waterstres veroorsaak, wat gelei het daartoe dat die dominante aroma in wyne vanaf druiwe op die Tukulu grond vars vegetatief was. Die Hutton grond het bestendig gebly in terme van opbrengs en wynkwaliteit in vergelyking met vorige seisoene. Daarteenoor het die Tukulu grond ‘n hoër opbrengs gelewer, maar met laer as gewoonlike wynkwaliteit. Die Avalon grond by Papegaaiberg het die hoogste grondwatermatrikspotensiaal behou tot die einde van die seisoen, heelwaarskynlik a.g.v. kapillêre aanvulling vanuit die ondergrond. Wingerdstokke op die Tukulu grond by Papegaaiberg het heelwat meer waterstres ondervind as op die ander drie gronde, veral later in die seisoen. Dit kan toegeskryf word aan ‘n kombinasie van faktore, die belangrikse daarvan die erge grondkompaksie vlak in die grond, wat worteldiepte en -verspreiding ernstig beperk het, wat op sy beurt nadelig is vir wingerdprestasie. Beide die grondwaterstatus en atmosferiese toestande het ‘n belangrike rol gespeel in die bepaling van die hoeveelheid waterstres wat die wingerdstok op verskillende stadiums ondervind het. Die lugtemperatuur en waterdampdruktekort was regdeur die seisoen laer by Helshoogte, die koeler terroir, as by Papegaaiberg, die warmer terroir. Gedurende blom was die Ψl laer vir wingerdstokke by Helshoogte as by Papegaaiberg, wat daarop wys dat daaglikse wingerdstok waterstatus hoofsaaklik deur die grondwaterinhoud bepaal was. Die verskil in wingerdstok waterstatus tussen die twee terroirs het geleidelik verminder totdat dit omgekeer was gedurende die na-oes periode toe Ψl in wingerdstokke by Papegaaiberg geneig het om laer te wees in vergelyking met die by Helshoogte. Die relatiewe lae voorsonop Ψl by Helshoogte het daarop gedui dat die wingerdstokke aan oormatige waterstres onderwerp was. Die wingerdstokke by Helshoogte het egter nie materiële waterstres (i.e. Ψl < -1.20 MPa) gedurende die warmste gedeelte van die dag ondervind nie, wat aandui dat gedeeltelike huidmondjiesluiting plaasgevind het om die ontwikkeling van oormatige waterstres te verhoed.Dit dui aan dat lae voorsonop Ψl waardes nie noodwendig impliseer dat wingerdstokke meer waterstres gedurende die warmste gedeelte van die dag sal ondervind nie, of visa versa. Dit sluit nie die moontlikheid uit dat negatiewe neweeffekte van gedeeltelike huidmondjiesluiting, soos ‘n vermindering in fotosintese, ‘n negatiewe effek kan hê op die wingerdstok se funksionering in die algemeen nie. Hierdie resultate stel voor dat die meting van daaglikse Ψl siklusse gedurende verskeie fenologiese stadia benodig word om die effek van terroir op die wingerdstok se waterstatus te verstaan en te kwantifiseer.

Thesis (MscAgric (Viticulture and Oenology))--Stellenbosch University, 2008.
The effect of various irrigation strategies on grapevine water relations during the berry ripening period was investigated in a Shiraz/Richter 99 vineyard. Comparisons between different irrigation strategies (full/seasonal, véraison+post véraison, post véraison and no irrigation) were made. During the day, the seasonally irrigated vines experienced less water stress than the deficit treatments. Non-irrigated vines seemed to maintain higher diurnal leaf water potentials. Lower leaf water potentials indicated lower water contents in the vegetative and reproductive tissue. Full irrigation seemed to stimulate primary shoot length. Longer water deficit induced earlier and more complete shoot maturation (reserve accumulation). Re-distribution of leaf area on the shoot may occur when vines are subjected to water deficit. Extended water deficit seemed to induce earlier and restricted water loss from vegetative tissue. The water relations were reflected in the berry size. Irrigation during ripening seemed to induce a continuation of berry water loss. Transpiration losses were apparently much higher in fully irrigated vines whereas stomatal control efficiently maintained water relations in non-irrigated vines. Water deficit seemed to have enhanced the soluble solid accumulation. Irrigation treatments did not seem to affect the titratable acid and pH. The post véraison irrigation in particular seemed to favour a wide window for harvesting. Irrigation at post véraison and especially véraison+post veraison seemed to have a greater effect on the synthesis and extraction of phenolics, anthocyanins and tannins in the berry skins. Different irrigation strategies may affect grapes in such a way that different wine styles are obtained.

Thesis (MScAgric (Viticulture and Oenology)--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: ‘Crimson Seedless’ is currently one of the most important and popular table grape cultivars produced in South Africa, and as such it is of great economic value for table grape producers. Major concerns with ‘Crimson Seedless’ is that it is prone to inadequate colouring, and with increased yields the berry size decreases. An additional difficulty is that methods used to increase berry size, further impede berry colouring. A plant growth regulator (PGR) commonly used in table grape production, to enhance colour formation, is ethephon (2-chloro-ethyl-phosphonic acid, 2-CEPA). In recent years significant research has been done on the effect of sunlight on anthocyanin production in grapes, although this has primarily been on wine grape cultivars. Currently, there is limited knowledge on the effect of sunlight on table grapes, and how this might influence their anthocyanin composition and content. The effect of ethephon on colour of grapes and other fruit have been extensively researched and well documented. However, the effect of ethephon on the anthocyanin composition of ‘Crimson Seedless’ is not well known. The current study aimed to explore the effect of sunlight (by matter of exclusion) and management practices, namely defoliation and ethephon application, on the anthocyanin profile and content of ‘Crimson Seedless’. Four different treatments were applied to two ‘Crimson Seedless’ vineyards, the first site located in Paarl, and the second in De Doorns. The treatments were: 1. Naturally exposed bunches, 2. Exposed bunches treated with ethephon, 3. Bunches kept in shade boxes, 4. Shaded bunches treated with ethephon. At the De Doorns site an additional defoliation treatment was superimposed over the above treatments. An HPLC technique was modified for the separation and detailed profiling of ‘Crimson Seedless’ anthocyanins and was used to analyse the effect of the reported treatments on the anthocyanin profile of berry skins. The predominant anthocyanin in ‘Crimson Seedless’ is peonidin-3-glucoside (Pn-gluc), and this was found to be significantly increased only by ethephon application, and was not altered by sunlight or leaf removal. The responses of the other anthocyanin types varied according to the respective treatments applied. However, a general observation was that ethephon application more consistently increased the concentration of anthocyanins in berry skins than did sunlight. Leaf removal had the least significant effect on anthocyanin concentration.
AFRIKAANSE OPSOMMIMG: ‘Crimson Seedless’ is tans een van die belangrikste en gewildste tafeldruif cultivars wat in Suid-Afrika verbou word en daarom is dit van groot ekonomiese waarde vir tafeldruifprodusente. ‘Crimson Seedless’ is egter daarvoor bekend dat dit te swak kleur (volgens uitvoer spesifikasies) en tweedens is die cultivar geneig om kleiner korrels te ontwikkel wanneer die oeslading vermeerder word. ‘n Addisionele probleem is dat die praktyke wat in die industrie gebruik word om korrels te vergroot ‘n verdere negatiewe impak op ‘Crimson Seedless’ se kleur ontwikkeling kan veroorsaak. Die plant-groei-reguleerder wat algemeen in tafeldruif verbouing gebruik word, ten einde beter gekleurde druiwe te produseer, is ethephon (2-chloro-ethyl-phosphonic acid, 2-CEPA). In die laaste paar jaar was daar baie navorsing gedoen oor die effek wat sonlig het op die antosianien produksie van druiwe, maar navorsing was gefokus op wyndruif cultivars. Huidiglik is daar beperkte tegniese kennis oor die effek wat sonlig op tafeldruiwe het, en hoe dit moontlik die antosianien samestelling en inhoud kan beïnvloed. Daar is ook reeds verskeie studies gedoen en data gepubliseer oor die invloed wat ethephon op die kleur het van druiwe en ander vrugte, maar die invloed wat ethephon op die antosianien samestelling van ‘Crimson Seedless’ het, is nie wel bekend nie. Die doel van hierdie studie was om die effek van sonlig (deur uitsluiting) en bestuurspraktyke (blaarverwydering en ethephon toediening) te bestudeer en hoe dit die antosianien samestelling van ‘Crimson Seedless’ beïnvloed. Vier verskillende behandelings is toegedien in twee ‘Crimson Seedless’ wingerde, die eerste proefperseel in die Paarl en die tweede proefperseel in De Doorns. Die behandelings was: 1. Natuurlik blootgestelde trosse, 2. blootgestelde trosse met ethephon, 3. Trosse met skadubokse omhul, 4. Skaduboks trosse met ethephon. By De Doorns is ‘n addisionele blaarverwydering proef bygebring. ‘n HPLC tegniek was aangepas om die antosianien samestelling en inhoud van ‘Crimson Seedless’ te bepaal, en om die effek van die behandelings te ondersoek. Die HPLC data het getoon dat peonodien-3-glukosied (Pn-gluc) die primêre antosianien in ‘Crimson Seedless’ is met die hoogste inhoud van al die antosianiene. Pn-gluc was betekenisvol beïnvloed deur ethephon toediening, terwyl die ander behandelings geen betekenisvolle effekte daarop gehad het nie. Die effekte wat die ander antosianiene gehad het, het gevarieer volgens die behandelings wat toegedien was. ‘n Algemene observasie was dat ethephon toediening die antosianien konsentrasie in ‘Crimson Seedless’ druiwe skille meer konsekwent vermeerder het as die sonlig blootstelling. Die blaarverwydering het die minste betekenisvolle effek op die antosianien inhoud van ‘Crimson Seedless’ gehad.

Thesis (MScAgric) -- University of Stellenbosch, 2002.
ENGLISH ABSTRACT: Fungi are an extremely diverse group of organisms and, by acting as pathogens, they can colonise various other organisms, including humans, plants and animals. The effect of this is usually detrimental, not only to agricultural crops and livestock, but also to human well-being. The extensive farming of crops and livestock requires persistent control of fungal populations, commonly through the use of chemical fungicides. However, the exclusive use of fungicides is no longer a sustainable practice, as a result of serious problems, such as increasing fungicide resistance in pathogen strains, the high costs of fungicides, as well as concern about the environment. The search by producers and scientists for alternative control measures is an ongoing process. The fungal cell wall consists of polysaccharides that not only playa role in protection of the fungi, but also in relaying signals for the invasion and infection of susceptible hosts. Chitin, a polysaccharide composed of N-acteylglucosamine (GleNAc) residues linked by P-1,4 glucosidic linkages, is one of the major components of the fungal cell wall, where it plays an important role in the apical growth of the vegetative hyphae. Chitinases (EC 3.2.1.14) are abundant proteins produced by a variety of microorganisms and plants and are necessary for the hydrolysis of the chitin polymer. During the invasion of many plant species by a pathogen, the production of a specific group of proteins, designated pathogenesis-related (PR) proteins that include chitinases, is induced as part of their defence response. Due to the facts that pathogenic fungi contain chitin in their cell walls and that plant chitinases are induced upon pathogen attack, chitinases have been confirmed as an integral and crucial part of the plant's natural defence response. Chitinases have increasingly been targeted to upregulate plants' endogenous disease resistance mechanisms through transgenic overexpression in a variety of hosts. Several species of fungi, including various Trichoderma spp., are potent biocontrol agents of plant pathogenic fungi and insects. The antagonistic activities of these biological control agents towards phytopathogens are based on the secretion of extracellular hydrolytic enzymes, such as cell wall-degrading chitinase enzymes. However, biological control is not restricted to naturally occurring biocontrol agents. Through the process of genetic transformation, other fungal or yeast species can be enhanced to produce their own chitinases or other antimicrobial substances more effectively in order to yield potent biocontrol agents. Various types of chitinases have been applied in the production of fungal resistant plants and some research has been done on the application of chitinases, from a variety of microorganisms, as biological control agents. In contrast, very little is known about the antifungal activity of the Saccharomyces cerevisiae chitinase enzyme, encoded by the CTS1-2 gene. The CTS1-2 gene was utilised in this study as a candidate for overexpression in both yeast and plant expression systems to analyse the ability of the encoding chitinase to inhibit fungal growth. The first objective of this study involved the high level expression and optimisation of the secretion of the CTS1-2 gene in S. cerevisiae to render recombinant yeast with enhanced antifungal abilities and with possible applications as a biocontrol agent to control plant pathogenic fungi. It was hypothesised that high-level expression and efficient secretion would be prerequisites in a biocontrol yeast strain. To this end, two strong promoters and terminators were included in the study and the secretion of the chitinase gene was evaluated by testing three different secretion signals. The secretion signals included: the native CTS1-2 secretion signal, the S. cerevisiae mating pheromone a-factor (MFa1) secretion signal, as well as the Trichoderma reesei f3-xylanase 2 (XYN2) secretion signal. The phosphoglycerate kinase 1 (PGK1) and alcohol dehydrogenase 2 (ADH2) promoters and terminators were employed to achieve high-level expression. The results obtained from the analysis of the recombinant yeasts showed that the PGK1 promoter-terminator constructs yielded high level CTS1-2-expressing and chitinase-producing strains of S. cerevisiae PRY488. The ability of the different secretion signals to efficiently secrete the overexpressed chitinase was analysed and it was found that the non-native secretion signals delivered significantly more protein to the extracellular environment. It was thus evident that the performance of the MFa1 and XYN2 secretion signals was superior to that of the native secretion signal. The antifungal activities of the recombinant chitinases produced by these constructs were tested in in vitro assays against Botrytis cinerea. The enzymes led to a significant reduction in hyphal development, caused by extreme structural damage to the hyphal tips, the hyphal cell walls as well as the ability of the fungus to form reproductive and survival structures, thereby confirming the antifungal abilities of this enzyme. The ADH2 promoter-terminator constructs yielded CTS1-2 transcripts, but no chitinase activity could be detected with any of these strains. The reasons for this still remain unclear. The second objective of this study was to assess the potential of the yeast chitinase gene to upregulate defence against fungal infection in planta. In order to elucidate this, the CTS1-2 gene was constitutively overexpressed in tobacco plants, targeting the chitinase both to the intra- and the extracellular environment. The results obtained showed that the transgenic tobacco lines regenerated in this study stably integrated the transgene, exhibiting transgene expression as well as the production of a biologically active yeast chitinase enzyme. The F, progeny were rigorously tested for resistance to B. cinerea, and both in vitro and in planta assays confirmed that the yeast chitinase increased the plant's tolerance to fungal infection; some of the lines showed disease resistance of 65 and 70%. The plants expressing an extracellularly targeted chitinase gene are still under evaluation. Interesting results are expected relating to the effect of the chitinase on the plant surface with regards to disease resistance to fungal pathogens. In conclusion, the combined set of results from both the yeast and plant overexpression studies has confirmed the strong antifungal effect of yeast chitinases. The yeast CTS1-2 chitinase could be instrumental in the development of a new generation of yeast strains with improved antifungal capabilities. This enzyme could also play an important role in genetic transformation technologies aimed at enhanced disease resistance.
AFRIKAANSE OPSOMMING: Swamme omsluit 'n uiterste diverse groep organismes wat mense, plante en diere deur patogeniese aksie kan koloniseer. Die uitkoms hiervan op landbougewasse, die veebedryf en menslike gesondheid is gewoonlik skadelik. Uitgebreide gewas- en veeboerderye benodig voortdurende beheer van fungiese populasies, tipies deur van chemiese swamdoders gebruik te maak. Die uitsluitlike gebruik van swamdoders is egter nie meer 'n lewensvatbare praktyk nie, hoofsaaklik as gevolg van probleme soos die opbou van weerstand van patogeniese rasse teen swamdoders, die hoë kostes van die middels, asook besorgheid oor die omgewing. Die soektog na alternatiewe beheermaatreëls deur produsente en wetenskaplikes bly 'n aaneenlopende proses. Die swamselwand bestaan uit polisakkariede wat nie net In rol in die beskerming van die swam speel nie, maar ook betrokke is in die oordrag van aanvals- en infeksieverwante seine in 'n vatbare gasheer. Chitien, 'n polisakkaried bestaande uit N-asetielglukosamien (GlcNAc) residu's gekoppel deur 13-1,4glukosidiese bindings, is een van die hoofkomponente van die swamselwand, waar dit 'n belangrike rol in die apikale groei van vegetatiewe hifes speel. Chitinases (EC 3.2.1.14) is proteïene wat oorvloedig deur 'n verskeidenheid van mikroërganismes en plante geproduseer word, waar hulle vir die hidrolise van die chitien polimeer noodsaaklik is. Tydens die infeksie van verskeie plantspesies deur In patogeen, word die produksie van 'n spesifieke groep proteïene, die sogenaamde patogeen-verwante (PR) proteïene wat chitinases insluit, as deel van die plant se verdedigingsreaksie geïnduseer. Die feit dat patogeniese swamselwande chitien bevat en dat plantchitinases tydens infeksie geïnduseer word, het daartoe gelei dat dit bevestig is dat chitinases In integrale en kritiese deel van die plant se natuurlike verdedigingsreaksie uitmaak. Chitinases word toenemend geteiken in pogings om die plant se intrinsieke siekteweerstandsmeganismes te verbeter deur transgeniese ooruitdrukking daarvan in 'n verskeidenheid van gashere. Verskeie swamspesies, insluitend verskillende Trichodenna-spesies, is kragtige bio-antagoniste van plantpatogeniese swamme. Die antagonistiese aksies van hierdie biologiese beheeragente teenoor fitopatogene is gebaseer op die uitskeiding van ekstrasellulêre hidrolitiese ensieme, soos die selwandverterende chitinase ensieme. Nietemin is biologiese beheer nie net tot bio-antagoniste wat natuurlik voorkom beperk nie. Deur die proses van genetiese transformasie kan ander swam- of gisspesies verbeter word om hul eie chitinases of ander antimikrobiese substanse meer effektief te produseer, wat aanleiding sal gee tot kragtige bio-antagoniste. Verskeie tipes chitinases is al in die produksie van swambestande plante ingespan en uitgebreide navorsing is gedoen op die toepassing van 'n reeks chitinases, afkomstig van 'n verskeidenheid van mikroërganismes, as biologiese beheeragente. In teenstelling is baie min bekend oor die antifungiese aktiwiteite van die Saccharomyces cerevisiae chitinase ensiem, wat deur die CTS1-2 geen ge-enkodeer word. Die CTS1-2-geen is in hierdie studie gebruik vir ooruitdrukking in beide gis- en plantuitdrukkingsisteme om die chitinase se vermoë om swamgroei te inhibeer, te ondersoek. Die eerste oorkoepelende oogmerk van hierdie studie het hoë-vlak uitdrukking en optimalisering van sekresie van die CTS1-2-geen in S. cerevisiae behels, met die toekomstige doelwit om 'n rekombinante gis met verbeterde antifungiese eienskappe en met moontlike toepassings as 'n bio-antagonis teen plantpatogeniese swamme te ontwikkel. Die hipotese was dat hoë-vlak uitdrukking en voldoende sekresie voorvereistes vir 'n bio-antagonisras is. Omdié rede is twee sterk promotors en termineerders by hierdie studie ingesluit en is die sekresie van die chitinase-geen geëvalueer deur drie verskillende sekresieseine te toets. Die sekresieseine sluit in: die wilde-tipe CTS1-2 sekresiesein, die S. cerevisiae paringsferomoon a-faktor (MFa1) sekresiesein, en die Trichoderma reesei p-xilanase (XYN2) sekresiesein. Die fosfogliseraat kinase 1 (PGK1) en alkohol dehidrogenase 2 (ADH2) promotors en termineerders is gebruik om hoë-vlak uitdrukking te dryf. Die resultate wat vanaf die analises van die rekombinante giste verkry is, het getoon dat die PGK1 promotor-termineerder konstrukte hoë-vlak CTS1-2-uitdrukkende en chitinase-produserende S. cerevisiae PRY488 rasse opgelewer het. Die vermoë van die verskillende sekresieseine om die ooruitgedrukte chitinase voldoende uit te skei, is geanaliseer, en daar is gevind dat die heteroloë sekresieseine aansienlik meer proteïene na die ekstrasellulêre omgewing geloods het. Dit was dus duidelik dat die MFa1 en XYN2 sekresieseine beter as die wilde-tipe sekresiesein presteer het. Die antifungiese aktiwiteit van die rekombinante chitinases wat deur hierdie konstrukte geproduseer is, is ook in in vitrotoetse teen Botryits cinerea getoets. Die teenwoordigheid van die ensieme het gelei tot 'n aansienlike afname in hife-ontwikkeling, veroorsaak deur ekstreme strukturele skade aan die hifepunte, die hifeselwande, asook die vermoë van die swam om voortplanting- en oorlewingstrukture te vorm. Die ADH2 promotor-termineerderkonstrukte het CTS1-2 transkripte vertoon, maar geen chitinase-aktiwiteite kon in hierdie konstrukte waargeneem word nie. Die redes hiervoor is tot op hede onbekend. Die tweede oogmerk van hierdie studie was om die potensiaal van die gischitinase om swaminfeksie in planta teë te werk, te ondersoek. Die CTS1-2-geen is konstitutief ooruitgedruk in tabakplante, waarin die chitinase na beide die intra- en ekstrasellulêre omgewing geteiken is. Resultate het getoon dat die geregenereerde transgeniese tabaklyne die transgeen stabiel geïntegreer het, transgeenuitdrukking vertoon en dat 'n biologies aktiewe chitinase-ensiem geproduseer is. 'n F1-generasie is aan strawwe toetse onderwerp om weerstand teen B. cinerea te ondersoek. Beide die in vitro en in planta toetse het bevestig dat die gischitinase die plant se verdraagsaamheid teenoor swaminfeksie verhoog het; sommige lyne het siekteweerstand van tussen 65 en 70% getoon. Die plante wat 'n ekstrasellulêre chitinase produseer, word steeds geëvalueer. Interessante resultate word verwag aangaande die effek van die chitinase op die plant se oppervlak met betrekking tot siekteweerstand teen swampatogene. Ten slotte, die gekombineerde stel resultate wat vanaf beide die gis- en plantuitdrukkingstudies verkry is, het die sterk antifungiese effek van gischitinases bevestig. Die gis CTS1-2 kan instrumenteel wees in die ontwikkeling van 'n nuwe generasie gisrasse met verbeterde antifungiese eienskappe. Die ensiem kan ook 'n belangrike rol in genetiese transformasietegnologieë, wat op verbeterde siekteweerstand gemik is, speel.

Thesis (MscAgric (Viticulture and Oenology))--Stellenbosch University, 2008.
Over the years, different techniques have been used to legally reduce the ethanol content of wines. Several physical processes are available for producing wines with less alcohol. Despite their efficacy, these treatments have a capital and operational cost influence. They can also affect the concentration of other wine components. On the other hand, vast amount of research has been conducted through genetic modification of wine yeast strains in order to reduce the ethanol yield of Saccharomyces cerevisiae by diverting sugar metabolism towards various byproducts. However, genetically modified yeasts are not currently accepted in most wine industries worldwide, including South Africa. Therefore, other approaches need to be envisaged. Commercial enzymes are commonly added during winemaking. Most enzymes essential for vinification naturally occur in grapes, but are inefficient under pH and sulphur levels associated with winemaking. Enzymes of fungal origin are resistant to such conditions. The most widely used commercial enzymes include pectinases, hemicellulases, glucanases and glycosidases. With the exception of glucanases, produced by Trichoderma harzianium, all the other enzymes are produced by Aspergillus niger. In this study, the possibility of using Gluzyme Mono® 10.000 BG (Gluzyme) (Novozymes, South Africa) to reduce the glucose content of synthetic grape must and grape must before fermentation in order to produce wine with a reduced alcohol content was investigated. Gluzyme is a glucose oxidase preparation from Aspergillus oryzae, currently being used in the baking industry. Glucose oxidase catalyses the oxidation of glucose to gluconic acid and hydrogen peroxide in the presence of molecular oxygen. Gluzyme was initially used in synthetic grape must where different enzyme concentrations and factors influencing its activity were investigated for its use in winemaking. The results showed that up to 0.5% v/v less alcohol were obtained using an enzyme concentration of 20 kU compared to the control. This reduction in alcohol was increased to 1 and 1.3% v/v alcohol at pH 3.5 and pH 5.5 respectively in aerated synthetic grape must using 30 kU enzyme. Secondly, Gluzyme trials were carried out using Pinotage grape must. Gluzyme treated wines after fermentation contained 0.68% v/v less alcohol than the control samples at 30 kU enzyme. Colour and volatile flavour compounds of treated wine did not differ significantly from the untreated samples. Lower free anthocyanin and total phenol concentrations in treated than control samples were observed, possibly due to the hydrogen peroxide oxidation which could have led to polymerisation. The present study has clearly demonstrated that Gluzyme may be used in winemaking to produce reduced-alcohol wine without affecting its colour and aroma compounds. The enzyme in its current form is however, not ideal for winemaking; other forms such as liquid or powder form should be considered if the enzyme is to be used under winemaking conditions. Future work should focus on evaluating the potential new form of the enzyme and studying the effects of Gluzyme in various grape must in semi-industrial scale. A tasting panel should also evaluate its impact on the organoleptic properties and the overall quality of the resulting wines.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Grapevine water status is considered to be the most important factor limiting plant growth and production in the Mediterranean zones. In these regions with limited summer rainfall and limited water resources for irrigation grapevines may experience water deficits for an extended period of time. The demand of water for agriculture is constantly increasing, and will continue to do so due to the rise in the world population and to the effects of climate change on rainfall and evaporative demand in these regions. The Western Cape wine region is also classified as Mediterranean and grapevines grown in this region are often exposed to water “stress” conditions due to high evaporative demand and low water availability in the soil. Plant water status of grapevines may dependent on, amongst other factors, the water potential of soil layers close to the root system, canopy size and evaporative demand. The canopy size of a grapevine can inherently be seen as a measure of grapevine vigour, and vigour variation among grapevines within a vineyard is a common phenomenon in the Western Cape. The importance of the contributions from several factors causing vigour variation within vineyards is still a subject of debate. This may be largely ascribed to the significant amount of variability in vineyards that researchers have to deal with during viticultural studies. However, the recent advances in remote sensing technology have established new methods to assess grapevine vigour variability. In the face of the recognized variation within vineyards and the importance of a sustained grapevine water status, for wine grape productivity and -quality, it is alarming to think that a vineyard block is generally managed as a homogeneous entity when it comes to irrigation scheduling. What is more alarming is the assumption that grape, juice and wine quality will be homogeneous throughout a vineyard block – even without irrigation. With this in mind, a study was conducted to study the interaction between grapevine vigour and grapevine water status within a commercial vineyard with variable vigour by implementing various irrigation regimes. Vigour variation was identified through multispectral aerial imagery and plant-based water status determinants were used to assess grapevine water status in plots of differing vigour within the vineyard. Soil water status was also assessed, and vegetative growth quantified to ultimately determine the variability in vigour and its possible contribution to the variability through the water status of the plant. Reproductive growth was monitored continually before evaluating the effect of water status and grapevine vigour on grape composition and subsequent wine quality. The various methods used to evaluate grapevine vigour showed good correspondence. Pruning mass measured at the end of the season confirmed leaf area measurement (main leaves and lateral leaves) during vegetative growth, and corresponded well, in terms of main vigour classifications with the NDVI images collected. Berry weight and volume responded to the various classifications, with a decrease in water deficits from one classification to the next accompanying an increase in berry weight and volume. Analyses of the berry composition and wines showed statistically significant differences between the classifications. This was found for sugar content per berry, total phenols, total red pigment, malic acid, nitrogen and pH for the grape juice analyses. Wine pH and total acidity also differed significantly.
AFRIKAANSE OPSOMMING: In die Mediterreense sones word plantwaterstatus beskou as `n hooffaktor wat groei en produksie van `n wingerdstok negatief beinvloed. In hierdie sones kan wingerdstokke vir lang periodes `n tekort aan water ervaar a.g.v `n tekort aan reënwater gedurende die somer en lae beskikbaarheid van besproeingswater. Die vraag na water vir landbou is ook konstant besig om toe te neem in dié sones en die tendens sal voorduur a.g.v die groei in die wêreldbevolking, die effek van klimaatsveranderig op reënvalpatrone en die hoë verdampingsfaktor. Die wingerd- en wynstreek van die Wes-Kaap word ook geklassifiseer as Mediterreens en wingerdstokke in hierdie streek ervaar dikwels waterspanning wat deur hoë evapotranspirasie en min beskikbare grondwater veroorsaak word. Van die faktore wat die waterstatus van `n wingerdstok bepaal is onder andere die waterpotensiaal van die grondlae rondom die wortelstelsel, die grootte van die wingerdlowerraamwerk en die evapotranspirasiebehoefte. Die omvang van `n wingerdstok se lower binne die prieel word beskou as `n aanduiding van wingerdstokgroeikrag en variasie in groeikrag tussen wingerdstokke is `n algemene verskynsel in die Wes-Kaap. Die rangorde, wat die effek van die verskeie faktore wat groeikragvariasie tussen wingerdstokke bepaal, word steeds gedebatteer. Die debat kan groottendeels toegeskryf word aan die beduidende hoeveelheid variasie tussen wingerde waarmee navorsers te doen kry in wingerdkundige studies. Hoewel, met onlangse vordering aangaande afstandswaarnemingstegnologie is daar nou nuwe metodes beskikbaar om wingerdgroeikrag te evalueer. Dit is kommerwekend om te dink dat `n wyndruifwingerd normaalweg as `n homogene eenheid bestuur word as dit kom by besproeiing. Veral met die wete dat groeikragvariasie tussen wingerde algemeen erken en aangeteken word, en dat volhoubare waterstatus van `n wingerdstok van kardinale belang is vir produksie en kwaliteit van wyndruiwe. Die aanname dat wyndruiwe, die sap- en ook wynkwaliteit homogeen sal wees regdeur `n wingerdblok is egter meer kommerwekkend. Na aanvang van dié denke is daar `n studie geloods om die interaksie tussen wingerdstokgroeikrag en wingerdstokwaterstatus te evalueer. Met die studie is verskeie besproeiingsregimes aangebring binne `n kommersiële wingerd wat interne groeikragvariasie tentoonstel. Groeikragvariasie was geïdentifiseer deur middel van multispektrale lugfotos terwyl die wingerdstok se waterstatus geëvalueer is met behulp van plantgebaseerde metings in die verskillende groeikragareas. Die waterstatus van die grond is geëvalueer tesame met die vegetatiewe groei van die wingerd sodat die groeikragvariasie en die invloed van die plantwaterstatus op die groeikrag bepaal kon word. Die reproduktiewe groei is deurlopend gemonitor voor die effek van wingerdstokwaterstatus en wingerdstokgroeikrag op druifsamestelling en wynkwaliteit bepaal is. Daar was `n goeie ooreenkoms tussen die verskeie metodes wat gebruik is om wingerdgroeikrag te bepaal. Snoeimassa aan die einde van die seisoen was ooreenkomstig met die blaaroppervakte (hooflootblare en sylootblare) wat tydens vegetatiewe groei gemeet is, en het ook goed korreleer, met die multispektrale lugfotos se hoof groeikragklassifikasie. Korrelgewig en -volume het reageer op die verskeie besproeiingsregimes, en daar was `n toename in korrelgewig en -volume saam met die afname in watertekort van een regime tot `n ander. Daar was beduidende verskille tussen die verskeie klassifikasies t.o.v. korrelsamestelling analise en wynevaluasie. Die suikerinhoud per korrel, totale fenole, totale rooi pigment, appelsuur, stikstof en pH het verskil in druiwesap analises. Die pH en suur van die wyne het ook beduidend verskil.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: One of the most striking qualities of wine is its tart, sour taste. The sensory perception of sourness is mainly attributed to the presence of hydrogen ions (protons) at high concentrations. Large amounts of weak carboxylic acids (organic acids) are the main sources of these ions within wine. Once wine enters a person's mouth, the dissociable protons of the weak organic acids within wine are partially neutralized or, in other words, titrated by the saliva secreted inside one's mouth. This explains why the duration and intensity of a wine's sourness is related to its titratable acidity content. The sour taste of wine is usually considered refreshing and it helps balance wine flavour. In fact, wines become watery when its titratable acidity content is too low. After alcoholic fermentation, the titratable acidity of wine will usually be less than that of the grape juice from which was made due to ethanol-induced precipitation of potassium bitartrate crystals and partial consumption of malic acid by fermenting wine yeasts. Occasionally however, increases in titratable acidity are observed during alcoholic fermentation. If wine is produced from grape juice with optimum levels of titratable acidity, unforeseen increases in titratable acidity during alcoholic fermentation can be detrimental to the quality of the final product. Although the net production of malic acid by wine yeasts contributes to increases in titratable acidity seen during grape juice fermentations, the production of succinic acid is regarded as the primary contributor. In fact, succinic acid accounts for approximately 90% of the non-volatile acids produced during fermentation of grape juice. Between 0.5 and 1.5 g/L succinic acid is normally found in wine, but higher concentrations thereof (up to 3.0 g/L) have been detected within certain red wines. Acidity adjustments should preferably be carried out before the onset of alcoholic fermentation to allow better integration of the added compound(s) and to ensure that conditions during fermentation favour the quality and microbial stability of the final product. In doing so unfortunately, winemakers run the risk of ending up with wines that may taste too sour if they are unable to accurately predict and take into consideration the amount of succinic acid produced during alcoholic fermentation. Knowledge with regard to the factors involved in succinic acid's production by fermenting wine yeasts is therefore required in order to manage the titratable acidity of wines more accurately. Ever since Louis Pasteur first noticed succinic acid amongst the by-products of alcoholic fermentation, attempts have been made to determine the metabolic pathways and factors involved in its production by fermenting wine yeasts. Up until now however, it remains unclear why wines sometimes end up with exceptionally high levels of succinic acid. For these reasons it was decided to investigate the possible causes of very high succinic acid concentrations within wine. Due to complexity of grape juice's chemical composition and the problems associated with sterilizing grape juice, fermentation experiments were conducted within a chemically defined grape juice-like medium. Succinic acid production by nine different industrial wine yeast strains was studied under various conditions with regard to the nutrient status of the synthetic grape juice, temperature and availability of molecular oxygen during alcoholic fermentation. The amount of succinic acid produced during alcoholic fermentation was found to depend on the yeast strain, fermentation temperature and chemical composition of the synthetic grape juice. Out of the nine commercial yeast strains selected for this study, strain WE372 produced the largest amount of succinic acid in synthetic grape juice at 28°C. Strain WE372 produced significantly smaller amounts of acetic acid than the other yeast strains of this study and very little acetic acid at 28°C, which indicated that strain WE372 may have less acetaldehyde dehydroganase activity than the other yeast strains of this study under the conditions tested. The effect this has on NAD: NADH balance is the probable cause for its ability to form more glycerol, succinic and malic acid than the other strains. Results from our study show that succinic acid production is influenced primarily by the metabolizable fraction of YAN, which we termed metabolically available nitrogen (MAN). Succinic acid production by fermenting yeasts will be favoured by moderate to high fermentation temperatures (20°C to 28°C) in grape juice with a nicotinic acid and/ or nicotinamide deficiency, high sugar content (200 g/L to 240 g/L), moderate amounts of metabolically available nitrogen (300 ± 50 mg/L MAN), the presence of flavonoids and large supplies of unsaturated long-chain fatty acids. Even higher concentrations of succinic acid were produced when oxygen was made available to fermenting yeasts by aerating the fermenting grape juice. Fermentation temperatures below 18°C, too much metabolizable nitrogen (> 450 mg/L MAN), very high concentrations of fermentable sugar (> 240 g/L), lipid deficiencies and a lack of pantothenic acid, thiamine, biotin or pyridoxine will decrease the amount of succinic acid produced fermenting yeasts.
No Afrikaans summary available.

Thesis (PhD(Agric))--Stellenbosch University, 2012
ENGLISH ABSTRACT: Field spectroscopy was used to study leaf composition and selected factors (including canopy growth manipulation and water status changes) that may impact on it in a Vitis vinifera cv. Shiraz vineyard, showing considerable variability in vigour. Temporal and spatial variability in leaf composition were incorporated into measurements by analysing leaves in different shoot positions and at different developmental stages during three different growing seasons. Irrigation and canopy manipulation treatments were also imposed in order to provide new insights into assessing the grapevine leaf and possibly also the canopy growth and ageing dynamics as well as pigment content, as a basis of executing a generally non-destructive measurement approach. Despite large climatic differences between the seasons, canopy size seemed of crucial importance in determining grapevine water relations in the grapevines from the different canopy manipulation treatments. Drastic compensation effects in terms of secondary shoot growth also followed the canopy reduction treatment. Despite this, canopy microclimate was apparently improved, considering the results from light measurements as well as the ripening dynamics in the reduced canopies. Reduced canopies also seemed to display a different canopy composition, in favour of secondary growth. This could have impacted positively on water use efficiency as well as ripening, due to higher photosynthetic efficiency of these leaves during the ripening stages. The reduced canopy treatments offered the possibility of attaining technological ripeness at an earlier stage and at comparatively lower potential alcohol levels. This study illustrated the relevance of considering the vegetative development of the grapevine, along with leaf ageing in the canopy, when conducting calibrated non-destructive measurements of leaf pigments, structure and water content. The relevance of using multivariate techniques in leaf spectroscopy was shown. This can be applied and simplified to aid in non-destructive leaf pigment, structure and water content estimation in future studies. Even with the general variation encountered in this vineyard, predictions of the major pigments in grapevine leaves were within acceptable error margins. Further work is required to improve the modelling of xanthophylls, which may require non-linear multivariate techniques. Logistical shoot growth modelling was used in leaf age estimation and classification, which made it possible to simplify statistical analysis of the leaf parameters mentioned. Practical application of the modelled and predicted parameters was shown for a specific period in season two by comparing the reaction of different treatments to developing water deficits. The results indicated that several parameters, with special mention of the carotenoid:chlorophyll ratio and chlorophyll a:b ratio, can be monitored on young and old leaves in the canopy in order to monitor developing water deficit stress. The modelled parameters, however, did not seem to be sensitive enough to allow specific prediction of predawn leaf water potential values. Specific leaf mass, equivalent water thickness, total specific leaf mass as well as leaf chronological age were successfully predicted from leaf spectral absorbance data, and this may be useful in future work on quantifying leaf adaptation to the micro-environment within the canopy.
AFRIKAANSE OPSOMMING: Veldspektroskopie is gebruik om blaarsamestelling en spesifieke faktore (insluitend lowergroei manipulasie en waterstatus veranderinge) wat ‘n impak kan hê in ‘n Vitis vinifera cv. Shiraz wingerd, met beduidende variasie in groeikrag, te ondersoek. Ruimtelike, asook tydsgebonde, variasie in blaarsamestelling is geïnkorporeer in die metings deur blare van verskillende lootposisies en vir verskillende ontwikkelingstadia gedurende drie verskillende groeiseisoene te meet. Besproeiings- en lowermanipulasie behandelings is ook uitgevoer om die dinamiek van blaar- en lowergroei, veroudering, asook pigmentinhoud te bestudeer binne die konteks van die uitvoering van ‘n nie-destruktiewe meetstrategie. Ondanks groot klimaatsverskille tussen die seisoene, blyk lowergrootteverskille belangrik te wees in die bepaling van wingerdstok-waterverhoudings in die verskillende lowermanipulasie behandelings. Drastiese kompensasiereaksies ten opsigte van sylootgroei is waargeneem in reaksie op die gereduseerde lowerbehandeling. Indien die resultate van ligmetings en druifrypwording in die gereduseerde lowerbehandeling in ag geneem word, is lowermikroklimaat egter steeds verbeter. Hierdie behandeling het oënskynlik ook veranderde lowersamestelling gehad, tot voordeel van sylootgroei. Dit kon moontlik ‘n positiewe invloed gehad het op waterverbruikseffektiwiteit asook druifrypwording, as gevolg van moontlike hoër fotosintetiese effektiwiteit van die blare gedurende die rypwordingstadia. Die gereduseerde lowerbehandeling het die moontlikheid gebied om tegnologiese rypheid by ‘n vroeër datum te bereik, met moontlike laer alkoholvlakke in die wyn. Hierdie studie het die belangrikheid beklemtoon om die vegetatiewe ontwikkeling van die wingerdstok in ag te neem wanneer gekalibreerde nie-destruktiewe metings van blaarpigmente, blaarstruktuur asook waterinhoud onderneem word. Die belang van multi-variant meettegnieke in blaarspektroskopie is aangetoon. Dit kan verder vereenvoudig word ter ondersteuning van niedestruktiewe meting van blaarpigment, -struktuur en -waterinhoudsbepaling in toekomstige studies. Selfs met die variasie wat in die wingerd voorgekom het, was die voorspellings van die vlakke van die belangrikste pigmente wat in wingerdblare aangetref word binne aanvaanbare foutgrense. Opvolgwerk is nodig om die modellering van xanthofil te verbeter, aangesien dit moontlik nielineêre multi-variant analise mag benodig. Logistiese groeimodellering is gebruik om blaarouderdom te bepaal en te klassifiseer, wat dit moontlik gemaak het om statistiese analise te vereenvoudig vir die genoemde blaarparameters. Die praktiese toepassing van die gemodelleerde en voorspelde parameters is aangetoon vir ‘n spesifieke gedeelte in seisoen twee, deur die reaksie van verskillende behandelings op toenemende watertekorte te bestudeer. Resultate het aangetoon dat verskeie parameters, met spesifieke klem op die karotenoïed:chlorofil verhouding, asook die chlorofil a:b verhouding, gemoniteer kan word op jong en ouer blare in die lower ten einde ontwikkelende waterstrestoestande te identifiseer. Die gemodelleerde parameters was egter klaarblyklik nie sensitief genoeg vir akkurate voorspelling van voorsonsopkoms-waterpotensiaalvlakke nie. Spesifieke blaarmassa, ekwivalente waterdikte, totale spesifieke blaarmassa, sowel as blaarouderdom kon suksesvol voorspel word deur gebruik te maak van absorpsieblaarspektroskopie, wat nuttig kan wees in toekomstige studies wat handel oor die kwantifisering van blaaraanpassing by die mikro-omgewing binne ‘n wingerdlower.

Thesis (MScAgric)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: The distinctive varietal flavour of wines is a combination of absolute and relative concentrations of chemical compounds. Volatile compounds are responsible for the odour of wine and non-volatiles cause the sensation of flavour. Accompanying these senses, a third, tactile, sense of ‘mouth-feel’ is recognizable. This forms the complete organoleptic quality of wine. Several hundred different compounds are simultaneously responsible for the odour release in wine, and since there is no real character impact compound, the aroma of wine can be described as a delicate balance of all these compounds. One of the most important groups of volatiles is the monoterpenes, which play a role in both aroma and flavour. This is especially significant for the Muscat varieties, but these flavour compounds are also present in other non-muscat grape varieties, where they supplement the varietal aroma. Monoterpenes occur in wine as free, volatile and odorous molecules, as well as flavourless non-volatile glycosidic complexes. The latter slowly releases monoterpenes by acidic hydrolysis, but the impact on varietal aroma is considered insufficient for wines that are consumed young. It is therefore important to supplement the release mechanism, in order to enhance the varietal aroma of the wine. The enzymatic hydrolysis mechanism functions in two successive steps: firstly, depending on the precursor, the glycosidic linkage is cleaved by α-L-arabinofuranosidase, α-L-rhamnosidase, β-D-xylosidase or β-D-apiosidase. The second step involves the liberation of the monoterpene alcohol by a β-glucosidase. This enzymatic hydrolysis does not influence the intrinsic aromatic characteristics of the wine, as opposed to acid hydrolysis. Pectolytic enzymes play an important role in cell elongation, softening of tissue and decomposition of plant material. These enzymes are used to improve juice yields, release colour and flavour compounds from grape skins, as well as improve clarification and filterability. Pectolytic enzymes work synergistically to break down pectins in wine. Protopectinase produce water-soluble and highly polymerised pectin substances from protopectin, it acts on non-methylated galacturonic acid units. Pectin methylesterase split methyl ester groups from the polygalacturonic chain. Polygalacturonase break down the glycosidic links between galacturonic acid units. Pectin and pectate lyases have a β-eliminative attack on the chain and it results in the formation of a double bond between C4 and C5 in the terminal residues. From the above it can be seen that enzymes play a pivotal role in the winemaking process. Unfortunately, in winemaking a lot of factors can influence the effects of enzymes. One possible factor in the wine medium is the presence of acidprotease, from yeast and/or fungal origin. This type of enzyme utilizes other enzymes as substrates and renders them useless. Pure enzyme preparations were used to study the interactions of a yeast acid-protease and a report activity (β-glucosidase) in vitro. A bottled wine and a buffer were used as in vitro conditions. Enzyme assays were performed to determine the relative activity over a number of days. The results indicated that even though both enzymes showed activity in both the media, the yeast protease did not have any significantly affect on the report activity. Subsequently wine was made from Sauvignon blanc grapes, with varying enzyme preparation additions. Enzyme assays were performed during the fermentation; and chemical, as well as sensory analysis were done on the stabilized wine. The results confirmed that the yeast protease did not have any significant affect on the report activity in these conditions. The protease’s inability to affect the report activity seems unlikely due to the fact that it is active at a low pH range and has been suggested as the only protease to survive the fermentation process. It seems possible that a winerelated factor, possibly ethanol, is responsible. Thus it seems that yeast protease does not threaten the use of commercial enzymes in the winemaking process in any significant way. Future work would entail more detailed enzyme studies of interactions between protease, both from yeast and fungal origin, and other report activities in specified conditions. The degradation capability could be directed towards unwanted enzyme activities that cause oxidation and browning of the must. The characterization of interactions between protease and β-glucosidase activities may hold key to producing wines with enhanced aroma and colour potential, as well as the elimination of unwanted enzyme activities.
AFRIKAANSE OPSOMMING: Die herkenbare kultivar karakter van wyn is ‘n kombinasie van absolute en relatiewe konsentrasies van verskeie chemiese komponente. Vlugtige komponente is verantwoordelik vir die geur, of aroma, van wyn en die nie-vlugtige komponente veroorsaak die sensasie van smaak. ‘n Derde, fisiese sensasie, die ‘mondgevoel’, is ook herkenbaar. Dit vorm die omvattende organoleptiese kwaliteit van die wyn. ‘n Paar honderd verskillende komponente is gelyktydig verantwoordelik vir die aroma vrystelling in wyn en omdat daar geen werklike karakter ‘impak’ komponent is nie, kan die aroma van wyn beskryf word as ‘n delikate balans van al die betrokke komponente. Een van die mees belangrike groepe vlugtige komponente is die monoterpene wat ‘n rol speel in beide aroma en smaak. Dit is veral belangrik by Muskaat kultivars, maar hierdie aroma komponente is ook teenwoordig in niemuskaat druif kultivars, waar hulle bydra tot die kultivar karakter en aroma. Monoterpene kom in wyn voor as vry, vlugtige en aromatiese molekules en in geurlose, nie-vlugtige glikosidies-gebonde komplekse. Die gebonde vorm word stadig vrygestel deur ‘n suurhidrolise, maar dit word as onvoldoende beskou vir wyne wat vroeg gedrink word. Dit is dus belangrik dat die vrystelling van geurstowwe verhoog word om die kultivar karakter van die wyn te versterk. Die ensiematiese hidrolise proses behels twee opeenvolgende stappe: eerstens, afhangende van die aard van die voorloper, word die glikosidiese verbinding deur α-L-arabinofuranosidase, α-Lramnosidase, β-D-xilosidase, of β-D-apiosidase gebreek. In die tweede stap word die monoterpeen-alkohol deur β-glukosidase vrygestel. Hierdie ensiematiese afbraak proses verander nie die intrinsieke aromatiese kenmerke van die wyn, soos met suurhidrolise die geval is nie. Pektolitiese ensieme speel ‘n fundamentele rol in selverlenging, sagwording en afbraak van plant materiaal. Hierdie ensieme word gebruik om sap opbrengs te verhoog, aroma en smaak komponente vry te stel uit die doppe, asook om sapverheldering en filtrasie te verbeter. Die pektolitiese ensieme werk op ‘n sinergistiese wyse om pektien in wyn af te breek. Protopektinase produseer wateroplosbare en hoogs gepolimeriseerde pektien uit protopektien, slegs uit niegemetileerde galakturoonsuur eenhede. Pektien metielesterase verwyder metielester groepe van die poligalakturoonsuurketting. Die glikosidiese bindings tussen galakturoonsuur eenhede word deur poligalakturonase afgebreek. Pektien- en pektaat-liase het ‘n β-eliminasie aanslag op die ketting en as gevolg daarvan word dubbelbindings tussen C4 en C5 in die terminale residue gevorm. Vanuit bogenoemde is dit dus duidelik dat ensieme ‘n kardinale rol speel in die wynbereidingsproses. Ongelukkig is daar ‘n verskeidenhied van faktore wat die werking van ensieme in die wynbereidingsproses kan beïnvloed. Een moontlike faktor is die teenwoordigheid van ‘n suur-protease, van fungisidiese en/of gis oorsprong, in die wynmedium, omdat dit ander ensieme as substraat kan benut en degradeer. Suiwer ensiem preparate is gebruik om die ensiem interaksie tussen ‘n gis suur-protease en ‘n verslag aktiwiteit (β-glukosidase) in vitro te ondersoek. ‘n Gebotteleerde wyn en ‘n buffer is gebruik om die in vitro kondisies na te boots. Relatiewe ensiem aktiwiteit is ontleed oor ‘n aantal dae. Beide die ensieme het aktiwiteit getoon in die media, maar gis protease het geen statisties beduidende invloed gehad op die aktiwiteit van die verslag ensiem nie. Daaropvolgend is wyn berei van Sauvignon blanc druiwe, met verskillende ensiempreparaat toevoegings. Die ensiemaktiwiteit is deurlopend tydens fermentasie gemeet. Na afloop van stabilisasie is chemiese, sowel as sensoriese ontledings op die wyn gedoen. Die resultate het bevestig dat gis protease, onder hierdie kondisies, geen beduidende invloed op die verslag aktiwiteit gehad het nie. Die protease se onvermoë om die verslag aktiwiteit beduidend te beinvloed blyk onwaarskynlik aangesien die suurprotease aktief is by lae pH vlakke en dit as die enigste protease voorgestel is wat die fermentasie proses kan oorleef. Dit blyk asof ‘n wyn-verwante faktor, moontlik etanol, hiervoor verantwoordelik kan wees. Dus hou protease geen gevaar in vir die gebruik van kommersiële ensieme in wynbereiding nie. Navorsing kan in die toekoms fokus op meer gedetailleerde ensiem interaksie studies tussen protease en ander ensiem aktiwiteite, in gespesifiseerde kondisies. Die degradasie kapasiteit kan moontlik aangewend word om ongewenste ensiem aktiwiteite, wat byvoorbeeld oksidasie en verbruining veroorsaak, te verminder. Die karakterisering van die interaksies tussen protease en β-glukosidase kan dus die sleutel wees tot die produksie van wyne met verhoogde aroma potensiaal, asook die eliminasie van ongewenste ensiematiese aktiwiteite.

Thesis (MSc)--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: In the field of agriculture, plant pathogens are a major concern because of the severe damage these organisms cause to crops yearly. Fundamental studies regarding plant pathogens and their modes of action made it possible for researchers in the field of molecular biology to investigate pathogens further on a molecular level. Botrytis cinerea, has been used to great effect as a model system to investigate various aspects regarding pathogenesis, also on a molecular level. Molecular research done on B. cinerea over the last few years has shown that the endopolygalacturonases (EPGs) of this fungus are key role players in pathogenesis. This hydrolytic enzyme family of six members, encoded by the Bcpg1-6 genes, are important in breaking down the complex cell wall polymers of host plants, enabling the fungus to penetrate its host sufficiently. It has been shown that both BcPG1 and 2 are crucial for virulence of B. cinerea. A leucine-rich repeat inhibitor protein situated in the cell wall of various plant species, the polygalacturonase-inhibiting protein (PGIP), has been proven to interact with and inhibit EPGs, and thus the necrotic actions of B. cinerea. From literature it was clear that specific data regarding individual interactions of fungal EPGs with PGIPs are lacking currently. Furthermore, most experiments regarding the effects of EPG as well as interaction and inhibition studies of EPGs and PGIPs, rely on in vitro methods, without the possibility to contextualize the results on an in vivo or in planta level. The scope of this study was to specifically address the issues of individual EPG:PGIP interactions and the use of possible in vivo methodology by using EPGs from a highly virulent South African strain of B. cinerea and the grapevine VvPGIP1 that has been previously isolated in our laboratory. This PGIP, originally isolated from Vitis vinifera cv Pinotage, has been shown to inhibit a crude EPG extract from this strain with great efficiency. The approach taken relied on heterologous over-expression of the individual Bcpg genes and the isolation of pure and active enzymes to evaluate the inhibition of the EPGs with VvPGIP1. The genes were all successfully over-expressed in Saccharomyces cerevisiae with a strong and inducible promoter, but active enzyme preparations have been obtained only for the encoding Bcpg2 gene, as measured with an agarose diffusion assay. The in vitro PGIP inhibition assay is also based on the agarose diffusion assay and relies on activity of the EPGs to visualize the inhibiting effect of the PGIP being tested. The active EPG2, however, was not inhibited by VvPGIP1 when tested with this assay. The EPG encoding genes from B. cinerea were transiently over-expressed also in Nicotiana benthamiana by using the Agrobacterium-infiltration technique. Transgene expression was confirmed by Northern blot analysis and EPG-related symptoms were observed five to eight days post-infiltration. Differential symptoms appeared with the various EPGs, providing some evidence that the symptoms were not random events due to the infiltration or a hypersensitive response. Moreover, the symptoms observed for EPG2 was similar to those that were reported recently by another group on the same host. In spite of the expression data and the clear symptoms that developed, active preparations, as measured with the agarose diffusion plate asay, could only be obtained for EPG2 again. In our search for a possible in vivo method to detect and quantify EPG activity and inhibition by PGIPs, we tested and evaluated a technique based on chlorophyll fluorescence to detect the effect of EPGs on the rate of photosynthesis. Our results showed that the over-expression of these genes reduced the rate of electrons flowing through photosystem II, indicating metabolic stress occurring in the plant. We used the same technique to evaluate possible interaction between VvPGIP1 respectively with BcPG1 and 2 and found that the co-expressing of the Vvpgip1 gene caused protection of the infiltrated tissue, indicating inhibition of EPG1 and 2 by VvPGIP1. For EPG2, the observed interaction and possible inhibition by VvPGIP1 is the first report to our knowledge of an interaction between this specific EPG2 and a PGIP. Moreover, to further elucidate the in planta interaction between VvPGIP1 and the EPGs from the South African B. cinerea strain, we tested for possible interactions by making use of a plant two-hybrid fusion assay, but the results are inconclusive at this stage. Previous studies in our laboratory have shown that several natural mutations exist between PGIP encoding genes from different V. vinifera cultivars. Based on this finding and the fact that these natural mutations could result in changes with regard to EPG inhibition and ultimately disease susceptibility, we isolated an additional 37 PGIP encoding genes from various grapevine genotypes, some of which are known for their resistance to pathogens. Combined, these results make a valuable contribution to understand plant pathogen interactions, specifically in this case by modeling the interactions of pathogen and plant derived proteins. The possibility to use in vivo methods such as chlorophyll fluorescence to follow these interactions on an in planta level, provides exciting possibilities to strenghten and contextualize in vitro results.
AFRIKAANSE OPSOMMING: Plantpatogene organismes veroorsaak jaarliks erge skade aan landbougewasse en word dus as ’n ernstige probleem in die landbousektor beskou. Diepgaande studies wat handel oor plantpatogene en hul metodes van infeksie het dit vir molekulêre bioloë moontlik gemaak om patogene nou ook op molekulêre vlak verder te bestudeer. Botrytis cinerea is baie effektief as modelsisteem gebruik om verskeie aspekte van patogenese verder te bestudeer, ook op ‘n molekulêre vlak. Molekulêre navorsing op B. cinerea, het getoon dat die endopoligalakturonases (EPGs) van dié swam kernrolbelangrik in patogenese is. Hierdie sesledige hidrolitiese ensiemfamilie word gekodeer deur die Bcpg1-6 gene en is belangrik vir die afbraak van die komplekse selwandpolimere van plantgashere, om suksesvolle gasheerpenetrasie te veroorsaak. Daar is aangetoon dat beide BcPG1 en 2 essensieël vir virulensie van die patogeen is. ’n Leusienryke-herhalings inhibitorproteïen wat in die selwand van verskeie plantspesies voorkom, die poligalakturonase-inhiberende proteïen (PGIP), het interaksie met en inhibeer EPGs en gevolglik ook die nekrotiserende aksies van B. cinerea. Uit die literatuur is dit duidelik dat spesifieke inligting aangaande individuele interaksies van fungiese EPGs met PGIPs tans nog ontbreek. Verder word daar op in vitro metodologie staatgemaak wannneer die effekte van EPGs asook die interaksie en inhibisie met PGIPs bestudeer word, sonder om die konteks van die in vivo- of in planta-omgewing in ag te neem. Die fokus van hierdie studie was om aspekte van individuele EPG:PGIP interaksies, asook die moontlike gebruik van in vivo metodologie te bestudeer deur EPGs, afkomstig van ’n hoogs virulente Suid-Afrikaanse ras van B. cinerea en die wingerd VvPGIP1, wat vroeër in ons laboratorium geïsoleer is, te gebrruik. Hierdie PGIP wat uit Vitis vinifera cv Pinotage geïsoleer is, inhibeer ’n kru EPG-ekstrak van bogenoemde ras baie effektief. Die benadering wat gevolg is het op die ooruitdrukking van die individuele Bcpg-gene in heteroloë sisteme staatgemaak en die gevolglike isolering van suiwer en aktiewe ensieme om EPG-inhibisie deur VvPGIP1 te beoordeel. Al die gene is suksesvol in Saccharomyces cerevisiae ooruitgedruk onder ’n sterk induseerbare promotor, maar volgens ’n agarose-diffundeerbare toets kon aktiewe ensiempreparate slegs vir die enkoderende Bcpg2 verkry word. Die in vitro PGIP-inhibisie toets is ook op die gemelde toets gebasseer en vereis EPG-aktiwiteit om die inhiberende effek van die PGIP, te visualiseer. Die aktiewe EPG2 is egter nie deur VvPGIP1 geïnhibeer met die aanleg van hierdie toets nie. Die EPG-enkoderende gene van B. cinerea is ook tydelik in Nicotiana benthamiana ooruitgedruk deur gebruik te maak van ’n Agrobacterium-infiltrasietegniek. Transgeenuitdrukking kon met die Noordelike kladtegniek bevestig word en EPG-verwante simptome is vyf tot agt dae na infiltrasie waargeneem. Verskillende simptome vir die verskillende EPGs is waargeneem, wat aanduidend is dat die simptome nie lukrake gevolge van die infiltrasies, of ’n hipersensitiewe respons is nie. Verder kon die simptome wat EPG2 vertoon het, gekorreleer word met dié wat onlangs deur ’n ander groep op dieselfde gasheer waargeneem is. Ten spyte van die ekspressiedata en die waargenome simptome, kon aktiewe ensiempreparate op die agarose-diffundeerbare toets, weereens slegs vir EPG2 waargeneem word. ’n Metode wat gebasseer is op chlorofilfluoressensie is getoets en geëvalueer as ’n moontlike in vivo metode om EPG aktiwiteit en inhibisie deur PGIPs waar te neem en te kwantifiseer. Die resultate het bevestig dat die ooruitdrukking van hierdie gene die elektronvloeitempo deur fotosisteem II verminder het wat ’n aanduiding is dat metaboliese stres in die plant heers. Dieselfde tegniek is gebruik om die moontlike interaksies tussen BcPG1 en 2 en VvPGIP1 te bestudeer en het aangetoon dat die mede-uitdrukking van die Vvpgip1-geen aanleiding gee tot ’n beskermende effek van die geinfiltreerde weefsel, wat aanduidend is van inhibisie van EPG1 en 2 deur VvPGIP1. In die geval van EPG2 is hierdie interaksie en moontlike inhibisie met ’n PGIP die eerste waarneming in die verband. In ’n verdere poging om die in planta-interaksie tussen VvPGIP1 en die EPGs van die Suid-Afrikaanse B. cinerea ras uit te klaar, is ’n plantgebasseerde twee-hibriede toets aangelê, maar geen klinkklare resultate kon verkry word nie. Vorige werk het bevestig dat verskeie natuurlike mutasies in PGIP-enkoderende gene, afkomstig van verskillende V. vinifera kultivars, voorkom. Hierdie resultaat en die feit dat hierdie mutasies verskille in EPG inhibisie en uiteindelik vatbaarheid vir siektes kan beïnvloed, het aanleiding gegee tot die isolering van ’n verdere 37 PGIP-enkoderende gene uit ‘n verskeidenheid druifplantgenotipes, sommige waarvan juis bekend vir hul weerstand teen patogene is. Die gekombineerde resultate wat in dié studie verkry is, maak ’n waardevolle bydrae tot die verstaan van plant-patogeeninteraksies, spesifiek met die modelering van interaksies van patogeen- en plantgebasseerde proteïene. Die moontlikheid om in vivo-metodes soos chlorofilfluoressensie te gebruik in in planta-analises, is besonder bemoedigend om in vitro-resultate te versterk en ook in konteks te plaas.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2011.
Includes bibliography.
ABSTRACT: The grapevine must constantly find a balance between two continually changing environments, the rhizosphere (i.e. soil) and the troposphere (i.e. macroclimate). The adaptations are extremely complex because they encompass complicated and interrelated processes that are not yet fully understood. In terms of water-use behaviour, differences between cultivars have been described in the literature. In this study, the water status and stomatal conductance of four cultivars (Shiraz, Grenache, Pinot noir and Chardonnay) grafted onto R99 were studied. Diurnal cycles of water status and stomatal conductance, from 07:00 to 19:00, were followed for a single day at the end of the 2009 season. The results showed that, at the end of the season, Shiraz was subjected to water stress conditions, losing leaves and showing symptoms of berry shrivelling. The other three cultivars had a much better canopy status and no symptoms of berry shrivelling were observed. Based on the canopy observations and a comparison of the curves of stem water potential (Ψs) and stomatal conductance (gs), it seems that Pinot noir and Chardonnay are closer to the water-use behaviour of Grenache noir, which is known as a “pessimistic” cultivar, than to Shiraz, which is an “optimistic” cultivar. A study of four plots each of Chardonnay/101.14 Mgt and Shiraz/101.14 Mgt was carried out in eight commercial vineyards in the Robertson region in order to investigate the relationship between soil and root morphology, and the influence thereof on canopy development and berry growth. These plots had different soil types. Important soil properties are reported to limit root growth, individually or as a combination of restrictions. It was found that the size of the root system of 101.14 Mgt is defined by soil physical and chemical properties. The roots of 101.14 Mgt under irrigation can grow to a depth of 100 cm or beyond if the soil physical and chemical properties allow it. Because the soil properties define the root system and the water storage/drainage, they greatly influence the plant water status, even under irrigation. In an arid zone like Robertson, irrigation is an important management tool. The balance between canopy growth before véraison and the ability of the root-soil system to maintain that canopy size during the ripening process is crucial in an area with a high evaporative demand. In this regard, not all the soil properties-root system combinations showed satisfactory performance in maintaining the canopy functioning, which affected berry sugar loading and berry volume. In another study that is presented, forty soil profiles were characterised in the Robertson valley. The root systems were considered as a product of the soil properties, and thus the morphology of the root systems was used as a starting point to group soils together. The importance of soil depth has been described well, thus the root systems were first classified according to rooting depth – into shallow and deep root systems. The deep root systems were then subdivided, creating two subgroups of high root density and low root density. The two extreme groups (i.e. shallow roots, and deep roots with high root density) have particularly different soil properties. The soil characteristics found in these extremes are represented up to certain point by families of the South African soil taxonomy, mainly due to the restrictive function of the B horizon. This restrictive function is related to soil properties that are taken into consideration in the South African soil classification and that are important for grapevine root growth, as well as the thickness of the described horizons and the physical and chemical differences between the horizons. Soil properties have an important influence on root morphology. Due to the fundamental role played by the root system in the overall plant functioning, soil properties are of critical importance. In an arid area, the low water pressure in the atmosphere and the high temperature greatly affect the plant water status. The soil-root system combination plays an important role in the ability of the root system to supply the plant with water during times of high evaporative demand. Different cultivars will react differently due to differences in transpiration control. The maintenance of an adequate water status will have an immense influence on canopy development and maintenance, and on normal and steady berry ripening. In this study it was found that not all the soil-root combinations can fulfil this satisfactorily. Thus, the grapevine balance determined by the combination of the soil-root-canopy complex and the influence of management techniques is extremely important for the favouring of a good canopy:root system ratio, a functional canopy throughout the season and a steady berry ripening curve.

Thesis (MScAgric)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: The production of quality wine is influenced by numerous factors of which grape quality is one of the most important factors. The production of quality wine, however, is not possible without good winemaking techniques and effective quality control. Critical control points (CCP) during the winemaking process must be identified to ensure optimum wine quality. Grape must is a complex medium that contains different micro-organisms which can be either beneficial or negative to wine quality, depending on the physical and chemical conditions that prevail in the must. Yeasts are responsible for alcoholic fermentation, lactic acid bacteria (LAB) for malolactic fermentation (MLF) and acetic acid bacteria (AAB) for the production acetic acid from ethanol. Yeasts and certain LAB can also produce acetic acid and thereby increasing the volatile acidity (VA) of wine. These micro-organisms can influence each other in complex fashions by competing for growth nutrients and by producing inhibitory substances. Most winemakers nowadays use commercial yeast strains to inoculate wine fermentations. This, however, does not assure a problem-free fermentation and cases of stuck and sluggish fermentations are annually reported worldwide. In these or most cases fermentation takes longer than 21 days to complete and the wine contains a residual sugar concentration of more than 4 g/L, which can be utilised by wine spoilage micro-organisms such as certain bacteria and other wild yeasts. Stuck and sluggish fermentations also increase the chances of oxidation due to the absence of the protective CO2 layer on the surface of the wine, which is formed during alcoholic fermentation. Another effect of stuck and sluggish fermentations is that valuable tank space is wasted due to the unexpected time consumption of these fermentation problems. Many factors during the winemaking process can be responsible for stuck and sluggish fermentations. In this thesis the different factors is discussed with the emphasis on the effect of the yeast strain. The way that certain yeast strains influence AAB and LAB numbers during fermentation and MLF through the production of inhibiting by-products such as medium chain fatty acids has not been investigated in detail in the past. Certain fungicides and pesticides that are used in vineyards to control pests (e.g. mildew) contain copper which can be inhibiting to yeast growth and alcoholic fermentation. Legal limits and withholding periods on these sprays are not always strictly obeyed and can lead to stuck and sluggish fermentations. This motivated us to evaluate the growth and fermentation activities of a selection of commercial wine yeasts in the presence of copper levels in the range of maximum legal limits. The effect of these commercial strains on the LAB and AAB numbers during alcoholic fermentation and MLF were also investigated. Our results showed that there was no significant difference on numbers of the AAB obtained from fermentations inoculated with different commercial wine yeast strains. However, with regards to the LAB numbers, one of the strains produced significantly more sulphur dioxide (SO2), which led to the inhibition of MLF in that wine. Our results further indicated which commercial yeast strains were capable of effectively fermenting high sugar musts and which strains were less effective. From the strains tested VIN13, N96 & L2056 were able to utilize fructose more effectively than NT50, RJ11 & D80. We could further distinguish between yeast strains that produced the lowest (VIN13 & RJ11) and the highest (WE372, NT50 & L2056) VA concentrations in must containing high sugar levels. Strains that were more tolerant against high copper levels were also identified. We tested six yeast strains in must with added copper (0.25 mM cu2+) in the form of CuSO4 .H2O. Three Cu2+-tolerant (D80, Collection Cepage Cabernet & NT50) yeast strains were distinguished from three less Cu2+-tolerant yeast strains (VIN13, NT112 & RJ11). This study made a valuable contribution in knowledge gained about commercially available wine yeast strains that can ferment effectively under certain stress conditions. Research such as this, where wine yeasts are evaluated to ferment more effectively during strenuous winemaking conditions, will be very beneficial to winemakers.
AFRIKAANSE OPSOMMING: Die produksie van gehalte wyn word deur verskillende faktore beïnvloed waarvan druifkwaliteit seker die belangrikste is. Die produksie van gehalte wyn is egter nie moontlik sonder goeie wynmaaktegnieke en effektiewe kwaliteitsbeheer nie. Kritieke kontrole punte (KKP) tydens die wynmaakproses moet dus geïdentifiseer word om sodoende ‘n verlaging in wynkwaliteit te vermy. Druiwemos het ‘n komplekse mikrobiologiese samestelling en bestaan uit verskillende mikroörganismes wat vooren nadelig vir wynkwaliteit kan wees, afhangende van die fisiese en chemiese toestande wat in die mos bestaan. Giste is verantwoordelik vir alkoholiese fermentasie, melksuurbakterieë (MSB) vir appelmelksuurgisting (AMG) en asynsuurbakterieë (ASB) vir die produksie van asynsuur vanaf etanol. Asynsuur word egter ook deur giste en MSB geproduseer en dra so by tot die vlugtige suurheid (VS) van ‘n wyn. Hierdie mikroörganismes kan mekaar op komplekse wyses beïnvloed deur o.a. te kompeteer vir voedingstowwe asook deur die produksie van inhiberende verbindings. Die meeste wynmakers maak gebruik van kommersiële gisrasse om alkoholiese fermentasies mee uit te voer. Gevalle van sogenaamde slepende en gestaakte alkoholiese fermentasies, waar suiker nie volledig na etanol en CO2 omgeskakel word nie, kom egter nog gereeld in die wynbedryf voor. In sulke gevalle neem die fermentasie gewoonlik langer as 21 dae om te voltooi met ‘n suiker konsentrasie van meer as 4 g/L wat in die wyn oorbly. Dit is nadelig vir wynkwaliteit aangesien dit nie net die kanse vir bederf deur bakterieë en giste verhoog nie, maar ook die kanse vir oksidasie verhoog a.g.v. die verlies van die beskermende CO2 lagie bo-oor die wyn. Hoe sekere gisrasse, ASB en MSB getalle gedurende fermentasie en AMG beïnvloed deur die produksie van inhiberende verbindings soos medium ketting vetsure en SO2, is ook nie baie in die verlede ondersoek nie. Sommige spuitstowwe wat gebruik word in die bekamping van swamsiektes bevat koper wat inhiberend kan wees vir gisgroei en alkoholiese fermentasie. Wetlike maksimum limiete en onthoudingsperiodes op spuitstofresidue word egter nie altyd gehoorsaam nie en kan lei tot slepende en gestaakte fermentasies. Dit het ons gemotiveer om ‘n seleksie van kommersiële gisrasse te evalueer in terme van gisgroei en fermentasie in die teenwoordigheid van kopervlakke naby die maksimum limiet. Ons resultate het gewys dat daar nie noemenswaardige verskille in AAB getalle tydens alkoholiese fermentasie tussen behandelings met verskillende kommersiële gisrasse was nie. Een van die gisrasse het wel noemenswaardig meer SO2 geproduseer wat gelei het tot inhibering van AMG in hierdie wyn. Ons het verder uitgewys watter kommersiële gisrasse instaat is om meer effektief in hoër suiker mos te fermenteer en watter van die rasse minder suksesvol was. Ons het ook rasse geïdentifiseer wat meer weerstandbiedend is teen hoë kopervlakke in mos en sodoende groter kans op ‘n suksesvolle fermentasie sal hê in mos wat koperresidue bevat wat afkomstig is van sekere spuitstowwe. Die effek van die ASB en MSB getalle gedurende fermentasie en AMG is ook ondersoek. Ons resultate het verder gewys watter kommersiële gisrasse instaat was om mos met hoë suikervlakke meer effektief te fermenteer. Vam die gisrasse wat getoets was het VIN13, N96 & L2056 fruktose meer effektief benut as NT50, RJ11 & D80. Ons kon verder onderskei tussen gisrasse wat die laagste (VIN13 & RJ11) en die hoogste (WE372, NT50 & L2056) vlakke van VS produseer in mos met hoë inisiële suikervlakke. Gisrasse wat meer tolerant was teen koperresidue in mos is ook geidentifiseer. Ons het ses gisrasse getoets in mos met bygevoegde koper (0.25 mM Cu2+) in die vorm van CuSO4 .5H2O. Daar is onderskei tussen drie Cu2+-tolerante (D80, Collection Cepage Cabernet & NT50) en drie minder Cu2+-tolerante gisrasse (VIN13, NT112 & RJ11). Hierdie studie lewer ‘n waardevolle bydrae in die invordering van kennis oor kommersieel beskikbare wyngisrasse wat meer effektief sal fermenteer onder sekere streskondisies wat in mos voorkom. Inligting soos hierdie is belangrik om die wynmaker se keuse uit die reeks bestaande kommersiële gisrasse te vergemaklik.

Thesis (PhD (Viticulture and Oenlogy))--University of Stellenbosch, 2008.
Includes bibliography and list of tables and figures.
ENGLISH ABSTRACT: Over the last decade research has shown the importance of small antimicrobial peptides in the innate immunity of plants. These peptides do not only play a critical role in the multilayered defense systems of plants, but have proven valuable in the engineering of disease resistant food crops towards the ultimate aim of reducing the dependency on chemical fungicides. As the lists of isolated and characterized peptides grew, it became clear that other biological activities, in addition to the antimicrobial capacity, could be linked to some of these peptides; these alternative activities could have important applications in the field of medicine. This has made the defensin encoding genes prime targets for the agricultural and medical biotechnology sectors. To this end we set out to evaluate South African flora for the presence of plant defensin sequences and to isolate plant defensin genes that might be useful in biotechnology applications. Moreover, by isolating and characterizing these novel peptides, also in an in planta environment and in interaction with fungal pathogens, important knowledge will be gained of the biological role and importance of the peptides in the plant body. The plant host targets were South Africa Brassicaceae species including indigenous species, as well as Vitis vinifera, as the most important fruit crop in the world and since no defensins have been isolated from this economically important crop plant. The Brassicaceae family has been shown to be abundant in defensin peptides and several of the best characterized peptides with potent activity have been isolated from this family. Based on initial activity screens conducted on selected South African Brassicaceae spp. we concluded that these spp. contain promising antifungal peptide activities, warranting further efforts to isolate the genes and encoding peptides and to characterize them further. The preliminary activity screens used a peptide-enrichment isolation strategy that favored the isolation of basic, heat-stable peptides; these properties are characteristic features of plant antimicrobial peptides. These peptide fractions showed strong antifungal activities against the test organisms. A PCR-amplification strategy was subsequently designed and implemented, leading to the isolation of 14 novel defensin peptide encoding genes from four South African Brassicaceae spp., including the indigenous South African species Heliophila coronopifolia. Amino acid sequence analysis of these peptides revealed that they are diverse in amino acid composition and share only 42% homology at amino acid level. This divergence in amino acid composition is important for the identification of new biological activities within closely related plant defensins. Single amino acid changes have been contributed with the divergent biological activities observed in closely related plant defensin peptides. Phylogenetic analysis conducted on the deduced amino acid sequences revealed that all the new defensins share a close relationship to other Brassicaceae members of the plant defensin superfamily and was furthest removed from the defensins isolated from the families Solanaceae and Poaceae. Classification analysis of these peptides showed that they belong to subgroup A3 of the defensin superfamily. A putative defensin sequence was also isolated from V. vinifera cultivar, Pinotage, and termed Vv-AMP1. Genetic characterization showed that only a single gene copy of this peptide is present within the V. vinifera genome, situated on chromosome 1. Genetic characterization of this peptide encoding gene within the Vitis genus showed that this gene has stayed conserved throughout the divergent evolution of the Vitis genus. Expression studies of Vv-AMP1 revealed that this gene is expressed in a tissue specific and developmentally regulated manner, being only expressed in grape berries and only at the onset of vèraison. Induction of Vv-AMP1 in grapevine leaf material could never be achieved through the external application of hormones, osmotic stress, wounding, or pathogen infection by Botrytis cinerea. Deduced amino acid analysis showed that Vv-AMP1 encoded for a 77 amino acid peptide consisting of a 30 amino acid signal peptide and a 47 amino acid mature peptide, with putative antifungal activity. The Vv-AMP1 peptide grouped with the subclass B type defensins, which have been documented to have both antifungal and antibacterial activities. The Vv-AMP1 signal peptide directed the green fluorescent protein (GFP) reporter gene to the apoplastic regions in cells with high levels of accumulation in the vascular tissue and the guard cells of the stomata. Recombinant Vv-AMP1 peptide was successfully purified from a bacterial host and shown to have a size of 5.495 kDa. Recombinant Vv-AMP1 showed strong antifungal activity at low concentrations against a broad spectrum of fungal pathogens, which included Verticillium dahliae (IC50 of 1.8 μg mL-1) and the necrotrophic pathogen Botrytis cinerea (IC50 of 12-13 μg mL-1). Antifungal activity of Vv-AMP1 did not induce morphological changes in fungal hyphae, but its activity was associated with induced membrane permeabilization in treated hyphae. Vv-AMP1 was successfully introduced into Nicotiana tabacum as confirmed by Southern blot analysis and 20 individual lines were generated. Genetic characterization confirmed the integration and expression of the gene in the heterologous tobacco environment. The peptide was under control of its native signal sequence which has been shown to direct its product to the apoplastic regions of cells. The transgenic lines were analyzed to determine the presence and activity of the grapevine defensin peptide. Western blot analyses of partially purified plant extracts detected a signal of the expected size in both the untransformed control and the transgenic lines. Comprehensive analysis of EST databases identified three highly homologous sequences from tobacco that probably caused the background signal in the control. These crude protein extracts were able to inhibit the growth of V. dahliae in vitro when tested in a microtiter plate assay, but the inhibition could not be conclusively linked to the presence of the transgenic peptide, since non-expressing transgenic lines, included as controls, also showed inhibition. Similar results were obtained with infection studies, clearly showing that despite successful integration and expression of the transgene, the peptides was either not functional in the heterologous environment, or perhaps unstable under the particular regulatory conditions. This peptide belongs to a subclass of peptides known for associated activities that might activate tight control by plant hosts if threshold levels are reached. These aspects need further investigation, specifically since it is in stark contrast to previous results obtained with defensins from a different subclass. This study has also yielded significant other related resources that would be instrumental for further possible biotechnology exploitation of some of the novel peptides, but also to provide genetic constructs and plant material that would be invaluable to address fundamentally important questions such as the regulation and mode of action of defensin peptides, specifically in interaction with pathogen hosts. The novel peptides have been transformed to various hosts, including grapevine and these transgenic populations are available to facilitate the next rounds of research into this extremely promising group of antifungal peptides.
AFRIKAANSE OPSOMMING: In die laaste dekade het navorsing die belangrike rol van klein antimikrobiese peptiede in plantweerstandsmeganismes beklemtoon. Hierdie peptiede speel nie alleenlik 'n belangrike rol in die komplekse lae van plantweerstandstelsels nie, maar het ook hulle ekonomiese potensiaal getoon in die manipulering van siekteweerstandbiedendheid in voedselgewasse met die oorkoepelende doel om landbougewasse minder afhanklik van chemiese spuitstowwe te maak. Soos wat die hoeveelheid geïsoleerde en gekarakteriseerde peptiede toeneem, het dit duidelik geword dat ander biologiese aktiwiteite, bykomend tot die antimikrobiese kapasiteit, met sommige van dié peptiede verbind kan word; hierdie alternatiewe aktiwiteite het belangrike toepassing in veral die mediese veld. Dit het die defensin-koderende gene kernteikens vir die landbou- en mediese biotegnologiesektore gemaak. In die studie is daar begin om die Suid-Afrikaanse blommeryk te evalueer vir die teenwoordigheid van plantdefensingene en om dié gene te isoleer wat van ekonomiese belang vir die biotegnologiebedryf kan wees. Deur die in vitro- én in planta karakterisering van die unieke plantdefensinpeptiede word daar gemik daarna om belangrike inligting in te win oor die biologiese rol van die peptiede binne die plantligggaam. Die plantgashere wat geteiken is sluit in die Suid-Afrikaanse Brassicaceae-spesies, insluitende inheemse spesies, asook Vitis vinifera, wat as die belangrikste vrugtegewas ter wêreld beskou word. Die Brassicaceae-familie is welbekend daarvoor dat dit 'n ryk bron van plantdefensinpeptiede is en verskeie van die bes gekarakteriseerde antifungiese defensinpeptiede is van dié familie afkomstig. Aanvanklike aktiwiteitstoetse het getoon dat die Suid-Afrikaanse Brassicaceae-spesies belowende antifungiese aktiwiteit toon, wat die verdere isolering en karakterisering van dié gene en hul peptiedprodukte regverdig. Die aanvanklike aktiwiteitstoetse het 'n selektiewe peptiedverrykingstrategie gevolg wat die isolering van basiese, hittestabiele peptiede bevoordeel het; hierdie eienskappe is baie kenmerkend van plant-antimikrobiese peptiede. Die peptiedfraksies wat met hierdie metode geïsoleer is, het sterk antifungiese aktiwiteit teen die toetsorganismes getoon. Die resultate het gelei tot die ontwikkeling en toepassing van 'n polimerasekettingreaksie-strategie, wat daartoe gelei het dat 14 nuwe defensingene van vier Suid-Afrikaanse Brassicaceae-genera, insluitend die inheemse spesie Heliophila coronopifolia, geïsoleer kon word. Afgeleide aminosuurvolgorde-analises van die nuwe defensinpeptiede het gewys dat hulle slegs 42% homologie het. Hierdie diversiteit in aminosuurvolgorde is belangrik vir die identifisering van nuwe biologiese aktiwitiete binne die groep van verwante peptiede. Navorsing het verder getoon dat enkel-aminosuurverskille bydra tot die diverse spektrum van biologiese aktiwiteite binne 'n groep van verwante defensinpeptiede. Filogenetiese analise van die aminosuurvolgordes het getoon dat al die nuwe defensinpeptiede 'n sterk verwantskap met plantdefensinpeptiede, wat van ander Brassicaceae-spesies geïsoleer is, toon. Daarteenoor het dit die kleinste verwantskap getoon met plantdefensinpeptiede wat van die Solanaceae- en Poaceae-families geïsoleer is. Klassifikasiestudies het bewys dat die nuwe peptiede saam met subgroep A3 van die plantdefensin-superfamilie groepeer. 'n Moontlike plantdefensingeen, genaamd Vv-AMP1, is ook van die V vinifera-kultivar, Pinotage, geïsoleer. Genetiese karakterisering het aangedui dat slegs 'n enkele kopie van die geen in die V. vinifera-genoom teenwoordig en op chromosoom 1 geleë is. Genetiese karakterisering van Vv-AMP1 binne die Vitus-genus het gewys dat die geen binne die genus evolusionêr gekonserveerd is. Uitdrukkingstudies van Vv-AMP1 het verder bewys dat die geen uitgedruk word op 'n weefselspesifieke, ontwikkelingsgekoppelde wyse, naamlik slegs in druiwekorrels en slegs tydens rypwording. Vv-AMP1-uitdrukking kon nooit geïnduseer word in wingerdblare deur die uitwendige toediening van hormone, osmotiese stres, wonding of patogeeninfeksie deur Botrtys cinerea nie. Ontleding van die afgeleide aminosuurvolgorde het gewys dat Vv-AMP1 kodeer vir 'n 77-aminosuurpeptied, wat uit 'n 30-aminosuurseinpeptied en 'n 47-aminosuur-aktiewe peptied met voorspelde antifungiese aktiwiteit bestaan. Die Vv-AMP1-peptied is gegroepeer met subgroep B van die plantdefensin-superfamilie, 'n subgroep wat vir beide antifungiese en antibakteriese aktiwiteit gedokumenteer is. Die Vv-AMP1-seinpeptied het die groen fluoressensie-indikatorproteïen (GFP) na die apoplastiese areas van die plantselle gelei, met hoë vlakke van lokalisering in die vaatbundelweefsel en sluitselle van die huidmondjies. Die rekombinante Vv-AMP1-peptied is suksesvol geproduseer en uit 'n bakteriese produksieras gesuiwer, en het 'n molekulêre massa van 5.495 kDa gehad. Die gesuiwerde peptide het by lae konsentrasies 'n sterk aktiwiteit getoon teen 'n breë spektrum van fungiese patogene, wat Verticllium dahliae (IC50 van 1.8 μg mL-1) en die nekrotrofiese patogeen, B. cinerea (IC50 van 12-13 μg mL-1), ingesluit het. Vv-AMP1-aktiwiteit het geen ooglopende morfologiese veranderinge in die fungi-hifes veroorsaak nie, maar hulle aktiwiteit is verbind met 'n verhoogde membraandeurdringbaarheid in behandelde fungi-hifes. Suksesvolle intergrasie van Vv-AMP1 in die Nicotiana tabacum-genoom is deur Southern-kladontledings bevestig en 20 individuele transgeniese lyne is ontwikkel. Genetiese karakterisering van die transgeniese lyne het gewys dat Vv-AMP1 suksesvol geïntegreer is en ook in die transgeniese tabakomgewing uitgedruk word. Die peptied is uitgedruk onder beheer van sy eie seinpeptied, wat die aktiewe produk na die apoplastiese areas van die plantselle teiken. Die transgeniese tabaklyne is ook ontleed om te bepaal of die wingerdpeptied suksesvol geproduseer word en sy aktiwiteit in die transgeniese omgewing behou. Western-kladanalise van semi-gesuiwerde plantproteïenekstrakte het 'n positiewe sein gelewer in beide die kontroleplante en die transgeniese plantlyne. Bestudering van tabakgeenuitdrukkings-databasisse het drie nukleotiedvolgordes opgelewer wat homologie met Vv-AMP1 toon en moontlik verantwoordelik kan wees vir die positiewe sein in die ongetransformeerde kontroleplante. Kru proteïenekstrakte van die transgeniese tabaklyne het in vitro-aktiwiteit teen V. dahliae getoon. Geen oortuigende ooreenkoms kon egter gevind word tussen V. dahliae-inhibisie en die teenwoordigheid van die transgeniese Vv-AMP1-peptied nie, aangesien kontroleplante wat Southern-klad-positief is, maar nie geenuitdrukking toon nie, ook inhibisie van V. dahliae veroorsaak het. Soortgelyke resultate is met infeksiestudies verkry. Alle resultate dui daarop dat, al is daar suksesvolle integrasie en uitdrukking van die geen in tabak verkry, dat die Vv-AMP1 peptied óf onaktief óf onstabiel in die transgeniese tabakomgewing is. Die peptied behoort aan 'n subgroep peptiede met aktiwiteite wat, sodra sekere vlakke van peptied oorskry word, die moontlik streng kontrole op proteïenvlak in die gasheerplant kan uitlok. Sekere aspekte van die studie sal verder bestudeer moet word, aangesien die data teenstrydig is met data wat verkry is met soortgelyke plantdefensinpeptiede wat aan 'n ander subgroep behoort. Die studie het baie hulpbronne gegenereer wat vir die biotegnologiesektor belangrik kan wees, veral op ekonomiese gebied. Verder is die geenkonstrukte en plantlyne wat ontwikkel is waardevol om fundamentele vrae rondom die regulering en meganisme van aksie van defensinpeptiede, spesifiek plantpatogeeninteraksie, te beantwoord. Die nuwe plantdefensingene is na verskeie gasheerplante, insluitende wingerd, getransformeer waar die transgeniese lyne die volgende rondte van navorsing oor die bestudering oor die belangrike groep van antifungiese peptiede, sal aanvul.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2006.
The assessments of grape and wine quality are complex issues and the wine industry needs more objective analysis of grape and wine quality. The standard quality assessment protocol for grading grapes at most wine cellars in South Africa is based on viticultural practices and the determination of chemical parameters such as ºBrix, pH and titratable acidity (TA). Grape juice indices calculated by formulae such as ºBrix/pH, TA/pH, ºBrix/TA, ºBrix x (pH)2 have been used in the past but these approaches have had limited success. It was shown that the total anthocyanins and total phenolics of red grapes correlate with wine quality and provide additional objective measures of grape quality. Most methods for the quantification of total anthocyanins and total phenolics are complex and time consuming and therefore not easily implemented in the routine laboratory environment. Fourier transform infrared spectroscopy (FT-IR) is widely used in South African laboratories for the routine quantification of wine and grape parameters but the commercial calibration models supplied for the quantification of grape total anthocyanins and phenolics are not satisfactory. The focus of this study was to develop new FT-IR calibration models for the quantification of total anthocyanins and phenolics of grapes and to use the generated data during a preliminary evaluation of the implementation of these parameters as part of the grape quality control protocol at a commercial winery in South Africa. The potential of Fourier transform infrared spectroscopy (FT-IR) for the rapid quantification of total anthocyanins and total phenolics in red grapes was investigated and evaluated for prediction accuracy with independent validation sets. The design of calibration sets aimed at capturing most of the variation due to vintage (2004 and 2005), cultivar (Cabernet Sauvignon, Merlot, Pinotage, and Shiraz) and sugar concentration. Best prediction accuracies were obtained for calibration sets using grapes from a single vintage or cultivar or approximately the same sugar concentration. The highest prediction accuracies were obtained for total anthocyanins calibration sets of grapes with sugar concentrations ≥ 23.5ºBrix (SEP = 0.13 mg/g; R2 validation set = 0.77) and for total phenolics calibration sets of grapes with sugar concentrations < 23.5ºBrix (SEP = 0.13 OD280/g; R2 validation set = 0.74). Strong correlations were found between the spectral data and the total anthocyanins (SEP = 0.12 mg/g; R2 validation set = 0.84) and total phenolics concentration data (SEP = 0.10 OD280/g; R2 validation set = 0.76) for 2005 Merlot calibration sets indicating that the FT-IR spectra captured most of the variation. Overall the RPD (ratio of the standard deviation of the reference data to the standard error of prediction) values of all calibration models were below 3 indicating that calibration models are fit for screening purposes. Spectroscopic absorbance at 280 nm is not specific enough for the quantification of total phenolics and the use of an alternative reference method such as high performance liquid chromatography (HPLC) will be considered in the future. Principal component analysis (PCA) revealed that the major sources of variation in the FT-IR spectra of grapes could be ascribed to vintage and grape sugar concentration and this had an effect on the accuracy of the analytical data generated when using FT-IR spectroscopy. This report is the first to our knowledge where FT-IR has been used for the quantification of total anthocyanins and phenolics of grapes. The evaluation of the reference laboratory protocol for the quantification of total anthocyanins and total phenolics in grapes were evaluated in Chapter 4 and emphasized the importance of meticulous laboratory practices to obtain reliable reference data for calibration purposes. This large scale investigation of the total anthocyanins and phenolics concentrations in grapes is the first of its kind in South Africa and a quantitative database containing analytical data of the anthocyanins and total phenolics concentrations of 692 grape samples representing a wide range of grape maturities of Vitis vinifera cultivars Cabernet Sauvignon, Merlot, Pinotage and Shiraz from the 2004 and 2005 vintages was established based on the reference values. The data were used in a preliminary investigation of the implementation of total anthocyanins and total phenolics concentrations as part of grape quality classification at a commercial South African winery (Chapter 5). The results showed that the total anthocyanins and total phenolics concentration in grapes increased with increasing grape maturity (measured as sugar concentration). ANOVA and post-hoc analysis (Bonferroni testing) revealed significant differences between the total anthocyanins and total phenolics concentrations of the four Vitis vinifera cultivars investigated. Grapes harvested earlier in the season had significantly higher (p≤0.05) total anthocyanins and total phenolics concentrations than grapes harvested later in the season. This implies that grapes harvested earlier in the harvest season could produce wines with higher quality. Grapes from regularly irrigated vineyards had lower total anthocyanins and total phenolics concentrations compared to dryland vineyards. The current grape grading system in use at the industrial cellar did not correlate well with the total anthocyanins and total phenolics concentrations of grapes which highlighted the need for the inclusion of more objective measures during grape grading. The information captured in the database can be used as a starting point to establish profiles of the typical anthocyanins and total phenolics of South African grapes and data from more vintages should be included and continually updated. These findings highlight the important contribution of the results obtained in this preliminary study for the incorporation of total anthocyanins and phenolics concentrations as objective parameters of grape quality. Finally multivariate data analysis of the FT-IR spectra revealed important information regarding factors (both physical and chemical) that contribute to the variation of the spectra. The main variation between the 2004 and 2005 samples can probably be interpreted in terms of the water content of the samples.

Thesis (MScAgric)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: Glutathione (GSH), a tripeptide consisting of glutamate, cysteine and glycine, is the most ubiquitous non‐protein intracellular thiol in a large variety of organisms, including plants, animals and fungi. The thiol moiety of the cysteine residue confers unique redox and nucleophilic properties. In plant cells, GSH fulfils an indispensible role in the antioxidant system, sulphur metabolism and detoxification of xenobiotics. Upon grape crushing, GSH is extracted into the juice where it exerts several protective effects during the vinification process. In must, it reacts with oxidized phenolic compounds to form the colourless grape reaction product (GRP) which limits must browning to a certain extent. During wine ageing, GSH impedes the decrease of important aroma compounds, including certain esters, terpenes and volatile thiols, while at the same time preventing the development of atypical ageing off‐flavours. GSH may also inhibit the yellowing of wine during ageing. It is thus evident that elevated GSH levels in wine, in particular white wine which is more sensitive to oxidation, may be highly valuable for wine quality. The reductive crushing and pressing of white grape varieties, which limits oxidation and the consequent incorporation of GSH into GRP, promotes higher GSH levels in the juice. The reductive handling of juice also limits the formation of oxidized glutathione (GSSG). However, during alcoholic fermentation and maturation, levels generally decrease as a result of assimilation by the yeast Saccharomyces cerevisiae, and inevitable oxidation that takes place during the vinification process. The principal focus of this study was to gain a better understanding of the fate of glutathione during alcoholic fermentation and to establish whether certain oenological applications could result in elevated wine GSH levels. The application studied, included choice of yeast strain, extended lees contact, nitrogen supplementation and supplementation with glutathione enriched inactive dry yeast preparations (GHS‐IDYs). In addition, the need for a rapid analytical method for the simultaneous quantification of both GSH and GSSG in must and wine which does not involve derivatization or require extensive sample preparation, led to the development of a novel UPLC‐MS/MS method. The method was also employed to determine intracellular GSH and GSSG contents of the yeast S. cerevisiae and was studied for the first time in winemaking conditions. It was shown that the GSH levels fluctuated during alcoholic fermentation, suggesting the uptake and release by yeast. At the end of alcoholic fermentation, levels were generally lower than those initially present in grape juice, but in some cases, concentration increases were also observed. This finding indicates that, in some cases, endogenously‐produced GSH may be secreted into must during alcoholic fermentation, contributing to higher GSH levels in wine. Albeit small, significant differences in GSH content could be seen in wines fermented with different yeast strains, implying that yeast strain may to a certain extent influence wine GSH levels. While the effects of lees ageing and nitrogen supplementation seem to be insignificant in contributing to higher GSH levels in wine, the supplementation of must with GSH‐IDYs could result in increased wine GSH levels, provided the supplementation is done early during fermentation. This study has broadened our knowledge of several oenological factors, influencing GSH levels in wine and provided a new baseline for future research studies.
AFRIKAANSE OPSOMMING: Glutatioon (GSH), ’n tripeptied bestaande uit glutamaat, sisteïen en glisien, is die mees algemene nieproteïenagtige intrasellulêre tiool in ‘n wye verskeidenheid organismes, insluitende plante, diere en fungi. Die tioolfunksiegedeelte van die sisteïenresidu verleen unieke redoks‐ en nukleofiliese eienskappe. GSH vervul ‘n onmisbare rol in die antioksidantsisteem, swaelmetabolisme en die ontgiftiging van xenobiotika in plantselle. Tydens die maal van druiwe word glutatioon in die sap geëkstraeer waar dit verskeie beskermende effekte tydens die wynbereidingsproses uitoefen. GSH reageer met geöksideerde fenoliese verbindings om die kleurlose druifreaksieproduk (DRP) te vorm wat die verbruining van mos in ‘n sekere mate beperk. GSH verminder ook die afname van belangrike aromaverbindings tydens wynveroudering, insluitende sekere esters, terpene en vlugtige tiole, terwyl dit terselfdertyd die vorming van atipiese verouderingswangeure belemmer. So ook kan GSH die vergeling van wyn tydens veroudering inhibeer. Dit is dus voor die hand liggend that verhoogde GSH‐vlakke in wyn, in die besonder witwyn, wat meer oksidasie‐sensitief is, van waarde kan wees vir wynkwaliteit. Die reduktiewe maal en pers van witdruifvariëteite wat oksidasie en gevolglike inkorporasie van GSH in DRP beperk, bevorder hoër GSH‐vlakke in sap. So ook beperk die reduktiewe behandeling van sap die vorming van geoksideerde glutatioon (GSSG). Gedurende alkoholiese gisting en veroudering neem GSH‐vlakke egter af as gevolg van assimilasie deur die gis, Saccaromyces cerevisiae, asook onvermydelike oksidasie wat gedurende die wynbereidingsproses plaasvind. Die hooffokus van die studie was om ‘n beter begrip van die lot van glutatioon tydens alkoholiese gisting te verkry en om vas te stel of sekere wynkundige praktyke verhoogde GSH‐vlakke in wyn tot gevolg kan hê. Die studie het gisraskeuse, verlengde gismoerkontak, stikstofaanvulling en aanvulling met glutatioon‐verrykte, onaktiewe droëgis ingesluit. Daarbenewens het die behoefte aan ‘n vinnige analitiese metode vir die gelyktydige kwantifisering van sowel GSH as GSSG in mos en wyn wat nie derivatisering of uitgebreide monstervoorbereiding vereis nie, gelei tot die ontwikkeling van ‘n nuwe UPLC‐MS/MS metode. Hierdie metode is ook gebruik om die intrasellulêre GSH‐ en GSSG‐inhoud van die gis S. cerevisiae te bepaal wat vir die eerste keer in wynbereiding bestudeer is. Daar is bewys dat GSH‐vlakke tydens alkolholiese gisting fluktueer, wat dui op die opname en vrystelling daarvan deur die gis. Die vlakke aan die einde van alkoholiese gisting was oor die algemeen laer as vlakke aanvanklik teenwoordig in die sap. In sommige gevalle is konsentrasietoenames egter ook waargeneem. Hierdie bevinding dui daarop dat intrasellulêr‐vervaardige GSH, in sommige gevalle, in die mos uitgeskei kan word, wat tot hoër GSH‐vlakke in wyn lei. Klein, dog beduidende verskille in GSHinhoud is waargeneem in wyne wat met verskillende gisrasse berei is, wat daarop dui dat gisras in ‘n sekere mate die GSH‐vlakke in wyn kan beïnvloed. Alhoewel die effek van gismoerveroudering en stikstofaanvulling onbeduidend is, kan die aanvulling van mos met glutatioon‐verrykte, onaktiewe droëgis tot verhoogde GSH‐vlakke in wyn lei, mits die aanvulling vroeg tydens alkoholiese gisting gedoen word. Hierdie studie verbreed ons kennis van verskeie wynkundige praktyke wat GSH‐vlakke in wyn beïnvloed en vorm ‘n nuwe basis vir toekomstige navorsingstudies.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: The table grape industry employ a wide range of viticultural management practices in order to produce the high quality grapes demanded by the export market. A common contributor to degrading the quality of table grapes is the occurrence of berry split, which not only has an unattractive visual effect, but also increases the berries’ susceptibility to infection by spoilage organisms. A number of environmental conditions such as rainfall and humidity, and/or agricultural practices, such as irrigation, and high density canopies, can lead to higher plant cell water content. This in turn, can increase the potential of berry split to occur. To date, the main method of berry split prevention has been the management of plant water status by; (i) regulating irrigation withdrawal times, and (ii) covering of canopies if rainfall is predicted prior to harvest. The aim of this study was to determine the effect that irrigation frequency, as induced by irrigation withdrawals; as well as boron (B) and calcium (Ca) treatments, applied as bunch directed sprays, have on pre- and post-harvest berry split. To this end, a newly released late ripening, white seedless cultivar, SoutherngrapeOne was chosen as a model cultivar as it has a high susceptibility to berry split. SoutherngrapeOne vines were subsequently subjected to a range of irrigation frequencies based on typical irrigation scheduling used in the table grape industry, which comprised of a low, medium and high frequency. The low frequency was duplicated in order to demonstrate the effect that a heavy irrigation, just before harvest may have on berry split. These treatments were further subdivided to investigate the effect that B and Ca may have on berry split. For the B treatment, four Solubor1 bunch directed sprays were applied from 8mm berry size to véraison. The Ca treatment consisted of Stopit R 2 and Caltrac R 3 bunch directed sprays applied over the same period. In addition, a combination of the B and Ca treatment were applied to investigate any possible interaction. To account for the effect of water as solvent in the B and Ca treatments, and the spraying effect, pure water as treatment was also evaluated. Control vines received no sprays. The applied irrigation treatments resulted in different plant water status conditions. Separate applications of B and Ca treatments resulted in a decrease of B and Ca content in the flesh respectively. The control and combination treatment, of B and Ca resulted in the same of B and Ca content in the flesh. Furthermore, none of the applied treatments resulted in an increase of either B or Ca content in the berry skin. It was found that the medium frequency irrigation resulted in the best irrigation strategy to prevent pre-harvest berry split. Surprisingly, all the subtreatments: B, Ca, and combination of B and Ca, resulted in an increased incidence of pre-harvest berry split when compared to the control group for the 2006/07 season. However, in the 2007/08 season only the B treatment resulted in an increase of pre-harvest berry split. Concerning post-cold-storage physiological disorders, Ca treatments appear to have reduced berry drop, but increased decay. In the 2006/07 season, the B treatment resulted in reduced post-cold-storage berry split, whereas Btreatment in the 2007/08 season had no effect. Both B and Ca should be considered as having the potential to increase the appearance of hairline cracking. Calcium treatment also led to an increase in decay which may have been as a result of the splitting it contributed to. Low frequency irrigation recieving irrigation before harvest was found to result in browner stems. Low irrigation frequencies decreased the cell size of the berry skin. The Ca treatment gave rise to thicker (weaker) cell walls, this morphological change may be responsible for the physiological disorders it caused. From these findings, it can be deduced that poorly managed irrigation, together with unnecessary application of B and/or Ca may result in an increase of berry split and other physiological disorders, with subsequent financial losses for the producer.
AFRIKAANSE OPSOMMING; Die tafeldruifindustrie maak gebruik van ’n wye reeks wingerdkundige praktyke ten einde die hoë gehalte druiwe te produseer wat die uitvoermark vereis. Korrelbars is ’n algemene verskynsel wat afbreek maak tot die gehalte van tafeldruiwe. Behalwe dat voorkoms van die druiwe benaadeel word, verhoog dit ook in vatbaarheid vir infeksie deur verrottingsveroorsakende swamme. Hoë reënval en humiditeit, sowel as wingerdkundige praktyke soos besproeiing en hoë lowerdigtheid, wat kan lei tot verhoogde waterstatus in plante, kan lei tot ’n toename in korrelbars. Daar word hoofsaaklik van twee metodes gebruik gemaak om korrelbars te beheer, naamlik die bestuur van plantwaterstatus deur; (i) beheer van besproeiingsontrekkingstye en (ii) bedekking van lowers indien reën voorspel word voor oestyd. Die doel van hierdie studie was om vas te stel wat die invloed van besproeiings frekwensies sowel as trosgerigte boor (B) en kalsium (Ca), spuitbehandelings, op voor- en na-oes korrelbars het. Die onlangs vrygestelde laat rypwordende, wit, pitlose kultivar, SoutherngrapeOne is gebruik, aangesien dit hoogs gevoelig is vir korrelbars. Stokke is aan verskillende besproeiings intervalle, soos tipies gebruiklik in die tafeldruifindustrie, blootgestel. Hierdie intervalle bestaan uit n’ lae, medium en hoë besproeiings frekwensie. Die lae besproeiings frekwensie is herhaal ten einde die invloed van besproeiing net voor oestyd op korrelbars te ondersoek. Die invloed van B- en Ca-behandeling op korrelbars is ook ondersoek. Vir die B-behandeling is vier Solubor1 trosgerigte spuite aangewend vanaf 8mm korrelgrootte tot deurslaan. Vir die Ca-behandeling is Stopit R 2 en Caltrac R 3 as trosgerigte spuite oor dieselfde tyd toegedien. Kombinasiebehandelings is ook aangewend om enige interaksie tussen B en Ca te ondersoek. Waterbehandelings is ook toegedien om die invloed van water as oplosmiddel van B- en Ca-behandelings sowel as die spuit-effek te ondersoek. Kontrole stokke is ook ingesluit en het geen spuitebehandeling ontvang nie. Die besproeiingsbehandelings het verskillende plantwater toestande tot gevolg gehad, B- en Ca-behandelings het gelei tot ’n afname in B- en Cainhoud in die vleis onderskeidelik. Die kombinasie en kontrole behandelings het eenderse hoeveelhede B en Ca in die vleis tot gevolg gehad. Geen van die aangewende behandelings gelei tot ’n toename in B- en Ca-inhoud in die dop nie. Die resultate toon dat medium besproeiings frekwensie die beste besproeiingsstrategie is om voor-oes korrelbars te voorkom. In vergelyking met die kontrole-behandeling in 2006/07, het B, Ca en die kombinasie van B en Ca, ’n toename in voor-oes korrelbars tot gevolg gehad. In die 2007/08 seisoen het slegs die B-toediening egter tot ’n toename in voor-oes korrelbars gelei. Kalsium behandelings het ’n afname in los-korrels, maar ’n verhoging in korrelbars tot gevolg gehad. In 2006/07, het B-toediening tot ’n afname in korrelbars na koelopberging gelei, maar in die 2007/08 seisoen het dit geen effek gehad nie. Beide B- en Ca-toediening het die potensiaal om haarlyn barste te veroorsaak. Kalsium toediening het bederf verhoog wat moontlik aan die hoër bars wat dit induseer toegeskryf kan word. Lae besproeiings frekwensie, het bruiner stingels veroorsaak, en ook gelei tot ’n afname van selgrootte in die dop. Die Ca-toediening het aanleiding gegee tot dikker selwande in die dop. Hierdie anatomiese veranderinge kan moontlik die rede wees vir die verhoging in fisiologise afwykings. Van hierdie bevindinge kan ons aflei dat swak bestuur van besproeiing, sowel as die onnodige aanwending van B en/of Ca, kan aanleiding gee tot ’n toename in korrelbars en ander fisiologiese afwykings, en dus finansiële verliese vir die produsent inhou.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Within-vineyard variability in vigour and water status commonly occurs in South African vineyards. Different soil types found over short distances are probably the main cause of vigour variability, while differences in grapevine water status are commonly induced by lateral water flow in the vineyard, blocked irrigation emitters and differences in soil water-holding capacity. These factors can cause heterogeneous ripening and differences in fruit quality between different parts of the vineyard, an aspect that needs to be avoided as far as possible in order to produce quality wines. Measurements of carbon isotope discrimination (CID) have proved to be a tool to assess grapevine physiology in order to study the effects of environmental parameters on leaf carbon dioxide (CO2) gas exchange and stomatal conductance (gs). Grapevine water deficit stress/strain in reaction to these environmental conditions can then be determined by observing the amount of 13C absorbed by plant material after discrimination of 13C has taken place, and this is influenced by the grapevine stress condition and can indicate water-use efficiency. In this study, the variability of grapevine water status and vigour was determined in order to quantify these parameters in different parts of the vineyard. Two separate trials were conducted, the first at Wellington, South Africa, where different irrigation regimes resulted in variability in grapevine water status between plots. The second trial was at Stellenbosch, South Africa, where plots were divided among different vigour classes and irrigation was applied in different quantities for different irrigation treatments. Within-vineyard variability in water status (Wellington and Stellenbosch) and vigour (Stellenbosch) were then quantified and the effects on some grapevine physiological parameters and berry composition were measured. The treatments in the Wellington trial led to differences in grapevine water status, which could be quantified by measurements of stem water potential (SWP) and leaf water potential (LWP). Soil variability also led to differences in grapevine vigour, which were quantified by measurements of pruning mass, leaf area and shoot length. The effect of the variability in grapevine water status on grapevine physiology was assessed by measuring CID, which was the main focus of the study. Other physiological measurements, such as gs and leaf and canopy temperature, were also conducted. The effect of these conditions on grape berry composition was also studied. In the Stellenbosch trial, soil water content, plant water status measurements (SWP, predawn LWP and LWP), physiological measurements (CID and gs) and berry size measurements were used to classify plots into water status treatments (“wet” and “dry” treatments). The effect of vigour differences was analysed separately from these treatments by using pruning mass as a covariate in the statistical analyses. The effect of vigour variability on the measurements was studied by looking at the effect of the covariate on the measurements, while shoot growth rate, shoot length and leaf area measurements were conducted as vegetative growth measurements. Differences in measurements were then studied between the treatments and between the vigour levels of the different plots. In the Wellington trial, plant water status was determined by irrigation, showing increased stress for treatments that received less irrigation. The differences in plant water status then caused differences in grapevine physiology between the treatments, leading to increased gs for increased irrigation. This of course influenced leaf internal CO2 and therefore CID, although CID was also clearly influenced by berry development. Berry size was influenced by irrigation, with larger berries found in wetter treatments, while berry chemical composition was influenced by the irrigation regime, with increased irrigation leading to increased pH and leading to trends showing increased total soluble solids and malic acid, and reduced total and tartaric acid and colour intensity. In the Stellenbosch trial, plots with higher vigour had increased shoot growth rate, longer shoots and increased leaf area, although topping influenced this. Wet treatment vines also showed slightly longer shoots and larger leaf areas. There were differences in soil water content between the wet and dry treatments, and this led to differences in plant water status. Vigour also influenced pre-dawn LWP, especially in the 2007 season, as higher-vigour vines struggled more to rehydrate through the night. Differences in plant water potential led to differences in grapevine physiology, with increased gs for vines from the wet treatment, while higher-vigour vines had slightly increased gs. The differences in gs led to gas exchange differences and therefore differences in CID, meaning that water status and vigour influenced CID. CID measurements illustrated the long term effect of water status on plant physiology, while measurements such as SWP illustrated the short term effects. CID measurements therefore proved to be accumulative over the season, in contrast to SWP measurements that were much more dependent on the current state of grapevine water status. Other physiological measurements showed that wet-treatment vines had higher photosynthetic rates and evapotranspiration and lower leaf temperatures, while higher-vigour vines had slightly increased evapotranspiration and decreased leaf temperatures. Wet-treatment vines had larger berries, while a higher vigour also led to slightly larger berries. Berry composition was influenced by treatment, where wet-treatment vines had increased pH and total soluble solids, while higher-vigour vines had increased juice pH and, in the 2008 season, decreased total soluble solids. Extremely stressed conditions did not show significant effects on plant water potential, but SWP measurements indicated slightly higher stress for the extremely stressed vines and LWP showed slightly less stressed conditions for these vines. Measurements of gs showed slightly lower values for the extremely stressed vines, while measurements of CID showed large significant differences, with the extremely stressed vines having measurements showing high stress. The measurement therefore indicated highly stressed conditions accurately, while other physiological measurements, such as photosynthetic rate, evapotranspiration and leaf temperatures, only showed trends and no significant differences. Measurements of stomatal conductance reacted to plant water status measurements throughout the diurnal measurement days, while CID only reacted slightly with gs changes during these days and was perhaps influenced more by berry chemical composition and development at this early stage of the season. Vigour and water status therefore influenced grapevine physiology, with a more direct effect by water status and an indirect effect by vigour due to microclimatic differences. This also influenced berry composition and therefore quality. In future studies, CID measurements should be done on juice from which organic acids have been removed in order to eliminate the effect of seasonal berry composition on the measurement. Measurements of CID proved to be an integrative, but sensitive, indicator of grapevine stress, especially at the end of the season. It might at best be useful as a post-harvest management tool for producers or grape buyers, especially for irrigation control, as has also been stated by Van Leeuwen et al. (2007).
AFRIKAANSE OPSOMMING: Binne-wingerd variasie in groeikrag en waterstatus is algemeen in Suid-Afrikaanse wingerde. Verskillende grondsoorte wat na aan mekaar voorkom, is seker een van die vernaamste oorsake van variasie in groeikrag, terwyl verskille in wingerdwaterstatus algemeen deur laterale watervloei in die wingerd, verstopte besproeiingspuite en verskille in grond waterhouvermoë geïnduseer word. Hierdie faktore kan aanleiding gee tot heterogene rypwording en verskille in vrugkwaliteit tussen verskillende dele van die wingerd, ‘n aspek wat so ver moontlik vermy moet word om kwaliteitwyne te kan produseer. Die meting van koolstof-isotoopdiskriminasie (KID) is bewys om as gereedskap te kan dien vir die assessering van wingerdfisiologie om die effekte van omgewingsparameters op blaar koolstofdioksied (CO2) - gasuitruiling en stomatale geleiding (gs) te bestudeer. Die stres/stremming as gevolg van ‘n watertekort in die wingerd in reaksie op hierdie omgewingstoestande kan dan bepaal word deur te kyk na hoeveel 13C deur die plantmateriaal geabsorbeer word ná 13C-diskriminasie plaasgevind het, en dít word deur die wingerdstrestoestande beïnvloed en kan ‘n aanduiding verskaf van die doeltreffendheid van waterverbruik. In hierdie studie is die variasie in wingerdwaterstatus en groeikrag bepaal om hierdie parameters in verskillende dele van die wingerd te kwantifiseer. Twee afsonderlike proewe is uitgevoer, die eerste by Wellington, Suid-Afrika, waar verskillende besproeiingsregimes gelei het tot verskille in die wingerdwaterstatus tussen persele. Die tweede proef was by Stellenbosch, Suid-Afrika, waar persele tussen verskillende groeikragklasse verdeel is en besproeiing in verskillende hoeveelhede vir verskillende besproeiingsbehandelings toegepas is. Binne-wingerd variasie in waterstatus (Wellington en Stellenbosch) en groeikrag (Stellenbosch) is toe gekwantifiseer en die effekte op sekere wingerd-fisiologiese parameters en korrelsamestelling is gemeet. Die behandelings in die Wellington-proef het gelei tot verskille in wingerdwaterstatus, wat deur metings van stamwaterpotensiaal (SWP) en blaarwaterpotensiaal (BWP) gekwantifiseer kon word. Grondverskille het ook gelei tot verskille in wingerdgroeikrag, wat deur metings van snoeimassa, blaaroppervlak en lootlengte gekwantifiseer is. Die effek van die variasie in wingerdwaterstatus op wingerdfisiologie is deur metings van KID bepaal wat die hooffokus van hierdie studie was. Ander fisiologiese metings, soos gs en blaar- en lowertemperatuur, is ook gedoen. Die effekte van hierdie toestande op die samestelling van die druiwekorrels is ook bestudeer. In die Stellenbosch-proef is grondwaterinhoud, metings van plantwaterstatus (SWP, voorsonopgang SWP en BWP), fisiologiese metings (KID en gs) en metings van korrelgrootte gebruik om die persele in waterstatusbehandelings (“nat” en “droë” behandelings) te verdeel. Die effek van verskille in groeikrag is apart van hierdie behandelings geanaliseer deur snoeimassa as ‘n kovariaat in die statistiese analises te gebruik. Die effek van groeikragvariasie op die metings is bestudeer deur ondersoek in te stel na die effek van die kovariaat op die metings, terwyl lootgroeitempo-, lootlengte- en blaaroppervlakmetings as metings van vegetatiewe groei uitgevoer is. Verskille in metings tussen die behandelings en tussen die groeikragvlakke van die verskillende persele is toe bestudeer. In die Wellington-proef is plantwaterstatus deur besproeiing bepaal, met verhoogde stres in behandelings waar daar minder besproeiing toegedien is. Die verskille in plantwaterstatus het dan verskille in wingerdfisiologie tussen die behandelings veroorsaak, wat gelei het tot ‘n verhoogde gs in die geval van verhoogde besproeiing. Dit het natuurlik ‘n effek op die interne CO2 van die blaar en dus op KID gehad, hoewel KID ook duidelik deur korrelontwikkeling beïnvloed is. Korrelgrootte is deur besproeiing beïnvloed, met groter korrels in die natter behandelings, terwyl die chemiese samestelling van die korrel deur besproeiingsregime beïnvloed is. Verhoogde besproeiing het pH verhoog en gelei na tendense wat verhoogde totale oplosbare vaste stowwe en appelsuur, en verminderde totale suur, wynsteensuur en kleurintensiteit getoon het. In die Stellenbosch-proef het persele met hoër groeikrag ook verhoogde lootgroeitempo, langer lote en verhoogde blaaroppervlak getoon, hoewel dit deur top beïnvloed is. Wingerdstokke van die nat behandeling het ook effe langer lote en groter blaaroppervlakke getoon. Daar was verskille in grondwaterinhoud tussen die nat en droë behandelings en dit het verskille in plantwaterstatus veroorsaak. Groeikrag is ook deur voor-sonopgang BWP beïnvloed, veral in die 2007-seisoen, aangesien stokke met hoër groeikrag meer gesukkel het om in die nag te rehidreer. Verskille in plantwaterpotensiaal het gelei tot verskille in wingerdfisiologie, met ‘n verhoogde gs vir stokke in die nat behandeling, terwyl stokke met hoër groeikrag ‘n effens verhoogde gs getoon het. Die verskille in gs het gelei tot verskille in gasuitruiling en dus verskille in KID, wat beteken dat waterstatus en groeikrag ‘n invloed op KID het. KID was meer verteenwoordigend van die langtermyneffekte van water status op plantfisiologie, terwyl metings soos SWP die korttermyneffekte weerspieël het. KID metings was dus akkumalatief oor die seisoen, terwyl SWP metings meer ‘n weerspieëling was van die huidige toestand van plantwaterpotensiaal. Ander fisiologiese metings het getoon dat stokke in die nat behandeling ‘n hoër fotosintesetempo en evapotranspirasie sowel as laer blaartemperature ondervind het, terwyl die stokke met hoër groeikrag effe verhoogde evapotranspirasie en verminderde blaartemperature getoon het. Stokke in die nat behandeling het groter korrels gehad, terwyl hoër groeikrag ook effens groter korrels veroorsaak het. Korrelsamestelling is deur die behandelings beïnvloed, met stokke in die nat behandeling wat verhoogde pH en totale oplosbare vaste stowwe getoon het, terwyl stokke met hoër groeikrag verhoogde pH van die sap en verminderde totale oplosbare vaste stowwe (laasgenoemde in die 2008-seisoen) gehad het. Uitermate toestande van stres het geen beduidende effekte op plantwaterpotensiaal getoon nie, hoewel SWP-metings effens hoër stres vir die uitermate gestresde wingerde getoon het en BWP effens minder gestresde toestande vir hierdie stokke getoon het. Metings van gs het effens laer waardes vir die uitermate gestresde stokke getoon, terwyl metings van KID groot noemenswaardige verskille getoon het, met die metings vir die uitermate gestresde wingerde wat hoër stres aangedui het. Dié meting het dus hoogs gestresde toestande akkuraat aangedui, terwyl ander fisiologiese metings, soos tempo van fotosintese, evapotranspirasie en blaartemperature net tendense en nie beduidende verskille aangedui het nie. Metings van stomatale geleiding het dwarsdeur die dae waarop daaglikse metings gedoen is op plantwaterstatusmetings gereageer, terwyl KID net effens met gs-veranderinge op hierdie dae gereageer het en moontlik meer deur die chemiese samestelling en ontwikkeling van die korrel in hierdie vroeë stadium van die seisoen beïnvloed is. Groeikrag en waterstatus het dus wingerdfisiologie beïnvloed, met ‘n meer direkte effek deur waterstatus en ‘n indirekte effek deur groeikrag as gevolg van mikroklimaatsverskille. Dit het ook korrelsamestelling en dus kwaliteit beïnvloed. In toekomstige studies moet KID-metings gedoen word op sap waarvan die organiese sure verwyder is om die effek van seisoenale korrelsamestelling op die meting uit te sluit. Metings van KID is getoon om ‘n integrerende, maar gevoelige, aanduider van wingerdstres te wees, veral aan die einde van die seisoen. Dit is ten beste miskien bruikbaar as naoesbestuursgereedskap vir produsente of druiwekopers, veral vir besproeiingsbeheer, soos ook reeds deur Van Leeuwen et al. (2007) aangedui is.

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2009.
ENGLISH ABSTRACT: Tannins and colour components in red wine are important quality parameters. These factors can be manipulated in the vineyard by grape growing techniques or in the cellar by different winemaking practices. Grape seeds make a significant contribution to tannin concentration in wine when compared to those from the skins and pulp. Tannins contribute to the ageing potential, organoleptic properties and stabilisation of red wine colour. The colour of a red wine is also influenced by malolactic fermentation, the biological process that transforms malic acid into lactic acid which normally leads to an increase in pH. The subsequent change in pH alters the anthocyanin equilibrium, the primary colour components in red wine. Oxygen contributes to the polymerisation of anthocyanins over time to form more stable pigments that are less sensitive to pH fluctuations and sulphur dioxide bleaching. Limited research has been done on the use of oxygen after alcoholic fermentation and the impact it has on the phenolic composition of red wines. Similarly, only a few studies have examined the impact of either the addition or removal of seeds to the phenolic composition of a red wine in combination with oxygen addition. Additionally, little published data seems to exist on the effect of different pHs on red wine’s phenolic and colour development after oxygen addition. In our results we have shown that it is possible to stabilise wine colour by adding supplementary seeds before alcoholic fermentation. This led to an increase in colour intensity in certain cases. Some red wines produced without seeds had significantly lower colour intensities. This clearly suggests that catechins and proanthocyanidins are extracted from seeds and contribute to wine colour as they combine with other pigments to stabilise wine colour. Spectrophotometric and HPLC analyses have shown that the total phenolic content increased with seed concentration. However, we have observed that a wine may possibly become saturated with phenols when supplementary seeds are added. Anthocyanin concentrations often decreased when oxygen was added, while polymeric phenols and polymeric pigments sometimes increased. When applying different oxygen dosages to a red wine on commercial scale with microoxygenation, it was found that monomeric anthocyanins decreased as more oxygen was added and this decrease in anthocyanins led to the formation of stable polymeric pigments. This was reflected in the significant increase in colour intensity for the wines receiving oxygen. Small differences were detected in the total phenol and tannin concentration for the control and oxygenated wines. However, some of these phenolic and colour differences disappeared during subsequent ageing of the wine. When making wines of different initial pHs, we observed that the colour density decreased as the pH increased. The application of oxygen reduced the decrease in colour during MLF, especially at a lower pH. The addition of oxygen did not result in significantly different polymeric pigment concentrations in the various pH treatments, although the results could have been different if the wines were aged. However, pH differences in the range between 3.4 and 4.0 did not significantly influence the phenol composition of the wines under our conditions. This study led to a better understanding on the effect of oxygen additions under different conditions on red wine's phenolic and colour composition.
AFRIKAANSE OPSOMMING: Tanniene en kleur komponente teenwoordig in rooiwyn is belangrike kwaliteit parameters. Hierdie faktore wat wynkwaliteit beïnvloed kan gemanipuleer word in die wingerd deur verskeie verbouingstegnieke toe te pas en in die kelder deur die toepassing van verskillende wynbereidingsmetodes. Die bydrae van sade tot die finale tannienkonsentrasie in rooiwyn is groot in vergelyking met dié van die pulp en doppe. Tanniene dra by tot die verouderingspotensiaal, organoleptiese eienskappe en die stabilisasie van die wynkleur. Die kleur van ‘n rooi wyn word ook beïnvloed deur appelmelksuurgisting (AMG), die biologiese proses wat appelsuur omskakel na melksuur en ‘n gevolglike toename in die pH van die wyn veroorsaak. Hierdie verandering in die pH van die wyn beïnvloed die antosianien ewewig, die primêre kleur komponente teenwoordig in rooiwyn. Suurstof dra by tot die polimerisasie van antosianiene oor tyd om meer stabiele kleur pigmente te vorm met ‘n hoër kleurintensiteit wat minder sensitief is teenoor pH veranderinge en die bleikingseffek van swaweldioksied. Beperkte navorsing is gedoen op die gebruik van suurstof na alkoholiese gisting en die impak daarvan op die fenoliese samestelling van ‘n rooiwyn. Slegs ‘n paar studies het die invloed van die verwydering of byvoeging van sade in kombinasie met suurstoftoediening op ‘n rooiwyn se fenoliese samestelling ondersoek. Dit wil voorkom of beperkte gepubliseerde data beskikbaar is oor die effek wat verskillende pH’s het op rooi wyn se fenoliese en kleurontwikkeling na suurstof byvoeging. Ons resultate het aangedui dat dit wel moontlik is om in sekere gevalle die kleur van ‘n rooiwyn te stabiliseer deur addisionele sade by te voeg voor alkoholiese fermentasie. Hierdie byvoeging het ‘n toename in kleurintensiteit tot gevolg gehad. Sekere wyne wat gemaak is sonder sade het ‘n kenmerkend laer kleur intensiteit gehad. Hierdie bevinding is ‘n duidelike bewys dat katesjiene en prosianidiene geëkstraheer word vanuit die sade en bydra tot wynkleur deurdat hulle met ander pigmente verbind om die kleur sodoende te stabiliseer. Spektrofotometriese en hoë druk vloeistof chromatografie (HDVC) analises het gewys dat die totale fenoliese konsentrasie neem toe met ‘n toename in saad konsentrasie. Daar is egter waargeneem dat ‘n wyn moontlik versadig kan raak met fenole wanneer addisionele sade bygevoeg word. Antosianien konsentrasies het meestal afgeneem wanneer suurstof bygevoeg is, maar polimeriese fenole en polimeriese pigmente het partykeer toegeneem. Met die toediening van verskillende suurstof dosisse tot ‘n rooiwyn op kommersiële skaal het ons bevind dat monomeriese antosianiene afneem wanneer meer suurstof bygevoeg word. Hierdie afname in antosianiene het egter gelei tot die vorming van stabiele polimeriese pigmente. Dié bevinding was gereflekteer in die toename in kleurintensiteit van wyne wat met suurstof behandel is. Klein verskille was waargeneem vir die totale fenol en tannien konsentrasies tussen die kontrole en wyne behandel met suurstof. Sekere van hierdie fenoliese kleur verskille het egter afgeneem tydens die daaropvolgende veroudering van die wyne. Wyne wat gemaak is met verskillende aanvanklike pH’s se kleurintensiteit neem af soos die pH toeneem. Die toediening van suurstof het die kleurverlies tydens AMG verminder, veral by ‘n laer pH. Die toediening van suurstof het nie verskillende polimeriese pigment konsentrasies by verskillende pH’s veroorsaak nie, maar ‘n verskil kon moontlik waargeneem word indien die wyne verouder was. pH verskille tussen 3.4 en 4.0 het egter nie die fenoliese samestelling van die wyne onder ons omstandighede beduidend beïnvloed nie. Hierdie studie het gelei tot meer kennis oor die effek van suurstoftoedienings onder verskillende kondisies op rooiwyn se fenoliese en kleursamestelling.

Thesis (MSc)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: The polysaccharides that are present in wine originate from the grapes, the fungi that grow on the grapes and from other microorganisms that come into contact with the must during winemaking. The grape-derived polysaccharides of most concern in winemaking are pectin, glucan and xylan that can be enzymatically degraded by pectinases, glucanases and xylanases, respectively. These are the main structural polysaccharides of the cell wall of the grape cell. Degradation of the cell walls will result in the separation and rupture of the grape cells, and cell wall-bound compounds will be released into the must. Treating the must with pectinase and macerating enzyme preparations can result in an increase in free-flow juice, an improvement in must clarification and filtration, and an increased extraction of phenols and tannins. The tannins that are extracted polymerise with anthocyanins in red wine during ageing, resulting in increased colour intensity and stability. Wine aroma is also influenced by enzyme treatment. The degradation of the cell wall contributes to the release of glycosidically-bound terpene or alcohol precursors from the berries. The hydrolysis of these precursors during fermentation can result in an improvement in aroma. It can thus be seen that it is possible to improve wine quality and processing by supplementing the endogenous enzymes that are present in the fermentation with commercial enzyme preparations. Commercial enzymes are typically crude fungal preparations. The majority of commercial pectinase and glucanase preparations are derived from Aspergillus and Trichoderma, respectively. Since the endogenous polysaccharase activity of Saccharomyces cerevisiae is very limited, the heterologous expression of specific polysaccharase genes in an industrial yeast strain can improve the winemaking process, resulting in a higher quality wine without the addition of expensive commercial enzyme preparations. Since only the desired enzymes are secreted by the recombinant strain, there will be no undesired sideactivities, which can be detrimental to wine quality. Several pectinase-, glucanaseand xylanase-encoding genes, cloned from a variety of organisms, have been expressed successfully in laboratory strains of S. cerevisiae. Attempts have also been made to construct industrial wine yeast strains that express these polysaccharase genes and secrete the encoded enzymes. Fermentation with some of these strains resulted in a decrease in total phenolics and turbidity, an increase in juice extraction, and alterations in the colour and aromatic profile of the resulting wines. In this study, four polysaccharide-degrading, recombinant wine yeast strains were constructed. The endo-β-1,4-xylanase gene, XYN2, and the endo-β-1,4-glucanase gene, end1, were previously cloned from the soft rot fungus Trichoderma reesei and the rumen bacterium Butyrivibrio fibrisolvens, respectively. These genes were subcloned into different expression cassettes which were used to construct the four integration plasmids. The recombinant plasmids contained the following gene cassettes: TEF1P-XYN2-ADH2T (plasmid pDLG29) ADH1P- MFα1S -end1-TRP5T (plasmid pDLG30) ADH1P-MFα1S-end1-TRP5T and ADH2P-XYN2-ADH2T (plasmid pDLG33), ADH1P-MFα1S-end1-TRP5T and YG100PXYN2- ADH2T (plasmid pDLG39). These four plasmids were then separately integrated into the ILV2 locus of the commercial wine yeast strain S. cerevisiae VIN13. Wine was made with the four strains constructed in this study, a pectolytic strain, VIN13[pPPK], a glucanase- and xylanase-secreting strain, VIN13[pEX], an untransformed VIN13 strain, and an untransformed strain with the addition of the commercial enzyme preparation Rapidase EX Colour. Microvinification experiments were carried out on Pinot noir, Ruby Cabernet and Muscat d’Alexandria wines. Fermentation with the polysaccharide-degrading strains resulted in significant improvements in juice extraction, colour intensity and stability, and in alterations in the aromatic profiles of the wines produced. Subject to the approval by the regulatory authorities and eventual consumer acceptance of the use of genetically modified organisms (GMOs) in fermented foods and beverages, it might be required that the GM status of the yeast that is used appears on the label. Currently, there is no robust technique available with which the use of GM yeast can be revealed in a finished wine because the yeast cells and their DNA are removed from or denatured in the wine during filtration and processing. One way with which the undeclared use of a GM yeast in winemaking could be exposed would be to compare the chemical profile of a suspect wine with that of non-GM wine. In order to explore this concept further, a secondary aim of this study was to investigate whether Fourier Transformation Infra Red (FT-IR) spectroscopy coupled with multivariate data analysis could distinguish between wines fermented with transgenic and non-transgenic yeast strains, or between wines fermented with different transgenic strains. The results showed that this method could be used to classify wines fermented with different yeast strains if fermentation with the strain resulted in a unique chemical profile in the resulting wine. This was a preliminary study and these findings were summarised as an addendum to the thesis.
AFRIKAANSE OPSOMMING: Die polisakkariede wat in wyn teenwoordig is, is afkomstig van die druiwe, die swamme wat op die druiwe groei en vanaf ander mikroörganismes wat tydens die wynmaakproses met die mos in aanraking kom. Die belangrikste druifpolisakkariede in wynbereiding is pektien, glukaan en xilaan, wat onderskeidelik deur pektinases, glukanases en xilanases afgebreek kan word. Hierdie is die vernaamste strukturele polisakkariede van ‘n druifsel se selwand. Die afbreking van die selwande veroorsaak dat die druifselle skei en skeur, met die gevolg dat die selwandgebonde verbindings in die mos vrygelaat word. Die behandeling van die mos met pektinase en versappingsensiempreparate kan tot ʼn toename in vry-afloopsap lei, sowel as ʼn verbetering in mosverheldering en -filtrasie en ʼn verhoogde ekstraksie van fenole en tanniene. Die tanniene wat geëkstraheer word, polimeriseer in rooiwyn tydens veroudering, en dit lei tot verhoogde kleurintensiteit en -stabiliteit. Wynaroma word ook deur ensiembehandeling beïnvloed. Die afbreking van die druifselwand dra by tot die vrylating van glikosidiesgebonde terpeen- en alkoholvoorlopers uit die korrels. Die hidrolise van hierdie voorlopers tydens gisting kan lei tot ʼn verbetering van die aroma. Dit is dus duidelik dat dit moontlik is om wynkwaliteit en wynbereiding te verbeter deur die endogene ensieme wat in die gisting teenwoordig is met kommersiële ensiempreparate te supplementeer. Kommersiële ensiempreparate is tipies ongesuiwerde swampreparate. Die meerderheid kommersiële pektinase- en glukanasepreparate word onderskeidelik vanaf Aspergillus en Trichoderma verkry. Aangesien die endogene polisakkaraseaktiwiteit van Saccharomyces cerevisiae baie beperk is, kan die heteroloë uitdrukking van spesifieke polisakkarase-gene in ʼn industriële gisras die wynbereidingsproses verbeter en lei tot ʼn hoër kwaliteit wyn sonder die byvoeging van duur kommersiële ensiempreparate. Omdat die verkose ensieme deur die rekombinante ras uitgeskei word, sal daar geen ongewenste newe-effekte teenwoordig wees wat ʼn nadelige effek op wynkwaliteit kan hê nie. Verskeie mikrobiese gene wat vir pektinases, glukanases en xilanases kodeer, is reeds voorheen uit ‘n wye verskeidenheid van organismes gekloneer en suksesvol in laboratoriumrasse van S. cerevisiae uitgedruk. Pogings is ook aangewend om industriële wyngisrasse te konstrueer wat hierdie polisakkarasegene uitdruk en hul enkodeerde ensieme uitskei. Gisting met sommige van hierdie rekombinante gisrasse het gelei tot ʼn afname in totale fenoliese verbindings en troebelheid, ʼn verhoging in sapekstraksie, en veranderings in die kleur en aromatiese profiel van die gevolglike wyne. In hierdie studie is vier polisakkaried-afbrekende, rekombinante wyngisrasse gekonstrueer. Die endo-β-1,4-xilanasegeen, XYN2, en die endo-β-1,4- glukanasegeen, end1, is voorheen reeds onderskeidelik vanaf die sagte vrotswam, Trichoderma reesei, en die rumenbakterium, Butyrivibrio fibrisolvens, gekloneer. Hierdie gene is in vier integrasieplasmiede in verskillende ekspressiekassette gesubkloneer. Die plasmiede het die volgende geenkassette bevat: TEF1P-XYN2- ADH2T (plasmied pDLG29) ADH1P- MFα1S -end1-TRP5T (plasmied pDLG30) ADH1PMFα1S- end1-TRP5T and ADH2P-XYN2-ADH2T (plasmied pDLG33), ADH1P-MFα1S end1-TRP5T and YG100P-XYN2-ADH2T (plasmied pDLG39). Hierdie vier plasmiede is toe afsonderlik in die ILV2-lokus van die kommersiële wyngisras, S. cerevisiae VIN 13, geïntegreer. Wyn is met hierdie vier gekonstrueerde gisrasse gemaak, die pektolitiese gisras, VIN13[pPPK], die glukanase- en xilanase-afskeidende gisras, VIN13[pEX], die ongetransformeerde VIN13-ras, en met ʼn ongetransformeerde VIN13 gis waarby die kommersiële ensiempreparaat, Rapidase EX Colour, bygevoeg is. Mikro-wynbereidingseksperimente is op Pinot noir-, Ruby Cabernet- en Muscat D’Alexandria wyne uitgevoer. Gisting met die polisakkaried-afbrekende gisrasse het gelei tot ʼn noemenswaardige verbetering in sapekstraksie, kleurintensiteit en kleurstabiliteit, asook in veranderinge in die aromatiese profiele van die geproduseerde wyne. Indien die gebruik van geneties gemodifiseerde organismes (GMOs) in gefermenteerde voedsel en drank deur die reguleringsowerhede goedgekeur en uiteindelik deur die verbruiker aanvaar sou word, sou dit vereis kon word dat die GMstatus van die wyngisgis op die etiket van die wynbottel aangebring word. Verpligte etikettering van GM-wyn sal metodes vereis waarmee die ‘nalentskap’ van GMgisselle in die finale produk geïdentifiseer en gemoniteer kan word. Tans is daar geen robuuste tegnieke beskikbaar waarmee die gebruik van GM-giste openbaar kan word nie, aangesien die gisselle en hul DNA tydens filtrasie en prosessering verwyder word. Een wyse waarop die onverklaarde gebruik van ‘n GM-gis in wynbereiding blootgestel sou kno word, is om die chemiese profiel van die verdagte wyn met dié van ‘n nie-GM-wyn te vergelyk. Ten einde hierdie konsep verder te ondersoek was ‘n sekondêre doelwit van hierdie studie om te bepaal of FT-IR (Fourier-transformasie-infrarooi) spektroskopie tesame met meervariante dataanalise gebruik kan word om te onderskei tussen wyne wat met transgeniese en nietransgeniese gisrasse gegis is, of tussen wyne wat met verskillende transgeniese rasse gegis is. Die resultate het aangedui dat hierdie metode gebruik kan word om wyne wat met verskillende gisrasse gegis is, te klassifiseer indien die betrokke gisras ʼn unieke chemiese profiel in die uiteindelike wyn veroorsaak het. Dit was egter ʼn voorlopige ondersoek en is as ʼn byvoegsel tot die tesis geskryf.

A three-year study was conducted at a commercial vineyard site in California’s Edna Valley AVA to evaluate the physiological and agronomical effects of the timing of cluster thinning on Pinot noir (clone 115) grapevines. Vines were thinned to one cluster per shoot at three selected time-points during the growing season (bloom, bloom + 4 weeks, bloom + 8 weeks), and fruit from each treatment was harvested and made into wine. Across all growing seasons, yield decreased 43% in thinned vines relative to un-thinned control vines. No effect of cluster thinning or interaction with growing season was found in vine shoot diameter, internode length, fruit zone light level, or cluster weight. Growing season significantly affected more fruit and wine parameters than did cluster thinning treatment, with interactions between treatment and growing season found in fruit Brix, titratable acidity, and anthocyanins, as well as wine anthocyanins and wine b* (yellow component). For example, bloom + 8 and bloom + 12 thinning treatments advanced Brix in 2017 but had no effect in 2018. Cluster thinning treatments increased berry anthocyanins by 43% in 2017 and by 103% in 2018 relative to the control. Similarly, cluster thinning increased berry total phenolics by 87% in 2017 and by 140% in 2018 relative to the control, with no significant differences found between the different thinning treatments. However, the levels of anthocyanins and total phenolics were generally not affected by cluster thinning treatment in the resulting wines. The fact that different cluster thinning treatments resulted in nil or minor effects on fruit and wine suggests that the vines tested were at or below a balanced crop load prior to the application of cluster thinning. Edna Valley AVA could likely support higher crop loads than 3.2 on the Ravaz index without negatively impacting fruit or wine composition and reducing crop load below that level is unlikely to increase fruit or wine quality.

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2006.
Under certain South African conditions, Redglobe develops a colour that is too dark and thus unacceptable for the Far Eastern markets. These markets require a pink colour instead of a dark red colour. The cultivation of grapes with an acceptable colour involves amongst other, canopy management practices. This generally includes the removal of leaves and/or lateral shoots. Hereby, the leaf area and the microclimatic conditions in the canopy are altered. The aim of this study was to test the usefulness of leaf and lateral shoot removal at different defoliation times after anthesis in order to obtain a pink coloured Redglobe crop. Other quality aspects, namely total soluble solids (TSS), total titratable acidity (TTA), berry mass and total yield, were also evaluated. A canopy management trial was conducted on six year old Redglobe vines with moderate vigour. The treatment design was a 2 x 3 x 4 factorial and involved two leaf removal (L) levels (L0 = 0% leaf removal; L33 = 33% leaf removal) in combination with three lateral shoot removal (LS) levels (LS0 = 0 % lateral shoot removal; LS50 = 50% lateral shoot removal; LS100 = 100% lateral shoot removal). Four defoliation times (DT) were selected: 36 (pea berry size), 69 (véraison), 76 (one week after véraison) and 83 (two weeks after véraison) days after anthesis (DAA). A total of 24 treatment combinations, replicated in four blocks, were applied. Generally, treatment combinations involving 33% leaf removal lowered the main shoot leaf area. Likewise, the lateral shoot leaf area was decreased by increasing levels of lateral shoot removal at any defoliation time. As expected, 33% leaf removal applied in combination with any level of lateral shoot removal, always resulted in a lower total vine leaf area compared to where 0% leaf removal was part of the treatment combination. Compensation reactions occurred and in this regard the main shoot leaf size increased due to 33% leaf removal applied at 1 week after véraison and 2 weeks after véraison. Treatment combinations involving lateral shoot removal increased the ratio of main shoot leaf area to the total leaf area. On the other hand, the main shoot leaf area percentage was lowered by the application of 33% leaf removal at 2 weeks after véraison compared to no leaf removal at the same defoliation time. It can therefore be assumed that the contribution of lateral shoot leaves to grape composition might have increased in cases where the main shoot leaf area was lowered at a later stage (e.g. 2 weeks after véraison). The bunches were visually evaluated and divided into classes from dark (class one) to light (class nine). This visual bunch evaluation showed that the mean bunch colour was in class three (lighter than class two) due to the defoliation time. The lateral shoot removal x leaf removal interaction resulted in a mean bunch colour that was in classes 2 and 3. However, within these classes, there was a tendency that bunch colour decreased for defoliation times later than pea berry size. The lateral shoot removal x leaf removal interactions showed that bunch colour was darker when the treatment combinations involved 0% leaf removal. The percentage of bunches with the desired colour was increased by application of the treatments at véraison, compared to the other defoliation times, and also with 50% lateral shoot removal and 100% lateral shoot removal compared to 0% lateral shoot removal. Biochemical analyses confirmed that increased levels of lateral shoot removal generally lowered the anthocyanin concentration regardless of defoliation time. A similar effect on TSS was observed, i.e. from véraison onwards, the application of 50% lateral shoot removal and 100% lateral shoot removal tended to lower TSS. The effect of these levels of lateral shoot removal at véraison was significant. The role of the lateral shoots in colour development and sugar accumulation is therefore emphasized. Furthermore, the special role that lateral shoots also play in berry development is illustrated in that berry mass tended to decrease when 100% lateral shoot removal in combination with 33% leaf removal and 100% lateral shoot removal in combination with 0% leaf removal were applied at véraison. This, together with the positive relationship obtained between grape colour and the lateral shoot leaf area:fruit mass ratio, accentuates the role of active leaf area during the ripening period. The possible effect of the microclimatic light environment on colour must also be considered. However, although the light intensity increased with increased levels of LS, the colour that was obtained was probably not associated with the differences in light intensity. It was found that it is possible to manipulate the colour of Redglobe grapes with defoliation treatments. However, the treatments that have a decreasing effect on grape colour also affected other quality parameters like TSS and berry size negatively. Although, it is possible to reduce the colour of Redglobe through the application of leaf and lateral shoot removal at different defoliation times, the question arises whether the treatment combinations used in this study are worthwhile to pursue because the mean bunch colour that was obtained was still too dark. However, it was possible to increase the percentage of bunches with the desired colour. Therefore, if such treatments are applied, it must be approached cautiously, keeping in mind that assimilate supply has to be sustained throughout the ripening period.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2009.
ENGLISH ABSTRACT: The total sale of brandy for 2007 in South Africa was R 7 300 000 000 and local statistics indicate that brandy is by far the most purchased spirit beverage. Sales of brandy even outweigh the total sales for whisky and the forecast for the estimated sales of brandy in the next five years is said to increase by 25%. It is therefore crucial to investigate those factors that influence the production of brandy as better understanding and control of these processes leads to the production of a brandy that is consistent and of premium quality. Many factors influence the final outcome of distillates. Of these factors, the distillation technique, the apparatus used for the purpose of distillation together with the low wine is of utmost importance as they influence the sensory profile and the chemical composition of the distillate. The effect of different variations of pot still designs on the chemical composition and the sensory profile of the resultant distillate was investigated. Five different Pot still variations were used and varied with regards to the design of their pot still head and swans neck apparatus. Two low wines were used for the purpose of distillations and were both from 2007 vintage. GCFID was used to identify the volatile compounds found in the distillates and together with Quantitative Descriptive Analysis (QDA) a profile of the distillates was produced which was used to differentiate between the different pot still variations and their effect on the final product. The data generated from the QDA sessions was subjected to Principal Component Analysis (PCA) and together with the chemical analysis a correlation between certain compounds and sensory attributes were found in the distillates. Distillate samples were also subjected to a sensory style classification system and were classified accordingly. The chemical composition of the two low wines prior to distillations differed significantly from each other with low wine one containing a larger amount of total esters and carbonyl compounds whilst low wine two contained a larger amount of total higher alcohols and acids. The distillates of low wine one also contained over all larger amounts of total esters and in the case of the distillates of low wine two, they contained larger amounts of higher alcohols and acids than low wine one. Variation one was based on the Alambic Charentais method of pot still design and it was found that only variation one influenced the chemical composition and the sensory profile of the distillates. This variation produced a distillate with a lower amount of total esters and more specifically ethyl acetate as well containing a lower intensity of the fruit and sweet associated caramel aromas and flavours. The esters, ethyl acetate and the ethyl esters of the long chained fatty acids were found to correlate with the sensory attributes known as fruit associated aroma, soapy aroma, and spicy aroma and therefore indicated that these compounds are responsible for these attributes. There were no correlations found between the chemical compounds, sensory attributes and sensory style classifications in the distillates of both low wine one and two. It was shown that the addition of certain esters, carbonyl compounds, higher alcohols and acids in specific ratios could alter the sensory classification of the distillates. Therefore the chemical composition and the sensory characteristics of distillates are largely dependent on the chemical composition of the low wine prior to distillation rather than the pot still design. Therefore, with further research it could be possible to predict the outcome of the chemical composition of the distillates by analyzing the chemical compounds found in the low wine prior to distillation.
AFRIKAANSE OPSOMMING: Die totale verkope aan brandewyn vir 2007 in Suid Afrika beloop R7 300 000 000 en statistiek wys dat brandewyn by verre die mees gesogte spiritus drank is. Verkope van brandewyn is selfs meer as die verkope van whisky en die voorspelling is dat die verkope van brandewyn met 25% gaan vermeerder in die volgende vyf jaar. Dit is dus belangrik om die faktore te ondersoek wat die produksie van brandewyn beïnvloed om sodoende die verstokingsproses te verstaan en te kontroleer om ‘n konsekwente kwaliteitsproduk op die mark te plaas. Baie faktore beïnvloed die finale produk. Faktore soos die distillasie tegnieke, die apperate wat gebruik word vir distillasie tesame met die rabatspiritus is van die uiterste belang aangesien dit die sensoriese profiel en die chemiese samestelling van die distillaat beïnvloed. Die effek van die verskillende variasies van potketelhelms op die chemiese samestelling van die distillate word ondersoek. Vyf verskillende helms met variasies in die swaannek ontwerp was gebruik. Twee verskillende rabatspiritus, van die 2007 oesjaar, was gebruik vir distillasie. GC-FID was gebruik om die vlugtige komponente van die distillate mee vas te stel. Kwantitatiewe Beskrywende Analise (QDA) is gebruik om ‘n profiel van die distillate op te stel wat weer gebruik is om te differensieer tussen die verskillende potketelhelm variasies en hulle effek op die finale produk. Die data wat deur die QDA sessies gegenereer was, is in die Vernaamste Komponent Analise (PCA) ingevoer en tesame met die chemiese analise is ‘n korrelasie tussen sekere komponente en die sensoriese analise van die distillate gevind. Distillaat monsters was ook aan sensoriese styl van klassifikasie onderwerp en is as volg daarvan geklassifiseer. Die chemiese samestelling van die twee rabatspiritus voor finale distillasie het betekenisvol van mekaar verskil ten opsigte daarvan dat die eerste rabatspiritus het hoë konsentrasies esters en karboniel verbindings gehad terwyl die tweede rabatspiritus meer hoë konsentrasies van sure en hoër alkohole gehad het. Die distillaat van die eerste rabatspiritus het ook hoë konsentrasies esters en karboniel verbindings gehad terwyl die distillaat van die tweede rabatspiritus weer hoë konsentrasies van sure en hoër alkohole gehad het. Variasie een is gebaseer op die Alambic Charentais van potketel ontwerp en daar is ook gevind dat hierdie variasie die enigste een was wat die chemiese samestelling en die sensoriese profiel van die distillate beïnvloed het. Hierdie variasie het ‘n distillaat geproduseer wat lae konsentrasies van totale esters, veral etielasetaat, sowel as laer intensiteit van vrugtige en soet geassosieerde karamel aromas en geure. Die esters, etielasetaat en etiel esters van die lang ketting vetsure, is gevind dat dit goed korreleer met die sensoriese eienskappe wat geassosieer word met vrugtige aromas, spesery-agtige aromas en seperige aromas. Daar is geen korrelasie gevind tussen die chemiese verbindings, sensoriese eienskappe en sensoriese styl van klassifikasie van distillate een en twee nie. Dit was ook bewys dat byvoeging van esters, karboniel verbindings, sure en hoër alkohole, in spesifieke verhoudings, die sensoriese eienskappe kan verander. Dus is die chemiese samestelling en sensoriese eienskappe van die distillate grootliks afhanklik van die chemiese samestelling van die rabatspiritus, voor die tweede distillasie, as wat dit afhanklik is van die potketelhelm ontwerp. Gevolglik, met verdere v navorsing, is dit moontlik om die uitkoms van die chemiese samestelling van die distillaat te voorspel deur die analise van die chemiese verbindings van die rabatspiritus te ontleed.

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2009.
ENGLISH ABSTRACT: Little scientific information regarding the effect of different geological parent materials on grapevine performance is currently available in South Africa. This aspect is of special significance for the Helderberg area, where parent material may change from granite to shale over a short distance. This results in shale- as well as granite-derived soils often occurring within the same vineyard. The objectives of this study were to (i) quantify the nutritional status and other soil properties of different parent materials (shale and granite) and overlying soils (ii) investigate the impact of geological differences in the soil on the vine nutritional status and certain vine parameters. This study was done over two seasons (2006/2007 and 2007/2008). Two Sauvignon blanc and two Cabernet Sauvignon vineyard blocks were selected at two different localities for each cultivar in the Helderberg area, South Africa. Shale- and granitederived soils were identified within each block. Kaolinite was the dominant mineral, whereas quartz and feldspar were sub-dominant. Traces of mica were also present in some shale- and granite-derived soils. Granite- contained significantly higher contents of coarse sand than shale-derived soils, whilst the opposite occurred in terms of fine sand. These differences affected the water holding capacity, in general making it higher in shale- than granite-derived soils. Shale-derived soils had higher concentrations of total K but granite-derived soils had a higher ability to release K as they contained higher concentrations of soluble K. The Q/I parameters, potential buffering capacity of K (PBCK) and equilibrium activity ratio of K (ARK) showed no consistent responses to geological differences. Potassium concentrations were higher in the leaf blades (obtained before harvest in 2007) from Sauvignon blanc grapevines on granite- than on shale-derived soils. Potassium concentrations in the Cabernet Sauvignon juice (obtained in 2007) tended higher in juice from granite- than from shale-derived soils. In 2008, K concentrations tended higher in juice from shale- than from granite-derived soils for both cultivars. The pH of the Sauvignon blanc juice (obtained in 2008) tended higher in juice from shale-than from granite-derived soils, thus corresponding with the K concentrations in the juice in this season. Nitrogen concentrations were higher in Cabernet Sauvignon juice (obtained in 2007) and Sauvignon blanc juice (obtained in 2008) from shalethan from granite-derived soils. In terms of vine water status, vines on granite-derived soils appeared more stressed than those on shale-derived soils in both seasons at one of the vineyards. In these Sauvignon blanc and Cabernet Sauvignon vineyards, the K nutritional status was not affected by geology in a consistent manner but there were some noticeable tendencies for a specific cultivar during certain seasons. On account of vines being planted on shale- and granite-derived soils within the same block, soil preparation was done similarly for both soils, and they were exposed to similar irrigation schedules, canopy management strategies and climatic conditions. Therefore, there is a high probability that all these practices may have negated the effect of geology on the K status of soils and especially on juice K concentration at the time of harvest. It was clear that seasonal differences and fertilisation affected the nutritional status of the vines more than geology.
AFRIKAANSE OPSOMMING: In Suid-Afrika is daar tans min wetenskaplike inligting oor die effek van verskillende geologiese moedermateriale op die prestasie van wingerd beskikbaar. Hierdie aspek is veral van belang in die Helderberg-area, waar moedermateriaal oor ‘n baie kort afstand van graniet na skalie kan wissel. Dit lei daartoe dat skalie-, sowel as granietgronde, dikwels binne dieselfde wingerd voorkom. Die doelwitte van die studie was om: (i) die voedingstatus en ander grondkundige eienskappe van die verskillende moedermateriale (skalie en graniet) en oorliggende gronde te kwantifiseer (ii) die impak van geologiese verskille in die grond op wingerd se voedingstatus en sekere wingerdkundige parameters, te ondersoek. Hierdie studie is oor twee seisoene (2006/2007 en 2007/2008) gedoen. Twee Sauvignon blanc en twee Cabernet Sauvignon wingerdblokke is geselekteer by twee verskillende lokaliteite vir elke kultivar in die Helderbergarea, Suid-Afrika. Beide skalie- en granietgrond is binne elke blok geïdentifiseer. Kaoliniet was die dominante mineraal, met kwarts en veldspaat sub-dominant, terwyl spore van mika ook in beide skalie- en granietgronde gevind is. Granietgronde het betekenisvol hoër hoeveelhede growwe sand bevat, terwyl skaliegronde meer fyn sand bevat het. Hierdie verskille het ‘n effek op waterhouvermoë gehad en daartoe gelei dat waterinhoude oor die algemeen hoër was vir skaliegronde. Skaliegronde het groter hoeveelhede totale K bevat, maar granietgronde se vermoë om K vry te stel was hoër, omdat hulle ‘n hoër konsentrasie oplosbare K bevat het. Die Q/I parameters, potensiële buffervermoë vir K (PBCK) en ewewig aktiwiteitsverhouding vir K (ARK), is nie op ‘n konsekwente wyse deur geologiese verskille beïnvloed nie. Vir die Sauvignon blanc wingerde was kalium konsentrasies in blaarskywe (gemonster voor oes in 2007) hoër vir graniet- as vir skaliegronde. Kalium konsentrasies in die sap vanaf Cabernet Sauvignon (gemonster in 2007) het hoër geneig vir granietgronde. In 2008 het die kalium konsentrasies, vir beide kultivars, hoër geneig in sap vanaf skaliegronde. Gedurende dié seisoen het die pH van sap ook hoër geneig vir Sauvignon blanc vanaf skaliegronde, wat dus ooreenstem met die K inhoud van die sap. Stikstof konsentrasies was hoër in sap vanaf skaliegronde vir Cabernet Sauvignon (2007) en vir Sauvignon blanc (2008). In terme van die wingerde se waterstatus, het stokke op die granietgrond, by een van die lokaliteite, geneig om gedurende beide seisoene onder groter stremming te wees op graniet as op skaliegrond. In hierdie Sauvignon blanc en Cabernet Sauvignon wingerde, is K voedingstatus nie op ‘n konsekwente wyse deur geologie geaffekteer nie, maar gedurende sommige seisoene was daar wel duidelike tendense vir ‘n spesifieke kultivar. Omdat die stokke binne dieselfde blok op skalie- en graniet gronde geplant is, was grondvoorbereiding eenders vir die twee grondtipes terwyl besproeiingskedule, lowerbestuur en klimaatstoestande ook identies was. Daar is dus ‘n hoë waarskynlikheid dat al hierdie faktore daartoe kon bygedra het dat die effek van geologie op die K status van van gronde versluier is, veral die effek op die K inhoud van sap teen oestyd. Dit was duidelik dat seisoenale klimaatsverskile en bemestingspraktyke ’n groter effek as geologie op die voedingstatus van die wingerd gehad het.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2011.
Includes bibliogaphy.
ENGLISH ABSTRACT: Sauvignon blanc wines have become increasingly popular in South Africa as it is a cultivar that can be easily manipulated in the vineyard and cellar to produce a range of wine styles. These wines are usually given aroma descriptors such as green pepper, grassy and asparagus; while other more tropical aromas include passion fruit and guava. These aromas are thought to be mainly caused by methoxypyrazines and volatile thiols. These compounds are known to be character impacting compounds of Sauvignon blanc and are present in the grapes in the aromatic form (methoxypyrazines) or as non‐aromatic precursors (thiols) that can be released by the yeast during fermentation. Other aroma compounds such as esters, higher alcohols, fatty acids and monoterpenes are compounds that could potentially influence the aroma bouquet of a wine significantly. These aroma compounds exist either as precursors in the grapes (monoterpenes) or arise due to yeast metabolism during fermentation (esters, higher alcohols, fatty acids) and often display fruity, floral and pleasant aromas. In the cellar, winemaking practices can be manipulated to a certain extent to achieve the desired wine style. Winemaking tools such as temperature, skin contact, pressing conditions, oxygen (O2), sulphur dioxide (SO2) and yeast strain are only a few factors influencing the outcome of a wine. In general, South African winemakers maintain a very reductive environment during Sauvignon blanc wine production by using inert gasses, thereby causing the production costs to increase. There is sufficient evidence to support the reductive handling of white wine, however there seems to be a lack of information as to why the must should be treated reductively before fermentation. The over all goal of this study was thus to investigate the effect of different O2 and SO2 additions to Sauvignon blanc must before settling, specifically focussing on the typical aroma compounds often found in these wines. Chapter 2 gives an overview of the oxidation reactions occurring in must (enzymatic oxidation) and wine (chemical oxidation). This chapter also reports the origin of the specific Sauvignon blanc aroma compounds and their reaction to different must and wine treatments with a focus on oxidation. Chapter 3 reports research results focussing on the effect of the different must treatments on the character impacting compounds of Sauvignon blanc wines, specifically the methoxypyrazines and the volatile thiols. The effect of the treatments on the polyphenols and glutathione content in the must and wine was also investigated. Oxidation in the absence of SO2 led to a decrease in glutathione and certain phenolic compounds in the must. In general, volatile thiols were protected against oxidation by SO2, even when O2 was present in the must. Methoxypyrazines concentrations were not significantly influenced by the treatments. Chapter 4 elucidates the effect of the treatments on other yeast and grape derived aroma compounds often found in Sauvignon blanc wines, such as the esters, higher alcohols, fatty acids and monoterpenes. In general, the effect of SO2 seemed to have the greatest influence on the produced aroma compounds. The results reported in this thesis could possibly change the way South African Sauvignon blanc musts are handled in future during the winemaking process. It is clear that O2 and SO2 management in the cellar is of critical importance for the winemaker to produce wines of high quality. Future work is important to fully understand the mechanisms and evolution of important aroma compounds of Sauvignon blanc wines during the winemaking process.
AFRIKAANSE OPSOMMING: Sauvignon blanc wyn aroma word gewoonlik beskryf met terme soos groen rissie, grasagtig en aspersie terwyl ander tropiese aromas soos grenadella en koejawel ook dikwels voorkom. Die manipulasie van Sauvignon blanc in die wingerd en in die kelder tydens wynmaak, gee die wynprodusent die vryheid om ‘n wye reeks wyn style te produseer. Dit maak Sauvignon blanc baie populêr in die Suid‐Afrikaanse wynindustrie. Die bogenoemde aromas word waargeneem in die wyn as gevolg van die teenwoordigheid van sekere aroma komponente genaamd metoksipirasiene en vlugtige tiole. Hierdie komponente lewer ‘n unieke bydrae tot die aroma samestelling van Sauvignon blanc wyne en kom voor in die druiwe in die aromatiese vorm (metoksipirasiene) of as nie‐aromatiese voorlopers (tiole) wat tydens alkoholiese fermentasie deur die gis vrygestel kan word. Komponente soos esters, hoër alkohole, vetsure en monoterpene kan ook ‘n potensiële bydra lewer tot die algehele aroma van Sauvignon blanc wyne en kom voor in die druiwe (monoterpene) of ontstaan as gevolg van gis metabolisme gedurende alkoholiese fermentasie (esters, hoër alkohole, vetsure). Hierdie geur komponente word dikwels beskryf as vrugtig, blomagtig en oor die algemeen aangenaam. Tydens wynmaak kan die wyn tot ‘n mate gemanipuleer word om ‘n spesifieke wynstyl te bekom. Hulpmiddels soos temperatuur, dopkontak, pers omstandighede, suurstof (O2), swawel dioksied (SO2) en gisras is slegs ‘n paar faktore wat die algemene uitkoms van ‘n wyn kan beïnvloed. Oor die algemeen word Sauvignon blanc in Suid‐Afrika baie reduktief behandel tydens wynbereiding. Dit vereis sekere hulpmiddels, soos die gebruik van inerte gas, wat die produksiekoste dikwels verhoog. Navorsing ondersteun die reduktiewe behandeling van wit wyn, maar dit wil voorkom asof daar ‘n tekort aan navorsing is wat die reduktiewe behandeling van die sap voor fermentasie regverdig. Die algemene doel van die studie is dus om die effek van verskillende O2 en SO2 byvoegings tot Sauvignon blanc sap (voor afsak) te ondersoek met die fokus op die tipiese aroma komponente wat in die wyn voorkom. Hoofstuk 2 lewer ‘n algemene oorsig van die tipes oksidasie reaksies wat voorkom in sap (ensiematiese oksidasie) en wyn (chemiese oksidasie). Spesifieke Sauvignon blanc aroma komponente word ook ondersoek in terme van die oorsprong van die komponente asook die reaksie wat plaasvind met verskillende mos en wyn behandelings, met ‘n fokus op oksidasie. In hoofstuk 3 word die effek van die verskillende mos behandelings op tipiese Sauvignon blanc aroma komponente, spesifiek metoksipirasiene en vlugtige tiole, ondersoek. Die effek van die behandelings op die polifenole en glutatioon inhoud in die mos en wyn word ook gerapporteer. Oksidasie van die sap in die afwesigheid van SO2, het ‘n afname in glutatioon en sekere polifenol konsentrasies veroorsaak. Dit wil voorkom asof die produksie van vlugtige tiole oor die algemeen beskerm word teen oksidasie indien daar genoegsame SO2 teenwoordig is. Hierdie effek word ondervind selfs as die sap met suursof versadig word. Die effek van die behandelings op die konsentrasies van metoksipirasiene was nie beduidend nie. Hoofstuk 4 rapporteer die effek van die behandelings op ander aroma komponente soos esters, hoër alkohole, vetsure en monoterpene. Oor die algemeen wil dit voorkom asof die effek van SO2 beduidend was en waarskynlik die grootste invloed op die produksie van hierdie aroma komponente het. Na aanleiding van die resultate bevind in hierdie tesis, is daar ‘n moontlikheid dat die manier waarop Sauvignon blanc wyne geproduseer word in Suid‐Afrika, moontlik kan verander in die toekoms. Vir die wynmaker om hoë kwaliteit Sauvignon blanc wyne te produseer, is O2 en SO2 bestuur in die kelder van kardinale belang. Verdere navorsing moet steeds gedoen word om die meganisme en evolusie van belangrike aroma komponente in Sauvignon blanc wyne tydens die wynmaakproses, ten volle te verstaan.

American Viticultural Areas (AVAs) are susceptible to small variations in climate and microclimates and are found within a narrow latitudinal range of prime climate conditions. These AVAs are geographically determined based on the best soil, climate, precipitation and temperature combinations for specific winegrape regions. As climate change continues to alter the local weather and the greater climate region of the Western United States, winegrape growing regions in Oregon are being affected. In an effort to determine what the pattern of change is, and compare previous studies of climate change using climate indices, a comparative study based in part on prior research was conducted. Using 800 meter resolution Parameter-elevation Regressions on Independent Slopes Model (PRISM) climate datasets, four individual climate indices were analyzed for statistical correlation with the climate data. These climate indices are: growing degree-days (GDD), the average growing season temperatures (GST), Huglin Index (HI) and the biologically effective degree-day (BEDD). Based on currently available data for this research, these climate indices were statistically analyzed during the years 2000 to 2010. A further avenue of research included a statistical analysis of the reported winegrape production, although this data was available only at an aggregated county-level. Results show that all four climate indices exhibit statistical significance, although the inclusion of the winegrape production data exhibited no statistical significance for many of the analyses, most likely due to subjective and aggregated data, few did result in significance with the climate indices. The research discussed here confirms the accuracy of the four climate indices and suggest that a longer time frame, coupled with less aggregated and subjective winegrape production data could produce interesting results in future research on the results of climate indices in winegrape growing regions.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: The quality of wine is influenced and determined by various factors, one of which includes the process of malolactic fermentation (MLF). MLF plays an integral role in the flavour and sensory profile of most red wines as well as some white wines like Chardonnay. This process is conducted by lactic acid bacteria (LAB), specifically of the genera Oenococcus, Lactobacillus, Pediococcus and Leuconostoc. Of these, Oenococcus oeni is best adapted to survive in the harsh wine environment. MLF is defined as the conversion of L-malic acid to L-lactic acid and carbon dioxide. The conversion of the dicarboxylic malic acid to the monocarboxylic lactic acid results in a decrease in acidity and an increase in pH, to give a softer mouthfeel and more favourable flavour profile. A further reason for conducting MLF in wine includes the improvement of microbial stability due to the removal of malic acid as a possible substrate for microorganisms. Recently, research focus has shifted to the ability of MLF and LAB to alter the aroma profile of wine via the production and/or modification of certain aroma compounds. In order for wine LAB to conduct MLF, they need to be able to survive the harsh and challenging wine environment. Conditions in South African wines are particularly challenging due to the long, hot ripening seasons resulting in high sugar concentrations which give high ethanol concentrations. Some LAB also struggle to adapt to an environment with high pH and low malic acid concentrations. These factors, combined with the use of sulphur dioxide, cause LAB to struggle in conducting and completing successful MLF. Many of the commercial starter cultures that are currently available contain LAB that have not been isolated from South African wine and are therefore not optimal for use under these challenging wine conditions. Oenococcus oeni is also the single LAB culture present in all commercially available starter cultures. The overriding goal of this study was to create a MLF starter culture containing a mixture of LAB cultures, namely O. oeni and Lactobacillus plantarum, which can successfully convert malic acid to lactic acid, ensure microbial stability, but also make a positive contribution to the wine aroma profile. Lactobacillus plantarum has previously been considered for possible use in a commercial starter culture. The LAB isolates used in this study were selected from the Institute for Wine Biotechnology culture collection as well as isolated from spontaneous MLF. The first objective was to characterise these LAB strains for important traits and for possible use as a MLF starter culture. A total of 23 strains were identified as O. oeni and 19 strains as Lb. plantarum. The identified strains were screened in a synthetic wine medium for their ability to convert malic acid to lactic acid. Based on the LAB strain performance in the synthetic wine medium, seven strains of both O. oeni and Lb. plantarum were selected. These 14 strains were screened for the presence of genes encoding for enzymes responsible for biogenic amine production and were found to contain none of the genes associated with the formation of histamine, tyramine or putrescine. The LAB strains were genetically screened for enzymes associated with aroma modification by LAB during MLF. The enzymes of interest that were screened for included β-glucosidase, esterase, protease and phenolic acid decarboxylase (PAD). The Lb. plantarum strains were found to possess more diverse enzymatic profiles related to aroma than O. oeni. The biggest differences were observed for the presence of β-glucosidase and PAD. The second objective was to perform small-scale fermentations with the individual LAB isolates. The individual isolates were evaluated in Pinotage and based on these results; three strains of each O. oeni and Lb. plantarum were selected for evaluation in mixed culture fermentations. The mixed cultures were evaluated in Pinotage, Shiraz and Cabernet Sauvignon in the 2008 vintage. As a third objective, the wines were also analytically and sensorially evaluated to investigate the changes in the aroma profile that could be attributed to the presence of the mixed LAB isolates. Based on the fermentation data as well as data pertaining to the aroma modification, three mixed cultures were selected for evaluation in the 2009 vintage in Pinotage, Cabernet Sauvignon and Chardonnay. The mixed cultures were able to successfully complete MLF in fermentation periods comparable to that of a commercial culture used as control. The different LAB cultures had distinct and diverse effects on the wine aroma profile. The O. oeni strain played a larger role in the ester concentration present after MLF, while the Lb. plantarum strain had a larger effect on the higher alcohol and volatile fatty acid concentration upon completion of MLF. The results generated by this novel study clearly indicate the potential of a mixed LAB starter culture for conducting MLF. The mixed cultures successfully completed MLF and made a positive contribution to the wine aroma profile.
AFRIKAANSE OPSOMMING: Die kwaliteit van wyn word beïnvloed en bepaal deur verskeie faktore en wynbereidings prosesse, wat die proses van appelmelksuurgisting (AMG) insluit. AMG speel ’n integrale rol in die sensoriese profiel van meeste rooiwyne, sowel as sommige witwyne soos Chardonnay. AMG word gedefinieër as die omskakeling van L-appelsuur na L-melksuur en koolstofdioksied. Hierdie omskakeling kan toegeskryf word aan die teenwoordigheid van melksuurbakterieë (MSB), spesifiek spesies van die genera Oenococcus, Lactobacillus, Pediococcus en Leuconostoc. Vanuit hierdie wyn MSB, is Oenococcus oeni die spesies wat die beste aanpas en oorleef onder stresvolle wyn kondisies. Die omskakeling van appelsuur, ’n dikarboksielsuur, na melksuur, ’n monokarboksielsuur, lei tot ‘n vermindering in suurheid en ’n verhoging in pH. Hierdie vermindering in suurheid gee ’n sagter en meer geronde mondgevoel aan die wyn en dra by tot ‘n meer aangename geurprofiel. ’n Verdere rede vir AMG in wyn is om mikrobiese stabiliteit te verseker deurdat appelsuur verwyder word as ’n moontlike koolstof substraat vir mikroörganismes. Onlangs het navorsing begin fokus op AMG en die vermoë van MSB om die aroma profiel van wyn te beïnvloed deur die produksie/modifisering van sekere aroma komponente. Vir MSB om AMG te kan deurvoer, moet hulle kan oorleef in die stresvolle wynomgewing. Wyntoestande in Suid-Afrika is veral uitdagend vir die oorlewing van mikroörganismes as gevolg van lang, warm somers wat lei tot ’n matriks met ’n hoë suikerkonsentrasie en wyn met ’n hoë etanolkonsentrasie. ‘n Omgewing met ‘n hoë pH en lae appelsuur konsentrasie, kan ook bydrae tot stresvolle kondisies vir MSB. Hierdie parameters, tesame met die gebruik van swaweldioksied, maak dit moeilik vir MSB om AMG te inisieer en te voltooi. Sommige van die kommersiële aanvangskulture wat tans beskikbaar is, bevat nie MSB wat onder Suid-Afrikaanse wyntoestande geïsoleer is nie en daarom is dit nie altyd optimaal vir gebruik nie. Oenococcus oeni is ook die enkele MSB kultuur wat in alle kommersiële kulture gebruik word. Die hoofdoelwit van hierdie studie was om ’n potensiële kommersiële aanvangskultuur te ontwikkel wat ‘n mengsel van MSB bevat. Hierdie aanvangskultuur moet AMG suksesvol kan voltooi, mikrobiologiese stabiliteit bevorder en steeds die wynaroma positief kan beïnvloed. Bakterierasse van O. oeni en Lb. plantarum is geselekteer vir gebruik in hierdie studie. Lactobacillus plantarum het reeds in vorige studies potensiaal getoon as ‘n moontlike aanvangskultuur. Die MSB isolate vir hierdie studie is geselekteer uit die Instituut vir Wynbiotegnologie se kultuurversameling en geïsoleer uit spontane AMG fermentasies. Die eerste doelwit was om hierdie MSB isolate te karakteriseer vir belangrike eienskappe en die moontlike gebruik as ’n kommersiële AMG aanvangskultuur. ‘n Totaal van 23 O. oeni en 19 Lb. plantarum isolate is geïdentifiseer. Hierdie isolate is in ’n sintetiese wynmedium geëvalueer vir hul vermoë om appelsuur na melksuur om te skakel. Op grond van hul reaksie in die sintetiese wynmedium, is sewe isolate van elk van die O. oeni en Lb. plantarum geselekteer. Hierdie 14 isolate is ondersoek vir die teenwoordigheid van die gene wat kodeer vir biogeenamien produksie en daar is gevind dat geen van die isolate enige van die biogeenamien gene wat ondersoek is, naamlik histamien, tiramien en putresien besit nie. Die MSB isolate is geneties ondersoek vir die teenwoordigheid van dié gene wat kodeer vir ensieme wat die aromaprofiel tydens AMG beïnvloed. Dié ensieme sluit β-glukosidase, esterase, protease, fenoliese suurdekarboksilase en sitraatliase in. Daar is gevind dat die Lb. plantarum isolate meer diverse ensiemprofiele as O. oeni besit. Die grootste verskille in die ensiemprofiele kan toegeskryf word aan die teenwoordigheid van β-glukosidase en fenoliese suurdekarboksilase. Die tweede doelwit was om kleinskaalse AMG fermentasies met die individuele MSB isolate uit te voer. Die individuele isolate is in Pinotage geëvalueer. Volgens hierdie resultate is drie isolate van elk van die O. oeni en Lb. plantarum geselekteer om in gemengde kulture getoets te word. Die gemengde kulture is in Pinotage, Shiraz en Cabernet Sauvignon in 2008 geëvalueer. As ’n derde doelwit is hierdie wyne ook analities en sensories geëvalueer om die veranderinge in die aromaprofiele as gevolg van die teenwoordigheid van die MSB te ondersoek. Op grond van die fermentasiedata, sowel as die data oor die aromaveranderinge, is drie gemengde kulture geselekteer vir evaluering in Pinotage, Cabernet Sauvignon en Chardonnay in 2009. Die gemengde kulture kon AMG suksesvol voltooi met fermentasietempo’s wat vergelykbaar was met dié van ‘n kommersiële AMG kultuur wat as kontrole gebruik is. Die verskillende MSB kulture het spesifieke en uiteenlopende uitwerkings op die wynaroma gehad. Die O. oeni isolaat in die gemengde kultuur blyk ‘n belangriker rol te speel in die esterkonsentrasie na AMG, terwyl die Lb. plantarum isolaat ’n groter effek het op die hoër alkohol en vlugtige vetsuurinhoud na AMG. Die resultate wat deur hierdie unieke studie gegenereer is, gee ’n aanduiding van die potensiaal van ’n gemengde MSB aanvangskultuur vir AMG. Die gemengde kulture kon AMG suksesvol voltooi en ‘n positiewe bydrae tot die aromaprofiel van die wyn lewer.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosach, 2010.
ENGLISH ABSTRACT: Cabernet Sauvignon, the most planted red wine cultivar in South Africa, is prone to vigorous growth with low yields. The aim of the study was to describe how Cabernet Sauvignon grapevines react to climate and irrigation within the Swartland region. Such knowledge would assist growers in decisions regarding long term as well as short term cultivation practices. This study was part of a larger project carried out by the Infruitec-Nietvoorbij institute of the Agricultural Research Council at Stellenbosch to determine effects of soil type and climate on yield and wine quality of Cabernet Sauvignon. The larger project was carried out in selected grape growing regions, i.e. Stellenbosch, Swartland, Lower Olifants River and Lower Orange River. Due to the proximity to the Atlantic Ocean, the study area in the Swartland region could be divided into two climatic regions for viticulture. Grapevines near Philadelphia closer to the ocean experienced less water constraints compared to those further inland near Wellington. Variation in stem water potential could also be related to soil water matric potential. Climate tended to have a more pronounced effect on the grapevine response to water constraints further inland than closer to the ocean. Vegetative growth, berry size and yield depended on water constraints experienced by the grapevines. In the warmer climate, severe constraints reduced yield. In the warmer climate, grapes started to ripen earlier than those in the cooler climate. Sugar concentration (mg/mL) was highest where grapevines experienced moderate water constraints. These seemingly balanced grapevines had the highest sugar accumulation, probably due to optimum photosynthesis and carbohydrate utilization. Low water constraints increased vegetative growth which could have been a sink for sugar loading. In addition to sugar loading, degree Balling (°B) increases could also have been due to a concentration effect where water constraints reduced berry volume. Therefore, °B is probably not a representative indicator of grapevine functioning. Anthocyanin biosynthesis, as quantified on a per berry basis, showed that sugar and anthocyanin could be co-regulated, with anthocyanin biosynthesis reaching a plateau when the sugar content per berry reached 200 mg/mL to 220 mg/mL. At véraison, the most intense grape colour occurred where grapevines experienced moderate water constraints, i.e. single drip line at Wellington and no irrigation at Philadelphia. However, at harvest grapes from the cooler climate tended to have more intense colour and higher phenolics, indicating that lower temperatures favoured anthocyanin biosynthesis. These results supported earlier findings that grapevine water status influences berry volume and dynamics of berry ripening. Water constraints tended to increase sensorial wine colour intensity, as well as wine fullness. Moderate water constraints at both localities resulted in the best sensorial wine quality. Yet there were indications that too severe water constraints could be detrimental to wine quality. Irrigation can be used to manipulate grapevine growth in warmer climates, but might be less effective in cooler climates. In warmer climates, moderate water constraints required to achieve balanced grapevine functioning can be obtained with single drip irrigation, but this might not be the case in cooler climates.
AFRIKAANSE OPSOMMING: Cabernet Sauvignon, die mees aangeplante rooiwynkultivar in Suid-Afrika, is geneig tot kragtige groei met lae opbrengste. Die doelwit van hierdie studie was om te beskryf hoe Cabernet Sauvignon-wingerdstokke reageer op klimaat en besproeiing in die Swartland-streek. Hierdie kennis sal kwekers help wanneer hulle besluite moet neem oor langtermyn sowel as korttermyn verbouingspraktyke. Hierdie studie was deel van ‘n groter projek deur die Infruitec-Nietvoorbij Instituut van die Landbounavorsingsraad op Stellenbosch om die effekte van grondtipe en klimaat op die opbrengs en wynkwaliteit van Cabernet Sauvignon te bepaal. Die groter projek is in geselekteerde wingerdverbouingstreke uitgevoer, nl. Stellenbosch, Swartland, Benede Olifantsrivier en Benede Oranjerivier. As gevolg van die nabyheid daarvan aan die Atlantiese Oseaan kon die studiegebied in die Swartland-streek in twee klimaatstreke vir wingerdbou verdeel word. Wingerdstokke by Philadelphia, wat nader aan die oseaan is, het minder waterstremming ervaar as dié verder binnelands naby Wellington. Veranderinge in stamwaterpotensiaal hou moontlik ook verband met die grondwater- matrikspotensiaal. Klimaat het ‘n groter effek op die reaksie van die wingerdstok op waterstremming verder binnelands as nader aan die oseaan. Vegetatiewe groei, korrelgrootte en opbrengs was afhanklik van die waterstremminge wat deur die wingerdstokke ervaar is. In die warmer klimaat het die ernstige stremminge opbrengs verminder. In die warmer klimaat begin druiwe vroeër ryp word as in die koeler klimaat. Suikerkonsentrasie (mg/ml) was die hoogste waar wingerde matige waterstremming ervaar het. Hierdie skynbaar gebalanseerde wingerdstokke het die hoogste suikerakkumulasie vertoon, moontlik as gevolg van optimum fotosintese en koolhidraatverbruik. Lae waterstremming het vegetatiewe groei verhoog, wat ook ‘n vraagpunt vir suikerlading kon wees. Benewens suikerlading kon verhogings in grade Balling (°B) ook moontlik aan ‘n konsentrasie-effek toegeskryf word in terme waarvan waterstremming die korrelvolume verminder het. °B is dus moontlik nie ‘n verteenwoordigende indikator van wingerdstokfunksionering nie. Antosianienbiosintese, soos gekwantifiseer op ‘n per-korrel basis, het getoon dat suiker en antosianien saam gereguleer kon word, en dat antosianienbiosintese ‘n plato bereik het wanneer die suikerinhoud per korrel 200 mg/mL tot 220 mg/mL was. By deurslaan het die mees intense druifkleur voorgekom waar die wingerdstokke matige waterstremming ervaar het, d.w.s. enkel druplyn op Wellington en geen besproeiing op Philadelphia. Teen oes was die druiwe in die koeler klimate egter geneig om meer intense kleur en meer fenole te bevat, wat aandui dat laer temperature antosianienbiosintese bevoordeel. Hierdie resultate ondersteun vroeër bevindings dat die waterstatus van die wingerdstok ‘n invloed op korrelvolume en die dinamika van korrelrypwording het. Waterstremming neig om die sensoriese wynkleurintensiteit te verhoog, asook die volheid van die wyn. Matige waterstremming op beide liggings het aanleiding gegee tot die beste sensoriese wynkwaliteit. Tog was daar aanduidings dat waterstremming wat te straf was, nadelig kon wees vir wynkwaliteit. Besproeiing kan gebruik word om wingerdgroei in warmer klimate te manipuleer, maar is moontlik minder effektief in koeler klimate. In warmer klimate kan die matige waterstremming wat benodig is vir gebalanseerde wingerdstokfunksionering, verkry word deur enkel drupbesproeiing, maar dit is moontlik nie die geval in koeler klimate nie.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2011.
Includes bibliography.
ENGLISH ABSTRACT: Grape and wine phenols consist of flavanols which is the building blocks for tannins. These building blocks are called monomers which consist of catechins, epicatechins, epigallocatechins and epicatechin-gallate. Tannin is important in wine as it contributes to bitterness, mouth feel (astringency) and maturation potential of the wine. Futhermore it has a health benefit as an antioxidant. Anthocyanins are responsible for the colour of red wine. The anthocyanins combine with tannins to form stable polymeric pigments. Due to the importance of tannins and anthocyanins in wine, it is imperitative that different winemaking techniques are used to extract as much of these components as possible and that the analysis is done quickly and accurately. The aim of this study was to evaluate different winemaking techniques and their extraction of tannins and anthocyanins into the wine. Too much tannin extraction can have a negative effect on the sensory quality of the wine. Therefore a second aim was to evaluate the mouth feel properties of a Shiraz wine. A third aim was to compare the two tannin precipitation methods in terms of time efficiency, repeatability and the ease of practice. To investigate the amount of tannin concentration extracted by different winemaking techniques, two cultivars (Cabernet Sauvignon and Shiraz) were used. These treatments included the addition of an enzyme during fermentation [E], cold maceration [CM], post maceration [PM] and the combination of cold and post maceration [CM+PM]. The grapes were harvested in two different climatic areas during the 2008 and 2009 vintages. The two climatic areas were classified according to the Winkler scale as a III (Morgenster) and a IV (Plaisir de Merle). The grapes were harvested at two different ripeness levels in order to evaluate the effect of the different winemaking processes on the extraction of tannins and anthocyanins. One harvest was before (LB) and the other after (HB) the commercial harvest. The results of this study showed significant differences in the phenolic composition of the wines. It was found that the warmer area showed higher tannin concentrations than the cooler area for both cultivars. In the 2008 Cabernet Sauvignon the CM extracted higher concentrations of tannin from the cooler area at both ripeness levels. In the warmer area, CM extracted the highest tannin concentration HB, but the CM+PM extracted the highest tannin concentration from Cabernet Sauvignon at the LB and CM at the HB of the warmer area. In 2009 the PM extracted the highest concentration of tannin at the lower ripeness level, while the E treatment extracted the highest concentration from the warmer area. In the cooler area the CM+PM extracted the highest concentration of tannin at a lower ripeness level, while there were no siginicant differences between the different treatments at the higher ripeness level. The highest anthocyanin concentration was found in the cooler area. The CM treatment was found to have no effect on anthocyanin extraction. Different methods are available to quantify the tannin concentration in wine. Two of the most popular tannin analytical methods are the bovine serum albumin (BSA) and the methyl cellulose precipitable tannin (MCP) methods. The BSA method is a very complex method which uses at least 3 times more reagents than the MCP method. The MCP method only analyzes tannins, while the BSA method analyzes tannins, monomeric pigments (MP), small polymeric pigments (SPP) and large polymeric pigments (LPP). In this study a good correlation was found between the two tannin precipitation methods (R2 – 0.88). There is controversy regarding the variability of these methods. Some scientists found that the two methods show a good correlation with HPLC, while others found that there was no such correlation between the precipitation methods and the HPLC. The MCP method had a practical advantage as it could be performed in half the time required for the BSA method. This has a significant impact in scenarios where a high sample throughput is required although it only measures total tannin. The phenolic composition and mouth feel of the wine was strongly influenced by the climatic area. In the warmer area the effect of tannin concentration on mouth feel was much less than in the cooler area. The wine made of riper grapes, was more grippy, bitter and numbing than the wines made from greener grapes. The E treatment was especially associated with a dry, grippy sensation.
AFRIKAANSE OPSOMMING: Druif en wyn fenole bestaan uit flavanole wat weer die boublokke is van tanniene. Hierdie boublokke, wat bekend staan as monomere, betsaan uit katesjiene, epikatesjiene, epigallokatesjiene an epikatesjien-gallaat. Tanniene is belangrik in wyn aangesien dit bydra tot bitterheid, mondgevoel (vrankheid) asook die verouderingspotensiaal van wyn. As antioksidante hou dit ook gesondheidsvoordele in. Antosianiene dra by tot die kleur van rooiwyn. Antosianiene kombineer met tanniene om meer stabiele polimeriese pigmente te vorm. As gevolg van die belangrikheid van tanniene en antosianiene is dit van uiterse belang dat verskillende wynmaak tegnieke gebruik word om ekstraksie in die wyn te bevoordeel en dat die analitiese metode so vinnig en akkuraat as moontlik gedoen word. Die eerste doel van hierdie studie was om die ekstraksie van tanniene en antosianiene deur middel van verskillende wynmaak tegnieke te evalueer. Te veel tanniene in die wyn kan negatiewe sensoriese kwaliteit tot gevolg het. Daarom is die tweede doel om die sensoriese kwaliteit van Shiraz wyn te evalueer. Die derde doel van hierdie studie was die twee tannien presipitasie metodes met mekaar te vergelyk in terme van die moeilikheidsgraad van die metode, tyd doeltreffendheid en herhaalbaarheid. Verskillende wynmaak tegnieke (ensiem byvoegings [E], koue maserasie [CM], verlengde dopkontak [PM] en ‘n kombinasie van koue maserasie en verlengde dopkontak [CM+PM]) is vergelyk ten opsigte van tannien en antiosianien ekstraksie. In 2008 en 2009 is twee kultivars (Cabernet Sauvignon en Shiraz) in twee verskillende klimatologiese areas gepars. Hierdie areas is geklassifiseer in die Winklerskaal as ‘n IV (Plaisir de Merle) en ‘n III (Morgenster). Om die effek van die verskillende wynmaak tegnieke op die ekstraksie van antosianiene en tanniene te vergelyk, is hierdie twee kultivars by twee verskillende rypheidsgrade geoes. Die eerste oes was net voor kommersiële oes (LB) en die tweede oes het net na kommersiële oes (HB) plaasgevind. Die 2009 Shiraz wyn is organolepties beoordeel om die effek van die verskillende wynmaak tegnieke op die wyn se mondgevoel te vergelyk. Die resultate van hierdie studie toon beduidende verskille in die fenoliese samestelling van die wyne. Dit is gevind dat die warmer area hoër tannien konsentrasies het as die koeler area. In 2008 het die CM+PM die meeste tanniene uit die Cabernet Sauvignon geëkstraheer by LB en die CM by HB in die warmer area. Die CM het in die koeler area meer tanniene geëkstraheer by beide die LB en HB rypheidsgrade. In 2009 het PM die meeste tanniene geëkstraheer by LB terwyl E die meeste tanniene geëkstraheer in die warmer area. In die koeler area het CM+PM die meeste tanniene geëkstraheer, terwyl geen van die behandelings ‘n effek gehad het by HB. Die meeste antosianien konsentrasie was in die koeler area gevind as in die warmer area. In beide 2008 (LB en HB) en 2009 (LB) het CM die meeste antosianiene geëkstraheer, terwyl geen behandeling ‘n effek gehad het by HB. Twee van die mees populêre tannien analitiese metodes is die BSA (bovine serum albumien) en die MCP (metielsellulose presipitasie) metodes. Die BSA metode is ‘n baie meer ingewikkelde metode waarvoor drie keer meer reagense gebruik word as vir die MCP metode. Maar waar die MCP net tanniene ontleed, ontleed die BSA metode tanniene, monomere (MP), klein polimeriese pigmente (SPP) en groot polimeriese pigmente (LPP). Dit help indien daar gekyk wil word na die evolusie van polimeriese pigmente. In hierdie studie is bevind dat daar ‘n redelike korrelasie (R2 – 0.88) tussen die BSA en MCP metode bestaan. Die herhaalbaarheid van die metodes het redelike kontroversie veroorsaak, waar sommige navorsers bevind het dat die BSA metode nie so herhaalbaar is soos eers bevind is nie. Die MCP metode het ’n praktiese voordeel aangesien dit in die helfde van die tyd van die BSA metode uitgevoer kan word. Dit het ‘n groot impak indien ‘n groot hoeveelheid monsters ontleed moet word. Die fenoliese samestelling en mondgevoel word sterk beïnvloed deur die klimatologiese area. In die warmer area was die effek van tannien konsentrasie op mondgevoel kleiner as in die koeler area. Die wyn van ryper druiwe het meer harder, verdowingseffek en bitter nasmaak gehad as by die wyn van groener druiwe. Die ensiem behandeling was meer geassossieerd met droë mond gevoel.

Thesis (MScAgric (Viticulture and Oenology. Wine Biotechnology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Necrotrophic fungi infect many economically important crop plants. This results in great losses in the agricultural sector world-wide. Understanding the nature by which plants respond to pathogens is imperative for genetically enhancing disease resistance in plants. Research tools have significantly contributed to our understanding of how the plant responds to pathogen attack, identifying an array of defence mechanisms used by plants upon attack. Many fungal pathogens secrete endopolygalacturonases (endoPGs) when infecting plants. These hydrolytic enzymes are inhibited by polygalacturonase-inhibiting proteins (PGIPs) associated with plant cell walls. PGIPs are well characterised and their current known functions are all linked to endoPG inhibition and the subsequent upregulation of plant defence pathways. Work on grapevine PGIPs have shown that apart from being efficient antifungal proteins, leading to protection of the plant against Botrytis cinerea when overexpressed, PGIPs might also have additional functions linked to cell wall strengthening. This working hypothesis formed the motivation of this study where a cinnamyl alcohol dehydrogenase (CAD) (1.1.1.195) gene was targeted for functional analysis in tobacco (Nicotiana tabacum). Some previous work and genetic resources obtained is relevant to this study, specifically previously characterized transgenic tobacco lines overexpressing the Vitis vinifera pgip1 (Vvpgip1) gene. These lines have confirmed PGIP-specific resistance phenotypes against B. cinerea, as well as increased levels of CAD transcripts in healthy plants. Moreover, preliminary evaluations indicated increased lignin levels as well as differential expression of several other cell wall genes in these overexpressing lines (in the absence of infections). In this study we generated a transgenic tobacco population, overexpressing the native CAD14 gene, via Agrobacterium transformations. The transgene was overexpressed with the Cauliflower Mosaic Virus promoter (CaMV 35Sp). The CAD transgenic population was analyzed for transgene integration and expression and showed active transcription, even from leaves that normally don’t express CAD to high levels. These lines, together with the untransformed control, and a representative transgenic VvPGIP1 tobacco line previously characterized with elevated expression of CAD were used for all further analyses, specifically CAD activity assays of stems and leaves, as well as whole plant infections with B. cinerea. CAD enzyme activity assays were performed on healthy uninfected plant lines, without inducing native CAD expression or resistance phenotypes (i.e. without Botrytis infection). CAD activity was detected in leaves and stems, but a statistically sound separation between the CAD population and the untransformed control was only observed in the stems. The CAD assays also confirmed previous results that indicated that CAD transcription was upregulated in the PGIP line in the absence of infection. Overall, in all plant lines the stems exhibited 10-fold higher levels of CAD activity than the leaves, but the transgenic VvPGIP1 line showed a further 2-3-fold increase in CAD activity in the stems, when compared to the untransformed control and the majority of the CAD overexpressing lines. Disease assessment by whole plant infections with B. cinerea of the CAD transgenic plants revealed reduced disease susceptibility towards this pathogen. A reduction in disease susceptibility of 20 – 40% (based on lesion sizes) was observed for a homologous group of transgenic lines that was statistically clearly separated from the untransformed control plants following infection with Botrytis over an 11-day-period. The VvPGIP1 transgenic line displayed the strongest resistance phenotype, with reduction in susceptibility of 47%. The reduction in plant tissue maceration and lesion expansion was most pronounced in the VvPGIP1 line compared to the CAD transgenic plants, while the CAD transgenic plants showed more reduction than the untransformed control. In combination, the data confirms that CAD upregulation could lead to resistance phenotypes. Relating this data back to the previously observed upregulation of CAD in the VvPGIP1-overexpressing lines, the findings from this study corroborates that increased CAD activity contributes to the observed resistance phenotypes, possibility by strengthening the cell wall. In conclusion, this study yielded a characterized transgenic population overexpressing the CAD14 gene; this overexpression contributed to increased RNA transcription compared to the untransformed control plant, increased CAD activity (most notably in the stems) and a disease resistance phenotype against Botrytis. These findings corroborates the current working hypothesis in our group that PGIPs might have a role in preparing the plant cell for attack by contributing to specific cell wall changes. The exact mechanisms are still currently unknown and under investigation. The transgenic lines generated in this study will be invaluable in the subsequent analyses where these various phenotypes will be subjected to profiling and accurate cell wall analyses.
AFRIKAANSE OPSOMMING: Nekrotrofiese swamme infekteer en beskadig verskeie ekonomies belangrike gewasse. Dit lei wêreldwyd tot groot verliese vir die landbousektor. Dit is noodsaaklik om te verstaan hoe plante reageer teenoor patogene, sodat die siekteweerstand van plante verbeter kan word. Navorsingshulpbronne het beduidend bygedra tot die kennis van plantreaksies tydens patogeniese aanvalle, en het sodoende ‘n reeks van weerstandmeganismes, wat die plant inspan tydens ‘n aanval, geïdentifiseer. Verskeie patogeniese swamme skei endopoligalakturonases (endoPGs) af tydens plantinfeksie. Hierdie hidrolitiese ensieme word geïnhibeer deur poligalakturonase-inhiberende proteïene (PGIPs) wat met die plantselwand geassosieerd is. PGIPs is goed gekarakteriseerd en al hulle huidiglik bekende funksies is gekoppel aan endoPG inhibisie en die daaropvolgende opregulering van plant weerstandspaaie. Navorsing op wingerd PGIPs het gewys dat, afgesien van die feit dat PGIPs goeie antifungiese proteïene is wat lei tot beskerming van die plant teen Botrytis cinerea wanneer dit ooruitgedruk word, PGIPs ook moontlik addisionele funksies verrig wat verwant is aan selwandversterking. Hierdie werkshipotese vorm die motivering vir hierdie studie waarin ‘n sinnamiel alkohol dehidrogenase (SAD) (1.1.1.195) geen geteiken is vir funksionele analise in tabak (Nicotiana tabacum). Vorige navorsing en genetiese hulpbronne daardeur verkry is belangrik vir hierdie studie, spesifiek die gekarakteriseerde transgeniese tabaklyne wat die Vitis vinifera pgip1 (Vvpgip1) geen ooruitdruk. Hierdie lyne besit bevestigde PGIP-spesifieke weerstandsfenotipes teen B. cinerea, sowel as hoër vlakke van SAD transkripte in gesonde plante. Voorlopige analises het ook aangedui dat hierdie ooruitdrukkende lyne hoër lignien vlakke het, asook differensiële uitdrukking van verskeie ander selwandgene (in die afwesigheid van infeksie). In hierdie studie is ‘n transgeniese tabakpopulasie gegenereer wat die natuurlike tabak SAD14 geen ooruitdruk, deur middel van Agrobacterium transformasie. Die transgeen is ooruitgedruk deur die Blomkool Mosaïek Virus promoter (CaMV 35Sp). Die SAD transgeniese populasie is geanaliseer vir transgeen integrasie en uitdrukking en het aktiewe transkriptering getoon, selfs in blare waar daar normaalweg nie hoë vlakke van SAD uitgedruk word nie. Hierdie lyne, die ongetransformeerde wilde-tipe kontrole sowel as ’n verteenwoordigende transgeniese VvPGIP1 tabaklyn wat vooraf gekarakteriseerd was met hoë SAD uitdrukking, is gebruik vir alle verdere analises, spesifiek SAD aktiwiteitstoetse in stingels en blare, asook heelplantinfeksies met B. cinerea. Aktiwiteitsanalises van die SAD ensiem is gedoen op gesonde ongeinfekteerde plantlyne, sonder om natuurlike tabak SAD uitdrukking of weerstandsfenotipes te induseer (dus sonder Botrytis infeksie). SAD aktiwiteit is waargeneem in beide die blare en stingels, maar ‘n statisties betekenisvolle skeiding is slegs gevind tussen die SAD populasie en die ongetransformeerde kontrole in die stingels. Die SAD toetse het ook vorige resultate bevestig wat aangedui het dat SAD transkripsie opgereguleer word in die PGIP lyn in die afwesigheid van infeksie. Die stingels het oor die algemeen ‘n 10-voudige vermeerdering in SAD aktiwiteitsvlakke getoon in vergelyking met die blare, maar die transgeniese VvPGIP1 lyn het ‘n verdere 2-3-voudige verhoging in SAD aktiwiteit gehad in die stingels ,in vergelyking met die ongetransformeerde kontrole en die meerderheid van die SADooruitdrukkende lyne. Siekteweerstand ondersoeke deur middel van heelplantinfeksies met B. cinerea van die SAD transgeniese plante, het verminderde vatbaarheid aangedui ten opsigte van hierdie patogeen. ‘n Afname in siekte-vatbaarheid van 20 – 40% (gebaseer op wondgroottes) is waargeneem vir ‘n homoloë groep transgeniese lyne wat statisties betekenisvol geskei kon word van die ongetransformeerde kontrole plante na infeksie met Botrytis in ‘n infeksietoets wat 11 dae geduur het. Die VvPGIP1 transgeniese lyn het die mees weerstandbiedende fenotipe gehad, met ‘n afname in siekte-vatbaarheid van 47%. Die afname in plantweefselafbreking en wondgrootte was die duidelikste in die VvPGIP1 lyn in vergelyking met die SAD transgeniese plante, terwyl die SAD transgeniese plante ‘n groter afname aangedui het as die ongetransformeerde kontrole. In kombinasie het die data bevestig dat SAD opregulasie kan lei tot weerstandbiedende fenotipes. Hierdie data, in vergelyking met die vorige bevinding van opregulasie van SAD in die VvPGIP1-ooruitdrukkende lyne, bevestig dat hoër SAD aktiwiteit bydra tot die waargenome weerstandbiedende fenotipes, moontlik deur versterking van die plantselwand. Ter afsluiting, hierdie studie het ‘n gekarakteriseerde transgeniese populasie wat die SAD14 geen ooruitdruk gelewer; hierdie ooruitdrukking het bygedra tot hoër RNA transkripsie in vergelyking met die kontrole, verhoogde SAD aktiwiteit (veral in die stingels) en siekteweerstandbiedende fenotipes teenoor Botrytis. Hierdie bevindinge ondersteun die huidige werkshipotese in ons groep dat PGIPs moontlik ‘n rol speel in die voorbereiding van die plantsel teen infeksie deur spesifieke selwandveranderinge te veroorsaak. Die spesifieke meganismes is steeds onbekend en word verder ondersoek. Die transgeniese lyne wat tydens hierdie studie gegenereer is, sal baie belangrik wees in opvolgende analises waar hierdie verskillende fenotipes gebruik kan word om die profiel van selwandkomponente, maar ook die akkurate selwandsamestelling te bestudeer.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Cabernet Sauvignon is the most planted red cultivar in South Africa and the second most planted red cultivar in the Olifants River region. The cultivar is prone to vigorous growth with low yields. Excessive irrigation could accentuate these cultivar characteristics. Considering the foregoing, the aim of the study was to describe how Cabernet Sauvignon will react to climate, soil type (texture) and irrigation within the Lower Olifants River wine region to enable growers to make the right decisions regarding long term as well as short term cultivation practices. This study is part of a project carried out by the ARC Infruitec-Nietvoorbij at Stellenbosch to determine the effects of soil type and atmospheric conditions on yield and wine quality of Cabernet Sauvignon in different grape growing regions of South Africa. Similar studies are being carried out in the Orange River, Stellenbosch and Swartland regions of South Africa. The Lower Olifants River region could be divided into three climatic regions. Furthermore, two climatic regions is evident regarding the formation of grape wine colour and aromas. Proximity to the Atlantic Ocean would play an important role in a cultivar establishment policy. The variation in stem water potential (ΨS) could be related to soil water status expressed in terms of matric potential (ΨM). In the case of sandy soils, ΨS decreased substantially more as the ΨM decreased compared to the sandy loam soils. The reason could be that the unsaturated hydraulic conductivity in sandy soils decreased more rapidly as the ΨM decreased compared to the heavier soils. Thus could explain why the grapevines in the sandy soils experienced more water stress than the ones in the sandy loam soils at a given ΨM. Climate had a strong influence on grapevine water status with grapevines nearer to the ocean experiencing less water stress compared to the ones further inland. This was especially true for grapevines in the sandy soils. Vegetative growth and yield of grapevines in the sandy soils were more sensitive to water deficits compared to the ones in the sandy loam soils. For deficit irrigated grapevines in the sandy soils, vegetative growth and yield decreased by ca. 30% when ca. 55% less water was applied from flowering to harvest. Yield reduction were ca. 15% with no or very little influence on vegetative growth with ca. 80% reduction in water applied from flowering to harvest for grapevines in the sandy loam soils. The influence of soil texture on wine quality and style were evident under intensive irrigation as well as over different climatic regions. Overall sensorial potential wine quality of grapevines in sandy soils tended to be higher compared to the ones in the sandy loam soils. Deficit irrigation tended to increase wine colour intensity, irrespective of soil texture. Furthermore, deficit irrigation in sandy loam soils tended to increase wine fullness and the berry characteristics of the wine. Berry characteristics of wines from the sandy soils tended to be higher compared to the ones from the sandy loam soils. Too severe water deficits in sandy soils could be detrimental to wine quality. Climate tended to have an influence on wine style of grapevines in the sandy soils with wines produced further away from the ocean tended to have higher berry characteristics. Irrigation management could be a powerful tool to manipulate the grapevine in sandy soils. For grapevines the sandy loam soils in addition to regulated deficit irrigation, additional canopy management practices could be needed to improve wine quality.
AFRIKAANSE OPSOMMING: Cabernet Sauvignon is die mees aangeplante rooidruif kultivar in die Suid-Afrikaanse wynbedryf. In die Olifantsriver streek is dit naas Shiraz, die tweede mees aangeplante rooidruif kultivar. Cabernet Sauvignon is bekend as ‘n groeikragtige skaamdraer. Indien oorbesproei word, kan hierdie potensiële nadelige eienskappe nog meer na vore tree. Die doel van die studie is om die invloed van die klimaat, grond en besproeiing op Cabernet Sauvignon se vegetatiewe groei, produksie en wyngehalte in die Benede Olifantsrivier streek te bepaal. Hierdie inligting kan produsente help om ingeligte kortsowel as langtermyn besluite te maak rakende die verbouing van Cabernet Sauvignon. Hierdie studie vorm deel van ‘n breër studie in die Suid-Afrikaanse wynbedryf, gedryf deur die Landbou Navorsingsraad (LNR) Infruitec-Nietvoorbij, Stellenbosch om die invloed van atmosferiese toestande en grond op die produksie en wyngehalte van Cabernet Sauvignon te bepaal. Soortgelyke projekte word uitgevoer in die Oranjerivier, Stellenbosch en Swartland wynstreke. Die Benede Olifantsrivier streek kan verdeel word in drie klimaatstreke op grond van temperatuurdata. In terme van die ontwikkeling van druifkleur en aromas, kan die streek verdeel word in twee klimaatstreke. Die afstand vanaf die Atlantiese Oseaan kan ‘n belangrike rol speel in die ontwikkeling van ‘n kultivarriglynplan vir die streek. Grondwaterstatus, uitgedruk as die matrikspotensiaal (ΨM), kan aanleiding gee tot variasie in middag blaarwaterpotensiaal (ΨS) lesings. Die ΨS van die sand gronde verlaag vinniger soos die ΨM verlaag invergelyke met die sandleem gronde. Dit kan moontlik wees as gevolg van die verskil in die grond onversadigde hidroliese konduktiwiteit. Sand gronde se hidroliese konduktiwiteit verlaag vinniger soos die ΨM verlaag, invergelyke met sandleem gronde. Dit verklaar waarom wingerde in sand gronde by dieselfde ΨM, meer waterspanning ondervind as wingerde in sandleem gronde. Klimaat het ‘n invloed op die waterstatus van die wingerdstok. Wingerde nader aan die see het minder waterspanning ondervind invergelyke met wingerde wat verder in die binneland geleë is. Dit was veral die geval met wingerde in die sand gronde. Vegetatiewe groei en produksie van wingerde in die sand gronde is meer sensitief vir waterspanning as wingerde in die sandleem gronde. Tekortbesproeiing in die sand gronde het die groei asook produksie met ongeveer 30% verlaag deur ongeveer 55% minder water toe te dien vanaf blom tot oes. In teenstelling daarmee is die produksie van wingerde in die sandleem gronde met ongeveer 15% verlaag met geen tot baie min verlaging in die groeikrag. Ongeveer 80% minder water is toegedien vanaf blom tot oes. Grondtekstuur kan wyngehalte en -styl beïnvloed ten spyte van intensiewe besproeiing en klimaatsverskille. Sensoriese potensiële wyngehalte van wingerde in die sand gronde was beter invergelyke met dié van die sandleem gronde. Die wyne vanaf die sand gronde het ook geneig om oor meer bessie intensiteit te beskik as wyne vanaf die sandleem gronde. Tekortbesproeiing neig om die wynkleur intensiteit te verhoog, ongeag van grondtekstuur. Tekortbesproeiing in die sandleem gronde kan ook die volheid van die wyne verbeter, asook die bessie intensiteit van die wyn verhoog. Te hoë waterspanning in die sand gronde kan wyngehalte nadelig beïnvloed. Klimaat kan ook die wynstyl vanaf sand gronde beïnvloed met wyne verder vanaf die see wat oor meer bessie intensiteit beskik as wyne nader aan die see. Beheerde tekortbesproeiing kan as ‘n kragtige hulpmiddel gebruik word om wingerde in die sand gronde te manipuleer. Vir wingerde in die sandleem gronde, addisioneel tot beheerde tekortbesproeiing en normale loofbestuurspraktyke, kan ekstra loofbestuurspraktyke bv. die verwydering van sylootlote, dalk nodig wees om wyngehalte te verbeter.

Fungal diseases pose significant challenges for grapevine producers in Kentucky due to the region’s abundant moisture and relative humidity. Methods to reduce fungicide application frequency would prove both economically and temporally valuable to producers. A field experiment was established in Bowling Green, KY in 2017 to investigate Bacillus mycoides isolate J (LifeGard) as a supplement to a fungicide program for systemic acquired resistance (SAR). Three fungicide treatment regimens were implemented consisting of a program modelled from the Midwest Fruit Pest Management Guide (2017) and an identical program supplemented with 140 g ha-1 LifeGard per application (both applied on 14 day intervals), a reduced frequency application every 28 days supplemented with 140 g ha-1 LifeGard, and an untreated control. Treatments were applied to 9-year-old French-Hybrid grapevines (cv. Chambourcin); each treatment was replicated 3 times in a randomized complete block design. All treatments were applied with a backpack sprayer delivering 150 L ha-1 at 2 Bar pressure. Canopy management, fertility, herbicide, and insect management were standardized across treatments and no supplemental irrigation was applied. Data collected included fruit yield, pH, ºBrix, and titratable acidity (TA). Data were analyzed with SAS PROC GLIMMIX; differences in means were determined at  < 0.05. Plots supplemented with B. mycoides had lower fruit pH than untreated plots but higher fruit pH than the traditional fungicide program. Treatment regime did not influence Brix, TA, or total yield; however, all treated plots yielded more high quality fruit than the untreated control.

A trial established in 2003 at Upper Moutere in Nelson, New Zealand, was used to evaluate the effect of mussel shells as reflective mulch on Vitis vinifera L. cv. Pinot noir vine performance and fruit and wine quality. Shell mulch had several effects on the environment and vine growth as well as grape and wine composition in the 2006/2007 season. Soil under mulch was cooler compared to un-mulched control, but buffered the extremes in temperatures. Fruiting zone temperature over shells was slightly higher during the day and cooler at night, showing no effect on mean hourly temperature. Shell mulch reflected greater amounts of UV-A, UV-B and PAR radiation into the fruiting zone. Shell reduced weed growth compared to control. Leaf petiole and blade samples showed higher amounts of calcium compared to control. Leaf SPAD values were higher in the shell treatment during veraison, previous and postharvest, but lower post budburst. While date of budburst was not affected by treatments, dates of flowering and veraison appeared to be slightly advanced over shells. Fruit set was similar between treatments and was considered poorer in shell bunches due to a larger population of seedless berries. Vine growth was not affected in terms of the number of nodes laid at pruning, flower cluster and shoot number pre shoot thinning, early shoot growth and lateral shoots development. Vigour was not increased by shells as demonstrated by pruning weights, canopy density and trunk circumferences being similar, though internode lengths in shell shoots were greater in 2007 and lower in 2006. Berry weights, bunch weights and vine yields were lower in shell than control, though greater berry numbers were recorded. There were slight differences between treatments in fruit and wine composition. Grape pH only varied in the middle of the sampling time, being higher the 2nd week and lower the 3rd week in shell grapes and TA was greater at harvest time. However, °Brix was only higher in shell grapes in the middle of the sampling period, being similar to control at veraison and harvest. Peduncle lignification was delayed at veraison as well as at harvest time. Shell must after crushing was greater in Brix but similar to control in pH and TA. Similarly, shell wines pre bottling showed higher alcohol and no differences for pH and TA. HPLC-DAD analyses of commercial-scale and microvin wines showed consistent differences of the individual flavonoid composition. Shell microvin wines were greater than control in quercetin and resveratrol. However, commercial shell wines were lower in epicatechin, gallic acid, resveratrol, and catechin than control. Leaf phenolic composition was also different between treatments. However, further analyses by HPLC-MS in wines as well as in leaves are necessary to identify individual compounds. Total anthocyanins and total phenolics were no different between treatments. Sensory analyses of microvin and commercial shell wines exhibited consistently lower levels of green and unripe tannins, and greater smoothness and complexity as well. Further analysis by GC-MS and HPLC-MS is warranted. Shell mulch improved sensory characteristics of the resulting wines.

Les vins qui apportent probablement le plus de plaisir et de magie aux amateurs sont très souvent les vins dits « de garde ». Ce sont des bouteilles que l’on conserve de nombreuses années et avec lesquelles on dépose l’espoir d’une grande dégustation. A l’ouverture, le dégustateur espérera découvrir la complexité et l’élégance du bouquet de vieillissement. Ce concept olfactif, mêlant de manière harmonieuse les odeurs produites au cours du vieillissement en bouteille à celles du vin jeune, est le cadeau qu’offre les grands vins aux épicuriens patients. L’expression du bouquet serait très intimement liée aux terroirs et au millésime, révélés par le passage du temps. Ce travail de thèse est essentiellement consacré à démonter et affiner cette connaissance empirique.Dans un premier temps, le dosage de plusieurs molécules liées à la composition de la baie de raisin, aux activités fermentaires et au vieillissement a été effectué dans 80 vins issus de « verticales » de différents crus du vignoble de Bordeaux. L’analyse des résultats montrent que les crus possèdent une identité moléculaire propre malgré la variabilité due aux millésimes et aux âges des vins. La présence de dérivés terpéniques, issus du métabolisme végétal, est fondamentale dans la construction de cette identité. Cette différenciation des crus montre l’importance de la composition du raisin, exprimant un terroir, sur l’identité des vins vieux.Dans un second temps, des paramètres du terroir (statut hydrique, statut azoté et cépage par exemples) ont été confrontés aux quantités des molécules ciblées pour l’étude et mesurées par des techniques de chromatographie en phase gazeuse. Ces travaux ont nécessité le développement d’outils analytiques ainsi que la mise en place de plans expérimentaux à diverses échelles (laboratoire à unités de production viticole). Enfin, dans l’objectif d’obtenir des outils susceptibles de nous aider à prédire le potentiel de garde et l’apparition du bouquet, l’étude des précurseurs des arômes du bouquet a été entamée. Parmi les composés qui influenceraient l’expression du bouquet, le sulfure de diméthyle (DMS) présente de multiples caractères organoleptiques. Il participe à la fois au caractère « fruits frais » et « truffe » des vins vieux. L’influence de paramètres viticoles sur les teneurs en précurseurs du DMS (pDMS) a été étudiée ainsi que la modulation de ses teneurs au cours de la vie du vin. Le DMS, par le biais de la mesure de son potentiel, est largement influencé par les conditions du millésime et par la phase de maturité du raisin. Cependant, les conditions fermentaires constituent un réel enjeu sur sa préservation au cours de la vinification et donc sa gestion à la mise en bouteille. Enfin, la libération du DMS au cours du vieillissement est dépendante de la matrice puisque des interactions avec les tanins du raisin interviennent. Dans ce cadre, le comportement du DMS au cours de l’acte de dégustation a été précisé. Le DMS pourrait donc être un bon marqueur à la fois du terroir, de la gestion des fermentations mais aussi du mode de service du vin vieux. Cette thèse aborde et ouvre de nombreuses questions sur l’origine moléculaire d’autres composés d’intérêt, l’importance des conditions fermentaires ou encore l’influence des conditions de la garde des vins sur le potentiel de vieillissement et le caractère aromatique des vins vieux
Among wines, those which impart the most pleasure and magic to amateurs are very often aged wines. These are bottles preserved for many years in the hope of a great tasting. Upon opening, the taster will expect to discover the complexity and elegance of the wine's aging bouquet. This olfactory concept, in which aromas produced during bottle aging blend harmoniously with those of the wine's youth, is the gift offered by great wines to epicureans. The expression of the bouquet is purported to be very closely linked to the terroir and the vintage, both revealed during the course of aging. This thesis work is essentially devoted to demonstrating and detailing this empirical knowledge.Firstly, quantitative analysis was carried out on several molecules linking the composition of grape berries to fermentation activities and to aging. Eighty wines of different vintages from several Bordeaux vineyards “Châteaux” were studied. Analysis of the results showed that the wines each have their own molecular identity despite variability due to vintage and age. The presence of terpene compounds, derived from plant metabolism, is fundamental in the construction of this identity. This differentiation of the wines at a molecular level shows the importance of grape composition and the expression of terroir on the identity of old wines.Secondly, the parameters of the terroir (water status, nitrogen status, and grape variety, for example) were compared to the concentrations of the aromas studied, which were measured using gas chromatography techniques. This work required the development of analytical tools as well as the designing of several experiments on various scales (from laboratory to wine production units). Finally, with the aim of obtaining tools that could help predict the aging potential and the emergence of the bouquet, an additional study focusing on the precursors of bouquet aromas was initiated. Among the compounds that influence the expression of the bouquet, dimethyl sulfide (DMS) has multiple organoleptic characteristics, contributing to the “fresh fruit” and “truffle” characters of old wines. The influence of wine-growing parameters on the levels of DMS precursors (pDMS) was studied. The modulation of its levels was also explored. DMS levels, determined through the measurement of its potential, are largely influenced by the conditions of the vintage and by the stage of ripeness of the grape. However, fermentation conditions constitute a real issue for its preservation during winemaking and therefore its management at bottling. The release of DMS during aging depends on the matrix since interactions with grape tannins take place. In that context, the behavior of the DMS during tasting was clarified. The DMS could therefore be a good marker both of the terroir and the management of fermentation, as well as the way of serving old wine. This thesis presents many questions on the molecular origins of other compounds of interest, and the influence of fermentation and wine aging conditions on the aging potential and aromatic character of old wines

Thesis (MSc (Wine Biotechnology))--University of Stellenbosch, 2009.
The PGU1 gene encodes an endo-polygalacturonase, an enzyme that degrades pectin. Although the presence and function of this gene is well characterized in Saccharomyces cerevisiae, its regulation is very complex and not yet fully understood. Yeast producing a highly active polygalacturonase (PG) during alcoholic fermentation could potentially improve filtration and turbidity and also enhance extraction of certain aroma compounds. This could replace the addition of expensive commercial enzyme preparations that often contain unwanted enzymes. The first objective of this study was to evaluate PGU1 expression in recombinant strains of S. cerevisiae that originally lacked the PGU1 gene. A functional PGU1 gene and its promoter were successfully re-introduced into their native position in the genomes of five wine strains. Three of these strains recovered PG activity while two did not transcribe the gene and subsequently lacked activity. The three strains that recovered activity were used in microvinification experiments to determine the effect of PG-producing yeast on the aroma profile of the wine. No significant differences were observed in the volatile compounds production between the recombinants and their respective wild types, but some tendencies arose, especially for the monoterpene geraniol. The second objective of this study was to analyze the PGU1 gene and promoter from Saccharomyces paradoxus RO88 (a strain that exhibits high PG activity) and to compare it to those of S. cerevisiae S288C in order to identify differences that could potentially be responsible for the difference in their PG activities. Comparison of the gene sequences revealed several amino acid differences, one of which was in the peptide secretion signal. Analyses of the promoters also indicated some potentially important differences. Furthermore, S. cerevisiae strain VIN13, RO88 as well as two interspecies hybrids (all displaying varying PG activities) were compared under winemaking conditions. Clear differences were observed for the production of certain compounds. RO88 and the hybrids produced higher concentrations of certain volatile compounds, although they were not strong fermenters. Two recombinants, each containing a PGU1-overexpressing plasmid (one with the PGU1 gene from S. paradoxus and the other from S. cerevisiae), were also used in vinification to determine the effects of the different PGU1 gene on the aroma profile of the wine. Unfortunately, the plasmids were unstable and lost during the fermentation. Nevertheless, some tendencies were observed that indicated possible higher production of certain compounds by the recombinants compared to their wild types. This study identified that regulation of the PGU1 gene differs between strains with different genetic backgrounds. Certain differences were observed in the PGU1 gene and promoter sequences between S. cerevisiae and S. paradoxus that could potentially be the reason for the difference in their PG activities. From an oenological point of view, the presence of PGU1 in the genome of a fermenting strain tends to increase the aromatic potential of wine. These results provide a good platform for further studies on the PGU1 gene.

The effects of varying levels of sulphur dioxide (SO₂) on the cold maceration process was investigated with Pinot noir (Vitis vinifera L.) wine. The effects of these varying levels on the wines composition and colour parameters were examined. Cold maceration is a technique whereby grapes are crushed and placed at low temperatures (4 - lO°C) in the presence 50 - 150 mgL⁻¹ SO₂. This process is believed to provide a medium for the extraction of water soluble phenolic compounds, rather than the alcoholic extraction employed in normal fermentations. The extraction of these phenolic compounds was monitored from the juice through to six months of bottle age. The changes were measured using both Spectrophotometric and High Performance Liquid Chromatographic (HPLC) procedures. Cold maceration wines were found to be not significantly different to the control wine in all compositional parameters other than titrateable acidity which was found to be less than the control for all the cold maceration wines. The unsulphured cold maceration wine was not significantly different from the control wine in any of the spectral measurements except natural degree of ionisation, in which it was higher, and total phenolics, in which it was lower. These results indicate that the cold maceration process alone does not alter the extraction of phenolic compounds. The HPLC analysis of the wine confirmed the spectral results indicating that their were no significant differences in the levels of extraction of anthocyanins. The sulphured cold maceration wines were significantly greater than the control in visible colour, colour density, total anthocyanins, natural degree of ionisation, ionised anthocyanins and total phenolics. These results followed similar patterns with wine ageing, at six months these wines were still significantly greater in all the measurements apart from natural degree of ionisation. The results for the sulphured cold maceration wines indicates that SO₂ is acting as a solvent for the extraction of phenolic compounds including anthocyanins. The 50 mgL⁻¹ SO₂ cold maceration wine had similar colour and phenolic content to the 100 mgL⁻¹ SO₂ cold maceration wine at bottling, at six months the 50 mgL⁻¹ SO₂ cold maceration wine still retained a similar colour to the 100 mgL⁻¹ SO₂ cold maceration wine but had vastly reduced anthocyanin content. This indicates that for the grapes utilised in this study the most appropriate level of addition at cold maceration would be 50 mgL⁻¹ of SO₂. With grapes of differing phenolic content the level of addition required will vary.